摘要
【目的】以条锈菌和白粉菌胁迫的普通小麦(Triticum aestivum L.)为研究对象,分析由可变剪切(alternative splicing,AS)形成的TaNAC结构变异转录本,同时分析TaNAC基因的microRNA调控位点,为进一步解析TaNAC基因通过转录后调控参与小麦响应真菌胁迫奠定基础。【方法】普通小麦兼抗种质N9134在被白粉菌和条锈菌分别侵染后,各8个时间点取样并混合,然后从混合样本中克隆得到大量TaNAC转录本。参考中国春小麦基因组注释信息(IWGSC RefSeq v1.1)进行比对,选择由可变剪切形成的TaNAC序列结构变异转录本,分析它们的序列结构特征。利用生物信息学软件和在线工具,对这些TaNAC结构变异转录本编码产物的功能结构域、高级结构、理化性质、亚细胞定位等特征和变异情况进行比对分析。同时,利用洋葱表皮细胞瞬时表达系统验证其中1对TaNAC结构变异转录本的亚细胞定位预测结果,并选取5组TaNAC基因的可变剪切序列结构变异转录本进行酵母转录自激活试验,研究序列结构变异对TaNAC基因转录调控活性的影响。此外,利用miRBase数据库收录的小麦中已报道的miRNAs和TaNAC基因,进行靶基因预测分析,建立小麦TaNAC家族成员与miRNAs的靶向关系。【结果】以条锈菌和白粉菌侵染的普通小麦兼抗种质N9134为材料,克隆得到的TaNAC转录本中的35条序列结构变异转录本由13个TaNAC基因可变剪切形成。通过分析发现,同一TaNAC基因由可变剪切形成的不同结构变异转录本的核酸序列结构存在差异,而且其对应编码产物的功能结构域、高级结构、理化性质和亚细胞定位等方面均会存在差异,同时也会表现出不同的转录调控活性;不同TaNAC基因的可变剪切方式存在差异,而且它们的结构变异转录本及其编码产物在结构特征、理化性质和转录调控活性等方面也均呈现出多样性的特征。通过分析TaNAC基因与其在编码序�
【Objective】In the present study,the common wheat(Triticum aestivum L.)was exposed to the stress of stripe rust and powdery mildew.Then the cloned TaNAC structural variation transcripts formed by alternative splicing were analyzed.And putative information of TaNAC genes regulated by miRNAs was annotated.It would shed light on the study of TaNAC genes in response to fungal stress of wheat at the post-transcriptional level.【Method】After common wheat resistance germplasm N9134 being infected stripe rust and powdery mildew respectively,the leaves were sampled at eight time points.Then a large number of TaNAC transcripts were cloned from the mixed sample pool.Referring to the wheat genome annotation of Chinese Spring(IWGSC RefSeqv1.1),the sequence structure characteristics of TaNAC structural variation transcripts formed by alternative cutting were revealed.Using bioinformatics software and online tools,the coding products derived from these TaNAC transcripts were compared and analyzed,including the functional domain,advanced structure,physical and chemical properties,subcellular localization and other characteristics.And then,one pair of TaNAC structural variation transcripts were selected to further verify the predicted subcellular localizations by onion epidermis transient expression system.Meanwhile,five groups of TaNAC transcripts were conducted transcriptional self-activation experiments in yeast.It was aimed to analyze the effects of structural variation,which caused by alternative splicing on transcriptional regulation activity.Additionally,using the miRBase database,the targeting relationship between TaNAC genes and tae-miRNAs was forecasted and established in wheat.【Result】In this study,35 TaNAC structural variation transcripts were formed by alternative splicing form 13 TaNAC genes,and they all were cloned from common wheat N9134 after infecting by stripe rust and powdery mildew.After analyzing,it was found that there were differences in the nucleic acid sequence structure of different structural
作者
吕士凯
马小龙
张敏
邓平川
陈春环
张宏
刘新伦
吉万全
LÜShiKai;MA XiaoLong;ZHANG Min;DENG PingChuan;CHEN ChunHuan;ZHANG Hong;LIU XinLun;JI WanQuan(College of Agronomy,Northwest A&F University/State Key Laboratory of Crop Stress Biology for Arid Areas,Yangling 712100,Shaanxi)
出处
《中国农业科学》
CAS
CSCD
北大核心
2021年第22期4709-4727,共19页
Scientia Agricultura Sinica
基金
陕西省重点研发计划(2019DLNY04-06)。
