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Effect of Plant Growth Promoting Rhizobacteria (pgpr) in Seed Germination and Root-shoot Development of Chickpea (cicer Arietinum L.) Under Different Salinity Condition

Mohammad Mosharraf Hossain
Department of Soil Science, Sher - e - Bangla Agricultural University, Dhaka - 1207, Bangladesh.

Keshob Chandra Das
SSO
Molecular Biotechnology Division, National Institute of Biote chnology, Savar - 1349, Dhaka, Bangladesh;

Sabina Yesmin
Mushroom Development Officer
Mushroom Development Institute, Savar -1340, Dhaka, Bangladesh.

Syfullah Shahriar
Department of Soil Science, Sher - e - Bangla Agricultural University, Dhaka - 1207, Bangladesh.

Abstract/ Executive Summary:

Plant growth promoting rhizobacteria (PGPR) are beneficial bacteria that colonize plant roots and enhance plant growth by a wide variety of mechanisms. Ten isolates of bacteria designated as SS01, SS02, SS03, SS04, SS05, SS06, SS07, SS08, SS09 and SS10 were successfully isolated and morphologically and biochemically characterized. Subsequently to investigate the effect of PGPR isolates on the growth of chickpea, a pot culture experiment was conducted in 2013 at National Institute Biotechnology, Bangladesh net house. Prior to seeds grown in plastic pots, seeds were treated with PGPR isolates and seedlings were harvested after 21 days of inoculation. All the isolates were gram negative in reaction, catalase positive, produced indole acetic acid (IAA) as well as performed phosphate solubilization, able to degrade cellulose and have the adaptability in wide range of temperature and showed positive growth pattern in medium. Most of isolates resulted in a significant increasing of shoot length, root length and dry matter production of shoot and root of chickpea seedlings. Application of PGPR isolates significantly improves the percentage of seed germination under saline conditions. The present study, therefore suggested that the use of PGPR isolates SS04, SS10 and SS08 as inoculants biofertilizers might be beneficial for chickpea cultivation in saline condition.

Keywords: Chickpea, Indole acetic acid, NaCl, PGPR

Location: National Institute of Biotechnology (NIB), Savar, Bangladesh.

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Chickpea

Objectives:

To determine the effect of plant growth promoting rhizobacteria strains that are compatible with chickpea.

Methodology:

Soil samples were collected from chickpea field of Tangail sadar upazila of Tangail district. After collection of soil samples, it was stored in plastic container, properly leveled and carried to the laboratory for further use. Ten grams of rhizosphere soil were taken into a 250 ml of conical flask and 90 ml of sterile distilled water was added to it. After serial dilution upto 10, an aliquot of this suspension was spread on the plates of nutrient (NA) agar medium (NH₄Cl 5.0 g; K₂HPO₄ 3.0 g; Na₂ SO₄ 2.0 g; KH₂PO₄ 1.0 g; NH₄NO₃ 1.0 g; MgSO₄, 7H₂O 0.1 g; glucose 2.0 g; distilled water 1 litre and pH 7.0±0.2) in the molecular microbiology laboratory of National Institute of Biotechnology (NIB), Savar, Bangladesh. After 3 days of incubation at 28°C, bacterial colonies were streaked to other NA agar plates and incubated at 28°C for 3 more days. Typical single bacterial colonies were observed over the streak. Well isolated single colonies were picked up and different characteristics of colonies such as shape, size, elevation, surface, margin, color, odor and pigmentation etc. were observed and recorded. All the morphological and biochemical characters of the isolates were determined based on Bergey’s Manual of Systematic Bacteriology. A loopful of bacterial culture from each isolates was diluted into a test tube containing 1 ml sterile distilled water and was vortexed for 2/3 minutes. A loopful suspension was then taken on a glass slide and smeared. The slide was air dried and fixed by heating on a Bunsen flame. The slide was flooded with crystal violet solution for 3 min. The slide was washed gently in flow of tap water and air dried. The slide was observed under microscope and recorded the shape. Motility of bacteria was observed by hanging drop method. A drop of suspension was taken on a cover slip. The cover slip was hanged on a hollow slide with vaseline. The slide was then observed under microscope to test the motility of bacteria. The culture of 10 isolates were streaked on NA agar plates and incubated at 10, 20, 28, 37 and 45°C and also in the NA plate with 0, 3.0, 6.0 and 12.0 mM NaCl solutions in the medium. The bacterial isolates were designated as SS01, SS02, SS03, SS04, SS05, SS06, SS07, SS08, SS09 and SS10. A single colony of bacterial culture was grown on nutrient broth medium. A loopful of the respective culture was transferred to the 100 ml of conical flask then incubated for 7 days on a rotary shaker in 80 rpm. The IAA production and phosphate solubilization were then examined according to the method. Phosphate solubilizing capacity of the isolates was estimated using Pikovskaya’s medium. The cellulose degradation test was performed by using carboxy methyl cellulose (CMC) agar plates according to the method. Seeds of chickpea were collected from pulse research centre of Bangladesh Agricultural Research Institute (BARI), Gazipur. Prior to germination, the chickpea seeds were surface sterilized in 3% H₂O₂ and then rinsed with distilled water. The seeds were then surface sterilized with 0.024% sodium hypochlorite for 2 minutes and rinsed thoroughly in ste rile distilled water. Seeds were inoculated by overnight soaking with suspensions of bacteria (approximately 10⁷ - 10⁸ cfu/ml). Seeds soaked in sterilized distilled water were used as the control. The soaked seeds with rhizobacterial isolates emerged with three different NaCl solutions which were derived from sterile distilled water by adding 0 (control), 3, 6 and 12 mM NaCl, respectively. Chickpea seeds were placed over the sterile filter paper into a petri dish and covered with tight fitting lid. Then the petri - plates were kept in an incubator maintaining the moisture and temperature of 28 ºC - 30 °C. Seed germination assay was laid out in Completely Randomized Design with three replications. Germinated seeds were recorded and discarded at 24 h interval over 10 days.An amount of 0.3 kg sand was placed into a pot. Ten PGPR inoculated seeds were sown at 4 to 5 cm depth of sand in each plastic pot and laid out in a completely randomized block design with three replications. The chickpea plants were harvested after 21 days of seed sowing through separating of plants from soil. Shoot length (cm plant^{- 1} ) and root length (cm plant^{- 1} ) and dry weight of shoots and roots of each plant were recorded after drying in an oven for 1 day at 70°C. The data was analyzed statistically by MS - STATC statistical program. The significance of differences between mean values was evaluated by DMRT according to the methods of Gomez and Gomez.

Source of Information: Res. Agric., Livest. Fish.3(1): 105-113, April 2016, ISSN: 2409-0603

Article Link (Online Journal): DOI: http://dx.doi.org/10.3329/ralf.v3i1.27864

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Results imply that PGPR are able to enhance the production of IAA, solubilization of phosphorus and resistance to pathogen and pests, thereby improving growth of chickpea plant. The use of PGPR as inoculants biofertilizers is an efficient approach to replace chemical fertilizers an d pesticides for sustainable chickpea cultivation in Bangladesh and other developing countries. Further investigations, including efficiency test under green house and field conditions needed to clarify the role of PGPR as biofertilizers that exert beneficial effects on plant growth and development.

Knowledge Management: Journal