草铵膦胁迫下小麦叶片转录组分析

doi:10.7668/hbnxb.20193925

摘要/Abstract

摘要：

为了挖掘小麦响应草铵膦胁迫下的关键基因,解析关键基因及其家族成员染色体分布、表达模式等,以冬小麦津农6号为研究对象,喷施不同浓度的草铵膦,观察其药后表型;提取胁迫处理0,3,9 h的小麦叶片总RNA,进行转录组测序,筛选关键基因并对其表达量、结构及表达模式进行分析。结果表明:转录组分析发现草铵膦胁迫诱导TraesCS4A02G044000基因上调表达,序列分析表明,该基因位于小麦第4同源基因A基因组。蛋白结构域分析可知,该基因编码蛋白存在1个信号肽、1个DUF568结构域和1个b561结构域。此外,在该基因家族成员中挖掘出12个同时含有DUF568和b561结构域的基因,位于第4,5,7同源群,其中位于第4同源群A、B、D染色体的TraesCS4D02G265000、TraesCS4B02G265100和TraesCS4A02G044000基因在草铵膦胁迫诱导下显著上调表达,该3个基因均编码细胞色素b561。编码细胞色素b561的TraesCS4A02G044000及其同源基因在草铵膦胁迫下显著上调,表明b561在小麦受到草铵膦胁迫后产生了积极的响应,参与了草铵膦在小麦体内的代谢。

关键词: 小麦, 草铵膦胁迫, 转录组分析, 基因定位及表达模式

Abstract:

In order to excavate the key genes in wheat which were resistant to glufosinate-ammonium stress and analyze family members chromosome distribution and expression pattern.Winter wheat Jinnong 6 was selected as the research object and different concentrations of glufosinate-ammonium was used in our study and observed symptoms of wheat after treatments.Total RNA was extracted from wheat leaves at 0,3 and 9 h after treatment.Transcriptomic sequencing technology was used to analyze and excavate the key genes in wheat.To confirm genes expression,protein structure of their family members and expression pattern.The results showed that TraesCS4A02G044000 was up-regulated induced by glufosinate-ammonium stress according to transcriptome analysis.Sequence analysis showed that TraesCS4A02G044000 was located in the 4th homologous gene A genome of wheat.The protein domain analysis showed that the protein encoded of TraesCS4A02G044000 had a signal peptide,a DUF568 and a b561.Twelve genes containing DUF568 and b561 domains were excavated from the genome family members,which were located in the 4th,5th and 7th homologous groups.The expression pattern analysis showed that TraesCS4D02G265000,TraesCS4B02G265100 and TraesCS4A02G044000 located in chromosomes A,B and D of the 4th homologous group which encoded cytochrome b561 were up-regulated obviously in wheat under glufosinate-ammonium stress. TraesCS4A02G044000 and its homologous genes encoding cytochrome b561 were significantly up-regulated under glufosinate-ammonium stress, indicated that b561 had a positive response to glufosinate-ammonium stress and participated in the metabolism of glufosinate-ammonium in wheat.

Key words: Wheat, Glufosinate-ammonium stress, Transcriptome analysis, Gene localization and expression pattern

于金萍, 张惟, 李琦, 白鹏华, 崔新仪, 刘亦学. 草铵膦胁迫下小麦叶片转录组分析[J]. 华北农学报, 2023, 38(3): 184-190. doi: 10.7668/hbnxb.20193925.

YU Jinping, ZHANG Wei, LI Qi, BAI Penghua, CUI Xinyi, LIU Yixue. Comparative Transcriptome Analysis of Wheat Leaf in Response to Glufosinate-ammonium Stress[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(3): 184-190. doi: 10.7668/hbnxb.20193925.

http://www.hbnxb.net/CN/Y2023/V38/I3/184

图/表 6

图1 小麦表型在不同浓度草铵膦处理下的变化

Fig.1 Phenotypes of wheat treated under glufosinate-ammonium at different concentrations

图2 TraesCS4A02G044000在草铵膦不同时间处理后的表达量不同字母表示在P<0.05水平差异显著。图5同。

Fig.2 FPKM of TraesCS4A02G044000 after treatment of glufosinate-ammonium at different time Different letters indicate significant difference at the level of P<0.05. The same as Fig.5.

图3 TraesCS4A02G044000基因及编码蛋白结构 A.TraesCS4A02G044000基因序列结构图;B.TraesCS4A02G044000基因编码蛋白结构域(蛋白结构域由信号肽、DUF568、b561组成)。

Fig.3 Structure of TraesCS4A02G044000 and encoding protein A. Sequence structure of TraesCS4A02G044000; B. Protein domain of TraesCS4A02G044000 encoded (protein domain consists of signal peptide, DUF568 and b561).

表1 TraesCS4A02G044000家族成员

Tab.1 Family members of TraesCS4A02G044000

基因ID Gene ID	染色体位置 Chromosomal location	起始位置/bp Start position	终止位置/bp End position
TraesCS5B02G278100	5B	463 991 797	463 994 199
TraesCS4D02G265500	4D	435 626 360	435 629 320
TraesCS4B02G265500	4B	536 894 334	536 896 673
TraesCS7A02G255600	7A	243 646 927	243 650 472
TraesCS4B02G265100	4B	536 117 250	536 120 707
TraesCS7B02G152000	7B	202 160 045	202 164 168
TraesCS4A02G044400	4A	37 006 387	37 010 128
TraesCS5A02G278800	5A	488 022 430	488 024 910
TraesCS4A02G044000	4A	36 611 408	36 614 890
TraesCS4D02G265000	4D	435 206 769	435 210 339
TraesCS7D02G254000	7D	230 814 306	230 817 980
TraesCS5D02G285700	5D	386 150 155	386 152 541

图4 TraesCS4A02G044000及其家族成员聚类分析

Fig.4 Cluster analysis of TraesCS4A02G044000 family members

图5 TraesCS4A02G044000家族成员在草铵膦胁迫处理下的基因表达模式分析

Fig.5 Gene expression pattern analysis of TraesCS4A02G044000 family members under glufosinate-ammonium stress

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