薺菜PIN3、PIN7同源基因在擬南芥中表達(dá)及其對(duì)形態(tài)建成的影響
發(fā)布時(shí)間：2018-08-06 19:12
【摘要】：PIN生長(zhǎng)素極性運(yùn)輸?shù)年P(guān)鍵基因,該基因家族在擬南芥中有8個(gè)成員,其中PIN3、PIN7主要參與側(cè)向的生長(zhǎng)素極性運(yùn)輸而對(duì)其形態(tài)建成起重要調(diào)節(jié)作用。本研究將薺菜中的同源性CbPIN3和CbPIN7基因轉(zhuǎn)化擬南芥,獲得了純合轉(zhuǎn)基因植株At(CbPIN3)和 At(CbPIN7),兩者雜交后獲得了薺菜兩個(gè)同源基因共轉(zhuǎn)化的擬南At(CbPIN3+CbPIN7).對(duì)幾種轉(zhuǎn)基因擬南芥形態(tài)發(fā)育的觀察,CbPIN3的轉(zhuǎn)入導(dǎo)致了較為顯著的心皮形態(tài)變異,而CbPIN7的轉(zhuǎn)基因擬南芥心皮與野生型擬南芥心皮形態(tài)差異較小,說(shuō)明在心皮形態(tài)發(fā)育過(guò)程中PIN3發(fā)揮更大的影響而PIN7作用較弱。兩者PIN3基因表達(dá)的定量分析也表明薺菜中PIN3具有更高的表達(dá)水平。在對(duì)At(CbPIN3)、At(CbPIN7)和At(CbPIN3+CbPIN7)擬南芥心皮形態(tài)更為細(xì)致的比對(duì)觀察發(fā)現(xiàn)At(CbPIN3+CbPIN4)的共轉(zhuǎn)入還導(dǎo)致了更嚴(yán)重的心皮變異和胚珠發(fā)育不良。說(shuō)明薺菜和擬南芥PIN3.PIN7在心皮發(fā)育過(guò)程中的表達(dá)是存在差異的。為了探明薺菜1IN3、PIN7同源基因在擬南芥中的表達(dá)規(guī)律,構(gòu)建了薺菜PIN3、PIN7啟動(dòng)子的GUS報(bào)告系統(tǒng)并轉(zhuǎn)化擬南芥,GUS檢測(cè)表明薺菜與擬南芥同源基因的表達(dá)模式基本一致,只存在著一定程度的表達(dá)水平差異。通過(guò)生物信息學(xué)對(duì)薺菜和擬南芥PIN7啟動(dòng)子進(jìn)行了元件比對(duì)分析后,將薺菜PIN7啟動(dòng)子中特異性包含的發(fā)育相關(guān)AuxRR-core、GARE、circadian元件分別進(jìn)行元件缺失并構(gòu)建成多個(gè)GUS報(bào)告基因,轉(zhuǎn)化擬南芥后的GUS檢測(cè)表明AuxRR-core、GARE、circadian元件的缺失導(dǎo)致了薺菜PIN7在多個(gè)組織中的表達(dá)出現(xiàn)變化。實(shí)驗(yàn)結(jié)果說(shuō)明薺菜與擬南芥PIN7在心皮形態(tài)建成的調(diào)控體系中存差異,特別是赤霉素調(diào)控體系可能在薺菜形態(tài)建成中發(fā)揮著更重要的作用,PINs的調(diào)控成為了赤霉素和生長(zhǎng)素信號(hào)交互作用的節(jié)點(diǎn)。外源赤霉素GA3處理幾種轉(zhuǎn)基因材料后,At(CbPIN7)擬南芥心皮較其他二者出現(xiàn)顯著的差異也印證了這一點(diǎn)。由此可以推論薺菜赤霉素調(diào)控體系與生長(zhǎng)素交互作用參與到植物發(fā)育事件中,從而使得薺菜的發(fā)育途徑和調(diào)控網(wǎng)絡(luò)產(chǎn)生出豐富的變異,最終這些變異與兩者基因水平上的微小差異相累加導(dǎo)致兩物種間的某些性狀或表型呈現(xiàn)非漸進(jìn)式進(jìn)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.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q943.2
本文編號(hào)：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.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q943.2
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 朱占偉;彭彥;趙燕;胡清云;張學(xué)文;;生長(zhǎng)素極性運(yùn)輸PIN基因在擬南芥和薺菜不同組織表達(dá)的定量分析[J];湖南農(nóng)業(yè)大學(xué)學(xué)報(bào)(自然科學(xué)版);2013年05期
2 薛皓月;徐桂霞;國(guó)春策;山紅艷;孔宏智;;擬南芥和琴葉擬南芥中MADS-box基因的比較進(jìn)化分析[J];生物多樣性;2010年02期
，本文編號(hào)：2168717
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