甘藍型油菜蠟質相關基因的克隆與表達分析
發(fā)布時間：2018-12-23 12:37
【摘要】：植物葉片中,表皮蠟質對植物的生長發(fā)育和適應外界環(huán)境起著重要作用。本研究針對擬南芥(Arabidopsis thaliana)已發(fā)表的蠟質相關基因信息,基于生物信息學的方法獲得以甘藍型油菜(Brassica napus)品種中雙11號為參考的10個蠟質相關基因的44個拷貝序列。然后,以在抽薹期前后葉片蠟質差異明顯的3對自然突變的甘藍型油菜(L1050,P144和W481)為材料,利用同源克隆的方法獲得這些拷貝成員在3對甘藍型油菜中的核苷酸和蛋白序列,通過多重比對分析發(fā)現(xiàn),甘藍型油菜蠟質基因(wax inducer 2,WIN2)家族中的Bn WIN2C01在核苷酸與氨基酸水平上均存在穩(wěn)定的突變位點。利用q RT-PCR分析基因Bn WIN2C01在甘藍型油菜突變體材料抽薹期前后葉片中的表達情況,結果表明該基因在表皮蠟質合成前后的葉片中達到極顯著差異水平(P0.01),說明Bn WIN2C01基因的特異性表達可能影響了葉片蠟質的合成。本研究將有助于闡明葉表皮蠟質形成過程的分子機理,為提高油菜抗逆性提供一定的參考依據。
[Abstract]:In plant leaves, epidermal wax plays an important role in the growth and development of plants and adaptation to the external environment. In this study, based on bioinformatics, we obtained 44 copy sequences of 10 waxy related genes of Arabidopsis thaliana (Arabidopsis thaliana) based on bioinformatics, which were referenced by Zhongshuang11, a Brassica napus (Brassica napus) variety. Then, three pairs of natural mutant Brassica napus (L1050P144 and W481), which had obvious difference in waxy quality of leaves before and after bolting, were used as materials. The nucleotide and protein sequences of these copy members in 3 pairs of Brassica napus were obtained by homologous cloning. The waxy gene (wax inducer 2 of Brassica napus was found by multiple alignment analysis. Bn WIN2C01 in the WIN2 family had stable mutation sites at nucleotide and amino acid levels. Q RT-PCR analysis was used to analyze the expression of Bn WIN2C01 gene in the leaves of Brassica napus mutant before and after bolting stage. The results showed that the gene reached a very significant difference level in the leaves before and after epidermal waxy synthesis (P0.01). These results suggest that the specific expression of Bn WIN2C01 gene may affect the waxy synthesis in leaves. This study will be helpful to elucidate the molecular mechanism of the formation of epidermis waxy and provide some references for improving the stress resistance of rape.
【作者單位】： 西南大學農學與生物科技學院/重慶市油菜工程技術研究中心;西南大學農業(yè)科學研究院;
【基金】：國家重點基礎研究發(fā)展計劃(973)項目(No.2015CB150201) 國家自然科學基金項目(No.31401412,No.U1302266和No.31571701) 國家科技支撐計劃項目(No.2013BAD01B03-12) 中央高�；究蒲袠I(yè)務費專項基金(No.XDJK2016B030) 西南地區(qū)強優(yōu)勢雜種優(yōu)勢利用項目(No.2016YFD0101300)
【分類號】：Q943.2;S565.4
本文編號：2389993
[Abstract]:In plant leaves, epidermal wax plays an important role in the growth and development of plants and adaptation to the external environment. In this study, based on bioinformatics, we obtained 44 copy sequences of 10 waxy related genes of Arabidopsis thaliana (Arabidopsis thaliana) based on bioinformatics, which were referenced by Zhongshuang11, a Brassica napus (Brassica napus) variety. Then, three pairs of natural mutant Brassica napus (L1050P144 and W481), which had obvious difference in waxy quality of leaves before and after bolting, were used as materials. The nucleotide and protein sequences of these copy members in 3 pairs of Brassica napus were obtained by homologous cloning. The waxy gene (wax inducer 2 of Brassica napus was found by multiple alignment analysis. Bn WIN2C01 in the WIN2 family had stable mutation sites at nucleotide and amino acid levels. Q RT-PCR analysis was used to analyze the expression of Bn WIN2C01 gene in the leaves of Brassica napus mutant before and after bolting stage. The results showed that the gene reached a very significant difference level in the leaves before and after epidermal waxy synthesis (P0.01). These results suggest that the specific expression of Bn WIN2C01 gene may affect the waxy synthesis in leaves. This study will be helpful to elucidate the molecular mechanism of the formation of epidermis waxy and provide some references for improving the stress resistance of rape.
【作者單位】： 西南大學農學與生物科技學院/重慶市油菜工程技術研究中心;西南大學農業(yè)科學研究院;
【基金】：國家重點基礎研究發(fā)展計劃(973)項目(No.2015CB150201) 國家自然科學基金項目(No.31401412,No.U1302266和No.31571701) 國家科技支撐計劃項目(No.2013BAD01B03-12) 中央高�；究蒲袠I(yè)務費專項基金(No.XDJK2016B030) 西南地區(qū)強優(yōu)勢雜種優(yōu)勢利用項目(No.2016YFD0101300)
【分類號】：Q943.2;S565.4
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1 譚晨;甘藍型油菜中基因表達的劑量效應及甘藍—黑芥附加系的創(chuàng)建[D];華中農業(yè)大學;2017年
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