Effect of different types of organic composting cultures on Trigonella foenum graecum plant- an invitro study.
Smitha N1, Deepthi H1, Nagarakshitha V K2, Sushritha V K2, Chidanandamurthy G3.
Corresponding author: deepthiharishrao@gmail.com
ABSTRACT
The project is mainly focused on the effect of composting cultures on Trigonella foenumgraecum plant. The market sample of compost consisted of Trichoderma viride, Phanerochaete chrysoporium, Pleurotus sp, Aspergillus sp. The Trichoderma viride compost culture treated seed shows more root length 6.55 cm, shoot length 16.69 cm, lead area 2.20cm2, seedling vigour index 2324 and shows high chlorophyll concentration 2.781 compared to Phanerochaete chrysoporium, Pleurotus species and Aspergillus species compost culture. Therefore, from the results it is concluded that among the four-compost culture used to grow Trigonella foenum-graecum plant, the Trichoderma viride compost culture treated seeds shows increased rate of growth of the plant, number of leaves, seedling vigour index and chlorophyll concentration as compared to other compost cultures.
KEYWORDS
Trigonella foenumgraecum, Phanerochaete chrysoporium, Pleurotus species, Trichoderma viride, Aspergillus species, organic composting.
INTRODUCTION
Soils are complex mixtures of minerals, air, water, organic matter and countless microorganisms that are the decaying remains of once-living things. It forms at the surface of land- it is the “skin of the earth”. Soil is capable of supporting plant life and is vital to life in earth (Acharya, 1946).
Composting is a process involving key microbiome that actively decomposes the degradable and putrescent organic waste under moist, self-heating and aerobic conditions and is a natural process characterized by microbial community successions. Microbial community structure is defined as the composition of the microbial community and the abundance of the members in a microbial community (Bharne et al., 2002)
During the process of composting, both bacteria and fungi that represent the microbial community structure of the composting environment are present and play an active role. The presence of different bacteria or fungi can either positively or negatively affect the entire composting process. Their diversity also suggests the composting mechanisms which takes place. Likewise, alteration in the choice and amount of initial organic matter can change the microbial communities and the composting output. Diversity and development of microbial population during composting are dependent on physical parameters such as oxygen, temperature, moisture content and nutrient availability. The physical parameters are crucial in understanding the activity of different groups of microorganisms that will indicate the quality of the final product of the composting process (Bhasme et al., 2006).
Usage of microorganisms in creating compost has been practiced from ancient times. All the agriculture wastes were processed by dumping in specific area and allowed to convert into manure by the natural process, even today it is followed in many areas, it is unscientific, time consuming and it may lead to accumulation of toxic chemicals in the manure. It also unsafe for the application of field as it may consist many pathogenic strains of microorganisms. This may hamper the yield of the farm resulting in the losses and spreading of the plant diseases. The area used for composting may also harbour consortia of microorganisms which may be pathogenic to humans and animals and may lead to disease spreading and development. Unscientific composting may lead to release of harmful gases which may contaminate the surroundings (Richardson et al., 2002).
The study was undertaken because the Compost Culture is formulated by Institute of Organic Farming, University of Agriculture Studies, Dharwad, Karnataka there is high usage of the compost in agricultural fields across Karnataka, analysis of this sample is important to understand the efficiency of the product and its application.
MATERIALS AND METHODS
The experiment was conducted in Shree Siddaganaga College of Arts Science and Commerce for Women, B H road Tumkur. The materials, procedures, and methodologies used are discussed below.
Study plant: For analysis T. foenum-graecum was selected for the following reasons; T. foenum-graecum is an N-fixing plant, thus reducing the need of nitrogen fertilizers for subsequent crops, It has low water requirements, It is leguminous plant, i.e. it has ability to form symbiotic relation with rhizobium, It usually takes 1 to 2 weeks for germination; hence it is easy for monitoring the pre plant effects, It requires less space and depth for growth, it is feasible for cultivating under potted conditions, The best average temperature to grow your plants is 18 to 24'C (64 to 75'F), so there was no thermal regulation required in growth site and seeds are easily available in local market.
Sample:
It was taken from ‘Consortia of Compost Culture’ formulated by ‘Institute of Organic Farming’, University of Agriculture Studies, Dharwad, Karnataka. Four separate composting cultures Aspergillus sps, Phanerochaete chrysoporium, Trichoderma viride, and Pleurotus sps were taken for the experiment.
Processing of sample:
Compost cultures are devoid of nutrients. Hence there is need for farm yard manure during application, cow dung serves as an excellent manure source. Cow Dung is a nutrient rich fertilizer, excellent growing medium for plants. It can be mixed into the soil or used as top dressing. Cow Dung Compost has balanced NPK content (Nitrogen, Phosphorus, and potassium) which is essential for plant growth. Moreover, it also eliminates harmful ammonia gas and pathogens, as well as weed seeds. The composted cow dung manure adds generous amounts of organic matter to the soil. The moisture-holding capacity of the soil can also be promoted by mixing this compost into soil. This allows sprinkling of water less frequently, because the roots of plants can use the additional water and nutrients from the manure whenever needed. Advantage of using this manure is that it increases the breakup of compacted soils through aeration in the soil. It has a tendency to convert nutrients into easily accessible forms so they can be slowly released without burning tender plant roots thus it is like protective shell to the delicate roots of the crop.
This is required for the activation of culture. The cultures available in the market are inactivated and are in the dehydrated state. They usually contain spores mixed with the transport media (peat or charcoal). It is important for reactivation of the cells. Mixing the mixture with cow dung and incubation facilitates the spore maturation and hyphal development Pramod et al., 2015).
Method of biofertilizer application
The activated mixture was then mixed 2000g of red potting soil and packed into the sterile pots respectively. The whole process was done in duplicates to minimise the errors and have more accurate results (Pramod et al., 2015).
Seeding:
The seeds of Trigonella foenum-graecum (Fenugreek) were taken from a local market, were used as explants. The seeds were washed thoroughly under tap water; surface sterilization was done using 70% ethanol. 10 seeds per pot were planted at equal spacing.
Seeds are not sterile; sperm sphere consist of variety of microorganisms which effect in the growth of plant. Hence it necessary to eliminate additional microorganism. 70% ethanol is best used disinfectant, when exposed eliminate most of the superficial organisms. After ethanol treatment it should be rinsed thoroughly with distilled water to avoid residual ethanol on seed surface (Pramod et al., 2015).
Growth:
The seeded plant was kept in well ventilated and lit up area. Periodic watering was done using distilled water. Distilled water is free from waterborne biological contaminants such as bacteria, viruses, organic and inorganic chemicals, heavy metals, volatile gases, cysts and other contaminants. Distilled water contains virtually no solids, minerals or trace elements. Growth was keenly observed and was noted from the time of sprouting till harvest. The seeds took 7-9 days for germination and took about 3 weeks for 8 leaf stage growth.
The length of each growth stage is greatly influenced by temperature, moisture, light (day length), nutrition and variety. The growth rate of the crop is closely related to the amount of solar radiation captured by the leaves. Nitrogen is a key component of chlorophyll, the green pigment in plants, so it's the critical nutrient when their energy is focused on growing stalks and foliage. At this stage the chlorophyll is at its matured form yet not in degradative stage, so corelation of the chlorophyll parameters indicates the further health of the plant and its development.
Following observation was done from the day of germination
Extraction was done by prior moistening of the soil. The whole plant including the root was carefully taken out from each pot (Pramod et al., 2015).
Measurements:
Plant height
The plant was recorded at the time of uprooting from the basal node of the plant to the base of the newly opened leaf and expressed in centimetre (Pramod et al., 2015).
Seedling vigour index
Vigour testing does not only measure the percentage of viable seed in a sample, it also reflects the ability of those seeds to produce normal seedlings under less than optimum or adverse growing conditions. Seed vigour is the sum of those properties which determines the potential level of activity and performance of seed during germination and seedling emergence.
Total plant length from shoot tip to the root tip, shoot length (from plumule to the tip), root length (from plumule to the root tip) and the leaf length was measured and seedling vigour was calculated using the formula.
Seedling vigour = (avg.root length+avg.shoot length) × % of seed germination
Leaf area
Leaf area was worked out by following the procedure
Leaf area cm2 of each leaf = L x W
Where, L = length of the leaf and W = maximum width of the leaf. The average leaf area of 10 plants were worked out (Pramod et al., 2015).
Chlorophyll estimation
RESULTS
The experiment was conducted in invitro condition. The test plant taken was Trigonella foenum-graecum (Fenugreek). It was inoculated with the compost cultures according to the specifications above. Following results were calculated according to five different parameters, Root length, shoot length, leaf area, seedling vigour, chlorophyll content.
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seeding (Day 0) |
Plant growth (Day 13) |
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Plant growth (Day 24) |
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Figure 1: Experimental plant growth pictures at different time intervals |
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Total plant length of the experimental plants
The total plant length of the plant after 24 days was recorded, in the observation the plants treated with Trichoderma viride showed maximum plant length (23.21±3.09) followed by Pleurotus sps (19.55±0.66), P. chrysoporium (18.73±2.33) and least plant length was observed in Aspergillus sps (17.24±1.78). The control group showed moderate plant growth (14.85±0.75) (Table. 01 & Figure 02, 03).
Table. 01: Total plant length (in cm) of experimental plants
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Sl Num. |
Control |
Aspergillus sps |
Phanerochaete chrysoporium |
Trichoderma viride |
Pleurotus sps |
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1 |
14.2 |
13.2 |
15.0 |
20.6 |
14.2 |
17.2 |
17.1 |
24.9 |
19.9 |
21.0 |
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2 |
15.7 |
14.9 |
15.0 |
19.0 |
21.0 |
20.0 |
23.3 |
20.2 |
19.3 |
19.3 |
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3 |
14.9 |
15.1 |
17.0 |
18.6 |
18.4 |
21.4 |
25.9 |
26.2 |
18.9 |
19.2 |
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4 |
15.6 |
14.4 |
16.0 |
16.5 |
17.0 |
19.8 |
26.8 |
24.5 |
19.8 |
19.7 |
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5 |
15.1 |
15.4 |
17.6 |
17.1 |
17.2 |
21.1 |
20.8 |
22.4 |
18.6 |
19.8 |
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Average |
14.85±0.75 |
17.24±1.78 |
18.73±2.33 |
23.21±3.09 |
19.55±0.66 |
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Total root length of the experimental plants
The total root length of the plant after 24 days was recorded, in the observation the plants treated with Trichoderma viride showed maximum root length (6.55±0.60) followed by P. chrysoporium (5.94±1.09), Pleurotus sps (5.93±0.36) and least root length was observed in Aspergillus sps (5.63±0.62). The control group showed moderate root growth (4.86±0.71) (Table. 02 & Figure 02, 03).
Table. 02: Total Root length (in cm) of experimental plants
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Control |
Aspergillus sps |
Phanerochaete chrysoporium |
Trichoderma viride |
Pleurotus sps |
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1 |
4.2 |
4.3 |
5.8 |
6.5 |
4.4 |
5.9 |
5.7 |
7.3 |
5.6 |
6.3 |
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2 |
6.2 |
5.7 |
5.3 |
5.4 |
5.6 |
7.4 |
6.9 |
6.7 |
5.8 |
5.8 |
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3 |
5.2 |
4.8 |
5.4 |
5.8 |
6.5 |
6.2 |
5.9 |
6.5 |
6.1 |
6.4 |
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4 |
4.9 |
3.9 |
5.8 |
5.9 |
4.8 |
5.2 |
7.3 |
5.9 |
6.4 |
5.9 |
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5 |
4.3 |
5.1 |
4.2 |
6.2 |
5.5 |
7.9 |
6.2 |
7.1 |
5.7 |
5.3 |
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Average |
4.86±0.71 |
5.63±0.62 |
5.94±1.09 |
6.55±0.60 |
5.93±0.36 |
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Total shoot length of the experimental plants
The total shoot length of the plant after 24 days was recorded, in the observation the plants treated with Trichoderma viride showed maximum shoot length (16.69±2.93) followed by Pleurotus sps (13.62±0.69), Phanerochaete chrysoporium (12.79±1.81) and least shoot length was observed in Aspergillus sps (11.55±1.82). The control group showed moderate shoot growth (9.99±0.64) (Table. 03 & Figure 02, 03).
Table. 03: total shoot length (in cm) of experimental plants
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Sl Num. |
Control |
Aspergillus sps |
Phanerochaete chrysoporium |
Trichoderma viride |
Pleurotus sps |
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1 |
10 |
8.9 |
9.2 |
14.1 |
9.8 |
11.3 |
11.4 |
17.9 |
14.3 |
14.7 |
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2 |
9.5 |
9.2 |
9.7 |
13.6 |
15.4 |
12.6 |
16.4 |
13.5 |
13.5 |
13.5 |
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3 |
9.7 |
10.3 |
11.6 |
12.8 |
11.9 |
15.2 |
20 |
19.7 |
12.8 |
12.8 |
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4 |
10.7 |
10.5 |
10.2 |
10.6 |
12.2 |
14.6 |
19.5 |
18.6 |
13.4 |
13.8 |
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5 |
10.8 |
10.3 |
13.4 |
9.9 |
11.7 |
13.2 |
14.6 |
15.3 |
12.9 |
14.5 |
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Average |
9.99±0.64 |
11.55±1.82 |
12.79±1.81 |
16.69±2.93 |
13.62±0.69 |
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Total leaf area of the experimental plants
The total leaf area of the plant after 24 days was recorded, in the observation the plants treated with Trichoderma viride showed maximum leaf area (2.20±0.13) followed by Phanerochaete chrysoporium (1.45±0.15), Pleurotus sps (1.23±0.17) and least leaf area was observed in Aspergillus sps (0.64±0.10). The control group showed moderate leaf area (0.53±0.05) (Table. 04 & Figure 02, 03).
Table. 04: Leaf area (cm2) of the experimental plants
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Sl Num. |
Control |
Aspergillus sps |
Phanerochaete chrysoporium |
Trichoderma viride |
Pleurotus sps |
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1 |
0.56 |
0.61 |
0.76 |
0.46 |
1.60 |
1.72 |
2.42 |
2.32 |
1.04 |
1.06 |
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2 |
0.45 |
0.58 |
0.54 |
0.74 |
1.32 |
1.38 |
1.98 |
2.16 |
1.23 |
1.12 |
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3 |
0.51 |
0.53 |
0.65 |
0.68 |
1.54 |
1.28 |
2.34 |
2.25 |
1.34 |
1.42 |
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4 |
0.59 |
0.49 |
0.78 |
0.52 |
1.24 |
1.52 |
2.14 |
2.21 |
1.02 |
1.24 |
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5 |
0.48 |
0.55 |
0.69 |
0.64 |
1.48 |
1.44 |
2.08 |
2.12 |
1.52 |
1.32 |
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Average |
0.53±0.05 |
0.64±0.10 |
1.45±0.15 |
2.20±0.13 |
1.23±0.17 |
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Figure 02: Graph representing the total plant length, root length, shoot length and leaf area of Trigonella foenum-graecum
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control pot |
Aspergillus sps inoculated pot |
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Trichoderma viride inoculated pot |
Pleurotus sps inoculated pot |
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P. chrysoporium inoculated pot |
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Figure 03: Plants measurement from different compost inoculated pots |
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Seedling vigour index
The seedling vigour was recorded for the experimental plant, in that, the plants treated with Trichoderma viride showed maximum seed vigour index (2324 seedlings) followed by Pleurotus sps (1955 seedlings), Aspergillus sps (1878 seedlings) and least leaf area was observed in Phanerochaete chrysoporium (1718 seedlings). The control group showed moderate leaf area (1485 seedlings) (Table. 05 & Figure 04).
Table.05: Seedling vigor index
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Organism |
Control |
Aspergillus sps |
Phanerochaete chrysoporium |
Trichoderma viride |
Pleurotus sps |
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Seedling vigor Index |
1485 |
1878 |
1718 |
2324 |
1955 |
Figure 04: Seedling vigor index of Trigonella foenum-graecum seed
Chlorophyll concentration in experimental plants
The chlorophyll content of the plant after 24 days was recorded, in the observation the plants treated with Trichoderma viride (2.781) showed maximum leaf area followed by Pleurotus sps (2.281), Phanerochaete chrysoporium (1.963), and least leaf area was observed in Aspergillus sps (1.584). The control group showed moderate leaf area (1.145) (Table. 06 & Figure 05, 06).
Table.06: Chlorophyll concentration
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Organisms |
O D Readings |
Chl a |
Chl b |
Total Chl |
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645 |
663 |
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Control |
0.44 |
0.32 |
0.472 |
0.857 |
1.145 |
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Aspergillus sps |
0.55 |
0.59 |
1.108 |
0.983 |
1.584 |
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Phanerochaete chrysoporium |
0.69 |
0.71 |
0.716 |
1.247 |
1.963 |
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Trichoderma viride |
1.25 |
1.39 |
1.213 |
2.303 |
2.781 |
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Pleurotus sps |
0.82 |
0.78 |
0.831 |
1.512 |
2.281 |
Figure 05: Graph represents the chlorophyll concentration of Trigonella foenum-graecum
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Figure 06: Extraction of chlorophyll from study plant inoculated with different compost cultures |
DISCUSSION
Organic wastes are biodegradable; thus, they can be treated with appropriate energy and resources. The natural process of aerobic decomposition of organic matter can be used as the basis for the controlled treatment of waste in a process referred to as composting. Composting has been successfully used as an organic waste reduction tool worldwide.
The process of composting has been widely studied and described. Interest in compost microbiota has increased significantly due to the fact that bacteria and fungi are the two main driving forces of composting, and efficient composting is dependent on the presence of a high microbial diversity. The methods previously used (culture-dependent or fingerprinting methods) did not allow for an accurate determination of microbial successions during composting. New sequencing methods now permit researchers to obtain information on the level of both dominant and minor species, at the family, genera or individual species level. It is necessary to add that very few publications analyse bacterial and fungal communities of the composts simultaneously. Understanding the changes in microbial communities during composting may improve reproducibility, performance, quality of the final compost as well as the evaluation of potential human health risks and choice of the optimal application procedure.
The decomposition of organic matter by composting is usually divided into four main stages that differ in physico -chemical conditions such as temperature–mesophilic, thermophilic, cooling and curing/maturation. We suggest that during these stages, microbial succession takes place, and strains that decompose increasingly recalcitrant organic matter are selected.
In some cases, a single waste type is not suitable for composting because it may have high concentrations of one type of nutrient, low or high pH or other factors. Co-treatment of organic wastes with compensative properties can be more efficient in terms of time, need of fertilizers or quality of the final product. Analysis of the interaction of different organic materials as a source of their own microbial community may help in understanding the microbial successions and to increase the effectiveness of the composting process. There are different opinions concerning microbial community formation in composts. One theory state that a microbial community is influenced more by physical location and conditions of composting, while another theory associates the presence and absence of microbes with temperature fluctuations. Supporters of a third theory believe that microbial composition is determined by the initial properties and micro biota of the composting material. Therefore, the final compost is re-colonized by the organisms abundant during the mesophilic stage and those able to survive the thermophilic stage in the form of spores.
Chlorophylls are the most important green pigments in plants for the photosynthetic process. Higher plants contain Chl a, Chl b, accessory pigments and several additional forms of chlorophyll. The Chl a and Chl b are the best known among five main types of chlorophyll and are most commonly found in all autotrophic organisms except pigment containing bacteria. Chl a has formula of C55H72O5N4Mg and the empirical formula of Chl b is C55H70O6N4Mg. Chl a usually appears blue green and Chl b is yellow-green. Both Chl a and Chl b pigments are associated with light harvesting processes, which are solely responsible for photosynthesis in higher plants.
Chlorophyll concentration in leaves is an indicator of plant health. The chlorophyll a:b ratio also indicates the developmental state of photosynthetic apparatus in plants. It has a determinative role in growth and development of higher plants. The chlorophyll content also indicates the photosynthetic capacity per unit area of the leaf that determines the rate of photosynthesis in the plant. Determination of chlorophyll content as an indirect method of estimating the productivity also provides a good understanding of the photosynthetic regime of plants. The chlorophyll content increases with leaf development and then decreases with the senescence phenomenon. The rate of photosynthesis is also higher in flowering and fruiting branches of sub-tropical fruit species in comparison to non-fruiting branches.
Effect of the health status and germination potential of the seed thus checking of seedling vigor index and it gives the health status of seed, it is one of the parameter which judge the quality of the seed and if there is a presence of microorganism and it affect the seedling vigor ,i.e., there will be disease in the root and shoot length of the plant which shows the effect of pathogen on seed.
Seedling Vigor is defined as the sum of the total properties of seed which determines the level of activity and performance of seed during germination and seedling emergence.
As per the result obtained the Trichoderma viride showed the highest values of the parameters, as compared to control, Phanerochaete chrysoporium, Pleurotus sp, Aspergillus sp.
Trichoderma sp have been widely used as plant growth enhancer by creating a positive environment and releases various types of secondary metabolites including growth hormones, end chitinase, proteolytic enzymes and improve the uptake of micronutrients to plants such as Cu, Zn, Fe, Na also helps in solubilization of phosphate in soil and available to plants.
CONCLUSION
Based on the results of the present experiment conducted to analyse the best compost culture to give good healthy plant and yield.
Among the four-compost culture used to grow Trigonella foenum-graecum plant, as compared to other compost cultures the Trichoderma viride compost culture treated seeds shows increased rate of growth of the plant, number of leaves, seedling vigour index and chlorophyll concentration.
Aspergillus sps compost culture treated seeds shows decreased rate of growth of the plant, number of leaves, seedling vigour index and chlorophyll concentration.
AKNOWLEDGEMENT
We are thankful to Dr Ashwathanarayana R, Department of Botany, SSCASCW, Tumkur for correcting our original research article.
REFERENCE
