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Isolation of Salt Tolerant Rhizobium and Their Characterization, Plant Growth Promoting and Symbiotic Performance on Oilseed Legume (2018–2019)

F. Alam
Soil Science Division, BARI, Joydebpur, Gazipur, Bangladesh

M.F.A. Anik
Soil Science Division, BARI, Joydebpur, Gazipur, Bangladesh

M. Rahman
Soil Science Division, BARI, Joydebpur, Gazipur Bangladesh

M.E. Ali
Soil Science Division, BARI, Joydebpur, Gazipur Bangladesh

S. Akhter
Soil Science Division, BARI, Joydebpur, Gazipur Bangladesh

Abstract/ Executive Summary:

A study was conducted at Soil Science Division, Bangladesh Agricultural Research Institute, during 2018-2019 with the objectives to isolate salt-tolerant rhizobia species from the coastal saline area of Bangladesh, characterize, plant-growth-promoting and symbiotic performance of those effective salt-tolerant bacteria on oilseed legumes. For isolating of salt-tolerant Rhizobium bacteria, 20 nodule samples were collected from soybean and groundnut roots from different fields of saline stress soils (Noakhali, Lakhmipur, Cox’s Bazar, Bhola, Patuakhali) of Bangladesh using global positioning system (GPS) record along with crop history. Nodules were collected aseptically, purified strains and preserved at 4°C for short time preservation and kept in -20^°C, for long time preservation. The characters of 20 bacterial isolates (10 isolates from soybean and 10 isolates from groundnut) were studied following the standard microbiological and biochemical methods. The cells of Rhizobium isolates were examined under microscope and found that the cells were rod shape, motile and gram-negative bacteria. The biochemical characters such as Congo red, Gram staining, salt tolerance were investigated. The isolates were observed to lack the ability to absorb Congo red from YEMA medium. The salt tolerance study was carried out on YEM agar plate with each modified by NaCl at 0, 2, 4, 8, 16, 32 and 48 dSm^-1. All isolated bacteria were tested by growing on a plate up to (48 dS m^-1) level. Seedlings were grown in a glass tube to check the infection ability of Rhizobium (strain SR1 to SR10 for soybean and strain GR1 to GR10 for groundnut) and found nodule formation in seedling roots within 30-50 days after inoculation.

Keywords: Isolation, Salt tolerant, Performance, Oilseed legume

Location: Soil Science Division, Bangladesh Agricultural Research Institute

Start Date: 00-00-2018

End Date: 00-00-2019

Program Area: Crop-Soil-Water Management

Commodity: Soil salinity, Bio fertilizer

Objectives:

To isolate, identify, characterize morphological, biochemical and PGPR of effective rhizobial strains and measure nodulation test of collected strains in respective crops.

Methodology:

Nodules from soybean and groundnut root (20 samples) were collected from different fields of saline soils (Noakhali, Lakhmipur, Cox’s Bazar, Bhola, Patuakhali) of Bangladesh using global positioning system (GPS) record along with crop history. Nodules were collected aseptically and crushed, streaked onto yeast extract mannitol agar (YEMA) plate and incubated at 30°C for 2-3 days. A final selection of the bacterial strains was done by comparing morphological (colony) characteristics and transferred to agar slants prepared with the corresponding plating media. The isolated strains were purified by the streak plate method. The purified strains were transferred to agar slants and preserved as stock culture in a refrigerator at 4⁰C. In addition, the culture was mixed with 50% (v/v) glycerol and kept at -20⁰C for preservation. The colony morphology of isolates was examined on YEM agar plates. After an incubation of 2-3 days at 30°C, individual colonies were characterized based on their colour, shape, appearance, motility, transparency, Congo red and Gram stain reaction (Aneja 2003). The bacterial strains were re-cultured in YEM broth at 30⁰C for 24-48 hours. The cell suspension was tenfold serial diluted to 10^–4 to 10^–8 before being used as bacterial samples. The salt tolerance study was carried out by spreading 0.1 ml of each sample on YEM agar plate with each modified by NaCl at 0, 2, 4, 8, 16, 32 and 48 dS m^-1. The plates were incubated at ambient temperature (28±2⁰C) for 3 days. The salt-tolerant bacteria were selected and preserved on a salt-containing medium. In order to check the infection ability of Rhizobium on soybean and groundnut seedlings roots, Rhizobium was grown in yeast extract mannitol (YEM) broth culture at 30^oC for 48h and 0.1 ml of this broth culture was added to the YEM agar slants tube (60 ml glass test tube, diameter 3 cm) having 1-2 seedlings and kept for 3-5 weeks at room temperature under aseptic condition and monitored nodulation infectivity. The nitrogen-free nutrient solution was supplied as a nutrient source to YEM agar slant tubes twice a week.

Source of Information: Annual Research Report 2018–2019, Soil Science Division, BARI, Joydebpur, Gazipur, Bangladesh

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The bacteria were isolated and identified from coastal saline areas of Bangladesh. From this study, twenty Rhizobium bacterial strains (10 strains for soybean and 10 strains for groundnut) were isolated, characterized and preserved. The bacteria were rod shape, gram-negative, motile and not absorbed congo red. All bacteria were salt-tolerant (?48 dS m^-1) and produced nodules in seedling roots.

Knowledge Management: Report/Proceedings