玉米MATE基因参与调控植物叶片衰老、下胚轴伸长与种子大小

发布时间：2018-06-18 01:58

本文选题：玉米 + MATE蛋白家族　；参考：《西北农林科技大学》2017年硕士论文

【摘要】：多药及有毒化合物外排家族(Mutidrug and Toxic Compund Extrusion,MATE)蛋白是阳离子次级转运蛋白,参与了植物生长发育阶段的众多生理功能调控。虽然越来越多MATE蛋白被鉴定发现,但对于MATE蛋白的了解还非常局限。关于玉米MATE蛋白家族的研究目前还非常少,因此本研究采用反向遗传学手段对玉米MATE蛋白家族ZmMATE884(GRMZM2G423884)基因进行研究。本研究主要取得如下结果。(1)本研究从玉米gDNA中克隆得到MATE基因ZmMATE884(GRMZM2G423884)。ZmMATE884与56个拟南芥MATE蛋白系统进化分析结果显示ZmMATE884与拟南芥9个MATE蛋白属于同一进化分支,具有较近亲缘关系。后续ZmMATE884与PfMATE蛋白同源比对结果显示二者高度同源,由PfMATE蛋白结构预测ZmMATE884蛋白同样具有12个跨膜结构域。(2)定量RT-PCR显示ZmMATE884在玉米B73植株根和叶中表达量相对较高。表明ZmMATE884具有有组织特异表达模式。亚细胞定位分析结果显示ZmMATE884蛋白与原液泡/次级内体、反式高尔基体/早期内体及高尔基体/内体上的特异标志蛋白有部分共定位,表明ZmMATE884蛋白分布于细胞内体形成转运过程中不同阶段的囊泡结构中。(3)通过农杆菌侵染技术构建ZmMATE884过表达系,观察表型显示过表达系植株较野生型开花明显提前、叶片的自然衰老加快、角果变小,过表达系植株整体矮化丛生。ZmMATE884过表达系在黑暗中萌发6 d、12 d和光下生长7 d时的下胚轴都明显短于野生型;外源添加蔗糖可以促进过表达系和野生型下胚轴的伸长,但过表达系下胚轴长度仍然短于野生型。表明ZmMATE884基因参与植物下胚轴伸长的负调控,且ZmMATE884基因对于下胚轴的调控与蔗糖属于不同调控通路。(4)ZmMATE884过表达突变体种子千粒重小于野生型,且种子长度也更小。表明ZmMATE884参与植物种子大小的调控。(5)ZmMATE884过表达系植物莲座叶衰老加快。碳饥饿诱导处理5 d和7 d后ZmMATE884过表达系植株明显黄于野生型,且总蛋白量也少于野生型。ZmMATE884过表达使叶片衰老提前。(6)ZmMATE884过表达可以将ABS3及其同源基因ABS4、ABS3L1和ABS3L2共同缺失突变体mateq表型回复至野生型,表明ZmMATE884与ABS3、ABS4、ABS3L1和ABS3L2基因具有相似功能。
[Abstract]:Mutidrug and toxin Compund ExtrusionmatEprotein is a cationic secondary transporter, which is involved in the regulation of many physiological functions in plant growth and development. Although more and more mate proteins have been identified, the understanding of mate proteins is still very limited. There are few studies on maize MATE protein family, so this study uses reverse genetics to study the ZmMATE884GRMZM2G423884) gene. In this study, we obtained the following results. 1) in this study, we cloned the MATE gene ZmMATE884GRMZM2G423884N. ZmMATE884 and 56 Arabidopsis thaliana MATE protein phylogenetic analysis showed that ZmMATE884 and Arabidopsis 9 MATE proteins belong to the same phylogenetic branch and have a close phylogenetic relationship. The result of homology comparison between ZmMATE884 and PfMATE protein showed that ZmMATE884 protein had high homology, and ZmMATE884 protein had 12 transmembrane domains. The quantitative RT-PCR showed that ZmMATE884 expressed relatively high in the roots and leaves of maize B73 plant. The results showed that ZmMATE884 had a tissue specific expression pattern. The results of subcellular localization analysis showed that ZmMATE884 protein was partially co-located with protonoplast / secondary endosome, transGolgi / early endosome and specific marker proteins on Golgi body / inner body. The results showed that ZmMATE884 protein was distributed in the vesicle structure of different stages of intracellular body formation and transport.) the overexpression line ZmMATE884 was constructed by Agrobacterium tumefaciens infection technique. The phenotypic observation showed that the over-expression line plants were earlier than wild type plants in flowering. The natural senescence of leaves was accelerated, the pods became smaller, and the Hypocotyl of the over-expressed lines was obviously shorter than that of wild type at 6 days of germination in darkness for 12 days and 7 days under light. Exogenous sucrose could promote the elongation of Hypocotyl of over-expression line and wild type, but the length of Hypocotyl of over-expressed line was still shorter than that of wild type. The results showed that ZmMATE884 gene was involved in the negative regulation of Hypocotyl elongation, and the regulation of ZmMATE884 gene on Hypocotyl and sucrose belonged to different regulation pathways. The 1000-grain weight of ZmMATE884 mutant was smaller than that of wild type, and the seed length was smaller. The results showed that ZmMATE884 was involved in the regulation of plant seed size. ZmMATE884 overexpressed line had accelerated senescence of rosette leaves. After 5 and 7 days of carbon starvation induction, ZmMATE884 overexpression lines were yellowish than wild-type plants. The over-expression of ZmMATE884 and ZmMATE884 could restore the mateq phenotype of ABS3 and its homologous gene ABS4, ABS3L1 and ABS3L2 to wild type, which indicated that ZmMATE884 had similar function with ABS3ABS4ABS3L1 and ABS3L2.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S513;Q943.2

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