In planta transformation of sorghum (Sorghum bicolor (L.) Moench) using TPS1 gene for enhancing tolerance to abiotic stresses

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• 作者：VARALAXMI YELLISETTY ; L. A. REDDY ; MAHESWARI MANDAPAKA
• 关键词：apical meristem ; sorghum ; in planta transformation ; TPS1 gene
• 刊名：Journal of Genetics
• 出版年：2015
• 出版时间：September 2015
• 年：2015
• 卷：94
• 期：3
• 页码：425-434
• 全文大小：5,617 KB
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• 作者单位：VARALAXMI YELLISETTY (1)
  L. A. REDDY (2)
  MAHESWARI MANDAPAKA (1)
  
  1. Division of Crop Sciences, Central Research Institute for Dryland Agriculture, Santhoshnagar, Hyderabad, 500 059, India
  2. Department of Genetics, Osmania University, Durgabhai Deshmukh Colony, Hyderabad, 500 007, India
  
• 刊物主题：Life Sciences, general; Microbial Genetics and Genomics; Plant Genetics & Genomics; Animal Genetics and Genomics; Evolutionary Biology;
• 出版者：Springer India
• ISSN：0973-7731

文摘

An in planta transformation protocol for sorghum (Sorghum bicolor (L.) Moench) using shoot apical meristem of germinating seedlings is reported in this study. Agrobacterium tumefaciens strain, LBA4404 with pCAMBIA1303 vector and construct pCAMBIA1303TPS1 were individually used for transformation. Since, the transgene is integrated into the cells of already differentiated tissues, the T₀ plants were chimeric and stable integration was observed in T₁ generation. ?-Glucuronidase (GUS) expression in the seedlings and spikelets of emerging cob was the first indication of transformability in T₀ generation which was further confirmed by PCR analysis using hpt and TPS1 gene-specific primers. Screening on 25 mg/L hygromycin combined with PCR analysis was used for selection of transformants in the T₁ generation. Transformation efficiencies ranged between 34-8% and 26-4% using pCAMBIA1303 vector and construct pCAMBIA1303TPS1, respectively. Molecular characterization of the T₂ transgenics using PCR, RT-PCR and Southern blot analyses further revealed the integration, expression and inheritance of the transgene. These results indicate the feasibility of the method to generate transgenics with pCAMBIA1303 vector and construct pCAMBIA1303TPS1. The abiotic stress tolerance of TPS1 transgenics developed in the present study was evident by the ability of the transformants to tolerate 200 mM NaCl as well as higher root growth and biomass. Keywords apical meristem sorghum in planta transformation TPS1 gene