通过RT-PCR与RACE技术从唐菖蒲(Gladiolus hybridus)品种‘Rose Supreme’球茎中克隆茉莉酸生物合成途径中的关键酶——12-氧-植物二烯酸还原酶(12-oxo-phytodienoic acid reductase,OPR)基因的1489bp全长cDNA序列,quantitative RT-PCR结果表明,GhOPR3在唐菖蒲叶、花、根、匍匐茎、新球茎和籽球中都表达,其中在籽球和匍匐茎中相对表达量较高;0.1~0.5mmol·L^-1的茉莉酸甲酯(Methyl jasmonate,MJ)处理后,提高了GhOPR3在球茎中的表达量和内源MJ含量;采用农杆菌介导侵染拟南芥花粉,进行GhOPR3基因的过表达分析,过表达拟南芥株系较野生型提高了耐盐性和抗旱性;提高了机械损伤后相关基因的表达水平和内源MJ含量。
OPR is a key enzyme in jasmonic acid biosynthesis pathway, a full-length cDNA named GhOPR3 was cloned in Gladiolus hybridus 'Rose Supreme' corms by RT-PCR and RACE. The results of quantitative RT-PCR showed that GhOPR3 gene was expressed in leaf, flower, root, stolon, corm and cormel, and the relatively high expression level of GhOPR3 was observed in cormel and stolon. Meanwhile, the expression level and the endogenous MJ content in corms steadily increased under MJ treatment with a raising concentration gradients from 0.1 mmol · L^-1 to 0.5 mmol · L^-1. GhOPR3 was overexpressed in Arabidopsis, which increased the salt tolerance and drought-resistant in Arabidopsis. After mechanical damage, the expression level of related resistance genes and the content of endogenous MJ were increased.