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Efficient Regeneration Potential is Closely Related to Auxin Exposure Time and Catalase Metabolism During the Somatic Embryogenesis of Immature Embryos in Triticum aestivum L

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Abstract

Regeneration of cultured tissue is a prerequisite of Agrobacterium- and biolistic-mediated plant transformation. In this study, an efficient protocol for improving wheat (Triticum aestivum L.) immature embryo regeneration was developed. Based on the statistical analysis of embryogenic callus induction efficiency, green spot differentiation efficiency, and plant regeneration efficiency from five wheat accessions, improved culture conditions were found to be more effective for embryogenic callus production than the traditional conditions. Using semi-quantitative reverse transcription polymerase chain reaction, a candidate gene, designated as TaCAT1, which encodes a catalase was identified to have a significant correlation with high-regeneration trait of wheat immature embryos. Three amino acid substitutions were found in TaCAT1 protein between high- and low-regeneration wheat accessions. Hydrogen peroxide content in the cultured calli increased from day 5 to 15, and then decreased sharply on day 20, followed by a second peak on day 25 during regeneration stage. Furthermore, a 3,500-bp 5′ flanking region upstream of the first codon ATG of TaCAT1 was isolated using inverse polymerase chain reaction. In silico, analysis revealed that the TaCAT1 promoter contained two regulatory motifs associated with responses to auxin.

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Acknowledgments

This research was financially supported by the National Natural Science Foundation of China (Grant No. 30971776) and the Ministry of Agriculture of China (Grant No. 2008ZX08010-004).

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Authors and Affiliations

  1. Key Laboratory of Crop Genetics and Breeding, Ministry of Agriculture, National Key Facility of Crop Gene Resources and Genetic Improvement, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, China

    Maoyun She, Guixiang Yin, Xing Li, Lipu Du & Xingguo Ye

  2. National Key Facility of Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, Zhong Guan Cun South St, Haidian District, Beijing, 100081, China

    Xingguo Ye

  3. Department of Plant Pathology, Kansas State University, Manhattan, KS, USA

    Jiarui Li

  4. Department of Agriculture & Food, Centre for Comparative Genomics, Murdoch University & Western Australian, Perth, WA, Australia

    Wujun Ma

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  1. Maoyun She
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Correspondence to Xingguo Ye.

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12033_2012_9583_MOESM1_ESM.pdf

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She, M., Yin, G., Li, J. et al. Efficient Regeneration Potential is Closely Related to Auxin Exposure Time and Catalase Metabolism During the Somatic Embryogenesis of Immature Embryos in Triticum aestivum L. Mol Biotechnol 54, 451–460 (2013). https://doi.org/10.1007/s12033-012-9583-y

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