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为了解龙眼生长素受体基因TIR1的功能,以龙眼胚性愈伤组织为材料,在转录水平上对TIR1(命名为Dl TIR1-3)及其启动子进行克隆和分析,并在转录后水平上研究TIR1与mi R393的相互调控关系.结果表明,Dl TIR1-3 c DNA全长2196 bp,包含ORF 1 716 bp,编码571个氨基酸(Gen Bank登录号:KR558761);生物信息学分析发现,Dl TIR1-3属于不稳定蛋白,不含有信号肽,含有跨膜螺旋结构,具有F-box保守结构域和富含异亮氨酸重复结构,且Dl TIR1-3与碧桃TIR1保持较高的同源性.染色体步移法克隆获得Dl TIR1-3启动子序列2909 bp(Gen Bank登录号:KR558763),该启动子不存在Cp G岛但含有大量光反应、激素应答、组织特异性调控、胁迫响应以及其他生长发育相关的作用元件.ps RNAtarget结合改良RLM-RACE验证表明mi R393能够裂解Dl TIR1-3从而抑制其m RNA转录;q PCR结果显示,Dl TIR1-3与mi R393相互平衡能够调节龙眼体胚形态建成及响应激素调控.由此可见,Dl TIR1-3通过转录水平的调控作用在龙眼生长发育、激素应答、组织分化及胁迫响应等过程中发挥重要功能,而在转录后水平上mi R393通过裂解Dl TIR1-3参与相关调控过程.
In order to understand the function of longan auxin receptor gene TIR1, TIR1 (named as Dl TIR1-3) and its promoter were cloned and analyzed at the transcriptional level using the embryogenic callus of longan. At the transcriptional level The relationship between TIR1 and miR393 was studied.The results showed that the full length of Dl TIR1-3c DNA was 2196 bp, containing 1 716 bp of ORF, encoding 571 amino acids (Gen Bank accession number: KR558761). Bioinformatics analysis showed that, Dl TIR1-3 is an unstable protein, does not contain a signal peptide, contains a transmembrane helix structure, has an F-box conserved domain and is isoleucine-rich repeat structure, and the Dl TIR1-3 and the Tachyphium TIR1 remain high (Gen Bank Accession No .: KR558763), the promoter does not exist CpG island but contains a large number of photoreactions, hormone response, tissue-specific regulation, stress Response and other growth and development-related components.ps RNAtarget combined with improved RLM-RACE showed that mi R393 could cleave Dl TIR1-3 and inhibit its m RNA transcription. QPCR results showed that the balance between Dl TIR1-3 and mi R393 could regulate Longan somatic embryo morphogenesis and response to hormone Therefore, Dl TIR1-3 plays an important role in the process of longan growth and development, hormone response, tissue differentiation and stress response by regulating the transcriptional level, while miR393 at the posttranscriptional level through the cleavage of Dl TIR1-3 Participate in the relevant regulatory process.