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以‘金煌’杧果(Mangifera indica L.‘Jinhuang’)为试材,通过RACE及PCR技术克隆得到叶绿素a/b结合蛋白CAB基因的cDNA全长,命名为MiCAB2(GenBank登录号:KT944730)。该基因全长为990 bp,含795 bp开放阅读框,编码246个氨基酸,与橄榄(Canarium album)亲缘关系最近。MiCAB2属于叶绿素a/b结合蛋白家族,其氨基酸序列中存在一个典型的捕光叶绿素a/b结合蛋白功能域、3个蛋白激酶C磷酸化位点和4个N–肉豆蔻酰位点。实时荧光定量PCR检测表明:MiCAB2在杧果各组织中均表达且表达量差异较大,在成熟叶片中表达量最高,在花、胚胎、果实中表达量仅为叶片的1/50~1/12,在根和茎中极低;在花芽分化至开花期表达量呈迅速上升后下降再上升趋势,在正常胚胎中呈下降后回升再下降趋势,而在败育胚胎中呈逐渐下降趋势;叶片在昼夜交替中,白天表达量明显高于夜晚,14时最高,22时最低。MiCAB2受不同催花药剂的诱导,乙烯利作用最为明显,硝酸钾次之,两者均为正向调控,而多效唑起负调控作用。
The full-length cDNA of CAB gene of chlorophyll a / b-binding protein was cloned by RACE and PCR with the primers ’MiCAB2’ (GenBank accession number: KT944730), using ’Mangifera indica L.’Jinhuang’ . The full length of this gene is 990 bp, contains 795 bp open reading frame and encodes a protein of 246 amino acids. It has the closest genetic relationship with Canarium album. MiCAB2 belongs to the family of chlorophyll a / b-binding proteins, and a typical light-harvesting chlorophyll a / b binding protein domain, three protein kinase C phosphorylation sites and four N-myristoyl residues exist in the amino acid sequence of MiCAB2. The results of real-time PCR showed that MiCAB2 was expressed in all tissues of ginkgo biloba and its expression was quite different. The expression level of MiCAB2 was the highest in mature leaves, and it was only 1/50 ~ 1 / 12 in roots and stems. The expression levels increased rapidly from the flower buds to the flowering stage, then decreased and then increased, decreased in normal embryos and then rose again and then decreased, but decreased gradually in aborted embryos. Leaf alternating at day and night, the daytime expression was significantly higher than the night, the highest at 14 o’clock, the lowest at 22 o’clock. MiCAB2 was induced by different drug-inducing agents, ethephon was the most obvious, potassium nitrate was the second, both were positive regulation, and paclobutrazol negatively regulated.