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The MYB80 Transcription Factor Is Required for Pollen Development and the Regulation of Tapetal Programmed Cell Death in Arabidopsis thaliana

机译:MYB80转录因子是拟南芥中花粉发育和调控细胞程序性死亡的必需条件。

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摘要

Arabidopsis thaliana MYB80 (formerly MYB103) is expressed in the tapetum and microspores between anther developmental stages 6 and 10. MYB80 encodes a MYB transcription factor that is essential for tapetal and pollen development. Using microarray analysis of anther mRNA, we identified 404 genes differentially expressed in the myb80 mutant. Employing the glucocorticoid receptor system, the expression of 79 genes was changed when MYB80 function was restored in the myb80 mutant following induction by dexamethasone. Thirty-two genes were analyzed using chromatin immunoprecipitation, and three were identified as direct targets of MYB80. The genes encode a glyoxal oxidase (GLOX1), a pectin methylesterase (VANGUARD1), and an A1 aspartic protease (UNDEAD). All three genes are expressed in the tapetum and microspores. Electrophoretic mobility shift assays confirmed that MYB80 binds to all three target promoters, with the preferential binding site containing the CCAACC motif. TUNEL assays showed that when UNDEAD expression was silenced using small interfering RNA, premature tapetal and pollen programmed cell death occurred, resembling the myb80 mutant phenotype. UNDEAD possesses a mitochondrial targeting signal and may hydrolyze an apoptosis-inducing protein(s) in mitochondria. The timing of tapetal programmed cell death is critical for pollen development, and the MYB80/UNDEAD system may regulate that timing.
机译:拟南芥MYB80(以前为MYB103)在花药发育阶段6和10之间的绒毡层和小孢子中表达。MYB80编码对于绒毡层和花粉发育至关重要的MYB转录因子。使用花药mRNA的微阵列分析,我们鉴定了myb80突变体中差异表达的404个基因。利用糖皮质激素受体系统,地塞米松诱导myb80突变体恢复MYB80功能后,79个基因的表达发生了改变。使用染色质免疫沉淀法分析了32个基因,其中3个被鉴定为MYB80的直接靶标。这些基因编码乙二醛氧化酶(GLOX1),果胶甲基酯酶(VANGUARD1)和A1天冬氨酸蛋白酶(UNDEAD)。所有三个基因均在绒毡层和小孢子中表达。电泳迁移率变动分析证实MYB80与所有三个靶标启动子结合,优先结合位点包含CCAACC基序。 TUNEL分析表明,当使用小分子干扰RNA沉默UNDEAD表达时,会发生过早的绒毛和花粉程序性细胞死亡,类似于myb80突变表型。 UNDEAD具有线粒体靶向信号,并且可以水解线粒体中的凋亡诱导蛋白。程序性细胞死亡的定时是花粉发育的关键,MYB80 / UNDEAD系统可能会调节该时间。

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