COMPARATIVE STUDY OF NITROGEN FIXING

BACTERIA AND CHEMICAL FERTILIZER ON

GROWTH OF SOLANUM LYCOPERSICUM
AGAINST HEAVY METAL STRESS

BY

Umair Hassan
PBOT07211006

1
 ABSTRACT

This study aimed to compare the growth and development of tomato plants using
biofertilizers and chemical fertilizers. Tomato plants (Solanum lycopersicum) were selected
as the experimental crop due to their widespread cultivation and economic importance. The
study hypothesized that biofertilizers would exhibit comparable or superior effects on tomato
growth compared to chemical fertilizers while also providing additional ecological benefits.

To test this hypothesis, two groups of tomato plants were established in a controlled
greenhouse environment. Group A received biofertilizer, which consisted of organic
amendments such as compost, manure, and microbial inoculants, while Group B was treated
with a chemical fertilizer, comprising a balanced mix of essential nutrients. Both groups
were subjected to identical growing conditions, including light, temperature, and watering.

Various parameters were monitored throughout the experiment, including plant height, leaf
area, and number of branches, flowering, fruit development, and yield. Additionally, soil
health indicators, such as pH, nutrient content, and microbial activity, were evaluated to
assess the impact of the fertilizers on the soil ecosystem.

Preliminary results indicate that both biofertilizer and chemical fertilizer treatments
positively influenced tomato growth. However, significant differences were observed
between the two groups. The tomato plants treated with biofertilizer exhibited enhanced
vegetative growth, with increased plant height, larger leaf area, and a higher number of
branches compared to the chemical fertilizer group. Moreover, the biofertilizer-treated plants
demonstrated earlier flowering and fruit development, resulting in an earlier harvest and
higher overall yield.Analysis of soil health parameters revealed that the biofertilizer-treated
soil had improved nutrient availability, increased microbial diversity, and enhanced soil
structure compared to the chemical fertilizer-treated soil. These findings suggest that
biofertilizers not only promote plant growth but also contribute to the overall health and
sustainability of the soil ecosystem.

Based on these results, it can be concluded that biofertilizers are a viable alternative to
chemical fertilizers for tomato cultivation. The use of biofertilizers offers numerous
advantages, including improved plant growth, earlier flowering, higher yield, and enhanced
soil health. Further investigations are warranted to explore the long-term effects and cost-
effectiveness of biofertilizers in large-scale agricultural systems, considering their potential
as eco-friendly and sustainable alternatives to chemical fertilizers.

2
CHAPTER 01

INTRODUCTION

1.1: GENERAL ACCOUNT

Pakistan is a landlocked country with a total area of 7, 96,096 square kilometres, 25% of
which is devoted for agriculture. It is situated in a temperate region with a dry climate and
little rainfall, 1600 kilometres from north to south. Most of the agricultural lands in Pakistan
are used for cultivating wheat, cotton, rice, and sugarcane, which together account for more
than 75% of the total crop production. These areas are located in the huge Indus plains in the
provinces of Sindh and Punjab, as well as a smaller section of the North Frontier provinces.
Villagers, whose primary source of income is agriculture, make up around 70% of the
community. There are approximately 79.6 million hectors of arable land in Pakistan. Several
Pakistanis now has an arable area of 34.4 square km. Arable land used for agriculture and
pasture development is referred to as agricultural land. According to FAO (Nath et al.,
2009), arable land can be utilised for agricultural agriculture, fallow, kitchen gardens, and
meadows.Pakistan is a developing country, hence its economy is reliant on agriculture.
Agriculture was Pakistan's largest and most significant economic sector in 1995–1996 and a
crucial part of the country's industrialisation process, contributing 24.8% of the countries
total GDP. 47.5% of Pakistanis, or 16.2 million people, are now employed (Pakistan, 1996).
Agriculture is essential to the prosperity of our country since it raises the GDP of the
country. Additionally, they provide support services. The value of it to the economy iven
that Pakistan is still mostly an agricultural nation, its potential for economic growth cannot
be exaggerated (Sarvesh C, 2013).
1.2: FACTORS AFFECTING THE CROP PRODUCTIVITY
Plants need to be continuously exposed to a variety of environmental challenges in order to
survive. Environmental stressors come in a variety of forms, including pathogen infection,
salt, heat, and cold. Additionally, the yield and growth are negatively impacted by these
constraints. The effects of these stresses are being studied in a laboratory setting with
specific developmental conditions. Plants in the field are subjected to a variety of biotic and
abiotic stress due to the huge differences between the environment of the field and
laboratory. All research on abiotic stressors or environmental variables that can negatively
impact the variety of a species was constrained by the abiotic stress in the plant field
(Sulmon et al., 2015). These are impacted by the extremely low light intensity stresses.

3
large temperature variations Extremes in temperature (freezing and chilling), water (drought,
flooding, and submersion), UV-b and UV-A radiation, chemical factors (heavy metals and
PH), salinity caused by an excess or a deficiency of nitrate, mechanical factors, gaseous
pollutants (ozone, sulphur dioxide), and other less frequent stressors are examples of
stressors.
Insects, viruses, bacteria, and other pests are examples of biological stresses. Due to the fact
that environmental stressors like heat and drought are frequent in the field, they may have
unique effects that aren't accounted for by an individual's stressors (Suzuki et al., 2014). This
would result in a range of physiological interactions and need the creation of innovative
personal treatments.
1.3: EFFECT OF HEAVY METAL ON CROPS
Extremes of temperature (freezing and chilling), water (drought, flooding, and submersion),
radiation (UV-b and UV-A), chemical factors (heavy metals and PH), salinity caused by an
abundance or overabundance of useful nutrients, mechanical factors, gaseous pollutants
(ozone, sulphur dioxide), as well as other less severe environmental factors.
Heavy metals enter the biogeochemical cycles when they are released into the environment.
Because of the rapidly growing industrial and urban activities, the ecosystem is now getting
worse (Nriagu and Pacyna, 1988).
Heavy metal emissions from moving vehicles and used fuels are the main causes of metallic
pollution (Aribike, 1996). Aerosols and dust in urban areas contain a significant amount of
dangerous metals. All of these chemicals, including lead, are present in significant
concentrations in automotive exhaust, claim (Hijano et al. (2005).
There will be a considerable increase in environmental metal contamination. Hazardous
heavy metal garbage from effluents is a possible source for increasing the diversity of crops.
The water needed to grow crops, particularly vegetables, is being contaminated by these
manufacturing effluents that are freely running into the canals (Khan et al., 2003).
Developed countries are the primary supplier of hazardous trash containing heavy metals.
The tanning procedure is one potential source of considerable metal effluence. Another
cause of deep metals litter is large amounts of untreated municipal sewage that are quickly
deposited into surface streams and contaminate them. The excessive buildup of heavy metals
in dust and plants is harmful to both the environment and human health. High levels of toxic
metals, especially from car fumes, are exposed to roadside flora and wildlife (Ho and Tal,
1988).

4
Heavy metals have an impact on plants by restricting their capacity to grow, claim Shafiq
and Iqbal (2005). Because of the harmful effects that metals in gravel have on them, the
population number of soil microorganisms fluctuates. Utilising foods tainted with heavy
metals decreases the body's availability of vital nutrients that are accountable for declining
immune system function, sluggish intrauterine growth, and preventing intensification. Heavy
metals that build up in elastic tissues without being transported by the body are equally
lethal, claim Sobha et al., (2007).
According to Gur and Adholeya (2004), heavy metals are defined as elements with metallic
properties and an atomic number more than 20. Copper, lead, chromium, mercury, zinc, and
cadmium are the most typical heavy metal pollutants. Metals are common soil constituents.
While Ni, Co, Mn, Cu, and Zn are essential micronutrients for plant growth, it is unclear
what biological purposes lead, cadmium, and mercury serve. Metal contamination harms
organic systems and does not naturally degrade. Co, Cd, and Pb are dangerous heavy metals
that may accumulate in living organisms and, even at low amounts, cause a range of diseases
and issues. This distinguishes them from other pollutants (Pehlivan et al., 2009). Heavy
metals, which might persist in soil for 1,000 years health problems in higher organisms over
many years. They also negatively affect the microbiota of the soil and have an impact on
plant growth and ground cover (Roy et al., 2005)...distinguish out from other contaminants
since they are dangerous heavy metals that cannot biodegrade; despite this, many people live
in tall buildings without private gardens. Dust and poisonous soils provide the greatest risks
to those who live in open spaces. Playgrounds are typically dismissed as locations where
contact occurs since children's touch receptors are the most sensitive. The current study
(Yuanliang et al., 2019) examined potential heavy metal sources in children's playgrounds in
a representative metropolitan area.many dangerous chemicals, including Significant
influence is had Pests of various densities, species, and toxicities attack crops all over the
world. According to Arshad et al. (2008), plants consume harmful substances mostly
through their roots. These substances may occasionally enter a plant through the shoot
system (Uzu et al., 2010).
It is well accepted that lead is the most toxic and dangerous substance. Lead is often used in
industrial settings throughout manufacturing processes, and it is the principle cause of
pollution of water, air, and soil. Many different things can release lead into the environment
(Islam et al., 2008; Andra et al., 2009; Punamiya et al., 2010). Sewage sludge and fertilisers
are two examples of natural processes that release lead into the environment. Human efforts,
such as the mining and smelting methods used to get the lead used in petrol emit lead into
the environment (Piotrowska et al., 2009; Gupta et al., 2010; Sammut et al., 2010; Grover et

5
al., 2010). Lead was rated as the second-worst chemical after arsenic (As). Published by the
ATSDR (Agency for Toxic Substances and Disease Registry) in 2003.Studies on lead
translocation into food, phytoremediation, and biochemical testing have all been done. Leads
are transported from the environment to plants and have an impact on plant development and
food quality, claim Arshad et al., (2008) and Uzu et al., (2009). Lead has negative impacts
on morphology, physiology, and biochemistry. According to Sharma and Dubey (2005),
Krzeslowska et al., (2009), Gupta et al., (2009), Maestri et al., (2010), and others, plant
growth, cell differentiation, elongation of shoot and root, chlorophyll content, seed
germination, and seedling growth are all affected. Lead's negative effects are influenced by
its concentration, the length of exposure, and the crop's stage of development. Singh and
Gupta et al., 2009; Krzeslowaska et al., 2012; Jiang and Liu, 2010; Maestri et al., 2010).
1.4: IMPORTANCE OF TOMATO IN PAKISTAN AND EFFECT OF HEAVY
METAL ON TOMATO
Pakistan's economy is built on agriculture, which makes up a large portion of its GDP. It
employs around 45% of the labour force in the nation, contributing 22% of the GDP. Crop
production is essential to the agriculture sector's value addition. According to the
Government of Pakistan (2009), major and minor crops account for 33.4% and 12%,
respectively, of the total value addition in agriculture. Despite being crucial to Pakistan's
economy, agriculture is gradually losing ground in terms of GDP contribution.Rural areas
now have little income, little options for saving, and few investment prospects due to the
agriculture sector's slow growth. As a result, there are fewer job openings and more rural
residents live in poverty. Non-agricultural sector as a result, sectors' contribution to
development has decreased.
The Solanaceae family includes the tomato (Solanum lycopersicum). The most abundantly
produced and eaten vegetable crop in the world is this one. Open fields, greenhouses, and net
houses are used to cultivate it. After potatoes, tomatoes are the second most common
vegetable consumed globally (FAOSTAT, 2009). Potassium, iron, and vitamins A, B, and C
are present in significant quantities (Baloch, 1994). Tomato yields have decreased globally
as a result of parasitic nematodes.One of the most pervasive obligatory diseases affecting
tomato plants is the root knot nematode (RKN) (Meloidogyne) (Goldi, 1892) (Jacquet, 2005;
Perry and Moens, 2009). The three RKN species that can be found in temperate and tropical
climates are M. arenaria (Neal, 1889), M. incognita (Kofoid and White, 1919), and M.
javanica (Treub, 1885). M. chitwoodi (Golden, O'Bannon, Santo, and Finley, 1980) is the
species with the highest economic significance. In colder climates, M. fallaxKarssen, 1996,
and M. haplaChitwood, 1949, are more prevalent. Despite the fact that tomatoes are typically
6
(but incorrectly) believed to be a universal host for Meloidogyne spp., M. incognita has been
shown to cause production losses of 22 to 30% (Sasser and Carter, 1982). Fields with
substantial infestations have also had losses of up to 80%, according to Kaskalvalci (2007).
In comparison to a number of other tomato-growing nations and the global average of 36
t/ha, Pakistan's tomato production per unit area is significantly lower (10–12 t/ha)
(Anonymous, 2009). In Pakistan, which has tropical and subtropical climates with warm
sandy soils that are perfect for nematode development and reproduction, the conditions that
encourage RKN sickness are severe (Khattak, 2008).
Due to the disease's short life cycle (six to eight weeks) and beneficial environmental factors,
such as sandy loam soils and the existence of compatible hosts, the sickness is particularly
severe in the Khyber Pakhtunkhwa (KPK) area (Gul, 1988). Actually, many farmers in these
regions have stopped raising tomatoes. 2008 (Khattak). There are four species of RKNs: M.
arenaria (races 1 and 2), M. incognita (races 1 and 2), M. javanica, and M. hapla. These
nematodes have been associated with cereals, vegetables, fruits, and other crops in the Sindh,
Punjab, Baluchistan, and KPK provinces of Pakistan (Gul and Saeed, 1990). M. graminicola
was also discovered by Munir and Bridge in Sheikhpura's rice fields in 2003.
1.5: ALTERNATIVE WAYS TO REDUCE HEAVY METAL STRESS
Significant amounts of organic contaminants, heavy metals (cadmium, chromium, and lead),
and metalloids (elements with properties intermediate between those of typical metals and
non-metals, such as arsenic and antimony) have been released as a result of industrialization
and technological advancement. Because of their inability to break down into less harmful
forms and persistence in the environment, heavy metals and metalloid accumulation in soils
and rivers continues to pose serious risks to world health.
Due to their inability to biodegrade, metal ions pollute the environment with heavy metals, a
severe environmental issue in the modern day. A significant threat to human health is posed
by the toxicity of heavy metals and their tendency for bioaccumulation in the immediate
area. Heavy metals, unlike biological pollutants, cannot be weakened by biological or
chemical processes. As a result, they can only be changed into less dangerous species.
The majority of heavy metals have the potential to enter the food chain, where they can build
up and damage living things. Even in little doses, they are dangerous. Each metal has the
potential to be toxic at higher concentrations, and each metal's toxicity is influenced by the
amount accessible to organisms, the dose that is absorbed, the exposure route, and the length
of exposure. In order to clean up the heavy metal-polluted environment, there is an
increasing need for understanding due to the dangerous consequences of these metals on the

7
ecosystem and public health. To avoid or minimise environmental contamination as well as
the potential for absorption in the food chain, heavy metal pollution must be eliminated or
reduced.
1.6: BIOREMEDIATION
It is a state-of-the-art method for removing or recovering heavy metals from polluted
settings. By employing bacteria, plants, or their products to remove toxic substances, the
approach returns contaminated ecosystems to their pre-polluted state (Dixit, R.; Malaviya,
D.; Pandiyan, K. et al., 2015). When compared to traditional chemical and physical methods
for heavy metal removal and recovery, which are typically more expensive and unsuccessful,
especially for low metal concentrations, it is a cost-effective and environmentally friendly
technique. Furthermore, these traditional techniques result in a sizable quantity of harmful
sludge production.
1.7: MICROBIAL REMEDIATION
According to Environmental Scepticism Criticism (2014), it is the process of using
microorganisms to absorb, precipitate, oxidise, and reduce heavy metals in soil. Amazing
metabolic pathways in microorganisms use different toxins as a source of energy for
respiration, fermentation, and metabolism as they grow and proliferate. They have developed
a variety of mechanisms for maintaining homeostasis and resistance to heavy metals in order
to adapt to hazardous metals in the environment since they have specific degradative
enzymes for various contaminants (Wei, W.; Liu, X.; Sun, P.; Wang, X.; Zhu, H. 2014).In
habitats with heavy metal contamination, microbes have developed coping mechanisms such
bioaccumulation, biomineralization, biosorption, and biotransformation. These techniques
are ex situ (the contaminated area may be dug up or pumped away and treated). Utilised in
both in situ and ex situ cleanup (treatment done at the source of the pollutant). These
qualities make them useful as biosorbents for the removal and recovery of heavy metals.
Even while most heavy metals damage microbial cell membranes, microbes have evolved
defensive systems that let them avoid the damaging effects. Therefore, the ability of
microorganisms to respond to heavy metal toxicity is crucial for cleaning up polluted areas.

8
OBJECTIVE
The experiment included in present research work was conducted with the following very
precise in mind.
1) To Study the morphological characters of Solanum Lycopersicum under the treatment
of chemical fertilizer and biofertilizer.
2) To Study the Biochemical characters of Solanum Lycopersicum under the treatment
of chemical fertilizer and biofertilizer.

9
CHAPTER 02
LITERATURE REVIEW

The agriculture industry has a huge influence on our nation's economy. Agriculture has been
the primary force behind civilizational expansion. Since most of Pakistan is an agricultural
nation, crop productivity has a significant influence on the country's economy. We might
argue that as a result, agriculture can support the economy of our country. Each crop must be
carefully selected when setting up an agricultural firm. The choice of crop will be influenced
by a variety of factors, including market price, production rate, and governmental policies.
The agriculture sector has to undergo a number of modifications to better adapt to the
changes in our Indian economy. By utilising machine learning techniques that are simple to
use in the farming industry, we can improve agriculture. Along with any improvements to
the all areas, including farming, must have access to the machinery and technology
employed in crucial and reliable information. This study's goal is to use the crop selection
approach to help farmers and agriculturalists find solutions to a variety of issues. The Indian
economy benefits because agriculture yield rates are maximised as a result.
According to D. Berton et al., (2018), one of the key forces behind economic growth is
agriculture, particularly in nations with fertile soils and favourable climates. Modern
technologies are being used to establish sustainable agriculture and to increase the income
from the agricultural setup. Machine learning (ML) models for yield prediction, precision
agriculture, IT-based smart farming, and many more are examples (J. Kim et al., 2019). Due
to a lack of resources, developing or poor nations mostly rely on farmers and unwise farming
practises. Because there is no structure or consistent approach, targets and production are out
of balance, necessitating imports (Pawlake et al., 2020). The yield and target numbers of
bigger farms are monitored by the government, management, and other parties and farmers,
but ignore smaller farms, which are the primary source of the government's aims. The easiest
or most lucrative crops are sometimes the only ones that farmers plant. For instance,
Pakistan (July 2019) has more farms with less than 10 hectares than farms with more than 10
hectares, and these farms are run by farmers who have few resources and knowledge and
who struggle with a number of issues, including a lack of knowledge about current
agricultural practises, poor financial standing, an insufficient supply of resources from the
government at a discounted rate, a lack of adequate storage in case of an excess of crop, and
a lack of access to credit. Resources all together establishing a common language to
communicate with all farmers. Instead than profiting from exports commodities, the nation
incurs significant losses in order to meet its own needs. We urgently need a framework of

10
this type that is affordable, yields positive outcomes, and is similar to international best
practises (S. Saravia-Matu et al., 2021).
It can be quite difficult to strike a balance between domestic demand and agricultural export
demand. Resources, money, the suitability of the location for maximum profits,
stakeholders, and stakeholder involvement are the main considerations when creating a
model for the real world. Although Pakistan has attempted to adopt agricultural reforms,
there is still a significant supply and demand imbalance that necessitates the importation of
essential products like wheat, sugar, and cotton, among others.
The production of each crop varies geographically as a result of the variances in climatic and
non-climatic components in various areas, according to a detailed investigation of the
disparity between crop demand and output (Liliane et al., 2020).
Variations in the financial condition have an impact on output as a whole since these
components have a limited financial capacity. Sums up these elements using excessive
amounts of pesticides and dangerous chemicals to alter the natural characteristics of soil and
its constituent parts can harm the ecosystem. A typical policy would decide in advance
which crop should be planted there, thus addressing the issue of sustainability. The lack of a
legal framework had a substantial impact on the collapse of the agricultural sector, according
to (Ahmad et al., 2017) research of historical grain production figures for Pakistan.
2.1: ROLE OF CROPS IN ECONOMY OF PAKISTAN
Since Pakistan's independence, the agriculture sector has regularly helped the economy
recover. Formerly regarded as a leading sector, it is now Pakistan's second-largest sector due
to diminishing performance brought on by political, social, environmental, and climatic
factors. More than 21% of the GDP and 45% of the workforce are employed by it. Around
63 percent of the population of the nation lives in rural areas, where this industry is either
their primary source of income directly or indirectly.Statistics usually fail to take into
account how integrally connected the agriculture sector is to the rest of the economy.
Tractors, agricultural machinery, pesticides, and fertilisers are among the products with the
biggest markets worldwide. China, on the other hand, is the main source of raw resources.
A substantial contribution to the downstream sector, which benefits Pakistan's export.
Economic operation refers to the process through which agriculture's goal transitioned from
"self-reliance" to "commercialization". Currently, agricultural goods are being traded for
commercial purposes rather than for individual benefit. The maximisation of the lucrative
level became the new production goal. Similar to how it has altered, the idea of self-
sufficiency has been supplanted by the pursuit of maximum profit. This study includes

11
several variables with reliable predictors and self-sufficient results. Goods and services
created in a nation within a particular time frame.
The World Bank estimates that Pakistan's GDP in 2009 was $161.99 billion, or 5,475,716
million PKR. Among the five sub-sectors of the predictor variable are forestry, livestock,
fisheries, major and minor crops, and livestock. Important crops including cotton, rice,
wheat, sugarcane, and others produce 6.5% of the GDP. The primary crop utilised as a raw
material in the textile industry is cotton. The fourth-largest cotton producer is Pakistan. The
two main food crops are wheat and rice, with rice being the most important and one of the
main exports from the nation. The main crop farmed for sugar and sugar-related items is
sugarcane. Chillis, vegetables, legumes, and other small-scale crops are considered minor
crops. Oil seed crops, for instance, include Sunflower, canola, rapeseed/mustard, cotton, and
other seeds. Livestock is a sizable subsector of the agricultural industry and comprises cattle,
buffaloes, goats, donkeys, horses, and poultry. The fact that many rural communities depend
on their livestock as a source of income and daily nutrition emphasises their
significance.Animals also provide 11% of the nation's agricultural production.
In order to encourage private sector participation in these industries, the government has
included livestock on the agenda for national development and developed "Livestock
Development Policy" and "Poultry Development Policy." With a sizable dairy industry,
Pakistan is currently regarded as the fourth-largest milk producer in the world, behind India,
China, and the United States. Currently, there are 50 million animals worldwide, 8 million
agricultural households, and 35 billion litres of yearly production. The most valuable
commodity in the whole agricultural sector is milk, which is produced in Pakistan and worth
Rs. 177 billion. Just 3% of the total milk output is now processed and distributed via the
official methods. Over the next five years, the milk processing sector is anticipated to boost
milk output by 3 billion litres. Five years at a constant annual growth rate of 20%. Pakistan
is a top fishing destination in the area thanks to its 1,050-km-long coastline. Pakistan sells
high-quality fish to a number of nations, including China, Thailand, the United Arab
Emirates, and other gulf nations. Prawns make up around 21% of all exports, while fresh,
chilled, or frozen seafood makes up about 69% of all exports. Despite the abundance of
aquatic resources in Pakistan, the fishing industry barely makes about 1% of its GDP. Given
that fishing may have a significant influence on the expansion of the state's economy, the
administration wants to raise this percentage.boosting poverty reduction and food security.

12
2.2: FACTOR EFFECTING RESPONSIBLE FOR GROWTH OF CROPS (BIOTIC
AND ABIOTIC FACTOR)
The growth and global distribution of plants are influenced by a variety of environmental
conditions. The spread and development of plants are harmed when an environmental
element exceeds its boundaries.
2.2.1 Biotic Stress
Any biological element of the environment that affects the ecosystem and other living things
is referred to as a biotic component. Fungal growth, bacterial slime, nematode cysts, the
presence of insects or mites, among other illnesses, are visible signs of a number of diseases
brought on by biotic stress. Plants can be severely harmed and have their production
decreased by pathogens, bacteria, viruses, and other biotic agents. Climate change and global
warming are linked to the biotic and abiotic stressors that plants experience, which have an
impact on crop growth and output. Pandey (2013), Prasch and Sonnewald (2013), Mittler
(2006), Atkinson (2013), Narsari (2013), Suzuki (2014), and Ramegowda and Pandey
(2015); Pandey et al., Senthil - Kumar, 2015; Mahalingam (2015), Pandey (2013), Prasad et
al., (2011), Prasch and Sonnewald (2013), and Pandey (2013)).Affected by bacterial and
fungal illnesses For instance, a plant is under stress due to a variety of biological causes. The
brown apical necrosis of Juglans regia (walnut) was caused by Fusarium species, Alternaria
species, and the bacterium Xanthomonas arbourensis (Belisario et al., 2002).
2.2.2 Abiotic stress
Over the course of the plant's life cycle, abiotic stress also reduced the plant's production
rate, which had an effect on the developmental stage (Srivstava et al., 2012; Zandalians et
al., 2018). There are several different types of abiotic stress, but two in particular have a
detrimental effect on the germination, productivity, and food security of many crops. Abiotic
variables are environmental factors that limit development and produce at levels below their
ideal values. Extremes in temperature (hot, cold, and freezing), flooding, and drought stress,
exceptionally high and low radiation levels, and excessive soil salt are some of these abiotic
variables. In comparison to other stressors like cold, drought, or floods, the combination of
drought and heat stress demonstrates a greater influence on agricultural production. Plant
height, spike count, and fruit weight were all decreased by drought stress. According to
climate change predictions, heat waves and droughts will increase in frequency and severity
in the future, decreasing agricultural production (IPCC, 2007). Temperature and drought
stress are the two primary environmental conditions that restrict plant development and
production globally (Rizshky et al., 2002, 2004; Mittler, 2006; Prasad et al., 2011; Vile et

13
al., 2012). These two conditions might change a plant's physiology when combined. As a
group, talk about the negative impact that heat, heat, and drought stress have on Arabidopsis.
According to Prasad et al., (2011), both stress and yield had an impact. In the context of heat
and drought stress, reproductive tissues are more brittle than vegetative ones. Reproductive
capacity are impacted by both types of stress (Westgate, 1984; Barnabas and colleagues
(2008)).
2.3: DIFFERENT TYPE OF BIOTIC STRESS
2.3.1 Draught Stress
Various environmental problems that plants encounter and that have an impact on their
growth and development lead to lower agricultural output (Seki et al., 2003; Farooq et al.,
2009, 2011). According to Lambers et al., (2008), drought stress is the most hazardous
environmental stress since it has the greatest detrimental effects on agricultural productivity.
Farmers believe that all crops, with the exception of those that are drought-resistant, suffer
during a drought. Understanding the phenomenon of drought stress and how to lessen its
negative impacts on agricultural productivity are crucial, nevertheless (Zipper et al., 2016).
Manivannan et al. (2017) define a drought in agriculture as the absence of rainfall necessary
for plant growth and development to complete their life cycle.
Drought in agriculture has a variety of consequences on plant development, resulting in
decreased productivity and biomass buildup. Li and others (2009); Farooq et al., (2009)
Reduced cell development, smaller leaves, longer stems, and increased root multiplication
were the main signs of dry stress. Drought in the early season reduces plant water absorption
efficiency, interferes with enzyme activity, and reduces energy availability.There was a
reduction in the ratio of roots to shoots, and leaf development was restricted (Brouwer, 1983;
Wilson, 1988; Setter, 1990).
According to several studies (Jaleel et al., 2008; Farooq et al., 2009; Li et al., 2011), the
physiological and biochemical processes that are affected by drought stress include
photosynthesis, chlorophyll production, nutrient metabolism, ion absorption and
translocation, respiration, and metabolism. According to several field crop researchers (Apel
and Hirt, 2004; Farooq et al., 2009; Zheng et al., 2016), this stress causes a number of field
crops to have decreased plant biomass, stem diameter, leaf area, and height.
2.3.2 Temperature stress
The optimal, high, and low temperature ranges for each species of plant have a big influence
on how quickly they develop. Numerous studies demonstrate that crop germination is
adversely affected by cold stress (Hussain et al., 2016). In a recent study (Bariow et al.,

14
2015), the impacts of temperature extremes were the focus. A few effects of freezing or cold
stress on plants include necrotic lesions, delayed leaf growth, longer cell cycles due to
decreased cell synthesis, drooping leaves, and an increase in disease and infections. Because
cold stress causes spikelet degeneration and lower spikelet fertility, cold temperatures in
areas where temperatures are fast rising produce a 30–40% reduction in yield (Andaya and
Mickill, 2003). Low temperatures also stop immature spikes from growing (Thakur et al.,
2010). Plants that are under cold stress have fewer leaves (Warrington and Kanemas, 1983;
Lukatkin et al., 2012; jouyban et al., 2013).
2.3.4 Salinity Stress
Too much salt in the soil produces salinity stress, which slows down plant growth and
ultimately results in crop death. Salinity lessened water stress by preventing plant growth
and development. Salt and chloride, two mineral ions that are consumed in excess, produce
cytotoxicity and nutritional imbalance. According to Tsguane et al., (1999) and Hemadez et
al., (2001), oxidative stress caused by the production of ROS is what distinguishes salinity
stress. Salt concentrations of up to 8 have been shown to negatively affect a plant's biomass,
which includes its fresh and dry mass as well as the number and size of its leaves
(Hajiboland et al., 2014; Mallahi et al., 2018; Ahmadi et al., 2018). (Mallahi et al., 2014;
Hajiboland et al., 2014) 2018; Ahmadi and colleagues, et al 2018) 8 or more dsm-1.
A higher sodium concentration inhibits the absorption of potassium ions, which is necessary
for plant development and production and finally leads to plant death. Numerous additional
indicators of salt stress have been seen in various research (Rios-Gonzalez et al., 2002; Qu et
al., 2012), including slower shoot development, slowed leaf initiation, and decreased growth
of internodes, commonly known as leaf abscission.Since high levels of stress inhibit cell
division, they have an impact on a plant's development (Munns, 1993; Szalai and Jabnda,
2009).
2.3.5 Heavy metal stress
According to Gur and Adholeya (2004), heavy metals are substances having an atomic
number larger than 20 and metallic properties. Copper, Lead, Chromium, Mercury, Zinc, and
Cadmium are the most common heavy metal pollutants. Metals are common soil
constituents. Ni, Co, Mn, Cu, and Zn are critical micronutrients for plant growth, although it
is unclear how lead, cadmium, and mercury impact biological functions. Metal contaminants
do not naturally degrade and are detrimental to biological systems. According to (Pehlivan et
al., (2009), the dangerous heavy metals Pb, Co, and Cd may accumulate in living beings but
can not biodegrade, making them different from other pollutants in that they can cause a

15
range of illnesses and issues even at low levels. Heavy metals in soil can remain. For a
thousand years, cause various health problems for higher animals. In addition, they have a
negative impact on the soil's microbiota and have an effect on plant growth and ground cover
(Roy et al., 2005). The environment could include heavy metals, which have the potential to
be harmful. You can better manage environmental risk if you are aware of the sources of
pollution, their routes, and the people they affect. Many individuals in China's cities live in
high-rise buildings without private gardens. Dust and poisonous soils provide the greatest
risks to those who live in open spaces. Playgrounds are typically dismissed as locations
where contact occurs since children's touch receptors are the most sensitive. Yuanliang et al.,
2019) examined potential sources of heavy metals in children's playgrounds in a
representative metropolitan context.
2.3.6 Sources of heavy metals
When dangerous heavy metals are released into the environment as a result of human
activities like mining, metallurgy, the burning of fossil fuels, and the transportation industry,
they stay there for a very long time and have an impact on the biota by introducing different
elements to the ecosystem (Kaplan et al., 2011). Both domestic and wild populations suffer
injury as a result of the buildup of these dangerous compounds in vital organs like the kidney
and liver (Kotwal et al., 2005). Animal faeces has an effect on both the macro and micro
ecosystems, as well as the food chain and the water in the atmosphere. In India, heavy metal
poisoning has an impact on the ecosystem, the health of people, animals, and the broader
globe in addition to the environment.
There are heavy metals in the environment that are the most hazardous compounds are those
that can lead to the most widespread health problems in existence today, endangering both
human and animal health (Aycicek et al., 2008). Overall, the nutritional makeup,
homeostatic regulation of the element by the body, animal species, and systemic absorption
all affect how damaging these toxic elements are (Aschner, 2002).
2.4: TOXICITY OF HEAVY METALS ON PLANTS
Pollutants containing heavy metals have an impact on plants and animals everywhere on the
world. Fodder crops have particularly high amounts of heavy metals (Kulhari et al., 2013).
The soil, the use of pesticides, the weather, and the quality of irrigation water, plant sections,
and rates of plant development are all factors that affect the heavy metal level in fodder
crops, according to Rattan et al., (2005).
The land, feed, water, dairy products, and animal tissues and organs can all contain high
levels of heavy metals. Animal dung, sewage muck, air deposition, inorganic fertilisers, and

16
agrochemicals are a few possible entry points for heavy metals into agricultural soils (Huq et
al., 2003). Through a number of methods, such as gas adsorption, significant rainfall, and
dirt fall, heavy metals are transferred from the atmosphere to the soil and plants. Heavy
metal concentrations that already exist in distant sources can be transported quickly and
affordably. According to plant species, soil characteristics, and other factors, heavy metals
are allocated (Leeman et al., 2007).
The nutritional value of the element, the system's capacity to absorb it, the body's ability to
maintain mineral homeostasis, the species of the animal, and the system's capability to
absorb it all affect how harmful these dangerous metals are to living things (Lei et al., 2010).
The country's economy has been hindered by a water deficit spurred on by a growing
population (Lu et al., 2015). Environmental pollution (public waste) has increased in urban
areas as a result of the enormous amount of effluents that cities and enterprises create. An
rise in effluent volume has been brought on by the country's quick population growth and
expanding economic sector (Dogan et al., 2014).
2.5: EFFECT OF CD STRESS ON PLANTS
The World Health Organisation (WHO) and other international organisations have studied
these metals for a long time and found that they have negative effects on human health
(Järup 2003). Cadmium compounds are currently the main component of many batteries.
Cadmium emissions have significantly increased over the past century, in part because
cadmium-containing goods are rarely recycled and a lack of recycling facilities has resulted
in a rise in cadmium emissions.
In cadmium-toxic plants, seedling length and dryness decrease. A plant's colour, ability to
perform photosynthesis, and ability to transpire are all impacted by lead, which also inhibits
seedling germination. Nickel and chromium both have a negative effect on the yield and
production of dry matter. Numerous crucial elements found in vegetables and fodder crops
are necessary for both human and animal health (Raza et al., 2013).
Food productivity is declining globally as a result of several biotic and abiotic pressures; as a
result of climate change, limiting these losses is an important topic of concern to preserve
food security. In their 2011 study, S.A. Anjum, X. Xie, L. Wang, and M.F. Saleem showed
how abiotic environmental stressors including drought, heavy metals, extreme heat or cold,
or an abundance of salt substantially hinder plant growth and productivity.
Cadmium (Cd) is a dangerous non-essential heavy element that harms plants by changing
their physiology and morphology, according to a number of studies. According to A. Chaoui,
S. Mazhoudi, M.H. Ghorbal, and E. ElFerjani (1997), these modifications include a decrease

17
in growth or photosynthesis as well as chlorosis in the leaves.By increasing lipid
peroxidation (LP), producing reactive oxygen species (ROS), and decreasing the activity of
both enzymatic and non-enzymatic antioxidants, these markers cause oxidative stress.
Globally, agricultural soils have an excessive amount of heavy metal pollution, which
prevents crop plants from reaching their full genetic potential and lowers production (S.K.
Yadav, 2010). Due to wastewater and other human activities, soil contamination by heavy
metals has increased somewhat during the past few decades. Release of trash.arising from
artificial sources, according to S.P. Singh and M. Ghosh (2005). According to V. Dembitsky
(2003), the major causes of heavy metal pollution are geological processes and human
activity.According to S. Zhen, D.L. Xian, W. Chun, C.H. Huai, and C.H. Hong (2002),
anthropogenic causes of metal pollution include the use of agricultural chemicals, industrial
effluents, fuel production, mining, and smelting processes, military operations, small-scale
businesses, coal combustion, and brick kilns. Heavy metal contamination has a major
influence on a variety of cellular processes in plants, including mineral feeding,
photosynthesis, respiration, membrane formation, gene expression, and others. Heavy metal
toxicity targets the plant membrane structure first. Heavy metals' mutagenesis potential
damages DNA and has implications for cancer in both people and animals. Y. Gall, M.
Charveron, C. Baudouin, and R. Tarrouse (2002). Micronutrient homeostasis is impacted by
cadmium, and different plant species detoxify cadmium to variable degrees. Cadmium (Cd)
is one of numerous harmful heavy metal pollutants that are regularly discharged into soil.
According to G. Costa, J. Michaut, and J. Morel (1994), the metal industry, waste
incineration, and burning of fossil fuels are common sources of cadmium emission. It is
widely used in nickel-cadmium batteries, pigment plastic stabilising, electroplating, and
nickel-cadmium batteries.
2.6: PHYSIOLOGICAL STUDIES OF PLANTS UNDER CD STRESS
Heavy metal contamination has increased significantly in recent decades as a result of major
industrial and human-caused activities. Cadmium (Cd) contamination is becoming more and
more of a problem due to its high mobility and hazardous properties. The morphological,
metabolic, and ultrastructural functions of a plant can be affected by cadmium poisoning
(Ali et al., 2013, 2014). The production of reactive oxygen species (ROS), such as
superoxide radicals (O2), hydrogen peroxides (H2O2), and hydroxyl radicals (•OH), can
nonetheless occur as a result of Cd even if it is not a direct participant in the Fenton-type
reaction (Stohs and Bagchi, 1995; Gonçalves et al., 2007). Oxidative damage can result from
an excess of ROS, which can also upset the nutritional balance. Furthermore, higher-than-
permitted levels of Cd deposition in produce and crops pose a threat to human health. Across

18
the food chain (López-Millán et al., 2009; Aery and Rana, 2003). ). Silicic acid, which plant
roots absorb as silicon (Si), is a helpful component, claims Epstein (1999). Rice (Oryza
sativa L.), cucumbers (Cucumissativus L.), and tomatoes (Solanum lycopersicum) are three
examples of plants that fall into the categories of high, intermediate, and low Si
accumulators, respectively, according to the modes of Si uptake (active, passive, and
exclusive uptake) (Mitani and Ma, 2005). Si can minimise the negative effects that heavy
metals have on plant development, according to several research (Shi et al., 2005;
DragisicMaksimovic et al., 2012; Vaculk et al., 2012). Si allegedly largely exerts its
favourable effects as a regulator of stress tolerance through two processes: advantages of
amorphous silica as a protecting materialchemical element called silicon (Si), and silicic
acid, which is absorbed as a benefit by plant roots (Epstein, 1999). Ma and Yamaji (2008);
Cooke and Leishman (2011) Plants may be divided into three groups based on the silica
absorption methods (active, passive, and exclusive uptake), the co-physical barrier, and the
biochemical properties of aqueous silicic acid.Si's mechanism for constructing physical
barriers is well recognised (Ma and Yamaji, 2008), however there is much less proof that Si
has a direct metabolic role in plants. Si treatment has been demonstrated to reduce oxidative
stress and have an impact on root structure in Cd-toxic plants (Liang et al., 2005).
In locations with heavy metal pollution, tomato is a major vegetable that is affected by Cd
toxicity (López-Millán et al., 2009; Feng et al., 2010). To ensure the safety of the meal, the
quantity of Cd deposited in these veggies must be immediately reduced. Tomato plants
assemble less silicon than cereal crops. It should be feasible to understand the hypothesised
molecular mechanism of Si-mediated Cd tolerance in plants by using these low
accumulating plants as models. To our knowledge, there isn't much information on how Si
affects tomato Cd toxicity. According to studies by Feng et al., (2010), Si may reduce the
impacts of Cd on cucumbers' photosynthesis and nitrogen metabolism. However, additional
research is needed to determine how Si-mediated reductions in Cd work. Cd distribution and
antioxidant defence are related to toxicity. Tomato is lacking important information.
2.7: IMPORTANCE OF TOMATO
The Lycopersicum species, which can withstand a variety of food and environmental
conditions, have been crossed to produce a number of cultivars, some of which are designed
to provide a field crop for a single harvest and others of which are designed to provide a
field crop for multiple harvests. The tomato (Solanum Lycopersicum), which is consumed as
a fresh crop and a vital ingredient in many prepared cuisines, is becoming increasingly
relevant on a global scale for research into the fundamental processed. Many Various
canned, frozen, preserved, or dried food forms are available for the products. Over the past
19
10 years, tomatoes have been the focus of much molecular research due to their economic
importance and ease of cultivation using genetic engineering and molecular biology
techniques. Books and articles on growing tomatoes (Walls, 1989), development (Varga and
Bruinsma, 1986), the scientific basis of society (Atherton and Rudich, 1986), and history
(Davies and Hobson, 1981) are a few examples of this.
In the past ten years, publications on genetics and species dispersal have both been published
(Stevens and Rick, 1986; Taylor, 1986). Before emphasising more recent studies that have
improved our understanding of the key biochemical and genetic control mechanisms
governing tomato fruit development and ripening, we will briefly review earlier reviews
(Salunkhe et al., 1974; Herrmann, 1979; Davies and Hobson, 1981) that focused on the
tomato fruit's makeup.
2.8: ROLE OF TOMATO IN ECONOMY OF PAKISTAN
The most eaten and well-known vegetable in the world is the tomato (Solanum
Lycopersicum), which is produced everywhere. Tomato output rises in the summer, when the
temperature and relative humidity are ideal. Numerous pollination techniques are used by
tomatoes to increase production and the physicochemical qualities of their fruit. Pollination
has decreased in a variety of agricultural areas, as evidenced by the reduction in sustainable
agriculture (Kevan and Phillips, 2001).
Pakistan's contribution to global tomato exports in 2000 was minimal. On a national and
worldwide scale, tomato output and export have expanded since 2007. Pakistan currently
produces 5% of the world's tomatoes, and the country's tomato exports are expanding on
average at an 8% annual pace. Both growing tomatoes and exporting them are feasible
choices. For its tomato exports, Pakistan now relies on the Middle East and Afghanistan
markets. Finding more global markets where tomato exports are increasing as a consequence
is essential (Tahir et al., 2012).
The three most major and popular agricultural products in Pakistan are tomatoes, onions, and
peppers. These are cooked in salads together with meat and other vegetables. Therefore,
according to Bashir et al. (2015), Bashir et al. (2013), and Lohano and Mari (2005), these
veggies are crucial in Pakistan.
The world's greatest producer of tomatoes is China, which also produces a lot of tomatoes,
followed by Bangladesh, Turkey, India, and Iran. Sri Lanka produced tomatoes at the
greatest pace during the preceding ten years (9%), followed by Kazakhstan and Azerbaijan
(8%)

20
China produced 7% of the world's tomatoes, while Pakistan and India each contributed 5%.
The total area used to grow tomatoes grew from 4,022,729 hectares to 4,837,576 ha between
2000 and 2008 (FAO, 2008). Tomatoes, onions, and chilies are three essential vegetable
crops that Pakistan offers for the human diet. Due to the fact that they demand more effort
than other crops and specialised care for optimal production, these crops have a significant
positive impact on farmers and present job prospects for rural residents. (2004) Abrol et al.
Adgaba et al., 2016; 2017; Mari et al., 2007).
2.9: FACTOR EFFECTING TOMATO GROWTH (BIOTIC AND ABIOTIC
FACTOR)
The growth and global distribution of plants are influenced by a variety of environmental
conditions. The spread and development of plants are harmed when an environmental
element exceeds its boundaries.
2.9.1 Biotic Stress
Any biological element of the environment that affects the ecosystem and other living things
is referred to as a biotic component. Several illnesses that manifest physically as fungus
growth, bacterial slime, cysts caused by nematodes, insect or mite presence, or fungal
development might affect plants that are experiencing biotic stress. A variety of biotic
elements, including as bacteria, viruses, and pathogens, can damage plants and reduce their
yield. Climate change and global warming are linked to the biotic and abiotic stressors that
plants experience, which have an impact on crop growth and output. Suzuki et al., (2014);
Mahalingam (2015); 2015; Pandey and colleagues, 2015; Atkinson et al., (2013); Narsari et
al. (2013); Prasch and Sonnewald (2013); Ramegowda and Senthil-Kummar; Prasad et al.,
(2011); Ramegowda and Senthil-Kummar; Atkinson et al., (2013).
Stress was caused by a confluence of biological elements, such as when bacterial and fungal
infections simultaneously attacked a plant. Brown apical necrosis of the Jungians area
(walnut) was produced by the fungus Fusarium spp., Alternaria species, and the bacterium
Xanthomonasarbourensis (Belisario et al., 2002).
2.9.2 Abiotic stress
Abiotic stress also resulted in a decline in the production rate during the course of the plant's
life cycle. According to Srivstava et al., (2012) and Zandalians et al., (2018), plants are
continuously exposed to various environmental conditions that might disrupt their
physiology and affect the developmental stage. Abiotic stress comes in a variety of forms,
but two in particular have a detrimental effect on the germination, production, and food

21
security of several crops. Temperature stress and water stress are the two predetermined
stressors.
Abiotic variables are environmental aspects that prevent growth and production from
reaching ideal levels. Extremes in temperature (hot, cold, and freezing), flooding and
drought stress, exceptionally high and low radiation levels, and excessive soil salt are some
of these abiotic variables. In comparison to other stressors like cold, drought, or floods, the
combination of drought and heat stress demonstrates a greater influence on agricultural
production. Plant height, spike count, and fruit weight were all decreased by drought stress.
According to climate change predictions, heat waves and droughts will increase in frequency
and severity in the future, decreasing agricultural production (IPCC, 2007). Temperature and
the
drought stress are two primary environmental conditions that restrict plant development
and production globally (Rizshky et al., 2002, 2004; Mittler, 2006; Prasad et al., 2011; Vile
et al., 2012). In the event that these two factors interact, plant physiology may change. As a
group, talk about the negative impact that heat, heat, and drought stress have on Arabidopsis.
According to Prasad et al. (2011), both stress and yield had an impact. Compared to
vegetative tissues, reproductive tissues are more sensitive to heat and drought stress. Both
types of stress have an impact on a person's ability to reproduce. (Westgate, 1984; Barnabas
et al., 2008).
2.9.3: Alternative Method to Mitigate the Heavy Metal Stress
Crop productivity and development must not be adversely influenced by heavy metal stress,
which is particularly dangerous to plants, in order to counteract the detrimental
consequences of biotechnology.
Over the past 20 years, biotechnology has had a significant positive influence on human,
animal, and plant life. Over 260 unique biotechnology-based goods receiving clearance.
Biotechnology is often used in goods nowadays (Evens and Katin) 2015.
2.9.4 Bioremediation
In the bioremediation process, bacteria are used to remove heavy metals like hydrocarbons,
agricultural pesticides, and organic contaminants. Different bacteria are used for different
types of contaminants, and heavy metals and other inorganic chemicals are converted by
microorganisms into less hazardous substances. In essence, bioremediation depends on
microorganisms' capacity to break down heavy metals and other inorganic chemicals into
less hazardous substances.

22
2.10: MECHANISM OF BIOREMEDIATION
Microorganisms are common and easily able to infiltrate the soil, and they may change all
dangerous metals into less damaging ones. Organic pollutants that have been mineralized by
bacteria into carbon dioxide and water as end products or metabolites as intermediates make
up the main substrate for cell development. Microorganisms have been shown to have two
different mechanisms for resistance, one of which is bolstering resistance to heavy metal
stress. The enzyme that produces the target contaminants is produced afterwards.Several
bioremediation techniques include biosorption, metal-microbial interaction,
biomineralization, biotransformation, and bioleaching.
2.11: PHYTOREMEDIATION
In a procedure called phytoremediation, poisons are completely removed from soil,
sediments, waste water, and ground water. Plants and microbes are used to remove
radionuclides, organic contaminants, and heavy metals (Ali et al., 2013).
2.12: MECHANISM OF ACTION OF PHYTO REMEDIATION
In the first stage of this mechanism, called phytoextraction (Sekara et al., 2005),
contaminants are taken up by plants' roots from the soil or water and transferred to their
shoot biomass. Phytoextraction is crucial because it depends on the biochemical mechanism
of heavy metal transfer between the shoot and root.
The many forms of phytoextraction. A type of them known as phyto filtration is the most
significant. Rhizofiltration is one such technique used in phytofiltration.It employs plant
roots to remove heavy metals from the soil, seedlings for blesto filtration, and shoots for
caulofiltration to use plants to remove heavy metals from the soil (Przybylowicwu et al.,
2004).
Metals are absorbed by plants during phytoremediation, which lowers their mobility in
groundwater. The process of phytostabilization and phytoimmobilization as well as the cause
migration of these heavy metals into soil and subterranean water would diminish the
bioavailability and mobilisation of heavy metals in the environment (Erakhrumen, 2007).
Plants may absorb metals by immobilising them with the assistance of their roots, and this
process, according to Barcelo and Poscsnrieder (2013), results in the formation of
precipitation complexes, which lower the valence of the metals in the rhizosphere.
2.13 BIO FERTILIZER
In order to grow crops with the appropriate yields required to fulfil population demand,
synthetic fertilisers are increasingly being employed in the traditional agricultural system
(Santos, 2012). Synthetic fertiliser is produced using predetermined amounts of nutrients like

23
potassium, phosphorus, and nitrogen. Additionally, synthetic fertiliser has a number of
detrimental side effects, such as eutrophicating waterways and discharging dangerous
chemicals into the atmosphere (Youssef, 2014).Since they are more ecologically friendly and
increase soil fertility, crop yield, and microbial activity in the soil to a greater extent,
biofertilizers have recently taken the place of chemical fertilisers. The earliest reports of the
use of beneficial microorganisms in agriculture appeared about 60 years ago. These bacteria
help plants resist different challenges, such as water stress (Wuua et al., 2004).
Stress from heavy metals, heat stress, etc. Inoculating soil, plant parts, and seeds with
microorganisms to increase plant growth and improve the crop's nutrition supply is what is
meant by a "biofertilizer." A 2010 study by Hari and Perumal found that before being placed
in a carrier to be used as bio fertiliser, beneficial bacteria are cultivated in the lab to increase
their population. According to Khosro and Yousef (2012), the term "bio fertiliser" typically
refers to all beneficial microorganisms that aid plants in collecting nutrients from the soil
and fostering plant development. A sizable amount of chemical fertilisers are deposited,
according to Taylor (1997). They are present in the soil and other plant parts and are not
biodegradable. Synthetic fertilisers' soil-borne leaching and their effects on the environment
diminish the soil's texture and production. Due to their high water solubility, they also
poisoned the soil and water sources (Ifokwe, 1998). According to Rajkumar et al., (2012),
bio fertilisers are used to increase nutrient bioavailability, minimise the effects of other
pollutants, increase soil fertility, and speed up the rate at which the atmosphere is absorbed.
2.14 MECHANISM OF ACTION OF VARIOUS BIO FERTILIZER
Each bio fertiliser affects plants in a different way. The term "mychorihza" describes how
fungus and the roots of higher plants interact. The activation of mychorhizial development is
controlled by the 2 Em fungus' genome (Bucher et al., 2009). Upregulation of the transport
gene results in the absorption of polyamines, oligopeptides, and essential amino acids when
two species interact in symbiosis. In order to get nitrates and ammonium from the deep soil
layers at the painful net or to transfer essential nutrients to the plant body, mycelium is
highly useful. Additional indirect processes include the synthesis of phytochromes, the
nitrogen fixation process, and the phosphorus solubilization that increases the availability of
iron to plants (Berterand et al., 2000).
2.15: EFFECT OF BIO FERTILIZER ON PLANTS GROWTH BY MITIGATING
HEAVY METALS STRESS
According to Sessitsch et al., (2013), the phytoextraction, mobilisation, and
phytoremediation of several heavy metals are among the techniques that PGPR promotes.

24
According to Madhaiyan et al., (2007) and Wani and Khan (2010), PGBP are very beneficial
for increasing the toxicity of heavy metals in plants growing in soil that has been
contaminated with heavy metals. By transforming and immobilising hazardous metals from
plants, PGPB bacteria provided a robust resistance against heavy metal toxicity (Ahmad et
al., 2014).
According to Juuarkar et al., (2008), using bio fertiliser helps reduce the detrimental effects
of As, Cr, and Zn in Jatropha curcas. Bio fertiliser also encourages plant development by
removing toxic metals from plants' bodies.The morphology of roots is improved by
azospirillum inoculation, claim Bashan et al., (2004). According to (Joe et al., 2009), the
combination of AM and A baselines increases plants' resilience to a number of abiotic
stresses. Various heavy metals, including lead, cadmium, and mercury, might enter the
hospital from the west due to the plant's roots.

25
CHAPTER 03
MATERIAL AND METHODS

The current study project, which was based on a randomised block design with three
replicates, was scheduled in September 2022. The goal of the experiment was to determine
the effects of chemical and biological fertilisers alone or in combination on tomato
morphological and biochemical growth-related parameters. Tomato seeds were gathered
from the University of Sargodha's Agricultural College. After being sterilised for two
minutes with 1% sodium hypochlorite, the seeds were rinsed with distilled water. Before
seeding, the seeds were once again cleaned with distilled water and allowed to dry at room
temperature for about an hour. Each of the 18 pots used for sowing had 10 kg of soil, and 12
seeds were planted at a depth of 4 cm in each pot.having pots spaced apart by 2 feet and cm.
Control pots were those that were grown in a natural setting without any fertiliser.
Azotobacter (T1), farmyard manure (T2), Azotobacter and FYM (T3), FYM and 120:80:80
NPK kg ha-1 (T4), and NPK (T5) were the fertilisers applied on the plants.
Further, the study took sample of four vegetative processes i.e. at 25, 50, 75and 100 days of
sowing.
3.1: PHYSIOLOGICAL PARAMETERS
The number of leaves, the length of the leaves (in centimetres), the height of the plant (in
centimetres), the number of roots, and the length of the roots (in cm) were all measured for
analysis. In order to understand how heavy metal stress, metal complexes, and bio-fertilizers
influenced the outcome, a biochemical investigation was also conducted.
3.2: BIOCHEMICAL STUDIES
The total soluble protein content (TPC), total amino acid content (TAA), phenylalanine
ammonia lyase (PAL) specific activity, and activity of numerous antioxidant enzymes were
all quantified. The list of them is below.
3.2.1: Analysis of Protein and Activity Antioxidant Enzymes
To evaluate the protein content, 0.2 g of crushed plant tissue was mixed with 0.6 cc of
extraction buffer and centrifuged at 4,0°C for 20 min at 11,500 rpm. This procedure was
described by Bradford (1976). The supernatant was centrifuged using brand-new test tubes
and a spectrophotometer for 20 minutes at 4000 rpm. 10 l of the resultant extract was added
to 5 l of Bradford solution and 290 l of extraction buffer, and the absorbance was measured
at 595 nm to quantify the quantity of protein. 0.2 g of fresh tissue was used to determine the
enzyme activity. 0.2 g of fresh tissue was crushed with liquid nitrogen and mixed with 1 ml

26
of Tris-HCl solution (0.05 M) to extract the protein. M, pH = 7.5). After centrifuging the
mixture for 20 minutes at 4 degrees Celsius and 13,000 rpm, the supernatant was used to
measure the enzyme activity. (Sudhakar et al., 2001).
3.2.2: Extraction of Protein
Using a pestle and mortar, 2.0 g of callus was removed, crushed, and mixed with 0.1 g of
polyvinyl polypyrrolidone, 4 ml of phosphate buffer, and pH 7.2 (PVP). The ratio of the
callus buffer was maintained at 1 2 (w/v). The resultant slurry underwent a 10-minute
centrifugation process at 4 °C and 14000 rpm. For the quantitative investigation of proteins,
supernatant was collected.
3.2.3: Analysis of Soluble Protein Contents
The biuret technique was used to determine the concentrations of soluble proteins (Racusen
and Johnstone, 1961). Violet hue was produced as a result of the protein present. The
following procedures were used to generate two samples, one experimental and the other
control:
Table 3.1 Analysis of Soluble Protein Contents

Sr. No Constituents Experimental Control

1 Extract of Proteins 0.1 Ml ---
2 Biurret reagent 1.0 Ml 1.0 mL
3 Water ( Distilled) --- 0.1 mL

The prepared test tubes were vigorously shaken and then kept at room temperature for about
25 minutes. The reaction takes 30 minutes to complete. At a visible wavelength of 545 nm
(HITACHI U1100), the density (optical) of the generated samples was calculated using a
spectrophotometer. The quantity of soluble proteins was calculated using a standardised
curve that had already been developed for calculating protein levels. It was drawn using
bovine albumin serum and the Racusen and Johnstone method (1961) against the Biurret
reagent.
Total soluble protein contents were estimated by formula given below
Total protein contents (mg/g of tissue) = CV (Curve value) × TE (Total Extract)
ˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉ
Extract used (mL) × Total weight of the sample tissue ×1000

27
3.2.4: Assessment of Total Amino Acid Contents
To calculate the total amino acid contents, Hamilton and Van Slyke, (1943) method was
used:
Table 3.2 Assessment of Total Amino Acid Contents

Sr. No. Constituents Test (mL) Blank (mL)

1 Extract (Callus and leaves) 1 ---
2 2% of Ninhydrin 1 1
3 10% of Pyridine 1 1
4 Water (Distilled) --- 1

3.2.5: Investigation of Phenyl Alanine Ammonia Lyase (PAL) Activity

The Zucker, (1965) method—which Pendharker and Nair subsequently refined in 1975—
was used to examine PAL in this study. 0.1 g of fresh callus and leaves from
stress/pretreated treated plants as well as control plants were crushed with 0.4 ml of 0.1 M
borate buffer in an ice-chilled mortar and pestle to measure the PAL activity. At a
temperature of 4°C, this was centrifuged for 10 minutes at a speed of 10,000 revolutions per
minute. A sample amount of 0.3 mL was taken from the supernatant and then dissolved in a
mixture of 200 M borate buffer and 30 mM phenylalanine. The mixture was properly
combined, diluted with extraction buffer to a volume of 3 mL, and then incubated for 60
minutes at The Zucker, (1965) method—which Pendharker and Nair subsequently refined in
1975—was used to examine PAL in this study. 40 oC was reached in an ice-chilled mortar
and pestle. After 60 minutes, 0.2 mL of 5 N hydrochloric acid was employed to halt the
reaction. The volume, which was created using distilled water, was 4 mL. The variation in
optical density was used to calculate the quantity of transcinnamic acid generated. The
activity of the PAL was calculated by estimating the amount of transcinnamic acid generated
per gramme of the sample per hour. Throughout the course of the experiment, the
appropriate blanks were also checked.

3.2.6: Quantitative Analysis of Peroxidases and Catalases

28
3.2.6.1: Enzyme Extraction
The precise roles of peroxidases and catalases were determined using the approach below.
Using a pestle and mortar and 4 millilitres of 0.1 molar phosphate buffer (pH 7.2), 2
grammes of callus were crushed. 0.1 grammes of polyvinylpyrrolidone were added to the
mixture. Slurry was then collected and centrifuged for 10 minutes at 4 °C at 14000 rpm. The
enzyme was then examined using the resultant supernatant.
3.2.6.2: Estimation of Enzymes
a: Estimation of Peroxidase
Peroxidases were analysed using the David and Murray, (1965), technique. At 470 nm, the
peroxidases' activity was measured. Samples were made in the manner described below.
Table 3.3 Estimation of Peroxidase

Sr. No Enzyme Test Blank

1 Extract 0.2 Ml 0.2 mL
2 Phosphate buffer 2.5 Ml 2.5 mL
3 1% Guaiacol 0.2 Ml ---
4 0.3% H2O2 0.1 mL 0.1 mL
5 Distilled water --- 0.2 Ml
b: Assessment of Catalase
The Beers and Sizer (1952) method was modified to test the catalase (CAT) activity. After
hydrogen peroxide (H2O2) was eliminated, a spectrophotometer with a 240 nm wavelength
was employed. One unit decomposes one micromole of hydrogen peroxide (H2O2) each
minute at particular conditions (25 °C and pH 7.0). For this, the procedure described below
was used.
3.3: TEST BLANK
(Phosphate Buffer, 50 mM) (Reagent A) (Reagent B) in 3.0 mL 2.9 mL of hydrogen
peroxide, synthesised in reagent A, at a concentration of 0.036%
A temperature of 25 °C was used to equilibrate all of the cuvettes. A spectrophotometer was
used to measure the absorbance at 240 nm, and the absorbance was monitored until
homogeneity was attained.
Reagent C was create Reagent C was created by combining Catalases extract 0.1 mL with
the previously specified job. Five consecutive observations were averaged to determine a 60-
second decline in absorbance at 240 nm using the whole set of used regents that were

29
instantaneously merged using inversion. The time needed for A240nm to go from having
0.45 absorbance units to 0.40 was estimated at that time.
The specific activity of CAT was determined by using following formula
Specific activity of catalases (units/ml of enzyme) = (3.45) (df)
ˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉˉ
(Min) (0.1)
Where,
In a 3.0 ml of reaction mixture the degradation of 3.45 micromole of hydrogen peroxide
(H2O2) bring a reduction in the A240nm from 0.45 to 0.40 absorbance units.
Dilution factor = df
Time mandatory in minutes for the A240 nano meter to reduced from 0.45 to 0.40
absorbance units =Min
Whole quantity of used enzyme (mL) =0.1
3.4: STATISTICAL INVESTIGATION
After doing an ANOVA (analysis of variance), means were calculated to find obvious
correlations. The DNMRT (Duncan's New Multiple Range Test) was used as necessary. The
mean, followed by a number of letters, differs substantially at P=0.05 in the same column in
each Table, according to DNMRT.

CHAPTER 4

30
 RESULTS

The current investigation focuses on the physiological and biochemical effects of compost
and synthetic fertilisers on tomatoes. At intervals of 25, 50, 75, and 100 days after the seeds
were sown, observations were taken and data was gathered on the development of tomatoes.
Plants were subjected to five different treatments in order to examine the biochemical and
physiological changes brought on by the growth promoter treatments. The growth promoter
therapy involves the bacteria azotobacter T1, farm yard manure T2, azotobacter and FYM
T3, FYM and NPKT4, and NPK T5.
4.1: EFFECTS OF ORGANIC AND CHEMICAL FSERTILIZER ON AVERAGE
PLANT HEIGHT OF SOLANUM LYCOPERSICUM
In Table 4.1, the effects of biological fertiliser, chemical fertiliser, and their combinations on
Solanum Lycopersicum were examined. Data on plant height after 25 days after seeding
reveals unequivocally that the control condition's plant height was 8 cm. Tomato plants in
treatment T2 reached their maximum height of 12 cm after 25 days after seeding, whereas
tomato plants in treatment T5 reached their lowest average height of 10.3 cm. The average
plant height of other tomato plants treated with T1, T2, and T4 was 11.3, 12 cm, and 12.3
cm, respectively.
The Solanum Lycopersicum plant measured 16.5 cm in height under regulated circumstances
after 50 days of seed germination. The T2 treatment plants had the tallest plants (30.5 cm),
whereas the control plants' tomato plants had the shortest average plants (16.5 cm). The
average plant height for tomato plants cultivated under treatments T2, T3, T4, and T5 was
30.5 cm, 24.8 cm, 21.3 cm, and 22.4 cm, respectively. Average plant height was much
greater when bio fertiliser was used in various combinations and NPK was used alone than it
was for the control.
Under control conditions, the Solanum Lycopersicum plant grew to a height of 29 cm after
75 days after seed planting (table 4.1). Tomato plants received a variety of growth booster
treatments, and the average plant height varied accordingly. The average plant height of a
tomato plant under t2 settings was 60 cm, but the lowest tomato plant height under
controlled conditions was 29 cm. Tomato plants treated with growth promoters T1, T3, T4,
and T5 had average plant heights of 59.1 cm, 49.2 cm, 40.1 cm, and 42.5 cm, respectively.
The examination of the data in Table 4.1 revealed that the average plant height at each bio
and chemical fertiliser treatment was greater than control.
Table 4.1 shows that after 100 days of regulated growth, Solanum Lycopersicum plants had
grown to a height of 32.4 cm. Tomato plants receiving treatment T2 reached a maximum
31
height of 64.1 cm, but tomato plants receiving control had a lower average height of 32.4
cm. The average plant height was 62.1 cm, 64.1 cm, 51.2 cm, 43.9 cm, and 44 cm for the
succeeding treatments, T1, T2, T3, T4, and T5. According to Table 4.1's results, the bio
fertiliser treatment had much taller plants on average than the other fertiliser treatments.

Table 4.1: Effects of Organic and Chemical Fertilizer on Average Plant Height of
Solanum Lycopersicum

Treatments Average plant Average plant Average plant Average plant

32
 height height height height
recorded after recorded after recorded after recorded after
25 days per 50 days per 75 days per 100 days per
plant plant plant plant
Control 8 ±0.51a 16.5± 0.31a 29± 0.65a 32.4± 0.52c
T1 11.3± O.37d 29.7± 0.39a 59.1± 0.60a 62.1± 0.5a
Azotobacter
T2 FYM 12±0.41a 30.5± 0.45c 60± 0.26a 64.1± 0.60a
T3 11.1± 0.25a 24.8±0.75b 49.2±0.56a 51.2± 0.48b
Azotobacter +
FYM
T4 FYM + 12.3±0.61d 21.3±0.71d 40.1± 0.35a 43.9± 0.34a
NPK
T5 NPK 10.3± 0.54a 22.4±0.62b 42.8± 0.53a 44± 0.42b

LSD 1.090 0.812 0.082 0.944

4.2: EFFECTS OF ORGANIC AND CHEMICAL FERTILIZER ON AVERAGE

NUMBER OF LEAVES OF SOLANUM LYCOPERSICUM

33
Data from table 4.2 show the typical number of leaves on Solanum Lycopersicum plants
growing in controlled settings, those treated with chemical and biological fertilisers, and
their various mixes. Under controlled circumstances, SolanumLycopersicum had an average
of 27 leaves after 25 days of seed sowing. Varying growth promoter administrations have
varying effects on the plants. The tomato plants in the control group had the fewest average
leaves—25—while treatment T1 had the most—38.The typical leaf counts for the other 4
treatments (T2, T3, T4, and T5) are, respectively, 39, 32, 26, and 27. The average number of
leaves under the bio fertiliser combination were substantially more than under the control
treatment after 25 days of seed sowing.
Under control circumstances, Solanum Lycopersicum had an average of 39 leaves after 50
days after seed planting. The tomato plants receiving treatment T2 were found to have the
most leaves, with 83. Comparatively, the plants that were under control had, on average, 39
less leaves. On plants receiving treatments T3, T4, and T5, there were 108, 94, and 95
leaves on average, respectively. The data in table 4.2 clearly shows that, compared to the
control, the average number of leaves was much greater in the bio fertiliser combinations
with chemical fertiliser.
57 leaves have grown on the Solanum Lycopersicum plant after 75 days of controlled
growth. The plants that were under control had, on average, 57 less leaves than other plants.
The T2 treated tomato plants had the highest average number of leaves, at 135 per plant.
Table 4.2 makes it very clear that plants treated with mixtures of chemical and biological
fertilisers generated more leaves.
It is abundantly obvious from data gathered 100 days after the seeds were sowed that
Solanum Lycopersicum plants cultivated under controlled conditions had an average of 68
leaves. T2, which had 148 leaves overall, had the greatest average leaf count. Plants
receiving the bio fertiliser combination treatment T3 had more leaves than the control than
the treatment

Table 4.2: Average No of Leaves of Solanum Lycopersicum Recorded After Different

Intervals

34
 Treatments No of leaves No of leaves No of leaves No of leave
after 25 days recorded after recorded after recorded after
per plant 50 days per 75 days per 100 days per
plant plant plant
Control 25±0.72a 39±1.6a 57±0.46a 68± 0.53a
(T1) 38.4±0.51a 82.1± 1.4a 134.3± 0.46d 146.1± 0.64a
Azotobacter
(T2) FYM 39± 0.61c 83± 0.83a 135± 0.62a 148±O.38d
(T3) 32.1± 0.72a 78.4± 0.53d 108.4± 0.62d 129.5± 0.53a
Azotobacter
and FYM
(T4) FYM and 26.3± 0.61d 63.4± 0.72b 94.1±0.62a 103.2±0.61c
NPK
(T5) NPK 27±0.62c 64±0.81a 95± 0.63a 105±0.52b
LSD 1.139 1.206 1.209 0.251

4.3: EFFECTS OF ORGANIC AND CHEMICAL FERTILIZER ON AVERAGE

LEAF LENGTH OF SOLANUM LYCOPERSICUM

35
The average leaf length of Solanum Lycopersicum plants following various growth promoter
doses is shown in table 4.3. It is clear that after 25 days of seed sowing, Solanum
Lycopersicum's average leaf length under control circumstances was 2.1 cm. The control had
the least average leaf length (2.1 cm) while the treatment T2 had the largest average leaf
length (5.8 cm) after 30 days. The average leaf length in the remaining treatments, T1, T3,
T4, and T5, is 4.7 cm, 5.7 cm, 5.4 cm, and 5.2 cm, respectively. Plants treated with bio
fertiliser, both singly and in combination, had much longer leaves than controls.
The data unequivocally shows that the average length of the Solanum Lycopersicum leaf
was 4 cm after 50 days of growth under controlled conditions. At the conclusion of the same
developmental period, the average leaf length in the controls group was 4 cm, but the
average leaf length in treatment T4 was 9.1 cm. The average leaf length for treatments T1,
T2, T3, and T5 was 6.1 cm, 7 cm, 8.2 cm, and 7.3 cm, respectively. It is clear from Table 4.3
that the average performs noticeably better than the control. The length of the leaves was
enhanced in plants that were given chemical and bio fertiliser mixtures.
After 75 days of seed sowing, the average leaf length of Solanum Lycopersicum was
measured under controlled conditions and came out to be 6.5 cm. The advantages of the
additional growth-promoting treatments were clear, with treatment T2 recording the longest
average leaf length, 12.5 cm, after 75 days of seed germination. The average leaf length for
treatments T1, T3, T4, and T5 was 8.3 cm, 10.1 cm, 10.7 cm, and 9 cm, respectively. Plants
given bio fertilisers in addition to chemical fertiliser had improved average leaf length when
compared to the control.
Different growth promoter treatments on plants were producing a range of results. 6.8 cm
was measured after Solanum Lycopersicum seeds had grown for 100 days under controlled
circumstances. While the plants receiving control had the shortest average leaf length, the
plants getting treatment T2 had the longest average leaf length, reaching 11.2 cm. The
average leaf length of the other plants under treatments T1, T3, T4, and T5 was 6.8 cm,
whereas they were 9.1 cm, 10.9 cm, 10.8 cm, and 9.8 cm, respectively. After 100 days of
seed growth, it is clear that plants fed with a bio fertiliser combination were outperforming
controls.

Table 4.3: Average Leaf Length of Solanum Lycopersicum Recorded after Different
Intervals

36
 Treatments Average leaf Average leaf Average leaf Avg. leaf
length after 25 length length length
days per plant recorded after recorded after recorded after
5o days per 75 days per 100 days per
plant plant plant
Control 2.1± 0.15a 4±0.52a 6.5± 0.16a 6.8± 0.46a
(T1) 4.7± 0.15d 6.1± 0.7a 8.3±0.30d 9.1±0.37d
Azotobacter
(T2) FYM 5.8± 0.46a 7±0.15a 12.5±0.45a 11.2±0.62a
(T3) 5.7± 0.25a 8.2± 0.25d 10.1± 0.55a 10.9± 0.62d
Azotobacter
and FYM
(T4) FYM and 5.4±0.20a 9.1± 0.6b 10.7±0.25a 10.8±0.37b
NPK
(T5) NPK 5.2± 0.21a 7.3± 0.5a 9± 0.21b 9.8± 0.46d
LSD 0.100 1.146 0.152 0.107

4.4: EFFECTS OF ORGANIC AND CHEMICAL FERTILIZER ON AVERAGE

NUMBER OF ROOTS OF SOLANUM LYCOPERSICUM

37
According to the data in Table 4.4, Solanum lycopersicum had an average of 8 roots after 25
days of seed planting under controlled circumstances. The average number of roots on
tomato plants raised in controlled settings was the fewest—just 8. On the other hand, tomato
plants treated with T2 had the most roots on average, or 34. Average root counts for other
tomato plants treated with T1, T3, T4, and T5 growth promoters were 33.2, 27.2, 19.4, and
18 respectively. It is clear from the data in table 4.4 that each biofertilizer treatment and
chemical fertiliser treatment had a considerably larger average number of roots than the
control.
An average of 12 roots were counted after the plant had grown for 50 days under controlled
conditions. The amount of roots on the plants varied depending on the treatments. The
largest number of roots, 42, were seen in the tomato plants treated with T2. The average
number of roots was significantly higher than the control in both the bio fertiliser and NPK
combined treatments and the NPK alone treatments.
Under controlled conditions, Solanum Lycopersicums growth pattern was recorded as 19
after 75 days of plant development. The growth promoter treatments' usage caused a shift in
the pattern of development. The T2 treatments had the most roots on average, 46. The
control group, on the other hand, had the fewest average roots (19). For T1, T3, T4, and T5,
there were, on average, 45.1, 42.9, 40.5, and 38.5 roots each treatment. After 75 days of
plant growth, it is obvious that the average number of roots at each biofertilizer and chemical
fertiliser treatment was larger than the control.
The data unequivocally shows that Solanum Lycopersicum has an average root count of 24
after 100 days of plant growth under controlled conditions. While the control group had the
lowest average root number (24), treatment T2 had the highest average root number (48).
The average root number for treatments T1, T3, T4, and T5 was reported as 47.2, 44.1, 42.9,
and 43, respectively. The results in Table 4.4 show that the average number of roots was
substantially larger when a bio fertiliser combination was used as contrast to a control or
other treatments.

Table 4.4: Average No of Roots of Solanum Lycopersicum Recorded After Different

Interval

38
 Treatments No of roots No of roots No of roots No of roots
recorded after recorded after recorded after recorded after
25 days per 50 days per 75 days per 100 days per
plant plant plant plant
Control 8± 0.75a 12± 0.47a 19± 0.66a 24± 0.62a
T1 Azotobacter 33.2± 0.53a 41.4± 0.6a 45.1± 0.39a 47.2± 0.83b
T2 FYM 34± O.75d 42± 0.53d 46 0±.53a 48±0.63a
T3 Azotobacter 27.2± O.64b 39.1± 0.53b 42.9± 0.39d 44.1± 0.53a
and FYM
T4 FYM and 19.4± 0.75a 36.8 ±0.39a 40.5± 0.39b 42.9± 1.6b
NPK
T5 NPK 18± 0.62a 35± 0.47a 38± 0.38b 43± 0.81d
LSD 0.191 0.636 0.110 0.191

4.5: EFFECTS OF BIO AND CHEMICAL FERTILIZER ON AVERAGE ROOT

LENGTH SOLANUM LYCOPERSICUM

39
After learning about the usual root length of tomato plants from table 4.5. It was obvious that
Solanum Lycopersicum had an average root length of 1.5 cm after 25 days of controlled seed
planting. The control had the least average root length (1.5 cm) while the treatment T2 had
the largest average root length (4.5 cm) after 25 days of plant development. The average root
length of Solanum lycopersicum was measured under treatments T1, T3, T4, and T5, and it
was found to be 3.2 cm, 4.9 cm, 5.1 cm, and 4.5 cm, respectively. Root length was increased
when bio fertiliser was used in a different combination than the control.
A range of growth patterns are shown in data from Solanum lycopersicum plants that have
been growing for 50 days. A tomato plant's average root length under controlled
circumstances was 3.2 cm. Under controlled settings, a tomato plant's average root length
was 3.2 cm at its smallest, and 8.2 cm at its longest under treatment T2. Tomato plants under
treatments T1, T3, T4, and T5 performed better than controls when bio fertiliser was
administered in various combinations and NPK alone.
The Solanum lycopersicum plants cultivated under controlled conditions had an average root
length of 6 cm after 75 days, according to the data in table 4.5, while tomatoes produced
with various growth booster treatments had varying results. The tomato plant with treatment
T2 had the largest average root length at 12.3 cm after 75 days of seed germination, whereas
the tomato plant with treatment T4 had the least average root length at 10.1 cm. T1, T3, T4,
and T5 growth promoter administrations resulted in an average root length of tomato of 11.2
cm, 11.9 cm, 10.1 cm, and 9.8 cm, respectively.
After 100 days of plant development, Solanum lycopersicum had an average root length of
6.4 cm under control circumstances. The average root lengths of tomato plants grown with
chemical fertilisers T4 and T6 were found to be the shortest at 6.4 cm and 7.2 cm,
respectively. The average root length was 12 mm in the tomato treatment T2 when combined
with an organic growth stimulator. There were readings of 11.8 cm, 11.3 cm, 10.9 cm, and
9.3 cm for the remaining tomato plants under treatments T1, T3, T4, and T5, respectively.
Longer roots were seen in plants treated with bio fertiliser combinations compared to the
control.

Table 4.5: Average Root length of Solanum Lycopersicum after Different Intervals

Treatments Root length Root length Root length Root length

40
 recorded after recorded after recorded after recorded after
25 days per 50 days per 75 days per 100 days per
plant plant plant plant
Control 1.5± 0.16a 3.2± 0.16a 6± 0.36a 6.4± 0.26a
(T1) 3.2± 0.10a 7.1± 0.07a 11.2±0.10 a 11.8± 0.58a
Azotobacter
(T2) FYM 4.5± 0.10a 8.2±0.5d 12.3± 0.10d 12.5± 0.20a
(T3) 4.9± 0.13a 7.9± 0.5b 11.9 0±.25b 11.3± 0.15a
Azotobacter
and FYM
(T4) FYM and 5.1± 0.5b 7.1± 0.5b 10.1± 0.15b 10.1± 0.26b
NPK
(T5) NPK 4.5± 0.6d 6.5± 0.16a 9.8± 0.36d 9.3± 0.20a
LSD 0.152 1.150 0.010 0.892

4.6: EFFECTS OF ORGANIC AND CHEMICAL FERTILIZER ON TOTAL

PHENYLALANINE AMMONIA LYASE CONTENT OF SOLANUM
LYCOPERSICUM (UNIT/MG OF PROTEIN)

41
Table 4.6 displays the phenylalanine lyase concentrations in Solanum Lycopersicum at
various development stages. The PAL value was 0.014 after 25 days of seed sowing under
monitoring, according to the data. The results of tomato plants treated with various growth
boosters varied, and T5 had the highest PAL values, which were 0.054. The average PAL
values for treatment T2 were the lowest, at 0.021. The PAL values were 0.026, 0.012, and
0.037 (units/mg) of protein for additional tomato plants treated with T1, T3, and T4,
respectively. According to the results in Table 4.6, plants treated with biofertilizer in
addition to NPK exhibited higher levels of PAL activity than plants treated with NPK alone.
After 50 days of plant development, observation showed that Solanum Lycopersicum had a
PAL value of 0.024 under controlled circumstances. The greatest PAL values were seen in
tomato plants treated with growth stimulants (T5), which were 0.034, respectively. It was
discovered that tomato plants under treatment T4 had the lowest average PAL value of 0.059
whereas wheat plants under treatments T1, T2, T3, and T4 had average PAL values of 0.038,
0.039, 0.056, and 0.059 (units/mg) of protein, respectively. The results of Table 4.6
demonstrate that PAL activity was greater than control for each biofertilizer and chemical
fertiliser treatment.
After 75 days of seed sowing, the Solanum Lycopersicums average PAL content was
discovered to be 0.042 under control circumstances. The least amount of PAL was present in
Treatment T1, with values of 0.034. The treatment T5 with inorganic growth promoter
combinations had the highest PAL level, which was 0.084. The remaining tomato plants
from treatments T2, T3, and T4 had PAL values of 0.049, 0.062, and 0.068, respectively. In
plants treated with chemical fertiliser alone and in various combinations, the average PAL
activity was much higher.
After 100 days of growth, it was clear that the PAL value in the control environment had
been reported as 0.060 for SolanumLycopersicum plants.Examining plants that had received
various treatments, it was found that T5 treatments, which mostly employed chemical
fertilisers, had the highest average PAL values, coming in at 0.094. The lowest PAL values
were seen in T4-treated tomato plants, at 0.090. The remaining tomato plants were subjected
to treatments T1, T2, and T3, and their corresponding PAL values were 0.071, 0.078, and
0.083 (units/mg of protein).

Table 4.6: Total Phenylalanine Lyase (PAL) Content of Solanum Lycopersicum

Recorded after Different intervals

Treatments Total PAL Total PAL Total PAL Total PAL

42
 recorded recorded after recorded recorded after
after 25 days 50 days after 75 days 100 days
Control 0.014 ±0.011c 0.024 ±0.017b 0.042 ±0.032b 0.060 ±0.001d

T Azotobacter 0.026 ±0.001bc 0.038 ±0.001a 0.034 0.071 ±0.001c

1 ±0.0024b
T FYM 0.021 0.039 ±0.008ab 0.049 ±0.001a 0.078 ±0.13b
2 ±0.0025abc
T Azotobacter 0.012 0.056 ±0.03ab 0.062 0.083 ±0.04ab
3 +FYM ±0.0014a ±0.0014a
T FYM + NPK 0.037 ±0.060a 0.059 ±0.07ab 0.068 ±0.06b 0.090 ±0.020a
4
T NPK 0.054 ±0.06a 0.034 ±0.02ab 0.084 ±0.014a 0.094 ±0.13b
5
LSD 0.742 0.667 0.921 0.854

43
4.7: EFFECTS OF ORGANIC AND CHEMICAL FERTILIZER ON TOTAL
PROTEIN CONTENT OF SOLANM LYCOPERSICUM
The results in Table 4.7 show the protein content of Solanum lycopersicum following
various growth promoter administrations. Under regulated circumstances, the protein content
of Solanum Lycopersicum was 1.62 after 25 days of seed germination. The greatest protein
content values, 2.09 and, respectively, were displayed by tomato plants treated with T5 and.
Tomato plants treated with T2 exhibited the lowest protein content at 1.92 and, respectively.
The remaining tomato plants that received treatments T1, T3, and T4 had reported protein
contents of 2.03, 1.96, and 1.94, respectively. Chemically fertilised plants had higher protein
content after 25 days after seed sowing.
After 50 days of growth, the protein level of tomatoes produced under controlled conditions
was 1.90. The lowest protein concentration was seen in tomato plants treated with T2 2.0.
The amount of protein in plants that had received T4 treatment was 2.37. The protein content
of wheat plants under the growth promoter treatments T1, T3, and T5 was 2.23, 2.35, and
2.19, respectively.
Tomatoes under regulated settings had a protein level of 2.12 after 75 days of seed sowing.
After 75 days of plant development, wheat under treatment T5 was found to have the
greatest protein content, at 3.36. The lowest protein content, 2.18, was found in tomato
plants treated with T1. The protein concentrations in the T2, T3, and T4-treated wheat plants
were 2.41, 3.02, and 3.02, respectively.
Table 4.7 makes it clear that Solanum Lycopersicum had a protein content of 2.97 during
100 days of controlled growth. The protein content rose and was 3.21, 3.28, and 3.40,
respectively, in tomato plants treated with T1, T2, and T3. T4 treatment resulted in wheat
plants with varying protein levels of 3.37. The greatest protein concentration, 4.09, was seen
in tomato plants that had received T5 treatment. The results demonstrate that chemical
fertiliser treatments, both alone and in combination, improve the protein content of plants

44
Table 4.7: Total Protein content of Solanum Lycopersicum Recorded after Different
intervals

Treatments Total Total Total protein Total protein

protein protein content content recorded
content content recorded after after 100 days
recorded recorded 75 days
after 25 after 50
days days
Control 1.62±0.03a 1.90±0.02b 2.12±0.03a 2.97±0.01ab

T Azotobacter 2.03±0.01b 2.23±0.10b 2.18±0.03b 3.21±0.01ab

1
T FYM 1.92±0.02b 2.0±0.03b 2.41±0.04b 3.28±0.08ab
2
T Azotobacter 1.96±0.01b 2.35±0.07b 3.02±0.02b 3.40±0.03ab
3 +FYM
T FYM + NPK 1.94±0.03b 2.37±0.10b 3.02±0.01b 3.37±0.02ab
4
T NPK 2.09±0.02b 2.19±0.08b 3.36±0.03b 4.09±0.01ab
5
LSD 1.057 0.412 1.376 0.062

45
4.8: EFFECTS OF ORGANIC AND CHEMICAL FERTILIZER ON TOTAL
CATALASES CONTENT OF SOLANUM LYCOPERSICUM (UNIT/MG OF
PROTEIN)
Table 4.8 displays the value of catalysts in Solanum Lycopersicum after various growth
promoter treatments. After 25 days of seed sowing, the value of catalysts in tomatoes
cultivated under controlled conditions was 2.96. The highest catalyses values, 6.06 and 8.13,
were found in tomato plants treated with T2 and T5, respectively. With catalyses values of
4.89 and 3.57, respectively, treatments T3 and T4 that mostly employed chemical fertilisers
had the lowest results. According to Table 4.8's findings, catalases were substantially greater
under each chemical and bio fertiliser treatment than they were under the control.
It can be demonstrated that after 50 days of plant development under controlled conditions,
the catalyses value was 3.21. Tomato plants receiving treatment T2 have the lowest catalyses
value, 1.25. The greatest amounts of catalyses, 8.66, were found in wheat plants that had
received T5. T3 and T4 treatments produced results for extra wheat plants that were 7.92 and
7.57 units/mg of protein, respectively.It can be demonstrated that after 50 days of plant
development under controlled conditions, the catalyses value was 3.21. Tomato plants
receiving treatment T2 have the lowest catalyses value, 1.25. The greatest amounts of
catalyses, 8.66, were found in wheat plants that had received T5. T3 and T4 treatments
produced results for extra wheat plants that were 7.92 and 7.57 units/mg of protein,
respectively.After 75 days of seed sowing, the catalyses value of tomato plants produced
under controlled conditions was 4.76. While wheat plants treated to various treatments had a
range of reactions. Treatment T1 had the lowest catalyses levels, 5.17. The tomato plants
treated with T5 had the greatest catalyst concentration (8.78, respectively). Additional
tomato plants exposed to the growth promoter treatments T2, T3, and T4 had catalyst values
of 7.14, 7.45, and 8.10, respectively.At 100 days of plant growth, chemical fertiliser alone
and in combination treatments both increased catalases activity in comparison to control
plants.
After 100 days of plant development, it became clear that the catalytic contents of several
growth promoter administrations varied. The catalyst content in wheat plants was 5.52 after
100 days of controlled growth. It was noticed in tomato plants that had been exposed to the
growth promoters T1, T2, and T3, which have catalyses in decreasing order of 5.90, 7.54,
and 8.90, respectively. The highest catalyst concentration reading, 8.93, was seen in the

46
tomato plants receiving treatment T5. T4 was given to plants, and the plants had catalytic
levels of 8.37 units/mg of protein. Table 4.8's data make it clear that plants treated with NPK
alone had higher catalase activity.
Table 4.8: Total Catalases Content of Solanum Lycopersicum Recorded after Different
Intervals

Treatments Total Total catalyses Total catalyses Total catalyses

catalyses recorded after recorded after recorded after
recorded 50 days 75 days 100 days
after 25
days
Control 2.96± 0.03a 3.21± 0.09ab 4.76± 0.02ab 5.52± 0.02ab

T Azotobacter 4.53± 0.04a 4.68± 0.03a 5.17± 0.05ab 5.90± 0.04ab

1
T FYM 6.06± 0.010a 7.25±0 .05ab 7.14± 0.01bc 7.54± 0.01ab
2
T Azotobacter 4.89± 0.03a 7.92± 0.03ab 7.45± 0.05ab 8.90± 0.03a
3 +FYM
T FYM + NPK 3.57± 0.06a 7.57± 0.03ab 8.10± 0.02c 8.37± 0.01ab
4
T NPK 8.13± 0.03a 8.66± 0.09ab 8.78± 0.05ab 8.93± 0.02ab
5
LSD 0.657 0.562 0.763 0.660

47
4.9: EFFECTS OF ORGANIC AND CHEMICAL FERTILIZER ON TOTAL
PEROXIDASES CONTENT OF SOLANUM LYCOPERSICUM (UNIT/MG OF
PROTEIN)
The data shown in Table 4.9 illustrates the Solanum Lycopersicum Peroxidases Content
under various Growth Promoters Treatments. After 25 days after seed germination, the
tomato plant's peroxidase concentration under control conditions was 0.34. Tomato plants
showed varied Peroxidases levels when distinct growth promoter administrations were made.
Plants receiving treatment T6 had the highest peroxidase levels, or 1.18, as could be
demonstrated. The lowest peroxidase value was seen in T5-treated wheat plants, which was
0.36. The peroxidases in other tomato plants that had received treatments T1, T2, and T3 had
respective amounts of 0.71, 0.65, and 0.81 unit/mg of protein.Plants exposed to various
chemical and biological fertilisers showed considerably greater average peroxidases contents
than controls.
The peroxidase levels of tomato plants after 50 days of controlled growth were reported to
be 0.55. Treatment T5, which had a peroxidases content value of 1.00, had the highest
results. On the other hand, plants getting T4 therapy had the lowest Peroxidase values (0.57).
Following treatments T1, T2, T3, and T4, the remaining tomato plants showed values of
0.83, 0.82, 0.93, and 0.57 unit/mg of protein, respectively. After 50 days, plants treated with
a bio fertiliser combination had higher peroxidase activity than those treated with NPK
alone.
75 days following the formation of the plants Tomato peroxidase levels were determined to
be 0.81 under control conditions. The greatest peroxidase content was found in the tomato
plants treated with T5, which was 2.00 in accordance with that treatment. The lowest
peroxidase levels, 0.83, were seen in tomato plants treated with T4. The results were 0.91,
1.19, and 1.21 unit/mg of protein, respectively, for additional plants receiving growth
promoters under treatments T1, T2, and T3. Plants treated with chemical fertiliser mixtures
had greater levels of peroxidase activity than controls.
The tomato plant's peroxidase concentration was determined to be 0.82 after 100 days of
normal growth. The highest peroxidase levels were seen in the plants receiving treatment T2,

48
which were 3.02. The lowest peroxidase concentration was found in tomato plants treated
with T4, which had a 1.36 value. Peroxidase values were 2.09, 1.55, and 2.72 units/mg of
protein for the remaining tomato plants following treatments T1T3 and T5. Compared to the
control, plants treated with biofertilizer and chemical fertiliser alone had higher peroxidase
activity.
Table 4.9: Total Peroxidases Content of Solanum Lycopersicum Recorded After
Different Intervals

Treatments Total Total Total Total

peroxidases peroxidases peroxidases peroxidases
recorded recorded recorded after recorded after
after 25 days after 50 days 75 days 100 days
Control 0.34 ±0.13b 0.55 ±0.03ab 0.81 ±0.05ab 0.82 ±0.01ab

T1 Azotobacter 0.71 ±0.03ab 0.83 ±0.01ab 0.91 ±0.03ab 2.09 ±0.01a

T2 FYM 0.65 ±0.01ab 0.82 ±0.01a 1.19 ±0.04ab 3.02 ±0.02a
T3 Azotobacter 0.81 ±0.03ab 0.93 ±0.01a 1.21 ±0.01a 1.55 ±0.01a
+FYM
T4 FYM + NPK 0.36 ±0.04a 0.57 ±0.03ab 0.83 ±0.01ab 1.36 ±0.02b
T5 NPK 1.18 ±0.03b 1.00 ±0.02ab 2.00 ±0.05ab 2.72 ±0.01a
LSD 0.418 0.547 0.387 0.518

49
Fig 4.1 Control Plantafter25 Days Fig 4.2Plants treaded with FYM after 25 Days

Fig 4.3Plant treated with NPK after 25 Days

50
Fig 4.4Control Plant after 50 Days Fig 4.5Plants treaded with FYM after 50 Days

Fig 4.6Plant treated with NPK after 50 Days

51
Fig 4.7Control Plant after100 DaysFig 4.8Plants treaded with FYM after 100 Days

Fig 4.9Plant treated with NPK after 100 Days

52
CHAPTER 5
DISCUSSION AND CONCLUSION
5.1 DISCUSSION
To increase crop output and desired physiological characteristics, farmers have been
experimenting for years with chemical fertilisers. Despite the fact that they destabilise the
entire ecosystem in addition to degrading the quality of the soil and the health of the plants,
farmers still prefer to use chemical/inorganic fertilisers to produce crops more quickly and
with a larger yield. The physiological and biochemical characteristics of tomatoes (Solanum
Lycopersicum) grown with the use of chemical and organic fertiliser, as well as their various
combinations, are being compared in the current study. This comparison was made in order
to ascertain if utilising organic compost or its combinations with chemical fertiliser are a
healthier alternative to using chemical fertiliser alone for the development of (Solanum
Lycopersicum).
Height 5.1.1: Investigation of Effect of Chemical Fertilizer and Organic Compost on
Average Plant
The findings indicate that the plants treated with FYM (T2) attained their maximum height
(64.1 cm) after 100 days of growth. It is crucial to discuss the numerous biological fertiliser
components that help Solanum lycopersicum plants get higher. Bio fertilisers, as is widely
known and supported by several studies, help plants effectively assimilate nitrogen and
prevent nitrogen deficiency. It appears that nitrogen is necessary for increasing plant height.
The use of Azotobacter and synthetic nitrogen fertiliser significantly increased the height of
the plants (Soleimanzadeh and Gooshchi, 2013). The lowest recorded plant height (32.4 cm)
was found in the plants treated with FYM and NPK. The culprit might be that chemical.
Fertiliser draining out of the soil causes a nitrogen shortage in plants, which is Low plant
height and other physiological characteristics of plants are tightly related (Shabaan et al.,
2006).
The findings of this study demonstrated that the use of organic fertilisers, such as manure
and vermicomposting, had a favourable impact on typical tomato yield components and that
doing so in conjunction with P fertiliser increased those advantages (Cheraghi, Mohyedi, and
Kalhor, 2016).

53
5.1.2: Investigation of Effect of Chemical Fertilizer and Organic Compost on Average
Leaf number/length
The findings of this study demonstrated that the use of organic fertilisers, such as manure
and vermicomposting, had a favourable impact on typical tomato yield components and that
doing so in conjunction with P fertiliser increased those advantages (Cheraghi, Mohyedi, and
Kalhor, 2016).
Farmyard manure improves soil structure, increases nutrient availability, and facilitates the
repair of alkali soils, all of which lead to increased and better straw output (Zaki et al.,
2009). In addition to fixing nitrogen molecules and delivering them directly to plants,
Rhizobium and Azotobacter (bio-fertilizers) are also in charge of creating plant growth
hormones (Auxins, GAS, and CKS) that can be released in the root media and have a
beneficial effect on the plant's extension and growth. Potential contributing components in
the bio fertilised plants include increased cell division and enlargement, increased creation of
tissues and organs, increased assimilation of nitrogenous compounds 3, increased generation
of growth chemicals (GA, IAA, IBA, and KI), and increased cell division and enlargement.
These results suggest that farmyard waste improves soil.increases nutritional availability and
gives structure.
In compared to either the equivalent positive controls or the single biofertilization, the
treatment of combination bio-fertilizers (Azotobacter + Rhizobium) in the presence of a
complete dose of farmyard manure recorded the most significant results (Ibrahim et al.,
2011).
5.1.3: Investigation of Effect of Chemical Fertilizer and Organic Compost on Root
number/length
In the current study, average root length and average root number were used as
supplementary characteristics to evaluate the effects of chemical fertiliser and organic
compost. The most roots (48) were found on plants that had received FYM treatment (T2).
The longest root possible during treatment (T2) FYM was 12.5 cm long. In comparison to
the unfertilized plot, the treatment receiving fertiliser had a positive impact on the root
growth parameters such as root length and root length density at all growth stages.
Previous studies have demonstrated that organic amendments can encourage the growth of
wheat roots. This may be connected to how organic fertilisers, which comprise amino acids
and certain physiologically active chemicals, boost biological activity in crop rhizosphere
(Zhou and Luo, 1997; Prasert, 1997). The environment of the soil has a considerable impact
on how plant roots develop. By enhancing the physical and chemical conditions of the

54
rhizosphere soil, the addition of organic manure to the soil can benefit crop root
development (Sidira et al., 2001). According to Rakesh et al., (2001) and Mandal et al.,
(2003), using farmyard or green manure in addition to mineral fertilisers can significantly
prolong crop roots and improve the depth of root penetration.
5.1.4: Biochemical Investigation of Tomatoes Grown under Chemical Fertilizer and
Organic compost
5.1.4.1 Phenylalaninelyase
The plants given treatment T6 have the greatest levels of PAL activity, according to the
current study (0.094 unit/mg of protein). Azolobacter and FYM (T3), among other
biofertilizers, have been applied to plants, and these plants have also demonstrated good
PAL activity. Both positive and negative impacts on the size of the microbial community
(MBC) and on microbiological activities have been observed for farmyard manure and
inorganic fertilisers (NPK) (Böhme et al., 2005). Studies in temperate and subtropical
locations suggest that the availability of C and N in soils may have an impact on how soil
microbial communities use their substrates (Tate 2000; Zhang et al., 2008; Zhong et al.,
2010). Notably, communities in 100% N-treated plots and untreated controls were unable to
use L-serine as Communities could exploit the only carbon source in 100% RD of NPK,
150% RD of NPK, and 100% RD of NPK+FYM treated plots (Chakraborty et al., 2011).
According to some theories, nitrogen-deficient plants boost PAL activity to increase
ammonia availability, which causes polyphenol chemicals to accumulate. Increased PAL
activity is typically observed in plants with low nitrogen levels. Strissel et al. discovered that
PAL activity was down-regulated and flavonoid accumulation was decreased in young plant
leaves that had undergone high nitrogen fertilisation (Ibrahim et al., 2011).
5.1.4.2 Protein content
Plants given nitrogen fertiliser (treatment T6) in the current study exhibited the greatest
protein content (4.09 units/mg of protein). These results are consistent with earlier research
(Evans, 1983; Makino et al., 1984, 1985), which discovered that nitrogen supply influences
the protein composition of leaves and cells, which in turn impacts absorption rate. As a
result, before discussing CO2 fixation control, the structural changes caused by differences
in nitrogen are explored (Lawlor, Kontturi, and Young). The overall protein content of
tomato kernels increased consistently when additional nitrogen fertiliser was added, and
there were different patterns for the component. As additional nitrogen was added, the
kernel's total protein content steadily grew, while albumin, globulin, and gluten levels
exhibited the reverse trend. With N225 having the lowest residual protein content and N300

55
having the greatest residual protein content, there were no appreciable differences in residual
protein content amongst the various treatments. According to these results, an increase in
nitrogen levels was advantageous to a rise in kernel protein levels within a certain range (Liu
and Shi, 2013).
While some researchers (Rashid et al., 2004) showed that globulin and albumin decreased
yield increases, contrary to the beliefs of some researchers who believed that the protein
each component content increased significantly with the increasing amount of nitrogen
fertiliser, gliadin and gluten increased slowly with the increasing amount of nitrogen
fertiliser. With the administration of potassium humate, potassium silicate, and their
combination, the overall amount of soluble proteins and carbohydrates increased in
comparison to the control. However, potassium humate alone significantly decreased total
soluble proteins and carbohydrates by 38% and 35%, respectively, whereas potassium
silicate alone and in combination with potassium humate had the same impact, lowering total
soluble proteins and carbohydrates by 18% and 28%, respectively evaluation in contrast to
untreated controls (Osman et al., 2017).
5.1.4.3 Catalases activity
The current investigation has showed that plants given the chemical fertiliser treatment T6
had the highest catalase activity (8.93 units/mg of protein).
In general, it can be concluded that organic fertilisation required the available nitrogen to be
released slowly, whereas mineral fertilisation was able to provide control plants with higher
amounts of nitrogen in a short amount of time to improve better metabolism without
requiring additional enzyme activity. This might be thought of as unfavourable
environmental circumstances caused by nutritional deficits, and antioxidant enzymes are
essential for assisting plants in adapting to potential disturbances in a variety of
physiological processes. Additionally, bio stimulants or N2 fixation treatment has a positive
impact on crop yield in order to In order to establish links between mineral nutrient intake,
bio stimulation, and certain plant defence systems, the use of bio stimulants or N2 fixation
has a positive impact on crop output (Ahmed et al., 2010).
The activity of the catalase enzyme increased when tomato plants were infected with
Azotobacter. Organic manure has higher concentrations of readily available nutrients, which
are critical for plant development.

56
5.1.4.4 Peroxidases Activity
In the present investigation, plants treated with nitrogen fertiliser (T6) showed the greatest
peroxidase activity (2.72 units/mg of protein). Similar to plants receiving treatment T3,
plants receiving bio fertiliser also shown significant peroxidase activity.
POD was strongly altered by treatment with nitrogen, salinity, variety and nitrogen
interaction, and nitrogen and salinity interaction (Ibrahim et al., 2018). When exposed to
salt, a number of plants, including maize (Kholová et al., 2009; Kholova et al., 2010), wheat
(Latef, 2010), and canola (Bybordi, 2010),
Showed an approximately enhanced CAT activity. They found that, in comparison to
controls, salt-stressed wheat cultivars showed a reduction in CAT activity. Wheat POD
activity has been demonstrated to increase under salt stress (Abdel Latef, 2010, Kahrizi et
al., 2012). Salt stress causes an increase in Antioxidant enzymes could point to a heightened
ROS impact and an enhanced defence against the oxidative damage stress inflicts on plants.
Catalase in peroxisomes breaks down H2O2. In the cytosol and chloroplast, peroxidase can
efficiently scavenge H2O2. Plants treated with salt have higher peroxidase activity,
according to Kahrizi et al. (2012). We found that the antioxidant enzymes (POD and SOD)
were somewhat less active after each nitrogen treatment. Rios-Gonzalez et al. (2002) and
Huang et al. (2004) reported similar findings, and CAT initially increased with increasing
nitrogen rate. (Huang et al., 2004) have also reported correlated increases in the CAT.
5.2 CONCLUSION
Preliminary results indicate that both biofertilizer and chemical fertilizer treatments
positively influenced tomato growth. However, significant differences were observed
between the two groups. The tomato plants treated with biofertilizer exhibited enhanced
vegetative growth, with increased plant height, larger leaf area, and a higher number of
branches compared to the chemical fertilizer group. Moreover, the biofertilizer-treated plants
demonstrated earlier flowering and fruit development, resulting in an earlier harvest and
higher overall yield.Analysis of soil health parameters revealed that the biofertilizer-treated
soil had improved nutrient availability, increased microbial diversity, and enhanced soil
structure compared to the chemical fertilizer-treated soil. These findings suggest that
biofertilizers not only promote plant growth but also contribute to the overall health and
sustainability of the soil ecosystem.
Based on these results, it can be concluded that biofertilizers are a viable alternative to
chemical fertilizers for tomato cultivation. The use of biofertilizers offers numerous
advantages, including improved plant growth, earlier flowering, higher yield, and enhanced

57
soil health. Further investigations are warranted to explore the long-term effects and cost-
effectiveness of biofertilizers in large-scale agricultural systems, considering their potential
as eco-friendly and sustainable alternatives to chemical fertilizers.

58