荠菜PIN3、PIN7同源基因在拟南芥中表达及其对形态建成的影响
发布时间：2018-08-06 19:12
【摘要】：PIN生长素极性运输的关键基因,该基因家族在拟南芥中有8个成员,其中PIN3、PIN7主要参与侧向的生长素极性运输而对其形态建成起重要调节作用。本研究将荠菜中的同源性CbPIN3和CbPIN7基因转化拟南芥,获得了纯合转基因植株At(CbPIN3)和 At(CbPIN7),两者杂交后获得了荠菜两个同源基因共转化的拟南At(CbPIN3+CbPIN7).对几种转基因拟南芥形态发育的观察,CbPIN3的转入导致了较为显著的心皮形态变异,而CbPIN7的转基因拟南芥心皮与野生型拟南芥心皮形态差异较小,说明在心皮形态发育过程中PIN3发挥更大的影响而PIN7作用较弱。两者PIN3基因表达的定量分析也表明荠菜中PIN3具有更高的表达水平。在对At(CbPIN3)、At(CbPIN7)和At(CbPIN3+CbPIN7)拟南芥心皮形态更为细致的比对观察发现At(CbPIN3+CbPIN4)的共转入还导致了更严重的心皮变异和胚珠发育不良。说明荠菜和拟南芥PIN3.PIN7在心皮发育过程中的表达是存在差异的。为了探明荠菜1IN3、PIN7同源基因在拟南芥中的表达规律,构建了荠菜PIN3、PIN7启动子的GUS报告系统并转化拟南芥,GUS检测表明荠菜与拟南芥同源基因的表达模式基本一致,只存在着一定程度的表达水平差异。通过生物信息学对荠菜和拟南芥PIN7启动子进行了元件比对分析后,将荠菜PIN7启动子中特异性包含的发育相关AuxRR-core、GARE、circadian元件分别进行元件缺失并构建成多个GUS报告基因,转化拟南芥后的GUS检测表明AuxRR-core、GARE、circadian元件的缺失导致了荠菜PIN7在多个组织中的表达出现变化。实验结果说明荠菜与拟南芥PIN7在心皮形态建成的调控体系中存差异,特别是赤霉素调控体系可能在荠菜形态建成中发挥着更重要的作用,PINs的调控成为了赤霉素和生长素信号交互作用的节点。外源赤霉素GA3处理几种转基因材料后,At(CbPIN7)拟南芥心皮较其他二者出现显著的差异也印证了这一点。由此可以推论荠菜赤霉素调控体系与生长素交互作用参与到植物发育事件中,从而使得荠菜的发育途径和调控网络产生出丰富的变异,最终这些变异与两者基因水平上的微小差异相累加导致两物种间的某些性状或表型呈现非渐进式进化。
[Abstract]:The key gene of polar transport of PIN auxin, which has eight members in Arabidopsis thaliana, is mainly involved in the polar transport of auxin and plays an important role in morphogenesis. In this study, homologous CbPIN3 and CbPIN7 genes from Shepherd's purse were transformed into Arabidopsis thaliana, and the homozygous transgenic plants at (CbPIN3) and at (CbPIN7) were obtained. Observation on morphological Development of several transgenic Arabidopsis thaliana the transformation of CbPIN3 resulted in significant carpel shape variation, while the difference of carpel morphology between CbPIN7 transgenic Arabidopsis and wild type Arabidopsis thaliana was small. The results showed that PIN3 played a more important role in the development of carpels and PIN7 played a weaker role. The quantitative analysis of PIN3 gene expression also showed that PIN3 had a higher expression level in Shepherd purse. A more detailed comparison of the carpel morphology of at (CbPIN3) at (CbPIN7) and at (CbPIN3 CbPIN7) in Arabidopsis thaliana showed that the cotransfer of at (CbPIN3 CbPIN4) also resulted in more severe carpel variation and hypoplasia of ovule. The results showed that there were differences in the expression of PIN3.PIN7 between Shepherd's purse and Arabidopsis thaliana during carpel development. In order to investigate the expression of PIN7 homologous gene in Arabidopsis thaliana, the GUS report system of PIN3PIN7 promoter was constructed and transformed into Arabidopsis thaliana Gus. The results showed that the expression pattern of PIN7 homologous gene in Shepherd's purse and Arabidopsis thaliana was basically the same. There is only a certain degree of expression level difference. The PIN7 promoters of Shepherd's purse and Arabidopsis thaliana were analyzed by bioinformatics. The developmental correlation AuxRR-corer-GAREEcircadian elements contained in the Shepherd's purse PIN7 promoter were deleted and several GUS reporter genes were constructed. The GUS analysis of Arabidopsis thaliana showed that the absence of AuxRR-Coreus gire circadian element resulted in the change of PIN7 expression in several tissues. The results showed that there were differences between Shepherd's purse and Arabidopsis PIN7 in carpel morphogenesis. In particular, the regulation system of gibberellin may play a more important role in the morphogenesis of Shepherd purse. The regulation of pins becomes the node of interaction between gibberellin and auxin signal. There were significant differences in carpel of Arabidopsis thaliana (CbPIN7) treated with exogenous gibberellin GA3. It can be inferred that the interaction between the regulation system of Gibberellin and auxin in Shepherd's purse is involved in plant development events, which makes the development pathway and regulatory network of Shepherd's purse produce rich variation. Ultimately, these variations and minor differences at the gene level lead to a non-progressive evolution of some traits or phenotypes between the two species.
【学位授予单位】：湖南农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2
本文编号：2168717
[Abstract]:The key gene of polar transport of PIN auxin, which has eight members in Arabidopsis thaliana, is mainly involved in the polar transport of auxin and plays an important role in morphogenesis. In this study, homologous CbPIN3 and CbPIN7 genes from Shepherd's purse were transformed into Arabidopsis thaliana, and the homozygous transgenic plants at (CbPIN3) and at (CbPIN7) were obtained. Observation on morphological Development of several transgenic Arabidopsis thaliana the transformation of CbPIN3 resulted in significant carpel shape variation, while the difference of carpel morphology between CbPIN7 transgenic Arabidopsis and wild type Arabidopsis thaliana was small. The results showed that PIN3 played a more important role in the development of carpels and PIN7 played a weaker role. The quantitative analysis of PIN3 gene expression also showed that PIN3 had a higher expression level in Shepherd purse. A more detailed comparison of the carpel morphology of at (CbPIN3) at (CbPIN7) and at (CbPIN3 CbPIN7) in Arabidopsis thaliana showed that the cotransfer of at (CbPIN3 CbPIN4) also resulted in more severe carpel variation and hypoplasia of ovule. The results showed that there were differences in the expression of PIN3.PIN7 between Shepherd's purse and Arabidopsis thaliana during carpel development. In order to investigate the expression of PIN7 homologous gene in Arabidopsis thaliana, the GUS report system of PIN3PIN7 promoter was constructed and transformed into Arabidopsis thaliana Gus. The results showed that the expression pattern of PIN7 homologous gene in Shepherd's purse and Arabidopsis thaliana was basically the same. There is only a certain degree of expression level difference. The PIN7 promoters of Shepherd's purse and Arabidopsis thaliana were analyzed by bioinformatics. The developmental correlation AuxRR-corer-GAREEcircadian elements contained in the Shepherd's purse PIN7 promoter were deleted and several GUS reporter genes were constructed. The GUS analysis of Arabidopsis thaliana showed that the absence of AuxRR-Coreus gire circadian element resulted in the change of PIN7 expression in several tissues. The results showed that there were differences between Shepherd's purse and Arabidopsis PIN7 in carpel morphogenesis. In particular, the regulation system of gibberellin may play a more important role in the morphogenesis of Shepherd purse. The regulation of pins becomes the node of interaction between gibberellin and auxin signal. There were significant differences in carpel of Arabidopsis thaliana (CbPIN7) treated with exogenous gibberellin GA3. It can be inferred that the interaction between the regulation system of Gibberellin and auxin in Shepherd's purse is involved in plant development events, which makes the development pathway and regulatory network of Shepherd's purse produce rich variation. Ultimately, these variations and minor differences at the gene level lead to a non-progressive evolution of some traits or phenotypes between the two species.
【学位授予单位】：湖南农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q943.2
【参考文献】
相关期刊论文 前2条
1 朱占伟;彭彦;赵燕;胡清云;张学文;;生长素极性运输PIN基因在拟南芥和荠菜不同组织表达的定量分析[J];湖南农业大学学报(自然科学版);2013年05期
2 薛皓月;徐桂霞;国春策;山红艳;孔宏智;;拟南芥和琴叶拟南芥中MADS-box基因的比较进化分析[J];生物多样性;2010年02期
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