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Biolistic transformation of cotton (Gossypium hirsutum L.) with the phyA gene from Aspergillus ficuum

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Abstract

Transgenic cotton with an increased level of phytase activity was generated from cotton (Gossypium hirsutum L.) cv. ND94-7 by subjecting shoot-apex explants to particle bombardment. These tissues were transformed with plasmid pC-KSA2300 carrying a selectable marker (for kanamycin) and a target gene (phytase, or phyA, from Aspergillus ficuum). Primary plants were regenerated in a medium containing 75 mg l−1 kanamycin. Of 1,534 shoot apices, 52 (3.4%) survived on this selection medium. Southern and Northern blot analyses confirmed that phyA was stably integrated and expressed in those primary transgenics. The progenies of the primary transgenic plants were found to have a 3.1- to 3.2-fold increase in root extracellular phytase activity, resulting in improved phosphorus (P) nutrition. Growth also was enhanced when they were supplied with phytate, and their P content was equivalent to that of wildtype plants supplied with inorganic phosphate. These results demonstrate that the expression of phyA in cotton plants improves their ability to utilize organic P in response to a deficiency.

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References

  • Brinch-Pedersen H, Sorensen LD, Holm PB (2002) Engineering crop plants: getting a handle on phosphate. Trends Plant Sci 7:118–125

    Article  PubMed  CAS  Google Scholar 

  • Bronson KF, Onken AB, Booker JD, Lascano RJ, Provin TL, Torbert HA (2001) Irrigated cotton lint yields as affected by phosphorus fertilizer and landscape position. Commun Soil Sci Plant Anal 32:1959–1967

    Article  CAS  Google Scholar 

  • Emani C, Garcia JM, Lopata-Finch E, Pozo MJ, Uribe P, Kim DJ, Sunilkumar G, Cook DR, Kenerley CM, Rathore KS (2003) Enhanced fungal resistance in transgenic cotton expressing an endochitinase gene from Trichoderma virens. J Plant Biotechnol 1:321–336

    Article  CAS  Google Scholar 

  • Findenegg GR, Nelemans JA (1993) The effect of phytase on the availability of phosphorus from myo-inositol hexaphosphate (phytate) for maize roots. Plant Soil 154:189–196

    Article  CAS  Google Scholar 

  • Finer JJ, McMullen MD (1990) Transformation of cotton (Gossypium hirsutum L.) via particle bombardment. Plant Cell Rep 8:586–589

    Article  Google Scholar 

  • George TS, Richardson AE (2008) Potential and limitations to improving crops for enhanced phosphorus utilization. In: White PJ, Hammond JP (eds) Ecophysiology of plant-phosphorus interactions. Springer SBM, Dordrecht, pp 247–270

    Chapter  Google Scholar 

  • Greiner R, Haller E, Konietzny U, Jany KD (1997) Purification and characterization of a phytase from Klebsiella terrigena. Arch Biochem Biophys 341:201–206

    Article  PubMed  CAS  Google Scholar 

  • Gu XF, Meng H, Qi G, Zhang JR (2008) Agrobacterium-mediated transformation of the winter jujube (Zizyphus jujuba Mill.). Plant Cell Tiss Org Cult 94:23–32

    Article  CAS  Google Scholar 

  • Hayes JE, Richardson AE, Simpson RJ (1999) Phytase and acid phosphatase activities in extracts from roots of temperate pasture grass and legume seedlings. Aust J Plant Physiol 26:801–809

    Article  CAS  Google Scholar 

  • Hayes JE, Simpson RJ, Richardson AE (2000) The growth and phosphorus utilisation of plants in sterile media when supplied with inositol hexaphosphate, glucose-1-phosphate or inorganic phosphate. Plant Soil 220:165–174

    Article  CAS  Google Scholar 

  • Keller G, Spatola L, Mccabe D, Martinell B, Swain W, John M (1997) Transgenic cotton resistant to herbicide bialaphos. Transgen Res 6:385–392

    Article  CAS  Google Scholar 

  • Khan T, Reddy VS, Leelavathi S (2010) High-frequency regeneration via somatic embryogenesis of an elite recalcitrant cotton genotype (Gossypium hirsutum L.) and efficient Agrobacterium-mediated transformation. Plant Cell Tiss Org Cult 101:323–330

    Article  Google Scholar 

  • Kumar M, Shukla AK, Singh H, Tuli R (2009) Development of insect resistant transgenic cotton lines expressing cry1EC gene from an insect bite and wound inducible promoter. J Biotechnol 140:143–148

    Article  PubMed  CAS  Google Scholar 

  • Lung SC, Chan WL, Yip W, Wang L, Yeung EC, Lim BL (2005) Secretion of beta-propeller phytase from tobacco and Arabidopsis roots enhances phosphorus utilization. Plant Sci 169:341–349

    Article  CAS  Google Scholar 

  • Maugenest S, Martinez I, Godin B, Perez P, Lescure AM (1999) Structure of two maize phytase genes and their spatiotemporal expression during seedling development. Plant Mol Biol 39:503–514

    Article  PubMed  CAS  Google Scholar 

  • Mudge SR, Smith FW, Richardson AE (2003) Root-specific and phosphate-regulated expression of phytase under the control of a phosphate transporter promoter enables Arabidopsis to grow on phytate as a sole P source. Plant Sci 165:871–878

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Murphy J, Riley JP (1962) A modified single solution method for the determination of phosphate in natural waters. Anal Chim Acta 27:31–36

    Article  CAS  Google Scholar 

  • Nitz I, Berkefeld H, Puzio PS, Grundler FMW (2001) Pyk10, a seedling and root specific gene and promoter from Arabidopsis thaliana. Plant Sci 161:337–346

    Article  PubMed  CAS  Google Scholar 

  • Parimalan R, Giridhar P, Ravishankar GA (2010) Enhanced shoot organogenesis in Bixaorellana L. in the presence of putrescine and silver nitrate. Plant Cell Tiss Org Cult. doi:10.1007/s11240-010-9865-7

  • Reichandt M, Rogers S (1994) Preparation of plant DNA using CTAB. In: Ausubel FM, Bernt R, Struhl K (eds) Current protocols in molecular biology. Sarah Greene, Brooklyn, pp 233–237

    Google Scholar 

  • Richardson AE, Hadobas PA, Hayes JE (2000) Phosphomonoesterase and phytase activities of wheat (Triticum aestivum L.) roots and utilisation of organic phosphorus substrates by seedlings grown in sterile culture. Plants Cell Environ 23:397–405

    Article  CAS  Google Scholar 

  • Richardson AE, Hadobas PA, Hayes JE (2001) Extracellular secretion of Aspergillus phytase from Arabidopsis roots enables plants to obtain phosphorus from phytate. Plant J 25:641–649

    Article  PubMed  CAS  Google Scholar 

  • Saeed NA, Zafar Y, Malik KA (1997) A simple procedure of Gossypium meristem shoot tip culture. Plant Cell Tiss Org Cult 51:201–207

    Article  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Press, New York, pp 672–690

    Google Scholar 

  • Satyavathi VV, Prasad V, Lakshmi BG, Sita GL (2002) High efficiency transformation protocol for three Indian cotton varieties via Agrobacterium tumefaciens. Plant Sci 162:215–222

    Article  CAS  Google Scholar 

  • Stalker DM, Kiser JA, Baldwin G, Coulombe B, Houck CM (1996) Cotton weed control using the BXNTM system. In: Duke SO (ed) Herbicide-resistant crops: agricultural environmental economic, regulatory and technical aspects. Lewis Publishers, New York, pp 93–105

    Google Scholar 

  • Tsai SF, Yeh SD, Chan CF, Liaw SI (2009) High-efficiency vitrification protocols for cryopreservation of in vitro grown shoot tips of transgenic papaya lines. Plant Cell Tiss Org Cult 98:157–164

    Article  CAS  Google Scholar 

  • Turner BL, Paphazy MJ, Haygarth PM, Mckelvie ID (2002) Inositol phosphates in the environment. Phil Trans R Soc B 357:449–469

    Article  PubMed  CAS  Google Scholar 

  • Xiao K, Harrison MJ, Wang ZY (2005) Transgenic expression of a novel M. truncatula phytase gene results in improved acquisition of organic phosphorus by Arabidopsis. Planta 222:27–36

    Article  PubMed  CAS  Google Scholar 

  • Zhang KW, Wang J, Hu XR, Yang AF, Zhang JR (2010) Agrobacterium-mediated transformation of shoot apices of Kentucky bluegrass (Poa pratensis L.) and production of transgenic plants carrying a betA gene. Plant Cell Tiss Org Cult 102:135–143

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by project No. 2009ZX08005-021B from the Ministry of Agriculture of China, and by the Natural Science Foundation of Hebei Province (No. C2006001034). The authors are grateful to Dr. Priscilla Licht for the critical reading of the manuscript.

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Authors and Affiliations

  1. North China Key Laboratory for Crop Germplasm Resources, Agricultural University of Hebei, 071001, Baoding, China

    J. F. Liu, X. F. Wang, Q. L. Li, X. Li, G. Y. Zhang & Z. Y. Ma

  2. College of Life Sciences, Nankai University, 300071, Tianjian, China

    M. G. Li

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  1. J. F. Liu
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  2. X. F. Wang
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  3. Q. L. Li
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  4. X. Li
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  5. G. Y. Zhang
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  7. Z. Y. Ma
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Correspondence to Z. Y. Ma.

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Liu, J.F., Wang, X.F., Li, Q.L. et al. Biolistic transformation of cotton (Gossypium hirsutum L.) with the phyA gene from Aspergillus ficuum . Plant Cell Tiss Organ Cult 106, 207–214 (2011). https://doi.org/10.1007/s11240-010-9908-0

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  • DOI: https://doi.org/10.1007/s11240-010-9908-0

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