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Isolation, Characterization and Selection of Rhizobium Strains for Cultivation of Sesbania Sp.

Md. Zakaria Hasan
CSISA-CIMMYT, Faridpur Hub

Md. Zahurul Islam
Dept. of Soil Science, Bangladesh Institute of Nuclear Agriculture, Mymensingh, Bangladesh

Musharraf Hossain Mian
Dept. of Soil Science, Bangladesh Agricultural University, Mymensingh

Mohammad Shah Alam
Dept. of Agriculture Extension, Ministry of Agriculture

Abstract/ Executive Summary:

The experiment was conducted to isolate Rhizobium form the root nodules of Deshi dhaincha (Sesbania aculeate) and African dhaincha (Sesbania rostrata), characterization of isolates for authentication, testing their performance as the inoculants and selection of better performed strains for future use as biofertilizer. Nineteen rhizobial strains were isolated from root nodule of dhaincha from 5 Agro Ecological Zones (AEZ 9, AEZ 11, AEZ 12, AEZ 13 and AEZ 28) of Bangladesh. The strains were characterized through morphological, biochemical, microscopic and growth observation. Experiment was conducted in bacteriologically controlled condition using sterile sand in plastic pot in glasshouse. Two varieties of Sesbania named Deshi dhaincha and African dhaincha were used as test crop varieties. Five plants were grown in each plastic pot. After 25 days of sowing plants were harvested and data on nitrogen content of shoot, shoot dry weight, root dry weight, nodule number and nodule dry weight of Sesbania cultivars in glass house were collected. Rhizobial inoculation had significant and positive effect on nitrogen content of shoot, shoot dry weight, root dry weight, nodule number and nodule dry weight for both varieties of Sesbania in sterile condition. The strain DhM3 recorded the highest nitrogen content of shoot, shoot dry weight, root dry weight, nodule number and nodule dry weight. The strain DhSk2 ranked second and DhSk4 ranked third in respect of growth, nodulation, nitrogen fixation and biomass production. Therefore, these three strains can be used for Rhizobium inocula production for cultivation of Sesbania but field level extended studies recommended.

Keywords: Dhaincha, AEZ, Rhizobium, Sesbania sp., Nodule and Biofertilizer

Location: BNF (Biological Nitrogen Fixation) Laboratory, Division of Soil Science, Bangladesh Institute of Nuclear Agriculture (BINA), Mymensingh.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Dhaincha

Objectives:

To isolate and characterize of rhizobia from Deshi and African dhaincha and the effect of isolated rhizobia inoculants on growth, nodulation and biomass production of Deshi dhaincha Sesbania aculeata and African dhaincha (Sesbania rostrata) plants.

Methodology:

Nodules of Sesbania plants were collected from five selected Agro Ecological Zones- AEZ 9: Old Brahmaputra Floodplain (BAU Farm, Mymensingh), AEZ 11: High Ganges River Floodplain (Ishurdi, Pabna), AEZ 12: Low Ganges River Floodplain (Faridpur Sadar), AEZ 13: Ganges Tidal Floodplain (BINA Substation, Satkhira) and AEZ 28; Madhupur Tract (Dhonbari, Tangail). Roots with intact nodules were carried out to BNF (Biological Nitrogen Fixation) Laboratory, Division of Soil Science, Bangladesh Institute of Nuclear Agriculture (BINA), Mymensingh.

Larger, healthy and mature nodules were selected. After separation nodules were washed in running tap water and transferred to sterile vial. Selected nodules were then immersed into 70% alcohol solution for 30 seconds and in 3% hydrogen peroxide solution for 3 minutes. Hydrogen peroxide treated nodules were then rinsed with sterile mild hot water for several times. Nodules were then cut transversely with sterile blade and the milky suspension directly from the cut nodule was streaked out on congo red yeast extract mannitol agar (CRYEMA) plates using inoculation needle under laminar flow hood. Plates were incubated for 7 days at 28º C temperature.
Colonies were characterized through a systematic series of tests such as cultural tests (i.e. shape, size, opacity, elevation, surface, margin, consistency, emulsifiability, colour odour etc.), microscopic tests (i.e. simple staining, gram staining, motility observation etc.), biochemical tests (i.e. Congo red absorption test, BTB test, Hofer’s alkaline broth test etc.) and nodulation tests.
Nodulation capacity of rhizobial isolates were determined by growing plants in plastic buckets containing 2 kg autoclaved sand (at 20 lbs pressure for 2 hours) in glass houses. Seeds of both Deshi dhaincha (Sesbania aculeata) and African dhaincha (Sesbania rostrata), were rinsed with concentrated Sulphuric Acid (H₂SO₄) for few seconds and immediately washed thoroughly by distilled water. Prior to sowing, seeds were treated with a thick Rhizobium suspension of each isolate. Ten seeds were planted in each bucket and thinned to keep five plants after germination. Sterilized seedling solution was added to each bucket at one day intervals.
Plants samples were collected from the plastic buckets at 25 days of sowing. From each buckets 3 randomly selected plants were carefully taken out so that no nodule was left in the soil. Roots were washed and the Nodules from the root system of each plant were separately collected and counted. Oven dry weights of nodules were also recorded.
Data obtained from the pot experiment were analyzed statistically by F-test to examine whether treatment effects were significant or insignificant. The treatment means were evaluated by Duncan’s New Multiple Range Test (DMRT).

Source of Information: Journal of Bioscience and Agriculture Research, Vol. 03 (02): 79-86, 2015, eISSN 2312-7945

Article Link (Online Journal): http://www.journalbinet.com/15v30210.html

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

All the strains increased nitrogen content of shoot, shoot dry weight, root dry weight, nodule number and nodule dry weight of Sesbania cultivars in glass of Deshi dhaincha (Sesbania aculeata) and African dhaincha (Sesbania rostrata) plants. The strain DhM3 produced the highest nitrogen content of shoot, shoot dry weight, root dry weight, nodule number and nodule dry weight. The strain DhSk2 ranked second and DhSk4 ranked third. So, the strains DhM3, DhSk2 and DhSk4 may be selected for producing inoculant for cultivation of Dhaincha in examined AES’s. But conclusions and perspectives are valid under the present experimental conditions. Further experiments are needed to be conducted in soils of different AEZ’s having wide variations in soil pH, organic matter and nutrient status for confirmation of the findings.

Knowledge Management: Journal