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Screening of Potential Bacterial Isolates as Specific Biofertilizer Agent for Mungbean Plants

Muhammad Tarek
Department of Microbiology, Primeasia University, 12 Kamal Ataturk Avenue, Banani, Dhaka-1213, Bangladesh

Md Imdadul Hoque
Bangladesh Institute of Nuclear Agriculture, BAU Campus, Mymensingh-2202, Bangladesh

Rubaiyat Binte Billah
Department of Microbiology, Primeasia University, 12 Kamal Ataturk Avenue, Banani, Dhaka-1213, Bangladesh

Abstract/ Executive Summary:

The Experiment was performed under glass house condition (24°8´ N 90°0´ E) with eight rhizobial strains namely MBR-3, MBI-5, MBI-19, MBM-4, MBM-8, MBP-10, MBB-3 and MBJ-7 obtained from mungbean rhizosphere of different locations in Bangladesh and BINA MB-1 (a registered biofertilzer for mungbean) was used as standard check along with un-inoculated control to test their potentiality under glass house condition for mungbean. Result revealed that the higher plant growth, biochemical parameters, seed yield attributes and seed yield were recorded in three bacterial isolates viz., MBI-5, MBB-3 and biofertilizer, BINA MB-1 with being the highest in MBI-5. Therefore, the isolate MBI-5 may be used as commercial biofertilizer after few more trials in the different mungbean growing areas of Bangladesh

Keywords: Bacterial isolates, Nodulation, Yield, Mungbean

Location: Bangladesh Institute of Nuclear Agriculture, Mymensingh, Bangladesh.

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Bio fertilizer, Mungbean

Objectives:

This study was undertaken to screen the BNF strains that are compatible with Vigna radiata and use as biofertilizer.

Methodology:

The present investigation was carried out in glass house at Bangladesh Institute of Nuclear Agriculture, Mymensingh, Bangladesh. The experimental material consists of one mungbean popular variety viz., Binamung-8 which were inoculated with 9 different rhizobial starins/isolates namely MBR-3, MBI-5, MBI-19, MBM-4, MBM-8, MBP-10, MBB-3 and MBJ-7 obtained from root nodules of mungbean plant of different locations in Bangladesh and BINA MB-1 (a registered biofertilzer for mungbean) was used as standard check along with un-inoculated control to test their potentiality under glass house condition for mungbean. Sixty five Bradyrhizobial strains were isolated from mungbean nodules at different locations of Bangladesh. Colony morphology of the each bacterial isolates was examined after 7 days of inoculation of each bacterial isolate on nutrient agar plate and stereoscopic microscope (Olympus, SZH 10) was used for better resolution of bacterial colony morphological characteristics. Gram staining was done to identify cell morphology. The collected strains were Gramnegative, colonies not exceeding 1 mm in diameter within 7 days in yeast extract-manitol medium at the optimal temperature of 25 to 28 °C and according to Jordan (1982), the strains were Bradyrhizobium. These strains were cultured and screened through laboratory experiments. For selection better strains, a mini pot experiment was setup in glass house following the standard protocol of Santos et al. (1999). The plants were harvested at 45 days after sowing and eight strains were selected and used in this study based on superior nodule number, plant height, shoot and root dry weight. Broth culture were prepared by taking a loopful of the respective Rhizobacterial isolates and transferred to the liquid medium of 100 ml conical flask which were incubated for 24 hours on a rotatory shaker. When the culture in the flask showed dense milky white growth, the broth cultures were ready for seed inoculation. Pots were filled with 8 kg sterilized sand and arranged in a Completely Randomized Design with 8 replications for each treatment. Four replications were used for nodulation count, growth and biochemical study. Before sowing, the seeds of mungbean were surface sterilized with 90 per cent alcohol for 30 second and followed by immersion in 32 per cent H₂O₂ for one minutes and ten times washing in sterile distilled water and then sown in sterile sand. Five treated seeds were sown in each pot at a depth of 3-4 cm. The pots were watered regularly with the tap water up to the retention capacity of the pot. After germination, 2 seedlings were maintained in each pot and 2ml of inoculum broth culture (108 bacteria/ml) of individual Bradyrhizobia was applied once below the surface of the root zone of the seedlings, in the afternoon. The pots were kept under vector free and optimal growth conditions [temperature of 28/25³C (day/night, with a standard deviation of ± 2.8°C) with air humidity 60-70%]. The plants were regularly supplied with one-fourth strength sterilized Long Ashton nutrient solution (Hewilt, 1966) twice a week until harvest. During present investigation, the influence of bacterial isolates on plant growth and biochemical parameters and at harvest seed yield and yield attributes were recorded. For growth parameters, two harvests were recorded at 40 and 55 days after sowing (DAS). The whole plants were oven drying at 80 ± 2°C for 72 hours and dry weights were recorded. The growth analysis was carried out following the formulae of Hunt (1978). Nodule number was recorded 2 times, at 30 and 50 DAS. Leaf chlorophyll, nitrate reductase and photosynthesis were determined at 55 DAS. Leg-hemoglobin was determined at 50 DAS. Leaf area of each sample was measured at 70 DAS (at maturity stage) by automatic leaf area meter (Model: LI 3000, LI-COR Biosciences, USA). Leaf chlorophyll was determined following the method of Yoshida et al. (1976). Leaf photosynthesis was measured by photosynthesis meter (LI 6400XT, LI-COR Biosciences, USA). Nitrate reductase activity was determined following the methods of Stewart and Orebamjo (1979). Leg-haemoglubin was determined by the Cyanmethaemoglobin method (Schifftmann and Lobel, 1970). Grain protein per cent was estimated by Micro-Kjeldhal method (AOAC, 1980). The yield contributing characters were recorded at harvest of each pot. All data were analyzed statistically as per the used design following the one way analysis of variance technique and the mean differences were adjusted with Duncan’s Multiple Range Test using the statistical computer package programme, MSTAT-C (Russell, 1986).

Source of Information: Res. Agric., Livest. Fish.7(2): 191-197, August 2020

Article Link (Online Journal): DOI: https://doi.org/10.3329/ralf.v7i2.48859

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Bacterial isolates treated plants improve growth, biochemical parameters and yield in mungbean than in uninoculated plant. Amongst nine tested bacterial isolates, MBI-5 performed the best regarding growth, morphological and biochemical parameters and yield contributing characters resulting the highest seed yield. This isolate may be used as commercial biofertilizer agent after few more trials.

Knowledge Management: Journal