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Research Article

volume 41 issue 5 (october 2018) : 771-777,   Doi: 10.18805/LR-389

Rhizobium sp.CCNWYC119: a single strain highly effective as biofertilizer for three different peas (Pigeon pea, Sweet pea and Chick pea)

A
Ali Mohammad Nushair
A
Ananda Kumar Saha
A
Abul Mandal
M
Md. Anisur Rahman
M
Moni Krishno Mohanta
M
Md. Ariful Hasan
M
Md Fazlul Haque
1Genetics and Molecular Biology Laboratory, Department of Zoology, University of Rajshahi, Rajshahi-6205, Bangladesh.

  • Submitted29-09-2017|

  • Accepted18-01-2018|

  • First Online 25-04-2018|

  • doi 10.18805/LR-389

Cite article:- Nushair Mohammad Ali, Saha Kumar Ananda, Mandal Abul, Rahman Anisur Md., Mohanta Krishno Moni, Hasan Ariful Md., Haque Fazlul Md (2018). Rhizobium sp.CCNWYC119: a single strain highly effective as biofertilizer for three different peas (Pigeon pea, Sweet pea and Chick pea). Legume Research. 41(5): 771-777. doi: 10.18805/LR-389.

ABSTRACT

Rhizobium spp. was isolated from root nodules of Pigeon pea (Cajanus cajan L.), Sweet pea (Lathyrus sativus L.), Chick pea (Cicer arietinum L.). The isolates ware rod shaped, aerobic, gram negative, motile and non-spore forming. Isolates were positive to Catalase, Citrate utilization, Urea hydrolysis, Congored, Nitrification, Oxidase, Triple sugar iron and MacConkey agar test. The isolates can ferment all nine sugars. Then, the isolates identified as Rhizobium spp. depending on above results were subjected to 16S rRNA sequencing for further confirmation and identification. Surprisingly, the isolates were same strain or member of same cluster of Rhizobium and identified as Rhizobium sp.CCNWYC119 strain based on 16S rRNA sequence (98% similarity). Then, different parameters of soil quality enrichment and plant growth viz. plant height; weight of pods and seeds; number, fresh and dry weight of nodules were studied to test the efficacy of the isolate as biofertilizer. Here, inoculant of Rhizobium sp. isolated from Pigeon pea was used as biofertilizer. The results showed the significant increase of nodulation, enrichment of soil of rhizosphere, plant growth and yield for all three types of inoculated peas as compared with non-inoculated control peas indicating that the isolated strain could be used as a common efficient biofertilizer for Pigeon pea, Sweet pea and Chick pea. It was also found that the isolate grew optimally at temperature 28°C and pH 7.0. Moreover, the isolate was sensitive to the higher concentration of NaCl (>1%) and to antibiotics- Mecillinam, Ciprofloxacin, Cotrimoxazole, Pefloxacin, Ceftazidime and Tetracycline.   

REFERENCES

  1. Aneja, K. R. (2003). Experiments in Microbiology Plant Pathology and Biotechnology, 4th edition, New age International Publishers, New Delhi,India.
  2. Appunu, C. and Dhar, B. (2006). Symbiotic effectiveness of acid-tolerant Bradyrhizobium strains with soybean in low pH soil. African J. Biotech. 5:842-845.
  3. Bhuiyan, M. A. H., Khanam, D., Khatun, M. R. and Hassan. M. S. (1998). Effect of molybdenum, boron and Rhizobium on nodulation, growth and yield of chickpea. Bull. Inst. Trop. Agric. 21: 1-7 
  4. Elsheikh, E. A.E and Wood, M. (1986).Soil biology. Biochem.21:883-887. 
  5. Erum, S. and Bano, A. (2008). Bariation in phytohormone production in Rhizobium strains at different altitudes of Northern areas of Pakistan. Int. J. Agric. Biol.10:536-540.
  6. Fernandes, P. I., Lima, A. A., Passos, S. R. and Gava, C. A. T. (2012). Phenotypic diversity and amylolytic activity of fast growing rhizobia from pigeonpea [Cajanuscajan (L.) Millsp.]. Braz. J. Microbiol. 43:1604-1612.
  7. Gangwar, S. and Dubey, M. (2012). Effect on N and P uptake by chickpea (Cicer arietinum L.) as influenced by micronutrients and biofertilizers. Legume Res. 35(2): 164 – 168
  8. Gupta, S. C. and Sahu, S. (2012). Response of chickpea to micronutrients and biofertilizers in vertisol. Legume Res. 35(3):248-251
  9. Hashem, F. M., Swelim, D. M., Kuykendall, L. D., Mohamed, A. I., Abdel-Wahab, S. M. and Hegazi, N. I. (1998). Identification and characterization of salt and thermo tolerant Leucaena modulating Rhizobium strains, Biol. Fert.Soil.27:335-341. 
  10. Janda, J. M. and Abbott, S. L. (2007). 16S rRNA Gene sequencing for bacterial identification in the diagnostic laboratory: Pluses, Perils, and Pitfalls, J. Clin. Microbiol. 45(9): 2761-2764.
  11. Jordan, D.C. (1984). Family III. Rhizobiacea Conn 1938. In: Bergey’s Manual of Systermnatic Bacteriology. Bol I (eds Krieg. N.R. and Holt.J.G) Willams and Wilkins Press, Baltimore, pp234-254.
  12. Khadraji, A. and Ghoulam, C. (2016). Effect of drought on growth, physiological and biochemicalprocesses of chickpea-rhizobia symbiosis. Legume Res. 40(1):94-99.
  13. Kucuk, C. M., Kivanc, M., Kinaci, E. (2006). Characterization of Rhizobium Sp. Isolated from Bean. Turk J. Biol. 30: 127-132.
  14. Kumawat, P.K., Tiwari, R.C., Golada, S.L., Godara, A.S., Garhwal, R.S. and Choudhary, R., 2013. Effect of phosphorus sources, levels and biofertilizers on yield attributes, yield and economics of black gram (Phaseolus mungo L.). Legume Res. 36(1): 70-73
  15. Lupwayi, N and Haque, I. (1994). Legume–Rhizobium technology manual environmental sciences division International Livestock center for Africa. Addis Ababa, Ethiopia. pp1-93.
  16. Mondal, M. M. A., Malek, M. A., Sattar, M. A., Puteh, A. B., Rafii, M.Y. and Ismail, M. R. Response of biofertilizer and urea on growth and yield in mungbean. Legume Res. 36 (5): 448 - 452
  17. Olivares, J., Bedmar, E.J. and Martinez, M. E. (1984). Infectivity of Rhizobium meliloti as affected by extra cellular polysaccharides.J. Appl.Bacteriol.56: 389-393. 
  18. Phillips, D.A. (1991). Flavonoids: Plant signals to soil microbes. Rec. Adv. Phytochem. 2: 1-33. 
  19. Ramana, V., Ramakrishna, M., Purushotham, K. and Reddy, K. B. (2010). Effect of bio-fertilizers on growth, yield attributes and yield of french bean (Phaseolus vulgaris L.). Legume Res. 33 (3): 178 - 183
  20. Sadowsky, M. J., Keyser, H. H. and Bohlool, B. B. (1983).Biochemical characterization of fast growing and slow growing rhizobia that nodulate soybeans.Syst. Bacteriol.33:716-722.
  21. Saha, A. K., Haque, M. F., Karmaker, S., and Mohanta, M. K. (2009). Antibacterial effects of some antiseptics and disinfectants. J. Life and Earth Science. 3: 19-21.
  22. Saha, A. K. and Haque, M. F. (2005). Effect of inoculation with Rhizobium on nodulation and growth of Bean, Dolchos lablab. J. Life Earth Science. 1(1) 71-74.
  23. Saitou, N. and Nei, M. (1987). The Neighbor- joining method: A new method for reconstruction Phylogenetic trees. Molecular Biology and Evolution. 4(4):406 -425.
  24. Santos, C.A.F.; Araujo, F.A. and Menezes, E.A. (2000). Yield performance of cowpea genotypes under irrigated and rainfed conditions in Petrolina and Juazeiro, Brazil. Pesquisa Agropecuária Brasileira. 35: 2229-2234.
  25. Stowers, M. I. and Eaglesham, A. R. J. (1983).A stem nodulating Rhizobium with physiological characterstics of both fast and slow growers. J.Gen. Microbiol.129: 3651-3655.
  26. Vincent, J. M. (1970). A Manual for the Practical Study of the Root Nodule Bacteria, IBP Hand Book No. 15, Blackwell Scientific publications, Oxford.
  27. Vitousek, P.M. (1997). Human alteration of the global nitrogen cycle: sources and consequences. Ecological Applications.7: 737 
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    Research Article

    volume 41 issue 5 (october 2018) : 771-777,   Doi: 10.18805/LR-389

    Rhizobium sp.CCNWYC119: a single strain highly effective as biofertilizer for three different peas (Pigeon pea, Sweet pea and Chick pea)

    A
    Ali Mohammad Nushair
    A
    Ananda Kumar Saha
    A
    Abul Mandal
    M
    Md. Anisur Rahman
    M
    Moni Krishno Mohanta
    M
    Md. Ariful Hasan
    M
    Md Fazlul Haque
    1Genetics and Molecular Biology Laboratory, Department of Zoology, University of Rajshahi, Rajshahi-6205, Bangladesh.

    • Submitted29-09-2017|

    • Accepted18-01-2018|

    • First Online 25-04-2018|

    • doi 10.18805/LR-389

    Cite article:- Nushair Mohammad Ali, Saha Kumar Ananda, Mandal Abul, Rahman Anisur Md., Mohanta Krishno Moni, Hasan Ariful Md., Haque Fazlul Md (2018). Rhizobium sp.CCNWYC119: a single strain highly effective as biofertilizer for three different peas (Pigeon pea, Sweet pea and Chick pea). Legume Research. 41(5): 771-777. doi: 10.18805/LR-389.

    ABSTRACT

    Rhizobium spp. was isolated from root nodules of Pigeon pea (Cajanus cajan L.), Sweet pea (Lathyrus sativus L.), Chick pea (Cicer arietinum L.). The isolates ware rod shaped, aerobic, gram negative, motile and non-spore forming. Isolates were positive to Catalase, Citrate utilization, Urea hydrolysis, Congored, Nitrification, Oxidase, Triple sugar iron and MacConkey agar test. The isolates can ferment all nine sugars. Then, the isolates identified as Rhizobium spp. depending on above results were subjected to 16S rRNA sequencing for further confirmation and identification. Surprisingly, the isolates were same strain or member of same cluster of Rhizobium and identified as Rhizobium sp.CCNWYC119 strain based on 16S rRNA sequence (98% similarity). Then, different parameters of soil quality enrichment and plant growth viz. plant height; weight of pods and seeds; number, fresh and dry weight of nodules were studied to test the efficacy of the isolate as biofertilizer. Here, inoculant of Rhizobium sp. isolated from Pigeon pea was used as biofertilizer. The results showed the significant increase of nodulation, enrichment of soil of rhizosphere, plant growth and yield for all three types of inoculated peas as compared with non-inoculated control peas indicating that the isolated strain could be used as a common efficient biofertilizer for Pigeon pea, Sweet pea and Chick pea. It was also found that the isolate grew optimally at temperature 28°C and pH 7.0. Moreover, the isolate was sensitive to the higher concentration of NaCl (>1%) and to antibiotics- Mecillinam, Ciprofloxacin, Cotrimoxazole, Pefloxacin, Ceftazidime and Tetracycline.   

    REFERENCES

    1. Aneja, K. R. (2003). Experiments in Microbiology Plant Pathology and Biotechnology, 4th edition, New age International Publishers, New Delhi,India.
    2. Appunu, C. and Dhar, B. (2006). Symbiotic effectiveness of acid-tolerant Bradyrhizobium strains with soybean in low pH soil. African J. Biotech. 5:842-845.
    3. Bhuiyan, M. A. H., Khanam, D., Khatun, M. R. and Hassan. M. S. (1998). Effect of molybdenum, boron and Rhizobium on nodulation, growth and yield of chickpea. Bull. Inst. Trop. Agric. 21: 1-7 
    4. Elsheikh, E. A.E and Wood, M. (1986).Soil biology. Biochem.21:883-887. 
    5. Erum, S. and Bano, A. (2008). Bariation in phytohormone production in Rhizobium strains at different altitudes of Northern areas of Pakistan. Int. J. Agric. Biol.10:536-540.
    6. Fernandes, P. I., Lima, A. A., Passos, S. R. and Gava, C. A. T. (2012). Phenotypic diversity and amylolytic activity of fast growing rhizobia from pigeonpea [Cajanuscajan (L.) Millsp.]. Braz. J. Microbiol. 43:1604-1612.
    7. Gangwar, S. and Dubey, M. (2012). Effect on N and P uptake by chickpea (Cicer arietinum L.) as influenced by micronutrients and biofertilizers. Legume Res. 35(2): 164 – 168
    8. Gupta, S. C. and Sahu, S. (2012). Response of chickpea to micronutrients and biofertilizers in vertisol. Legume Res. 35(3):248-251
    9. Hashem, F. M., Swelim, D. M., Kuykendall, L. D., Mohamed, A. I., Abdel-Wahab, S. M. and Hegazi, N. I. (1998). Identification and characterization of salt and thermo tolerant Leucaena modulating Rhizobium strains, Biol. Fert.Soil.27:335-341. 
    10. Janda, J. M. and Abbott, S. L. (2007). 16S rRNA Gene sequencing for bacterial identification in the diagnostic laboratory: Pluses, Perils, and Pitfalls, J. Clin. Microbiol. 45(9): 2761-2764.
    11. Jordan, D.C. (1984). Family III. Rhizobiacea Conn 1938. In: Bergey’s Manual of Systermnatic Bacteriology. Bol I (eds Krieg. N.R. and Holt.J.G) Willams and Wilkins Press, Baltimore, pp234-254.
    12. Khadraji, A. and Ghoulam, C. (2016). Effect of drought on growth, physiological and biochemicalprocesses of chickpea-rhizobia symbiosis. Legume Res. 40(1):94-99.
    13. Kucuk, C. M., Kivanc, M., Kinaci, E. (2006). Characterization of Rhizobium Sp. Isolated from Bean. Turk J. Biol. 30: 127-132.
    14. Kumawat, P.K., Tiwari, R.C., Golada, S.L., Godara, A.S., Garhwal, R.S. and Choudhary, R., 2013. Effect of phosphorus sources, levels and biofertilizers on yield attributes, yield and economics of black gram (Phaseolus mungo L.). Legume Res. 36(1): 70-73
    15. Lupwayi, N and Haque, I. (1994). Legume–Rhizobium technology manual environmental sciences division International Livestock center for Africa. Addis Ababa, Ethiopia. pp1-93.
    16. Mondal, M. M. A., Malek, M. A., Sattar, M. A., Puteh, A. B., Rafii, M.Y. and Ismail, M. R. Response of biofertilizer and urea on growth and yield in mungbean. Legume Res. 36 (5): 448 - 452
    17. Olivares, J., Bedmar, E.J. and Martinez, M. E. (1984). Infectivity of Rhizobium meliloti as affected by extra cellular polysaccharides.J. Appl.Bacteriol.56: 389-393. 
    18. Phillips, D.A. (1991). Flavonoids: Plant signals to soil microbes. Rec. Adv. Phytochem. 2: 1-33. 
    19. Ramana, V., Ramakrishna, M., Purushotham, K. and Reddy, K. B. (2010). Effect of bio-fertilizers on growth, yield attributes and yield of french bean (Phaseolus vulgaris L.). Legume Res. 33 (3): 178 - 183
    20. Sadowsky, M. J., Keyser, H. H. and Bohlool, B. B. (1983).Biochemical characterization of fast growing and slow growing rhizobia that nodulate soybeans.Syst. Bacteriol.33:716-722.
    21. Saha, A. K., Haque, M. F., Karmaker, S., and Mohanta, M. K. (2009). Antibacterial effects of some antiseptics and disinfectants. J. Life and Earth Science. 3: 19-21.
    22. Saha, A. K. and Haque, M. F. (2005). Effect of inoculation with Rhizobium on nodulation and growth of Bean, Dolchos lablab. J. Life Earth Science. 1(1) 71-74.
    23. Saitou, N. and Nei, M. (1987). The Neighbor- joining method: A new method for reconstruction Phylogenetic trees. Molecular Biology and Evolution. 4(4):406 -425.
    24. Santos, C.A.F.; Araujo, F.A. and Menezes, E.A. (2000). Yield performance of cowpea genotypes under irrigated and rainfed conditions in Petrolina and Juazeiro, Brazil. Pesquisa Agropecuária Brasileira. 35: 2229-2234.
    25. Stowers, M. I. and Eaglesham, A. R. J. (1983).A stem nodulating Rhizobium with physiological characterstics of both fast and slow growers. J.Gen. Microbiol.129: 3651-3655.
    26. Vincent, J. M. (1970). A Manual for the Practical Study of the Root Nodule Bacteria, IBP Hand Book No. 15, Blackwell Scientific publications, Oxford.
    27. Vitousek, P.M. (1997). Human alteration of the global nitrogen cycle: sources and consequences. Ecological Applications.7: 737 
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