摘要
本实验采用RT-PCR和RACE的方法,我们从辣椒叶片中克隆到3-磷酸甘油醛脱氢酶β亚基基因(CaGAPB)的全长序列,基因编码区共1 359 bp,编码452个氨基酸。生物信息学分析显示,该基因编码的蛋白分子量为48.2 kD,等电点6.52;进化树分析表明CaGAPB与同为茄科的番茄、马铃薯、烟草以及葡萄科的葡萄GAPB处于同一进化枝上,表现出较近的亲缘关系,其氨基酸序列同源性达到95%以上;荧光定量分析发现,CaGAPB基因可以被低温处理显著诱导表达,而其他的一些胁迫(NaCl,机械损伤,PEG,甘露醇)和信号分子(乙烯利,H_2O_2,SA)处理均会抑制其表达,同时非生物胁迫DC3000侵染对其表达无影响。综上所述,CaGAPB可能是同辣椒的低温抗性相关,其具体功能有待于进一步的研究。
In this study, the glyceraldehyde-3-phosphate dehydrogenase 13 subunit gene （named CaGAPB） was cloned from leaf tissues of pepper （Capsicum annuum L.） through the reverse transcription polymerase chain amplification （RT-PCR） and rapid amplification of cDNA ends （RACE） methods. It contained an open reading frame of 1 359 bp, encoding a protein of 452 amino acids. Bioinformatics analysis showed that the molecular weight of the protein encoded by this gene was 48.2 kD and the pI value was 6.52. Phylogenetic tree analysis indicated that the CaGAPB and GAPBs of tomato, potato, tobacco, and grape were at the same clade and showed a closer relationship. The amino acids sequence similarities of them were over 95%. Quantitative RT-PCR analysis found that the mRNA level of CaGA PB could be significantly induced by low temperature treatment, while inhibited by others stress （NaC1, mechanical damage, PEG, D-marmitol） and signaling molecules （ethephon, SA, H202）. Meanwhile, the abiotic stress DC3000 infection had no obvious influence on its expression. In summary, the CaGA PB was possibly involved in the cold stress response of pepper, the specific function of which might need further studies.
作者
李严曼
欧阳孟真
孙守如
朱磊
Li Yanman ,Ouyang Mengzhen ,Sun Shouru, Zhu Lei(College of Horticulture, Henan Agricultural Universitiy, Zhengzhou, 45000)
出处
《分子植物育种》
CAS
CSCD
北大核心
2018年第5期1382-1389,共8页
Molecular Plant Breeding
基金
国家青年科学基金青年科学基金项目(31301768)
河南省高等学校重点科研项目(16A210031)共同资助
关键词
3-磷酸甘油醛脱氢酶β亚基
辣椒
基因克隆
荧光定量PCR
The glyceraldehyde-3-phosphate dehydrogenase beta subunit, Pepper, Gene cloning, Real-timequantitative PCR
