黑毛雪兔子通气组织发育相关基因ShCTR1的克隆及表达

黑毛雪兔子(Saussurea inversa Raab-Straube)是一种典型的高山植物,具有发达的通气组织。该研究以黑毛雪兔子为材料,利用同源克隆和RACE技术克隆了通气组织形成相关基因(ShCTR1),并对其进行序列分析、系统进化分析、表达分析和亚细胞定位分析,以探讨该基因的功能及其与通气组织的关系。结果表明:(1)成功克隆获得ShCTR1基因cDNA全长为2891 bp(NCBI登录号为:ON081649),包含2550 bp的开放阅读框,编码849个氨基酸,理论等电点5.90,分子式C_(4066)H_(6417)N_(1159)O_(1268)S_(43),为疏水蛋白,无跨膜结构。(2)系统进化树分析显示,黑毛雪兔子ShCTR1与菜蓟(Cynara cardunculus L.)CcCTR1的氨基酸序列相似度最高;非编码区序列分析发现ShCTR1基因含有大量的光响应元件,表明ShCTR1基因可能参与对紫外胁迫的响应。(3)实时荧光定量分析显示,ShCTR1基因在根、茎和叶中均有表达,且在根中的表达量最高;在紫外、低温和低氧胁迫下,ShCTR1基因的表达量均上调,证明ShCTR1基因参与了对以上3种环境胁迫的响应。(4)亚细胞定位显示,ShCTR1主要分布于细胞核。研究推测,黑毛雪兔子ShCTR1基因可能在通气组织的形成以及对逆境胁迫的响应中具有重要作用。

Cloning and Expression Analysis of Aerenchyma Development Related Gene ShCTR1 in Saussurea inversa Raab-Straube

Saussurea inversa Raab-Straube is a typical alpine plant, having developed aerenchyma is the most representative structural feature. This study cloned the aerenchyma related gene (ShCTR1) with S. inversa using the homologous cloning and RACE technology, and by means of sequence analysis, phylogenetic analysis, expression analysis and subcellular location analysis, explored the function of the gene and its aerenchyma. The results indicated that: (1) the ShCTR1 gene cDNA was 2 891 bp (NCBI accession number: ON081649) and contains 2 550 bp open reading frame encoding 849 amino acids, the theoretical isoelectric point is 5.90, with the molecular formula C₄₀₆₆H₆₄₁₇N₁₁₅₉O₁₂₆₈S₄₃. It was a hydrophobic protein with no transmembrane structure. (2) Phylogenetic tree analysis showed that S. inversa ShCTR1 had high similarity with CcCTR1 of Cynara cardunculus L., Sequence analysis of non-coding regions identified ShCTR1 containing a large number of photoresponse elements, suggesting that ShCTR1 may be involved in the response to UV stress. (3) Quantitative real-time fluorescence analysis showed that ShCTR1 were expressed in roots, stems and leaves, with the highest expression in roots, and its expression was upregulated under UV, cold and hypoxia stresses, which demonstrated that the ShCTR1 was involved in the response to the above three stresses. (4) Subcellular localization revealed that ShCTR1 was predominantly distributed in the nucleus. It had speculated that ShCTR1 may have an important role in the formation of aerenchyma as well as in response to adversity stress.