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利用RNA-Seq技术研究小麦芒长发育相关基因

发表时间:2016-08-15  浏览量:1598  下载量:488
全部作者: 李丹丹,高亚男,鲍印广,王洪刚,李兴锋
作者单位: 山东农业大学农学院;山东农业大学作物生物学国家重点实验室
摘 要: 芒是小麦穗部重要的形态和光合器官,位于5A染色体长臂的小麦B1基因抑制小麦穗中下部的芒长发育。本研究以一对芒长存在差异的近等基因系SN051-1(有芒 b1b1)和SN051-2(无芒B1B1)及其F1(无芒B1b1)、F2有芒植株(b1b1)和无芒植株(B1B1、B1b1)为研究材料,利用RNA-Seq技术对芒的发育相关基因进行研究。转录组测序结果共得到Clean data总碱基数为48.06 Gbp,Q30均在85.01%以上;在亲本SN051-1(有芒 b1b1)和SN051-2(无芒B1B1)以及F1(无芒B1b1)中共获得180个共同差异表达基因(differentially expressed genes,DEGs),其中在无芒材料中表达上调的有56个差异基因,表达量下调的有134个差异基因;引入F2(无芒B1B1、B1b1,有芒b1b1)结果得到22个共同差异表达基因,其中在无芒材料中表达上调的有4个差异基因,表达量下调的有18个差异基因;对在无芒材料中表达上调的4个差异基因进行实时荧光定量PCR(real time fluorescence quantitative PCR, qRT-PCR),结果表明c18073.graph_c0、c55090.graph_c0和c73118.graph_c0等3个基因的基因表达模式与RNA-Seq结果基本相符。
关 键 词: 作物育种学与良种繁育学;小麦;芒;近等基因系;RNA-Seq;实时荧光定量PCR
Title: Comparative transcriptional analysis of two near-isogenic wheat lines of awn-inhibitor gene B1 using RNA-Seq
Author: LI Dandan, GAO Yanan, BAO Yinguang, WANG Honggang, LI Xingfeng
Organization: College of Agriculture, Shandong Agricultural University; State Key Laboratory Crop Biology, Shandong Agricultural University
Abstract: Awn is one of the most morphological characteristics of wheat and also acts as high effective organ for photosynthesis in spike. Gene B1, located on the wheat 5A chromosome, is the most obvious gene that inhibits awn-length development in wheat. In this study, two near-isogenic lines (NILs) with the only difference of awn-length, line SN051-1 (awned, b1b1) and line SN051-2 (awnless, B1B1), and their F1 hybrid and F2 populations were used for RNA-Seq analysis, in order to study the molecular mechanism of awn development. Approximately 48.06 Giga base pairs (Gbp) of clean data were generated, the Q30 values were higher than 85.01%. A total of 180 differentially expressed genes (DEGs) were identified by comparisons of the RNA-Seq data between SN051-2 vs. SN051-1 and F1 hybrid vs. SN051-1, with 56 up-regulated and 134 down-regulated genes, respectively. And 22 DEGs were identified by pairwise comparison with F2 data, with 4 up-regulated and 18 down-regulated genes, respectively. Real time fluorescence quantitative PCR (qRT-PCR) results proved that expression profiles of c18073.graph_c0, c55090.graph_c0 and c73118.graph_c0 in SN051-1 young spike is much higher than that in SN051-2 young spike, which was constant with the RNA-Seq data.
Key words: plant breeding science, improved variety thremmatology; common wheat; awn; near isogenic lines; RNA-Seq; real time fluorescence quantitative PCR
发表期数: 2016年8月第15期
引用格式: 李丹丹,高亚男,鲍印广,等. 利用RNA-Seq技术研究小麦芒长发育相关基因[J]. 中国科技论文在线精品论文,2016,9(15):1530-1540.
 
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