Analysis of Greenhouse Experiments

Greenhouse experiments

Validation of the decomposed rice straw by using the above mentioned isolates was done in a greenhouse experiment. Seeds were placed in sterile autoclaved soils filled in 12 inch x 12 inch pots (each pot having 2.5 kg of autoclaved soil and 500g of decomposed rice straw by using respective Trichoderma strains) and incubated under ambient conditions (25°C). For control treatment, seeds were sown in the pots having equal amount of soil as in treated pots and 500g of decomposed rice straw without application of Trichoderma strain.

Germination and early vigour

The germinability of the seeds in greenhouse was examined each day to calculate mean germination time till 8th days of sowing. On 8th day, germination was determined as the ratio of the number of normal seedlings to the total number of seeds and expressed as percentage. On day eight of germination, ten normal seedlings were selected randomly in each treatment from all replications for measurement of shoot and root lengths. Seedlings used for recording seedling length from each replication were subsequently weighed after removing the cotyledons and the fresh weight was expressed in gram per 10 seedlings.

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These seedlings were subsequently dried in an hot air oven at 55±1°C for 72 h and the dry weight was expressed in mg per 10 seedlings.

• Vigour index of the coated seeds were assessed based on germination percentage; seedling length and seedling dry weight.
• Vigour index was calculated by using the formula given by Abdul-Baki and Anderson, (1973)
• Vigour index -I = Germination (%) ? Seedling length (cm)
• Vigour index -II = Germination (%) ? Seedling dry weight (mg)

Determination of chlorophyll content

Chlorophyll content of the plants grown with decomposed rice straw by using respective Trichoderma strains in greenhouse was determined by following the procedure of Porra (2002).

Chopped fresh leaf tissue of 0.1 g was transferred to a capped measuring tube containing 25 ml of 80% acetone and kept inside a refrigerator at 40C for 48 h. Measurements were made using a spectrophotometer (Model- Evolution 300 UV-VIS; Double Beam Spectrophotometer, Thermo Fisher Scientific) at a wavelength of 663 nm and 645 nm to calculate chlorophyll a and chlorophyll b content, respectively.

Growth promotion

In-vitro experiment was carried out to check the effect of decomposed rice straw by using respective Trichoderma for evaluation of growth promotion by taking two direct seeded rice varieties in Kharif season of 2018 (i.e."Naveen"). Observations on health of plant (i.e. shoot length, root length, fresh shoot weight, dry shoot weight, fresh root weight, dry root weight) was recorded after 45 days and grain weight, yield/hill and yield/plot were recorded after harvesting.

Plant stress related enzyme analysis:

Expression of stress related enzymes (i.e. catalase, peroxidase and superoxide dismutase) produced by the plants grown with decomposed rice straw by using respective Trichoderma strains were analyzed according to Nounjana et al., (2012).

Statistical Analysis

Statistical analysis were performed by using the Statistical Analysis Software (SAS) of Indian Agricultural Statistics Research Institute (IASRI), New Delhi through the portal www.iasri.res.in/sscnars/-. Percentage of Seed germination data were ARCSINE transformed. The other seed quality parameters were analyzed without any transformation. All the data were subjected to one-way classified analysis of variance (ANOVA) and means of treatments were compared based on Tukey's honestly significant difference test (HSD) at 0.05 probability level using SAS. All the experiments repeated at least three times.