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基于病毒诱导基因沉默对小麦中六个与春化作用相关基因的功能验证

Virus-induced gene silencing-based functional verification of six genes associated with vernalization in wheat.

作者信息

Feng Ya-Lan, Wang Ke-Tao, Ma Chao, Zhao Yong-Ying, Yin Jun

机构信息

National Engineering Research Centre for Wheat, Henan Agricultural University, Zhengzhou 450002, China; Collaborative Innovation Center of Henan Grain Crops, Henan Agricultural University, Zhengzhou 450002, China; National Key Laboratory of Wheat and Maize Crop Science, Henan Agricultural University, Zhengzhou 450002, China.

Collaborative Innovation Center of Henan Grain Crops, Henan Agricultural University, Zhengzhou 450002, China; College of Life Science, Henan Agricultural University, Zhengzhou 450002, China.

出版信息

Biochem Biophys Res Commun. 2015 Mar 20;458(4):928-33. doi: 10.1016/j.bbrc.2015.02.064. Epub 2015 Feb 21.

Abstract

Vernalization requirement is an important characteristic in crop breeding. Wheat is a widely grown crop in the world that possesses enormous economic significance. To better understand the gene networks in vernalization process, we performed a high-throughput RNA sequencing analysis comparing the transcriptomes of spring and winter wheat cultivars, with and without vernalization (unpublished data). In this study, we selected six unigenes (CL14010, CL12788, CL176, Unigene 16777, CL8746 and Unigene10196) from our transcriptome analysis based on their expression differences to further characterize their function. Transient silencing of the six unigenes individually were achieved through virus-induced gene silencing (VIGS) using BSMV vector. The period from germination to spike differentiation were recorded and compared between plants underwent VIGS silencing and the control. Our result showed that VIGS of the six unigenes significantly shortened the period from seedling to double ridge (DR) stage. Resulting in SD period ranging from 59.8 ± 0.60 to 65.8 ± 0.48 days, compared to 85.0 ± 0.73 days in the control. The results indicated that these six unigenes function as suppressors in vernalization process and silence or down-regulation of these genes promoted flower development in wheat. Further characterization of these six unigenes and their function in vernalization and flowering control is needed.

摘要

春化需求是作物育种中的一个重要特征。小麦是世界上广泛种植的作物,具有巨大的经济意义。为了更好地理解春化过程中的基因网络,我们进行了高通量RNA测序分析,比较了春小麦和冬小麦品种在春化和未春化情况下的转录组(未发表数据)。在本研究中,我们根据六个单基因(CL14010、CL12788、CL176、Unigene 16777、CL8746和Unigene10196)在转录组分析中的表达差异来进一步表征它们的功能。通过使用BSMV载体的病毒诱导基因沉默(VIGS)分别实现了这六个单基因的瞬时沉默。记录并比较了进行VIGS沉默的植株和对照植株从发芽到穗分化的时间。我们的结果表明,这六个单基因的VIGS显著缩短了从幼苗到二棱期的时间。导致二棱期从59.8±0.60天到65.8±0.48天不等,而对照为85.0±0.73天。结果表明,这六个单基因在春化过程中起抑制作用,沉默或下调这些基因可促进小麦的花发育。需要对这六个单基因及其在春化和开花控制中的功能进行进一步表征。

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