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Current IssueEditorial BoardOnline First ArticlesArchive

Research Article

volume 54 issue 6 (december 2020) : 781-786,   Doi: 10.18805/IJARe.A-5578

Extraction of Indole-3-acetic Acid from Plant Growth Promoting Rhizobacteria of Bamboo Rhizosphere and Its Effect on Biosynthesis of Chlorophyll in Bamboo Seedlings

B
Bishnu Maya K.C.
D
Dhurva Prasad Gauchan
S
Sanjay Nath Khanal
S
Sharmila Chimouriya
J
Janardan Lamichhane
1Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel, Nepal.
Cite article:- K.C. Maya Bishnu, Gauchan Prasad Dhurva, Khanal Nath Sanjay, Chimouriya Sharmila, Lamichhane Janardan (2020). Extraction of Indole-3-acetic Acid from Plant Growth Promoting Rhizobacteria of Bamboo Rhizosphere and Its Effect on Biosynthesis of Chlorophyll in Bamboo Seedlings. Indian Journal of Agricultural Research. 54(6): 781-786. doi: 10.18805/IJARe.A-5578.

ABSTRACT

Background: Indole-3-acetic acid (IAA), a principal phytohormone, controls several crucial physiological processes of plants. It ameliorates plant growth by stimulating cell elongation, root initiation, seed germination and seedling growth. Alteration of IAA level by plant growth promoting rhizobacteria leads to varied impacts on plant growth and development.
Methods: Soil samples were collected from bamboo (Bambusa tulda, B. nutans subsp. cupulata, B. balcooa and Dendrocalamus strictus) rhizosphere. Altogether five bacterial isolates were screened by serial dilution method and subjected to biochemical analysis. The isolate BUX1 with high IAA production capacity was optimized for IAA production. IAA was partially purified and quantified from the bacterial extract by thin layer chromatography (TLC). The influence of extracted bacterial IAA on chlorophyll biosynthesis in bamboo seedlings of B. tulda was compared with uninoculated control plants.
Results: Biochemical analysis revealed that all the isolates belonged to genus Bacillus which were found capable of producing IAA. During optimization, BUX1 isolate produced 99.13 µg ml-1 of IAA at 37°C, pH 7, 3 mg l-1 concentration of L-tryptophan and 150 rpm agitation rate after 192 hour of incubation. The Rf value of the bacterial IAA during TLC was identical to that of standard IAA (0.425) indicating that IAA was present in crude extract of Bacillus (BUX1). The influence of bacterial IAA on chlorophyll biosynthesis in bamboo seedlings was significant in comparison to uninoculated plants. Therefore, this isolate could be a prospective candidate to be employed as biofertilizer. 

REFERENCES

  1. Adhikari, D., Kaneto, M., Itoh, K., Suyama, K., Pokharel, B.B., Gaihre, Y.K. (2012). Genetic diversity of soybean-nodulating rhizobia in Nepal in relation to climate and soil properties. Plant and Soil. 357: 131-145. DOI: 10.1007/s11104-012-1134-6.
  2. Ahmed, A. and Hasnain, S. (2010). Auxin-producing Bacillus sp.: Auxin quantification and effect on the growth of Solanum tuberosum. Pure and Applied Chemistry. 82: 313-319. DOI: 10.1351/PAC-CON-09-02-06.
  3. Arnon, D.I. (1949). Copper enzymes in isolated chloroplasts. Polyphenol oxidase in Beta vulgaris. Plant Physiology. 24: 1-15. 
  4. Asghar, H.N., Zahir, Z.A., Arshad, M., Khaliq, A. (2002). Relationship between in vitro production of auxins by rhizobacteria and their growth promoting activities in Brassica juncea L. Biology and Fertility of Soils. 35: 231-237. DOI: 10.10 07/s00374-002-0462-8.
  5. Barillot, C.D., Sarde, C.O., Bert, V., Tarnaud, E., Cochet, N. (2013). A standardized method for the sampling of rhizosphere and rhizoplan soil bacteria associated to a herbaceous root system. Annals of Microbiology. 63: 471-476. DOI: 10.1007/s13213-012-0491-y.
  6. Biswas, S., Kundu, D.K., Mazumdar, S.P., Saha, A.R., Majumdar, B., Ghorai, A.K., Ghosh, D., Yadav, A.N., Saxena, A.K. (2018). Study on the activity and diversity of bacteria in a New Gangetic alluvial soil (Eutrocrept) under rice-wheat- jute cropping system. Journal of Environmental Biology. 39: 379-86. DOI: 10.22438/jeb/39/3/MRN-523.
  7. Garrity, G.M., Bell, J.A., Lilburn, T.G. (2004). Taxonomic outline of the prokaryotes. Bergey’s manual of systematic bacteriology. Springer, New York, Berlin, Heidelberg.
  8. Gul, A., Salam, A., Afridi, M.S., Bangash, N.K., Ali, F., Ali, M.Y., Khan, S., Mubeeen, R. (2019). Effect of urea, bio-fertilizers and their interaction on the growth, yield and yield attributes of Cyamopsis tetragonoloba. Indian Journal of Agricultural Research. 53: 423-428. DOI: 10.18805/IJARe.A-395. 
  9. Hafeez, F.Y., Yasmin, S., Ariani, D., Rahman, M., Zafar, Y., Malik, K.A. (2006). Plant growth promoting bacteria as biofertilizer. Agronomy for Sustainable Development. 26: 143-150. DOI: 10.1051/agro:2006007.
  10. Hallmann, J. and Berg, G. (2006). Spectrum and population dynamics of bacterial root endophytes. In Microbial root endophytes Springer, Berlin, Heidelberg. pp. 15-31.
  11. Hussain, A. and Hasnain, S. (2011). Interactions of bacterial cytokinins and IAA in the rhizosphere may alter phytostimulatory efficiency of rhizobacteria. World Journal of Microbiology and Biotechnology. 27: 2645-2654. DOI: 10.1007/s11274-011-0738-y.
  12. Kang, S.M., Radhakrishnan, R., You, Y.H., Joo, G.J., Lee, I.J., Lee, K.E., Kim, J.H. (2014). Phosphate solubilizing Bacillus megaterium mj1212 regulates endogenous plant carbohy drates and amino acids contents to promote mustard plant growth. Indian Journal of Microbiology. 54: 427-433. DOI: 10.1007/s12088-014-0476-6.
  13. Kunwar, V.S., Chimouriya, S., Lamichhane, J., Gauchan, D.P. (2018). Isolation and characterization of phosphate solubilizing bacteria from rhizosphere of coffee plant and evaluating their effects on growth and development of coffee seedlings. Bio Technology: An Indian Journal. 14: 173.
  14. Malhotra, M. and Srivastava, S. (2008). Organization of the ipdC region regulates IAA levels in different Azospirillum brasilense strains: molecular and functional analysis of ipdC in strain SM. Environmental Microbiology. 10: 1365-1373. DOI: 10.1111/j.1462-2920.2007.01529.x.
  15. Meliani, A. Bensoltane, A., Benidire, L., Oufdou, K. (2017). Plant growth-promotion and IAA secretion with Pseudomonas fluorescens and Pseudomonas putida. Research and Reviews: Journal of Botanical Sciences. 6: 16-24.
  16. Mirza, M.S., Ahmad, W., Latif, F., Haurat, J., Bally, R., Normand, P., Malik, KA. (2001). Isolation, partial characterization and the effect of plant growth-promoting bacteria (PGPB) on micro-propagated sugarcane in vitro. Plant and Soil. 237: 47-54.
  17. Mohite, B. (2013). Isolation and characterization of indole acetic acid (IAA) producing bacteria from rhizospheric soil and its effect on plant growth. Journal of Soil Science and Plant Nutrition. 13: 638-649. DOI: 10.4067/S0718-95162 013005000051. 
  18. Nadeem, S.M., Zahir, Z.A., Naveed, M., Arshad, M. (2009). Rhizobacteria containing ACC-deaminase confer salt tolerance in maize grown on salt-affected fields. Canadian Journal of Microbiology. 55:1302-1309. DOI: 10.1139/W09-092.
  19. Nehra, V., Saharan, B.S., Choudhary, M. (2014). Potential plant growth promoting activity of Pseudomonas fluorescens sp. isolated from cotton (Gossypium hirsutum) crop. Indian Journal of Agricultural Research. 48: 97-104. DOI: 10.5958/ j.0976-058X.48.2.017.
  20. Pathak, R. Paudel, V. Shrestha, A., Lamichhane, J., Gauchan, D.P. (2017). Isolation of phosphate solubilizing bacteria and their use for plant growth promotion in tomato seedling and plant. Kathmandu University Journal of Science, Engineering and Technology. 13:61-70.
  21. Puri, R.R., Dangi, S.R., Dhungana, S.A., Itoh, K. (2018). Diversity and plant growth promoting ability of culturable endophytic bacteria in Nepalese sweet potato. Advances in Micobiology. 8: 734-761. DOI: 10.4236/aim.2018.8904.
  22. Rahman, A., Sitepu, .I.R., Tang, S.Y., Hashidoko, Y. (2010). Salkowski’s reagent test as a primary screening index for functionalities of rhizobacteria isolated from wild dipterocarp saplings growing naturally on medium-strongly acidic tropical peat soil. Bioscience, Biotechnology and Biochemistry. 74: 2202-2208. DOI: 10.1271/bbb.100360.
  23. Ruangsanka, S. (2014). Identification of phosphate-solubilizing bacteria from the bamboo rhizosphere. Science Asia. 40: 204-211. DOI: 10.2306/ 1513-1874.2014.40.204.
  24. Sandeep, C., Thejas, M.S., Patra, S., Gowda, T., Raman, R.V., Radhika, M., Mulla, S.R. (2011). Growth response of ayapana on inoculation with Bacillus megaterium isolated from different soil types of various agroclimatic zones of Karnataka. Journal of Phytology. 3: 13-18.
  25. Sergeeva, E., Hirkala, D.L., Nelson, L.M. (2007). Production of indole-3-acetic acid, aromatic amino acid aminotransfe rase activities and plant growth promotion by Pantoea agglomerans rhizosphere isolates. Plant and Soil. 297: 1-13. DOI: 10.1007/s11104-007-931.
  26. Sharma, S., Verma, P.P., Kaur, M. (2014). Isolation, purification and estimation of IAA from Pseudomonas sp. using high-performance liquid chromatography. Journal of Pure and Applied Microbiology. 8: 3203-3208.
  27. Shrivastava, U.P. (2013). Characterization of diazotrophic rhizobacteria under various conditions. International Journal of Applied Sciences and Biotechnology. 1: 110-117. DOI: 10.3126/ijasbt.v1i3.8607.
  28. Spaepen, S., Vanderleyden, J. Remans, R. (2007). Indole-3-acetic acid in microbial and microorganism-plant signaling. FEMS Microbiology Reviews. 31:425-448. DOI: 10.1111/j.1574-6976.2007.00072.x.
  29. Sridevi, M. and Mallaiah, K.V. (2007). Bioproduction of indole acetic acid by Rhizobium strains isolated from root nodules of green manure crop, Sesbania sesban L. Iranian Journal of Biotechnology. 5: 178-182. 
  30. Suresh, A., Ramesh, M., Reddy, S.R. (2014). Fluorescent pseudomonads contribute to the enhanced growth and yield of rice cultivated under system of rice intensification (SRI). Indian Journal of Agricultural Research. 48: 287-293. DOI: 10.5958/0976-058X.2014.00662.3.
  31. Tan, X., Calderon-Villalobos L.I.A., Sharon, M., Zheng, C., Robinson, C.V., Estelle, M., Zheng, N. (2007). Mechanism of auxin perception by the TIR1 ubiquitin ligase. Nature. 446: 640-645. DOI: 10.1038/nature05731.
  32. Tsavkelova, E.A., Klimova, S.Y., Cherdyntseva, T.A., Netrusov, A.I. (2006). Microbial producers of plant growth stimulators and their practical use: a review. Applied Biochemistry and Microbiology. 42:117-126. DOI: 10.1134/S000368 3806020013.
  33. Weisskopf, L., Heller, S., Eberl, L. (2011). Burkholderia species are major inhabitants of white lupin cluster roots. Applied Environmental Microbiology. 77: 7715-7720. DOI: 10.1128/ AEM.05845 -11.
  34. Zahid, M. (2015). Isolation and identification of indigenous plant growth promoting rhizobacteria from Himalayan region of Kashmir and their effect on improving growth and nutrient contents of maize (Zea mays L.). Frontiers in Microbiology. 6: 1-10. DOI: 10.3389/fmicb. 2015. 00207.
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    Research Article

    volume 54 issue 6 (december 2020) : 781-786,   Doi: 10.18805/IJARe.A-5578

    Extraction of Indole-3-acetic Acid from Plant Growth Promoting Rhizobacteria of Bamboo Rhizosphere and Its Effect on Biosynthesis of Chlorophyll in Bamboo Seedlings

    B
    Bishnu Maya K.C.
    D
    Dhurva Prasad Gauchan
    S
    Sanjay Nath Khanal
    S
    Sharmila Chimouriya
    J
    Janardan Lamichhane
    1Department of Biotechnology, School of Science, Kathmandu University, Dhulikhel, Nepal.
    Cite article:- K.C. Maya Bishnu, Gauchan Prasad Dhurva, Khanal Nath Sanjay, Chimouriya Sharmila, Lamichhane Janardan (2020). Extraction of Indole-3-acetic Acid from Plant Growth Promoting Rhizobacteria of Bamboo Rhizosphere and Its Effect on Biosynthesis of Chlorophyll in Bamboo Seedlings. Indian Journal of Agricultural Research. 54(6): 781-786. doi: 10.18805/IJARe.A-5578.

    ABSTRACT

    Background: Indole-3-acetic acid (IAA), a principal phytohormone, controls several crucial physiological processes of plants. It ameliorates plant growth by stimulating cell elongation, root initiation, seed germination and seedling growth. Alteration of IAA level by plant growth promoting rhizobacteria leads to varied impacts on plant growth and development.
    Methods: Soil samples were collected from bamboo (Bambusa tulda, B. nutans subsp. cupulata, B. balcooa and Dendrocalamus strictus) rhizosphere. Altogether five bacterial isolates were screened by serial dilution method and subjected to biochemical analysis. The isolate BUX1 with high IAA production capacity was optimized for IAA production. IAA was partially purified and quantified from the bacterial extract by thin layer chromatography (TLC). The influence of extracted bacterial IAA on chlorophyll biosynthesis in bamboo seedlings of B. tulda was compared with uninoculated control plants.
    Results: Biochemical analysis revealed that all the isolates belonged to genus Bacillus which were found capable of producing IAA. During optimization, BUX1 isolate produced 99.13 µg ml-1 of IAA at 37°C, pH 7, 3 mg l-1 concentration of L-tryptophan and 150 rpm agitation rate after 192 hour of incubation. The Rf value of the bacterial IAA during TLC was identical to that of standard IAA (0.425) indicating that IAA was present in crude extract of Bacillus (BUX1). The influence of bacterial IAA on chlorophyll biosynthesis in bamboo seedlings was significant in comparison to uninoculated plants. Therefore, this isolate could be a prospective candidate to be employed as biofertilizer. 

    REFERENCES

    1. Adhikari, D., Kaneto, M., Itoh, K., Suyama, K., Pokharel, B.B., Gaihre, Y.K. (2012). Genetic diversity of soybean-nodulating rhizobia in Nepal in relation to climate and soil properties. Plant and Soil. 357: 131-145. DOI: 10.1007/s11104-012-1134-6.
    2. Ahmed, A. and Hasnain, S. (2010). Auxin-producing Bacillus sp.: Auxin quantification and effect on the growth of Solanum tuberosum. Pure and Applied Chemistry. 82: 313-319. DOI: 10.1351/PAC-CON-09-02-06.
    3. Arnon, D.I. (1949). Copper enzymes in isolated chloroplasts. Polyphenol oxidase in Beta vulgaris. Plant Physiology. 24: 1-15. 
    4. Asghar, H.N., Zahir, Z.A., Arshad, M., Khaliq, A. (2002). Relationship between in vitro production of auxins by rhizobacteria and their growth promoting activities in Brassica juncea L. Biology and Fertility of Soils. 35: 231-237. DOI: 10.10 07/s00374-002-0462-8.
    5. Barillot, C.D., Sarde, C.O., Bert, V., Tarnaud, E., Cochet, N. (2013). A standardized method for the sampling of rhizosphere and rhizoplan soil bacteria associated to a herbaceous root system. Annals of Microbiology. 63: 471-476. DOI: 10.1007/s13213-012-0491-y.
    6. Biswas, S., Kundu, D.K., Mazumdar, S.P., Saha, A.R., Majumdar, B., Ghorai, A.K., Ghosh, D., Yadav, A.N., Saxena, A.K. (2018). Study on the activity and diversity of bacteria in a New Gangetic alluvial soil (Eutrocrept) under rice-wheat- jute cropping system. Journal of Environmental Biology. 39: 379-86. DOI: 10.22438/jeb/39/3/MRN-523.
    7. Garrity, G.M., Bell, J.A., Lilburn, T.G. (2004). Taxonomic outline of the prokaryotes. Bergey’s manual of systematic bacteriology. Springer, New York, Berlin, Heidelberg.
    8. Gul, A., Salam, A., Afridi, M.S., Bangash, N.K., Ali, F., Ali, M.Y., Khan, S., Mubeeen, R. (2019). Effect of urea, bio-fertilizers and their interaction on the growth, yield and yield attributes of Cyamopsis tetragonoloba. Indian Journal of Agricultural Research. 53: 423-428. DOI: 10.18805/IJARe.A-395. 
    9. Hafeez, F.Y., Yasmin, S., Ariani, D., Rahman, M., Zafar, Y., Malik, K.A. (2006). Plant growth promoting bacteria as biofertilizer. Agronomy for Sustainable Development. 26: 143-150. DOI: 10.1051/agro:2006007.
    10. Hallmann, J. and Berg, G. (2006). Spectrum and population dynamics of bacterial root endophytes. In Microbial root endophytes Springer, Berlin, Heidelberg. pp. 15-31.
    11. Hussain, A. and Hasnain, S. (2011). Interactions of bacterial cytokinins and IAA in the rhizosphere may alter phytostimulatory efficiency of rhizobacteria. World Journal of Microbiology and Biotechnology. 27: 2645-2654. DOI: 10.1007/s11274-011-0738-y.
    12. Kang, S.M., Radhakrishnan, R., You, Y.H., Joo, G.J., Lee, I.J., Lee, K.E., Kim, J.H. (2014). Phosphate solubilizing Bacillus megaterium mj1212 regulates endogenous plant carbohy drates and amino acids contents to promote mustard plant growth. Indian Journal of Microbiology. 54: 427-433. DOI: 10.1007/s12088-014-0476-6.
    13. Kunwar, V.S., Chimouriya, S., Lamichhane, J., Gauchan, D.P. (2018). Isolation and characterization of phosphate solubilizing bacteria from rhizosphere of coffee plant and evaluating their effects on growth and development of coffee seedlings. Bio Technology: An Indian Journal. 14: 173.
    14. Malhotra, M. and Srivastava, S. (2008). Organization of the ipdC region regulates IAA levels in different Azospirillum brasilense strains: molecular and functional analysis of ipdC in strain SM. Environmental Microbiology. 10: 1365-1373. DOI: 10.1111/j.1462-2920.2007.01529.x.
    15. Meliani, A. Bensoltane, A., Benidire, L., Oufdou, K. (2017). Plant growth-promotion and IAA secretion with Pseudomonas fluorescens and Pseudomonas putida. Research and Reviews: Journal of Botanical Sciences. 6: 16-24.
    16. Mirza, M.S., Ahmad, W., Latif, F., Haurat, J., Bally, R., Normand, P., Malik, KA. (2001). Isolation, partial characterization and the effect of plant growth-promoting bacteria (PGPB) on micro-propagated sugarcane in vitro. Plant and Soil. 237: 47-54.
    17. Mohite, B. (2013). Isolation and characterization of indole acetic acid (IAA) producing bacteria from rhizospheric soil and its effect on plant growth. Journal of Soil Science and Plant Nutrition. 13: 638-649. DOI: 10.4067/S0718-95162 013005000051. 
    18. Nadeem, S.M., Zahir, Z.A., Naveed, M., Arshad, M. (2009). Rhizobacteria containing ACC-deaminase confer salt tolerance in maize grown on salt-affected fields. Canadian Journal of Microbiology. 55:1302-1309. DOI: 10.1139/W09-092.
    19. Nehra, V., Saharan, B.S., Choudhary, M. (2014). Potential plant growth promoting activity of Pseudomonas fluorescens sp. isolated from cotton (Gossypium hirsutum) crop. Indian Journal of Agricultural Research. 48: 97-104. DOI: 10.5958/ j.0976-058X.48.2.017.
    20. Pathak, R. Paudel, V. Shrestha, A., Lamichhane, J., Gauchan, D.P. (2017). Isolation of phosphate solubilizing bacteria and their use for plant growth promotion in tomato seedling and plant. Kathmandu University Journal of Science, Engineering and Technology. 13:61-70.
    21. Puri, R.R., Dangi, S.R., Dhungana, S.A., Itoh, K. (2018). Diversity and plant growth promoting ability of culturable endophytic bacteria in Nepalese sweet potato. Advances in Micobiology. 8: 734-761. DOI: 10.4236/aim.2018.8904.
    22. Rahman, A., Sitepu, .I.R., Tang, S.Y., Hashidoko, Y. (2010). Salkowski’s reagent test as a primary screening index for functionalities of rhizobacteria isolated from wild dipterocarp saplings growing naturally on medium-strongly acidic tropical peat soil. Bioscience, Biotechnology and Biochemistry. 74: 2202-2208. DOI: 10.1271/bbb.100360.
    23. Ruangsanka, S. (2014). Identification of phosphate-solubilizing bacteria from the bamboo rhizosphere. Science Asia. 40: 204-211. DOI: 10.2306/ 1513-1874.2014.40.204.
    24. Sandeep, C., Thejas, M.S., Patra, S., Gowda, T., Raman, R.V., Radhika, M., Mulla, S.R. (2011). Growth response of ayapana on inoculation with Bacillus megaterium isolated from different soil types of various agroclimatic zones of Karnataka. Journal of Phytology. 3: 13-18.
    25. Sergeeva, E., Hirkala, D.L., Nelson, L.M. (2007). Production of indole-3-acetic acid, aromatic amino acid aminotransfe rase activities and plant growth promotion by Pantoea agglomerans rhizosphere isolates. Plant and Soil. 297: 1-13. DOI: 10.1007/s11104-007-931.
    26. Sharma, S., Verma, P.P., Kaur, M. (2014). Isolation, purification and estimation of IAA from Pseudomonas sp. using high-performance liquid chromatography. Journal of Pure and Applied Microbiology. 8: 3203-3208.
    27. Shrivastava, U.P. (2013). Characterization of diazotrophic rhizobacteria under various conditions. International Journal of Applied Sciences and Biotechnology. 1: 110-117. DOI: 10.3126/ijasbt.v1i3.8607.
    28. Spaepen, S., Vanderleyden, J. Remans, R. (2007). Indole-3-acetic acid in microbial and microorganism-plant signaling. FEMS Microbiology Reviews. 31:425-448. DOI: 10.1111/j.1574-6976.2007.00072.x.
    29. Sridevi, M. and Mallaiah, K.V. (2007). Bioproduction of indole acetic acid by Rhizobium strains isolated from root nodules of green manure crop, Sesbania sesban L. Iranian Journal of Biotechnology. 5: 178-182. 
    30. Suresh, A., Ramesh, M., Reddy, S.R. (2014). Fluorescent pseudomonads contribute to the enhanced growth and yield of rice cultivated under system of rice intensification (SRI). Indian Journal of Agricultural Research. 48: 287-293. DOI: 10.5958/0976-058X.2014.00662.3.
    31. Tan, X., Calderon-Villalobos L.I.A., Sharon, M., Zheng, C., Robinson, C.V., Estelle, M., Zheng, N. (2007). Mechanism of auxin perception by the TIR1 ubiquitin ligase. Nature. 446: 640-645. DOI: 10.1038/nature05731.
    32. Tsavkelova, E.A., Klimova, S.Y., Cherdyntseva, T.A., Netrusov, A.I. (2006). Microbial producers of plant growth stimulators and their practical use: a review. Applied Biochemistry and Microbiology. 42:117-126. DOI: 10.1134/S000368 3806020013.
    33. Weisskopf, L., Heller, S., Eberl, L. (2011). Burkholderia species are major inhabitants of white lupin cluster roots. Applied Environmental Microbiology. 77: 7715-7720. DOI: 10.1128/ AEM.05845 -11.
    34. Zahid, M. (2015). Isolation and identification of indigenous plant growth promoting rhizobacteria from Himalayan region of Kashmir and their effect on improving growth and nutrient contents of maize (Zea mays L.). Frontiers in Microbiology. 6: 1-10. DOI: 10.3389/fmicb. 2015. 00207.
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