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花芽的发育是花品质形成与调控的生物学基础,温度是影响蝴蝶兰(Phalaenopsis hybrida)花芽分化与发育乃至花朵开放的重要因素。为鉴定低夜温诱导蝴蝶兰花梗芽分化和发育相关基因,以蝴蝶兰兄弟女孩品种为材料,利用抑制削减杂交技术,构建低夜温诱导的花梗芽与营养生长期顶叶的正向差减文库。PCR验证后,挑取300个阳性单克隆进行测序和分析,共获得207条非冗余序列(Gen Bank登录号:JK720764~JK720970)。这些基因覆盖了宽广功能范围的开花相关基因,提供了从营养生长期向生殖生长期转变相关分子机理方面的有益信息。Real-timePCR检测了具不同功能的23个基因在营养生长期顶叶和不同大小花梗芽中的表达水平。这些基因的表达量变化趋势不同,但在一定大小花梗芽中的表达量较营养生长期顶叶均有上调。与在营养生长期顶叶中的表达量相比,flowering locusT(FT)、APETALA1(AP1)、茉莉酮酸酯-O-甲基转移酶(JMT)基因和NADH泛醌氧化还原酶基因在各阶段花梗芽中的表达量均显著上调,表明这4个基因可能在低夜温诱导蝴蝶兰花梗芽分化和发育中起重要作用。本研究为深入探究低夜温诱导蝴蝶兰开花的分子机理提供了重要的基础资料。
The development of flower buds is the biological basis for the formation and regulation of flower quality. Temperature is an important factor that affects the flower bud differentiation and development of Phalaenopsis hybrida and even the opening of flowers. In order to identify the genes involved in the differentiation and development of Phalaenopsis flower buds induced by low night temperature, the phallophyte buds induced by low night temperature and the positive difference of vegetative vegetative parietal lobe were constructed by using the suppression subtractive hybridization technique Less library. After PCR validation, 300 positive clones were picked for sequencing and analysis, and a total of 207 non-redundant sequences (Gen Bank accession number: JK720764 ~ JK720970) were obtained. These genes cover a broad range of flowering related genes and provide useful information on the molecular mechanisms involved in the transition from vegetative to reproductive growth. Real-time PCR detected the expression of 23 genes with different functions in the vegetative phase of the parietal lobe and different size of the pedicel. The expression of these genes showed different trends, but the expression level in peduncle buds of certain size was higher than that in vegetative phase. Flowering locusT (FT), APETALA1 (AP1), jasmonic acid ester-O-methyltransferase (JMT) genes and NADH ubiquinone oxidoreductase genes were significantly different in the The expression levels of these four genes were significantly up-regulated in the stage of pedicle buds, indicating that these four genes may play an important role in the differentiation and development of Phalaenopsis buds induced by low night temperature. This study provides important basic information for further exploring the molecular mechanism of low temperature-induced temperature on the flowering of Phalaenopsis.