小串联模拟靶标(Short tandem target mimic,STTM)技术是一种新开发的miRNA功能研究方法。Tae-miR9677作为一种新发现的在小麦穗部特异性高表达的miRNA,其功能至今未知。为了进一步探索Tae-miR9677的功能,构建了Ubiqutin(UBI)启动子启动...小串联模拟靶标(Short tandem target mimic,STTM)技术是一种新开发的miRNA功能研究方法。Tae-miR9677作为一种新发现的在小麦穗部特异性高表达的miRNA,其功能至今未知。为了进一步探索Tae-miR9677的功能,构建了Ubiqutin(UBI)启动子启动的Tae-miR9677 STTM过表达载体,并通过基因枪介导法对小麦品种绵阳19幼胚愈伤组织进行转化。结果表明,3 683个愈伤组织经过PPT(Phosphinothricin)筛选,最终分化获得42株再生植株;利用特异性引物进行PCR检测,鉴定出8株T0代阳性植株。展开更多
miR166作为重要的转录后调节因子,在植物的生长发育和对逆境胁迫的反应中扮演着重要的角色。利用PmiREN、Ensembl Plants数据库、MEGA-X、DNAMAN软件以及RNAfold web server、WebLogo和psRNA Target在线网站对小麦miR166(Tae-miR166)基...miR166作为重要的转录后调节因子,在植物的生长发育和对逆境胁迫的反应中扮演着重要的角色。利用PmiREN、Ensembl Plants数据库、MEGA-X、DNAMAN软件以及RNAfold web server、WebLogo和psRNA Target在线网站对小麦miR166(Tae-miR166)基因家族的进化特性和表达模式进行分析。在PmiREN数据库中搜索到19个Tae-miR166基因家族成员,染色体定位发现Tae-miR166成员定位于14条染色体上。序列比对发现Tae-miR166的成熟体序列高度保守。进化分析结果表明,Tae-miR166基因家族成员分别处于4个进化分支。靶基因预测结果表明,Tae-miR166的靶基因包括Ⅲ类同源异型结构域亮氨酸拉链蛋白、β半乳糖苷酶和钙依赖性蛋白激酶等。转录组数据分析表明Tae-miR166家族19个成员在小麦6个组织中都有表达,在籽粒和穗中表达量最高。荧光定量PCR结果表明,Tae-miR166家族15个成员在镉、干旱和低温胁迫处理后的表达模式存在差异,说明Tae-miR166基因家族在植株抵御非生物胁迫时发挥着重要作用。本研究为进一步阐明Tae-miR166基因家族的功能提供了理论依据。展开更多
Previous studies have revealed the miR164 family and the miR164-targeted NAC transcription factor genes in rice(Oryza sativa)and Arabidopsis that play versatile roles in developmental processes and stress responses.In...Previous studies have revealed the miR164 family and the miR164-targeted NAC transcription factor genes in rice(Oryza sativa)and Arabidopsis that play versatile roles in developmental processes and stress responses.In wheat(Triticum aestivum L.),we found nine genetic loci of tae-miR164(tae-MIR164 a to i)producing two mature sequences that downregulate the expression of three newly identified target genes of TaNACs(TaNAC1,TaNAC11,and TaNAC14)by the cleavage of the respective mRNAs.Overexpression of tae-miR164 or one of its target genes(TaNAC14)demonstrated that the miR164-TaNAC14 module greatly affects root growth and development and stress(drought and salinity)tolerance in wheat seedlings,and TaNAC14 promotes root growth and development in wheat seedlings and enhances drought tolerance,while tae-miR164 inhibits root development and reduces drought and salinity tolerance by downregulating the expression of TaNAC14.These findings identify the miR164-TaNAC14 module as well as other taemiR164-regulated genes which can serve as new genetic resources for stress-resistance wheat breeding.展开更多
文摘miR166作为重要的转录后调节因子,在植物的生长发育和对逆境胁迫的反应中扮演着重要的角色。利用PmiREN、Ensembl Plants数据库、MEGA-X、DNAMAN软件以及RNAfold web server、WebLogo和psRNA Target在线网站对小麦miR166(Tae-miR166)基因家族的进化特性和表达模式进行分析。在PmiREN数据库中搜索到19个Tae-miR166基因家族成员,染色体定位发现Tae-miR166成员定位于14条染色体上。序列比对发现Tae-miR166的成熟体序列高度保守。进化分析结果表明,Tae-miR166基因家族成员分别处于4个进化分支。靶基因预测结果表明,Tae-miR166的靶基因包括Ⅲ类同源异型结构域亮氨酸拉链蛋白、β半乳糖苷酶和钙依赖性蛋白激酶等。转录组数据分析表明Tae-miR166家族19个成员在小麦6个组织中都有表达,在籽粒和穗中表达量最高。荧光定量PCR结果表明,Tae-miR166家族15个成员在镉、干旱和低温胁迫处理后的表达模式存在差异,说明Tae-miR166基因家族在植株抵御非生物胁迫时发挥着重要作用。本研究为进一步阐明Tae-miR166基因家族的功能提供了理论依据。
基金financially supported by the National Natural Science Foundation of China(32072003and 32072059)the Key Research and Development Program of Shaanxi Province,China(2021NY-079)。
文摘Previous studies have revealed the miR164 family and the miR164-targeted NAC transcription factor genes in rice(Oryza sativa)and Arabidopsis that play versatile roles in developmental processes and stress responses.In wheat(Triticum aestivum L.),we found nine genetic loci of tae-miR164(tae-MIR164 a to i)producing two mature sequences that downregulate the expression of three newly identified target genes of TaNACs(TaNAC1,TaNAC11,and TaNAC14)by the cleavage of the respective mRNAs.Overexpression of tae-miR164 or one of its target genes(TaNAC14)demonstrated that the miR164-TaNAC14 module greatly affects root growth and development and stress(drought and salinity)tolerance in wheat seedlings,and TaNAC14 promotes root growth and development in wheat seedlings and enhances drought tolerance,while tae-miR164 inhibits root development and reduces drought and salinity tolerance by downregulating the expression of TaNAC14.These findings identify the miR164-TaNAC14 module as well as other taemiR164-regulated genes which can serve as new genetic resources for stress-resistance wheat breeding.
