巴西橡胶树叶绿体型果糖-1,6-二磷酸酶基因的全长cDNA克隆与表达分析

利用RACE技术从巴西橡胶树中克隆到一个叶绿体型FBPase基因,命名为HbcpFBPase。该基因的c DNA全长为1 512 bp,包含1 209 bp的开放阅读框,编码一个由402个氨基酸残基组成的蛋白,蛋白分子量大小约为43.88 ku,理论等电点为6.64。多重序列比对表明该基因为叶绿体型果糖-1,6-二磷酸酶。Target P软件预测HbcpFBPase在叶绿体中的概率是0.961。实时荧光定量PCR分析表明,HbcpFBPase基因在雌花中表达量最高,雄花及种子次之;此外,机械伤害和割胶以及多种植物激素如乙烯利ET及植物生长素2,4-D可使HbcpFBPase基因下调表达。研究结果对进一步研究橡胶树光合作用碳固定、以及深入研究光合作用与胶乳合成之间的关系提供理论参考。

Cloning and Expression Analysis of a Chloroplast Fructose-1,6-Bisphosphatase Gene in Hevea brasiliensis

Fructose 1,6 - bisphosphatase(fructose-1,6-bisphosphatase, FBPase, EC 3.1.3.11)is a regulatory key enzyme for dark reactions of photosynthesis in green plants. In this study, the full-length cDNA of a putative chloroplast FBPase gene from Hevea brasiliensis was cloned by using rapid amplification of cDNA ends(RACE)technology, and named as HbcpFBPase. The nucleotide sequence of HbcpFBPase cDNA was 1 512 bp long, which contained an open reading frame(ORF)of 1 209 bp, and predicted a peptide of 402 amino-acids s with a molecular weight of 43.88 ku and a theoretical pI of 6.64. Multiple sequence alignment indicated that HbcpFBPase were grouped together with other known chloroplast FBPase enzymes. TargetP software predicting showed HbcpFBPase had a probability of 0.961 localized in the chloroplast. Real-time PCR analysis showed that HbcpFBPase gene was most highly expressed in the female flower, and then in male flower and seeds. Furthermore, HbcpFBPase expressions were found to be down regulated by wounding, tapping and several phytohormones(ethylene and auxin). This study will be beneficial to further studies of rubber tree carbon fixation of photosynthesis, as well as the relationship between photosynthesis and rubber latex production.