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Genetic transformation of the bast fiber plant ramie (Boehmeria nivea Gaud.) via Agrobacterium tumefaciens

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Abstract

Hypocotyls and cotyledons of three ramie (Boehmeria nivea Gaud.) cultivars (Zhongzhu No.1, Zhongsizhu No.1 and NC01), important plants for textile fiber, were pre-cultured on callus-inducing medium for 1 day before co-cultivation for 2 days with Agrobacterium tumefaciens strain LBA4404 harboring the plasmid pGBI4ABC carrying two insect resistance (CryIA and CpTI), gus, and neomycin phosphotransferase (npt II) genes. Calli were induced from both hypocotyl and cotyledon explants grown on a kanamycin selection medium. Regenerated shoots were obtained after two cycles of culture and transferred to rooting medium. Kanamycin-resistant plantlets were rooted in 2 weeks, and then transplanted to soil. Transgenic plants were subsequently confirmed by polymerase chain reaction, Southern blot hybridization, and GUS assays. More than 100 transgenic plants carrying insect-resistance genes were produced. A transformation frequency of 8.8 to 10.3% was obtained using hypocotyls as explants, which was higher than all previously reported transformation frequencies. The whole protocol, from transformation recovery of plants grown in soil, was completed within 2–4 months. Therefore, a simple, efficient, and robust Agrobacterium tumefaciens -mediated transformation system for ramie has been developed.

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Abbreviations

BA:

Benzyladenine

GA3 :

Gibberellic acid

GUS:

β-Glucuronidase

TDZ:

Thidiazuron (N-phenyl-N′-1,2,3-thidiazol-5-urea)

IAA:

Indoleacetic acid

NAA:

α-Naphthaleneacetic acid

References

  • Boulter D, Gatehouse A, Hilder V (1989) Use of cowpea trypsin inhibitor (CpTI) to protect plants against insect predation. Biotechnol Adv 7:489–497

    Article  CAS  PubMed  Google Scholar 

  • Charity JA, Holland L, Donaldson SS, Grace L, Walter C (2002) Agrobacterium-mediated transformation of pinus radiata organogenic tissue using vacuum-infiltration. Plant Cell Tiss Organ Cult 70:51–60

    Article  CAS  Google Scholar 

  • Cui H, Guo S (1998) Construction of plant expression vectors harboring two insecticidal genes and their expression in tabacco. J Agric Biotechnol 1:7–13

    Google Scholar 

  • Duncan D (1995) Multiple range and multiple F tests. Biometrics 11:1–42

    Article  Google Scholar 

  • Dusi D, Dubald M, de Almeida E, Caldas L, Gander E (1993) Transgenic plants of ramie (Boehmeria nivea Gaud.) obtained by Agrobacterium mediated transformation. Plant Cell Rep 12:625–628

    Article  CAS  Google Scholar 

  • Goda K, Sreekala M, Gomes A, Kaji T, Ohgi J (2006) Improvement of plant based natural fibers of toughening green composites—effect of load application during mercerization of ramie fibers. Composites Part A 37:2213–2220

    Google Scholar 

  • Gulati A, Schryer P, Mchughen A (2002) Production of fertile transgenic lentil (Lens culinaris Medik) plants using particle bombardment. In vitro Cell Dev Biol Plant 38:316–324

    Article  CAS  Google Scholar 

  • Guo SD, Ni WC, Xu QF (1995) Expression vector of fused insecticidal protein gene and transgenic plants. Patent number : ZL 95119563.8, C12 N 15/32. 1995, 12.28

  • Guo QQ, Chen JR, Yang RF, Hu RS (1998) Callus induction and shoot regeneration from leaves of Ramie (Boehmeria nivea Gaud). China’s Fiber Crops 20:1–4

    Google Scholar 

  • Guo S, Cui H, Xia L, Wu D, Ni W, Zhang Z, Zhang B, Xu Y (1999) Development of bivalent insect-resistant transgenic cotton plants. Scientia Agricultura Sinica 32(03):1–7

    Google Scholar 

  • Huang JS, Mo RD (1981) De novo shoot organogenesis from hypocotyls of ramie (Boehmeria nivea Gaud). Acta Bio Exp Sin 14:111–114

    Google Scholar 

  • Huang JS, Mo RD, Ye CJ (1980) Research note on in vitro plant regeneration from cotyledon and hypocotyls of ramie (Boehmeria nivea Gaud.). Guangxi Agric Sci 7:27

    Google Scholar 

  • James C (2007) Global status of commercialized biotech/GM crops: 2007. ISAAA Briefs No 37, international service for the acquisition of agri-biotech applications, Ithaca, NY, 2007

  • Jefferson J, Kavanagh T, Bevan M (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6(13):3901–3907

    CAS  PubMed  Google Scholar 

  • Li F, Cui J, Liu C, Wu Z, Li F, Zhou Y, Li X, Guo S, Cui H (2000) The study of insect-resistant transgenic cotton habouring double-gene and its insect-resistance. Scientia Agricultura Sinica 33(1):46–52

    CAS  Google Scholar 

  • Liu F, Liu Q, Liang X, Huang H, Zhang S (2005) Morphological, anatomical, and physiological assessment of ramie [Boehmeria Nivea (L.) Gaud.] tolerance to soil drought. Genet Resour Crop Evol 52:497–506

    Article  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Pan CL, Li SC, Li YJ (1995) Factors influencing plant regeneration efficiency of ramie (Boehmeria nivea Gaud). China’s Fiber Crops 17:1–6

    Google Scholar 

  • Wang B, Peng DX, Liu LJ, Sun ZX, Zhang N, Gao SM (2007a) An efficient adventitious shoot regeneration system for ramie (Boehmeria nivea Gaud) using thidiazuron. Botanical Studies 48:173–180

    Google Scholar 

  • Wang B, Peng DX, Sun ZX, Zhang N, Xing XL (2007b) Regeneration of transgenic ramie plants expressing green fluorescent protein mediated by Agrobacterium tumefaciens. Acta Agronomica Sinica 33(10):1606–1610

    CAS  Google Scholar 

  • Wang B, Peng D, Sun Z, Zhang N, Gao S (2008) In vitro plant regeneration from seedling-derived explants of ramie [Boehmeria nivea (L.) Gaud]. In vitro Cell Dev Biol Plant 44:105–111

    Google Scholar 

  • Xu D, Xue Q, McElroy D, Mawal Y, Hilder VA, Wu R (1996) Constitutive expression of a cowpea trypsin inhibitor gene, CpTi, in transgenic rice plants confers resistance to two major rice insect pests. Mol Breeding 2:167–173

    Article  CAS  Google Scholar 

  • Yi ZL, Li X, Jiang JX, Wang ZC, Liu QB (2006) The establishment of regeneration system and obtaining the insect-resistant transgenic plants of ramie (Boehmeria nivea L.). China Fiber Crops 28(2):61–66

    Google Scholar 

  • Zhao J, Fan X (1997) Gene pyramiding: an effective strategy of resistance management for Helicoverpa armigera and Bacilluss thuringiensis. Resist Pest Manag 9(2):19–21

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Acknowledgments

This research was supported by National Key Technology R&D Programs of the China (2006BAD06B03) and Special Funds for Public Welfare Industries of the China (nyhyzx07-018).The authors wish to thank Prof Xinbo Chen, Prof Chengbin Xiang and Dr. Michael K Deyholos for critical review of the manuscript. Xiongfeng Ma wishes to thank Ms. Xueyan Zhang and Ms. Song Yun for their helpful work.

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Authors and Affiliations

  1. Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, 348 West Xianjiahu Road, 410205, Changsha, China

    Xiongfeng Ma, Chunming Yu, Shouwei Tang, Yanzhou Wang, Aiguo Zhu, Siyuan Zhu & Heping Xiong

  2. Biotechnology Research Center, Chinese Academy of Agricultural Sciences, 12 Zhongguancun, South Street, 100081, Beijing, China

    Sandui Guo & Rui Zhang

  3. Institute of Cotton Research, Chinese Academy of Agricultural Sciences, West Huanghe Street, 455000, Anyang, China

    Xiongfeng Ma

Authors
  1. Xiongfeng Ma
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  2. Chunming Yu
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  3. Shouwei Tang
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  4. Sandui Guo
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  5. Rui Zhang
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  6. Yanzhou Wang
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  7. Aiguo Zhu
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  8. Siyuan Zhu
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  9. Heping Xiong
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Correspondence to Heping Xiong.

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Ma, X., Yu, C., Tang, S. et al. Genetic transformation of the bast fiber plant ramie (Boehmeria nivea Gaud.) via Agrobacterium tumefaciens . Plant Cell Tiss Organ Cult 100, 165–174 (2010). https://doi.org/10.1007/s11240-009-9633-8

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  • DOI: https://doi.org/10.1007/s11240-009-9633-8

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