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Enhancement of in vitro growth of Swertia chirayita Roxb. Ex Fleming co-cultured with plant growth promoting rhizobacteria

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Abstract

Micropropagated plants usually show high mortality rate when transferred to the field that depends upon different biotic and abiotic factors. Also, a weak root system of the plants arises due to the absence of rhizospheric microflora. In this work, in vitro co-culturing of Swertia chirayita shoots with the plant growth promoting rhizobacteria (PGPR) isolated from the indigenous rhizospheric soil samples was performed. The rhizospheric isolates of fluorescent Pseudomonas and Azotobacter isolated from rhizosphere of S. chirayita from different high altitude regions of Uttarakhand were co-cultured with micropropagated shoots of S. chirayita on plant tissue culture medium and the effect on in vitro caulogenesis and rhizogenesis along with different phytohormones (alone and in combination and tryptophan precursor) was tested. A significant plant growth promotion was observed which not only increased the survival rate (up to 90 %), but also the multiplication rate at low auxin concentration compared to noninoculated plantlets. Both strains showed tryptophan dependent synthesis on MS basal medium supplemented with 200 μM l-tryptophan with maximum IAA production by Azo2-6 (1.5 μg/ml IAA in 96 h) and best phosphate solubilization efficiency of 126.31 %. Azo2-6 showed the best growth promotion on MS supplemented with 0.25 mg/l BAP compared to the control. The co-culturing of PGPR resulted in sevenfold to eightfold shoot multiplication and pronounced degree of vigor in the plantlets. The pots transferred plants inoculated with Azo-2-6 and PS-2 showed the maximum survival rate. A new technique of in vitro biotization in plant tissue culture was devised that helps in screening of positive plant–microbe interaction under in vitro conditions which can be suitably under the field condition for plant growth promotion.

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Abbreviations

PGPR:

Plant growth promoting rhizobacteria

Azo:

Azotobactor

PS:

Pseudomonas

BAP:

6 Benzyl aminopurine

IAA:

Indole-3 acetic acid

IBA:

Indole butyric acid

NAA:

Naphthalene acetic acid

LB:

Luria britani

Trp+ :

with Tryptophan

Trp :

without Tryptophan

References

  • Agarwal T, Gupta AK, Patel AK, Shekhawat NS (2014) Micropropagation and validation of genetic homogeneity of Alhagi maurorum using SCoT, ISSR and RAPD markers. Plant Cell Tissue Organ. doi:10.1007/s11240-014-0608-z

    Google Scholar 

  • Ali B, Hasnain S (2007) Potential of bacterial indoleacetic acid to induce adventitious shoots in plant tissue culture. Lett Appl Microbiol 45(2):128

    Article  CAS  PubMed  Google Scholar 

  • Asghar HN, Zahir ZA, Arshad M (2004) Screening rhizobacteria for improving the growth, yield & oil content of canola (Brassica nappus). Aust J Agr Res 55:187–194

    Article  Google Scholar 

  • Barka E, Belarbi A, Hachet C, Nowak J, Audran JC (2000) Enhancement of in vitro growth and resistance to gray mould of Vitis vinifera co-cultured with plant growth-promoting rhizobacteria. FEMS Microbiol Lett 186(1):91–95

    Article  CAS  PubMed  Google Scholar 

  • Bertrand H, Nalin R, Bally R, Marel JCC (2001) Isolation and identification of the most efficient plant growth promoting bacteria associated with canola. Biol Fertil Soils 33:152–156

    Article  Google Scholar 

  • Cattelan AJ, Hartel PG, Fuhrmann JJ (1999) Screening of plant growth promoting rhizobacteria to promote early soybean growth. Soil Sci Soc Am J 63:1670–1680

    Article  CAS  Google Scholar 

  • Chandrasekar B, Bajpai MB, Mukherjee SK (1990) Hypoglycaemic activity of Swertia chirata (Roxb ex Flem) Karst. Indian J Exp Biol 28:616–618 PMid: 2272647

    CAS  PubMed  Google Scholar 

  • Chaudhuri RK, Pal A, Jha TB (2007) Production of genetically uniform plants from the nodal explants of Swertia chirayita Buch.-Ham. ex Wall.—an endangered medicinal herb. In vitro Cell Dev Plant 43(5):467–472

    Article  CAS  Google Scholar 

  • Chaudhuri RK, Pal A, Jha TB (2008) Conservation of Swertia chirata through direct shoot multiplication from leaf explants. Plant Biotechnol Rep 2:213–218

    Article  Google Scholar 

  • Chopade BA, Huddedar SB, Shete AM, Tilekar JN, Gore SD, Dhavale DD (2008) Plasmid encoding IAA and method thereof. US Pat 7(341):868

    Google Scholar 

  • Dave A, Bilochi G, Purohit SD (2003) Scaling-up production and field performance of micropropagated medicinal herb ‘Safed Musli’ (Chlorophytum borivilianum). In Vitro Cell Dev Plant 39:419–424

    Article  Google Scholar 

  • Dobbelaere S, Vanderleyden J, Okon Y (2003) Plant growth promoting effects of dizotrophs in the rhizosphere. Crit Rev Plant Sci 22:149–197

    Article  Google Scholar 

  • Govindarajan M, Soon-Wo K, Han-Yeon W (2007) Isolation, molecular characterization and growth promoting activities of endophytic sugar cane diazotroph Klebsiella sp. GR9. World J Microbiol Biotechnol 23:997

    Article  CAS  Google Scholar 

  • Iqbal Z, Lateef M, Kha MN, Jabbar A, Akhtar MS (2006) Antihelmintic activity of Swertia chirata against gastrointestinal nematodes of sheep. Fitoterapia 77:463–465

    Article  PubMed  Google Scholar 

  • Joshi P, Dhawan V (2007) Axillary multiplication of Swertia chirayita (Roxb. Ex Fleming) H. Karst., a critically endangered medicinal herb of temperate Himalayas. In vitro Cell Dev Plant 43:631–638

    Article  CAS  Google Scholar 

  • Karnwal A (2009) Production of indole acetic acid by fluorescent Pseudomonas in the presence of l-tryptophan and rice root exudates. J Plant Pathol 91:61–63

    CAS  Google Scholar 

  • Khalid A, Arshad M, Zahir ZA, Khaliq A (1997) Potential of plant growth promoting rhizobacteria for enhancing Wheat yield. J Anim Plant Sci 7:53–56

    Google Scholar 

  • Khalid A, Arshad M, Zahir ZA (2004) Screening plant growth promotin rhizobacteria for improving growth and yield of wheat. J Appl Microbiol 96:473

    Article  CAS  PubMed  Google Scholar 

  • Kravchenko LV, Borovkov AV, Pshikril Z (1991) Possibility of auxin synthesis by association-forming nitrogen-fixing bacteria in the rhizosphere of wheat. Microbiology 60:647–650

    Google Scholar 

  • Kravchenko LV, Azarova TS, Makarova NM, Tikhonovich IA (2004) The effect of tryptophan present in plant root exudates on the phytostimulating activity of rhizobacteria. Microbiology 73:156–158

    Article  CAS  Google Scholar 

  • Martin KP (2003) Rapid in vitro multiplication and ex vitro rooting of Rotula aquatica Lour., a rare rhoeophytic woody medicinal plant. Plant Cell Rep 21:415–420

    CAS  PubMed  Google Scholar 

  • Mukherjee S, Sur A, Maiti BR (1997) Hepatoprotective effect of Swertia chirata on rat. Indian J Exp Biol 35:384–388

    CAS  PubMed  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Planta 19:473–497

  • Narumiya S, Takai K, Tokuyama T, Noda Y, Ushiro H, Hayaishi O (1979) A new metabolic pathway of tryptophan initiated by tryptophan side chain oxidase. J Biol Chem 254:7007–7015

    CAS  PubMed  Google Scholar 

  • Omay SH, Schmidt WA, Martin P (1993) Indoleacetic acid production by the rhizosphere bacterium Azospirillum brasilense Cd under in vitro conditions. Can J Microbiol 39:187–192

    Article  CAS  Google Scholar 

  • Pant M, Bisht P, Gusain MP (2010) De novo shoot organogenesis from cultured root explants of Swertia chirata Buch.-Ham. ex Wall.: an endangered medicinal plant. Nat Sci 8(9):244–252

    Google Scholar 

  • Patten C, Glick BR (1996) Bacterial biosynthesis of indole-3-acetic acid. Can J Microbiol 42:207–220

    Article  CAS  PubMed  Google Scholar 

  • Patten CL, Glick BR (2002) Regulation of indole acetic acid production in Pseudomonas putida GR12-2 by tryptophan and the stationary-phase sigma factor RpoS. Can J Microbiol 48(7):635

    Article  CAS  PubMed  Google Scholar 

  • Spaepen S, Vanderleyden J, Remans R (2007) Indole-3-acetic acid in microbial and microorganism-plant signalling. FEMS Microbiol Rev 31:425–448 

  • Sarwar M, Frankenberger WT (1994) Influence of l-Tryptophan and auxins applied to the rhizosphere on the vegetative growth of Zea mays L. Plant Soil 160:97–104

    Article  CAS  Google Scholar 

  • Sharma V, Srivastava N, Kamal B, Dobriyal AK, Jadon VS (2013) Effects of Additives in Shoot Multiplication and Genetic Validation in Swertia chirayita Revealed through RAPD Analysis. Plant Tiss Cult Biotech 23(1):11–19

    CAS  Google Scholar 

  • Sushil R, Prakash S, Bhargava P, Shukla S (2009) Antipyretic Potential of Swertia chirata Buch Ham. Root Extract. Sci Pharm J 77:617–623

    Article  Google Scholar 

  • Thomas J, Ajay D, Raj Kumar R, Mandal AKA (2010) Influence of beneficial microorganisms during in vivo acclimatization of in vitro-derived tea (Camellia sinensis) plants. Plant Cell Tiss Org 101:365–370

    Article  Google Scholar 

  • Vincent KA, Mary MK, Molly H (1992) Micropropagation of Kaempferia galanga L.—a medicinal plant. Plant Cell Tiss Org 28:229–230

    Article  CAS  Google Scholar 

  • Wang L, An L, Hu Y, Wei L, Li Y (2009) Influence of phytohormones and medium on the shoot regeneration from leaf of Swertia chirata Buch.-Ham. ex Wall. in vitro. Afr J Biotechnol 8(11):2513–2517

    CAS  Google Scholar 

  • Wawrosch C, Maskay N, Kopp B (1999) Micropropagation of the threatened Nepalese medicinal plant Swertia chirata Buch.-Ham. ex Wall. Plant Cell Rep 18:997–1001

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to express their gratitude to The Management, Sardar Bhagwan Singh (P. G.) Institute of Biomedical Sciences & Research, Balawala, Dehradun for providing research facilities to conduct the work and The Director, Uttarakhand State Council for Science and Technology, Dehradun for providing the financial assistance.

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Correspondence to Vikash Singh Jadon.

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Sharma, V., Kamal, B., Srivastava, N. et al. Enhancement of in vitro growth of Swertia chirayita Roxb. Ex Fleming co-cultured with plant growth promoting rhizobacteria. Plant Cell Tiss Organ Cult 121, 215–225 (2015). https://doi.org/10.1007/s11240-014-0696-9

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