论文部分内容阅读
以玉米自交系豫82为材料,同源克隆了与水稻q LTG3-1基因同源的玉米Zmq LTG3-1基因。序列分析发现,该基因c DNA序列全长606bp,开放阅读框441bp,编码包含147个氨基酸的蛋白。该蛋白含有AAI_LTSS超蛋白家族的LTP(含8个半胱氨酸)保守结构,分子量为14.16k Da,等电点是8.3,与高粱、水稻的相似度分别为92%和88%。荧光定量RT-PCR分析结果表明,Zmq LTG3-1在玉米胚中的表达量最高,其次是茎尖,在叶、根等器官中的表达量较低;随着胚萌发时间的延长,其表达量逐渐增加,在萌发24h时呈现最高丰度表达,之后表达丰度与萌发时间呈现规律性下降。亚细胞定位结果表明,该基因所编码的产物被定位在细胞膜上,是一跨膜蛋白。通过农杆菌介导的花序侵染法将过表达载体OE-Zmq LTG3-1和空载体p CAMBIA1304转入拟南芥,在诸如干旱、高温和盐等非生物胁迫下,与对照相比,T3转基因植株的成活率显著提高,说明Zmq LTG3-1基因对于抵抗非生物胁迫发挥了重要作用。
The maize Zmq LTG3-1 gene homologous to the q LTG3-1 gene was cloned from maize inbred line Yu-82. Sequence analysis showed that the cDNA sequence of this gene was 606 bp in length and contained 441 bp of open reading frame (ORF) encoding a protein of 147 amino acids. The protein contains conserved LTP (containing 8 cysteines) AAI_LTSS superfamily proteins with a molecular weight of 14.16 kDa and an isoelectric point of 8.3. Similarity with sorghum and rice is 92% and 88%, respectively. Fluorescence quantitative RT-PCR analysis showed that the expression level of Zmq LTG3-1 was the highest in maize embryos, followed by the shoot tips, and lower in leaves, roots and other organs. With the extension of germination time, the expression of Zmq LTG3-1 was higher Gradually increased and showed the highest abundance expression at 24 hours of germination, followed by a regular decline in expression abundance and germination time. Subcellular localization results show that the gene encoded by the product is located in the cell membrane, is a transmembrane protein. The over-expression vector OE-Zmq LTG3-1 and the empty vector p CAMBIA1304 were transformed into Arabidopsis by Agrobacterium-mediated inflorescence inoculation. Under abiotic stress such as drought, high temperature and salt, compared with the control, T3 The survival rate of transgenic plants was significantly increased, indicating that Zmq LTG3-1 gene plays an important role in resisting abiotic stress.