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Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (05): 691-700.doi: 10.3724/SP.J.1006.2017.00691

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Glu-B1 Silencing Influences Protein Body Formation and Expression of Genes Regulating Synthesis and Processing of Seed-Storage Protein in Somatic mutant Wheat AS208

LIU Hui-Yun1,**,WANG Wan-Qing2,**,LI Xin1, WANG Ke1,WANG Long3,DU Li-Pu1,YANG Yue-Ming2,*,YE Xing-Guo1,*   

  1. 1 Institute of Crop Science, Chinese Academy of Agricultural Sciences / National Key Facility for Crop Gene Resources and Genetic Improvement, Beijing 100081, China; 2 College of Life Science, Capital Normal University, Beijing 100048, China; 3 Institute of Genetics and Developmental Biology, Chinese Academy of Sciences / State Key Laboratory of Plant Cell and Chromosome Engineering, Beijing 100101, China
  • Received:2016-04-24 Revised:2016-11-03 Online:2017-05-12 Published:2016-12-02
  • Contact: Yan yueming, E-mail: yanym2004@163.com; Ye Xingguo, E-mail: yexingguo@caas.cn
  • Supported by:

    This study was supported by the National Natural Science Foundation of China (31371621).

Abstract:

High-molecular-weight-glutenin subunits (HMW-GSs) are important components of seed storage proteins in grains of wheat (Triticum aestivum L.), which determine wheat dough elasticity and processing quality. Using AS208, a wheat somatic variation line with silenced 1Bx20 and 1By20 at Glu-B1 locus, and its originating cultivar Lunxuan 987, we studied the present status of Glu-B1, protein body (PB) formation progress, and expression trends of four kinds of genes (BiP, Dof, PDI, and SPA) related to glutenin synthesis and accumulation at different stages of grain development. Southern blotting analysis revealed that AS208 had two specific bands less than its control parent Lunxuan 987. The fluorescence in situ hybridization (FISH) assay showed six and four chromosomes with signals in Lunxuan 987 and AS208, respectively. These results prove that the Glu-B1 locus has been deleted from the genome of AS208. Compared to Lunxuan 987, AS208 showed similar size and shape of seed PB during PB formation under a transmission electron microscope (TEM). A total of 12 transcription-factor or molecular-chaperone genes were subjected to qRT-PCR assay. These genes were related to glutenin coding genes expression, storage proteins accumulation and processing, PB assembling and transportation. Seven out of 12 genes had simialr expression patterns in AS208 and Lunxuan 987, and eight genes were expressed more in AS208 than in Lunxuan 987, especially the expression levels of Bip, PDI-1, and PDI-5 were 1.5–2.0 folds higher in AS208 than in Lunxuan 987. Our results suggested that Glu-B1 silencing had no significant effect on the formation of PB during wheat grain development, but partially spurred the expression of some genes regulating the synthesis and processing of seed-storage proteins. As a result, via a negative feedback regulation pathway, the protein content in seed and the shape and size of PB remain in similar levels between AS208 and its originating cultivar.

Key words: Wheat, High molecular weight glutenin subunits, Protein body, Transcription factor, Molecular chaperones

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