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小麦中低温诱导的Wcs120家族基因的染色体定位及耐寒性表达调控

Chromosome mapping of low-temperature induced Wcs120 family genes and regulation of cold-tolerance expression in wheat.

作者信息

Limin A E, Danyluk J, Chauvin L P, Fowler D B, Sarhan F

机构信息

Crop Development Centre, University of Saskatchewan, Saskatoon, Canada.

出版信息

Mol Gen Genet. 1997 Feb 27;253(6):720-7. doi: 10.1007/s004380050376.

Abstract

Low-temperature (LT) induced genes of the Wcs120 family in wheat (Triticum aestivum) were mapped to specific chromosome arms using Western and Southern blot analysis on the ditelocentric series in the cultivar Chinese Spring (CS). Identified genes were located on the long arms of the homoeologous group 6 chromosomes of all 3 genomes (A, B, and D) of hexaploid wheat. Related species carrying either the A, D, or AB genomes were also examined using Southern and Western analysis with the Wcs120 probe and the WCS120 antibody. All closely related species carrying one or more of the genomes of hexaploid wheat produced a 50 kDa protein that was identified by the antibody, and a Wcs120 homoeologue was detected by Southern analysis in all species. In the absence of chromosome arm 6DL in hexaploid CS wheat no 50 kDa protein was produced and the high-intensity Wcs120 band was missing, indicating 6DL as the location of Wcs120 but suggesting silencing of the Wcs120 homoeologue in the A genome. Levels of proteins that cross-reacted with the Wcs120 antibody and degrees of cold tolerance were also investigated in the Chinese Spring/Cheyenne (CS/CNN) chromosome substitution series. CNN chromosome 5A increased the cold tolerance of CS wheat. Densitometry scanning of Western blots to determine protein levels showed that the group 5 chromosome 5A had a regulatory effect on the expression of the Wcs120 gene family located on the group 6 chromosomes of all three hexaploid wheat genomes.

摘要

利用中国春(CS)品种双端体系列的蛋白质免疫印迹和Southern印迹分析,将小麦(普通小麦)中Wcs120家族的低温(LT)诱导基因定位到特定染色体臂上。已鉴定的基因位于六倍体小麦所有3个基因组(A、B和D)的6号同源群染色体的长臂上。还使用Wcs120探针和WCS120抗体通过Southern印迹和蛋白质免疫印迹分析了携带A、D或AB基因组的相关物种。所有携带六倍体小麦一个或多个基因组的近缘物种都产生了一种能被该抗体识别的50 kDa蛋白质,并且通过Southern分析在所有物种中都检测到了一个Wcs120同源基因。在六倍体CS小麦中缺失6DL染色体臂时,不产生50 kDa蛋白质且高强度的Wcs120条带缺失,这表明6DL是Wcs120的定位,但暗示A基因组中Wcs120同源基因的沉默。在中国春/夏延(CS/CNN)染色体代换系中,还研究了与Wcs120抗体发生交叉反应的蛋白质水平和耐寒程度。CNN的5A染色体提高了CS小麦的耐寒性。对蛋白质免疫印迹进行光密度扫描以确定蛋白质水平,结果表明5号染色体的5A对位于所有三个六倍体小麦基因组6号染色体上的Wcs120基因家族的表达具有调节作用。

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