橡胶树类萌发素蛋白基因HbGLP01的克隆及其表达分析

Cloning and Expression Analysis of Germin-like Gene HbGLP01 from Rubber Tree

类萌发素蛋白(GLPs)是一类重要的胁迫响应蛋白,参与植物的多种生理调控过程。试验前期通过cDNA-AFLP技术,从橡胶树转录组中筛选到一个与麻风树JcGLP(XM_012225699.1)高度同源的差异表达片段。在此基础上,通过基因预测和PCR扩增克隆到一个GLPs类基因HbGLP01。该基因包含2个外显子和1个内含子,cDNA全长696nt,编码231个氨基酸,预测蛋白分子量和等电点分别为24.8ku和5.54。同源比对表明,该蛋白与麻风树JcGLP1-13蛋白的同源性最高,达到81%。聚类分析研究表明,该蛋白与JcGLP1-13、胡杨PeGLP1-13及三角叶杨PtGLP等聚为一个分支。qRT-PCR分析发现,多主棒孢病菌侵染能诱导橡胶树HbGLP01基因的上调表达,其中高抗棒孢霉落叶病品种IAN873在侵染后24h时表达量达到峰值,高感品种PR107在48h后达到峰值,而且在IAN873中的表达峰值显著高于PR107。

Germin-like proteins（GLPs）, one type of stress response proteins, play an important role in many kinds of plant physiological regulation processes. One differential expression fragment was screened from rubber tree transcriptome with cDNA-AFLP technology in previous work, which showed a high homology with JcGLP（ XM012225699.1）from Jatropha curcas. Due to gene forecast and PCR results, one GLPs gene was cloned and named HbGLP01 in our research, which had two exons and one intron. Sequence analysis revealed that the CDS length of HbGLP01 was 696 nt, encoding a protein of 231 amino acids with an average molecular weight of 24.8 ku, and a p I value of 5.54. The protein of Hb GLP01 exhibited the highest sequence similarity of 81% to JcGLP1-13 from J. curcas. The phylogenetic analysis showed the amino acids sequence clustered in the same clade with JcGLP1-13, PeGLP1-13 from Populus euphratica and Pt GLP from Populus trichocarpa. The expressions of HbGLP01 were up-regulated when rubber tree was infected by Corynespora cassiicola, which was detected by q RT-PCR. For highly resistant cv.IAN87 to Corynespora leaf fall disease, the expressions peak value of HbGLP01 was emerged at 24 h after inoculation,while 48 h for highly sensitive cv. PR107. The peak value in IAN87 was significantly higher than in PR107.