Institute of Biotechnology, National Chung Hsing University, Taichung, Taiwan 40227 ROC.
J Exp Bot. 2014 Feb;65(2):621-39. doi: 10.1093/jxb/ert412. Epub 2013 Dec 9.
ANTHER INDEHISCENCE FACTOR (AIF), a NAC-like gene, was identified in Arabidopsis. In AIF:GUS flowers, β-glucuronidase (GUS) activity was detected in the anther, the upper parts of the filaments, and in the pollen of stage 7-9 young flower buds; GUS activity was reduced in mature flowers. Yellow fluorescent protein (YFP)+AIF-C fusion proteins, which lacked a transmembrane domain, accumulated in the nuclei of the Arabidopsis cells, whereas the YFP+AIF fusion proteins accumulated in the membrane and were absent in the nuclei. Further detection of a cleaved AIF protein in flowers revealed that AIF needs to be processed and released from the endoplasmic reticulum in order to function. The ectopic expression of AIF-C caused a male-sterile phenotype with indehiscent anthers throughout flower development in Arabidopsis. The presence of a repressor domain in AIF and the similar phenotype of indehiscent anthers in AIF-C+SRDX plants suggest that AIF acts as a repressor. The defect in anther dehiscence was due to the down-regulation of genes that participate in jasmonic acid (JA) biosynthesis, such as DAD1/AOS/AOC3/OPR3/OPCL1. The external application of JA rescued the anther indehiscence in AIF-C and AIF-C+SRDX flowers. In AIF-C+VP16 plants, which are transgenic dominant-negative mutants in which AIF is converted to a potent activator via fusion to a VP16-AD motif, the anther dehiscence was promoted, and the expression of DAD1/AOS/AOC3/OPR3/OPCL1 was up-regulated. Furthermore, the suppression of AIF through an antisense strategy resulted in a mutant phenotype similar to that observed in the AIF-C+VP16 flowers. The present data suggest a role for AIF in controlling anther dehiscence by suppressing the expression of JA biosynthesis genes in Arabidopsis.
花粉囊开裂因子(AIF)是在拟南芥中鉴定的 NAC 样基因。在 AIF:GUS 花中,β-葡萄糖醛酸酶(GUS)活性在花药、花丝的上部和 7-9 期幼花蕾的花粉中被检测到;在成熟花中 GUS 活性降低。缺乏跨膜结构域的黄色荧光蛋白(YFP)+AIF-C 融合蛋白在拟南芥细胞的核中积累,而 YFP+AIF 融合蛋白则在膜中积累,在核中不存在。对花中切割的 AIF 蛋白的进一步检测表明,AIF 需要被加工并从内质网中释放出来才能发挥作用。AIF-C 的异位表达导致拟南芥整个花发育过程中花粉囊开裂不完全的雄性不育表型。AIF 中存在阻遏结构域,以及 AIF-C+SRDX 植物中花粉囊开裂不完全的表型相似,表明 AIF 作为一种阻遏物发挥作用。花药开裂的缺陷是由于参与茉莉酸(JA)生物合成的基因如 DAD1/AOS/AOC3/OPR3/OPCL1 的下调。JA 的外源施加挽救了 AIF-C 和 AIF-C+SRDX 花中的花粉囊开裂不完全。在 AIF-C+VP16 植物中,AIF 通过融合到 VP16-AD 基序而转化为有效的激活物,转化为转基因显性负突变体,花药开裂得到促进,DAD1/AOS/AOC3/OPR3/OPCL1 的表达上调。此外,通过反义策略抑制 AIF 导致与 AIF-C+VP16 花中观察到的类似的突变表型。本数据表明,AIF 通过抑制拟南芥中 JA 生物合成基因的表达来控制花粉囊开裂。