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目的筛选灰毡毛忍冬Lonicera macranthoides基因表达分析的内参基因,研究其Mads-box基因家族AGL15基因(Lm AGL15)的时空表达特性。方法克隆灰毡毛忍冬内参基因18 S r RNA、Ubiquilin、Actin和Efl-β的基因片段,评价了4个基因在不同部位叶、茎和花蕾,以及4个基因在灰毡毛忍冬生长不同时期的稳定性,筛选内参基因,分析Lm AGL15基因的时空表达特性。结果 18 S r RNA表达最稳定,适宜作为内参基因,叶片、茎的Lm AGL15基因相对表达量较低,花蕾相对表达量较高。花蕾不同发育时期,Lm AGL15的相对表达量呈现逐渐降低的变化,20 d后Lm AGL15的相对表达量最低,然后依次是花蕾初期(15 d)、青绿色花蕾期(10 d)、绿白色花蕾期(5 d)。结论 18 S r RNA是适宜的内参基因。叶片、茎和花蕾中,Lm AGL15的相对表达量差异明显。花蕾发育不同时期,Lm AGL15的相对表达量呈逐渐降低的变化,与花蕾开放变化规律相似。
Objective To screen the internal control gene of Lonicera macranthoides gene expression in Lonicera macranthoides and study the spatial and temporal expression characteristics of its AGL15 gene (Lm AGL15). Methods The gene fragments of 18 S rRNA, Ubiquilin, Actin and Efl-β were cloned and the expression profiles of four genes in leaves, stems and buds in different organs and four genes were evaluated in different periods of growth of Lonicera macranthoides The stability of Lm AGL15 gene was analyzed. The spatial and temporal expression characteristics of Lm AGL15 gene were analyzed. Results The most stable expression of S r RNA was found, which was suitable as an internal control gene. The relative expression level of Lm AGL15 gene in leaves and stems was lower, and the relative expression level of buds was higher. The relative expression of Lm AGL15 showed a decreasing trend at different developmental stages of buds, and the relative expression of Lm AGL15 was the lowest at 20 days, followed by the initial stage of flower bud (15 d), the stage of green bud (10 d) Period (5 d). Conclusion 18 S r RNA is a suitable internal control gene. Among leaves, stems and buds, the relative expression level of Lm AGL15 was significantly different. The relative expression of Lm AGL15 in different stages of flower bud development decreased gradually, which was similar to that of flower bud opening.