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首页> 外文期刊>Acta Horticulturae >Kalanchoe x houghtonii SSH and microarray analysis to screen genes involved in vivipary.
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Kalanchoe x houghtonii SSH and microarray analysis to screen genes involved in vivipary.

机译：Kalanchoe x houghtonii SSH和微阵列分析以筛选参与活动的基因。

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Vivipary, referred here as the formation of novel complete plantlets on mature organs, has been reported in many families as an asexual propagation strategy. In K. x houghtonii (Crassulaceae), viviparous plantlets are formed on leaf margin notches in response to a long day photoperiod and their appearance follow a basipetal fashion. To identify genes involved in this process, suppression subtractive hybridisation libraries (SSH) were prepared. Two hundred c-DNA clones were classified and grouped into 14 functional categories according to Goldberg database (http://estdb.biology.ucla.edu/PcEST/). Six hundred thirty sequences (200 SSH library, 48 database Kalanchoe genus, 382 other species database genes) were used as probes for microarray analysis according to the CombiMatrix technology and a 4x2K Custom ArrayTM was synthesized. RNA was extracted from margin of leaves at 7 stages of development before buds emission (5 to 50 mm) during long-day photoperiod (permissive conditions). Three replications for each sample were prepared. From double strand cDNAs antisense RNAs (a RNA) were synthesized and amino-allil-UTP incorporated and coupled with Alexa FluorReg. 647. Microarray was hybridized according to CombiMatrix protocol. Data were extracted with CombiMatrix Microarray Imager software and exported into Microsoft Excel for computing of mean, median and standard deviation. Person's correlation was computed and data normalized. After background removal, probes were reduced to 484. "Fold change" method (FC=2) was used to compare different levels of gene expression of samples. Significance Analysis of Microarrays Statistic (SAM method) generated 263 significant modulated genes with a False Discovery Rate (FDR) of 5%.
机译：子房，在这里被称为在成熟器官上形成新的完整小苗，在许多家庭中已被报道为无性繁殖策略。在 K。 x houghtonii （景天科）中，响应于一整天的光周期，在叶缘凹口上形成了胎生小植株，它们的外观呈基底状。为了鉴定参与该过程的基因，制备了抑制消减杂交文库（SSH）。根据Goldberg数据库（http://estdb.biology.ucla.edu/PcEST/），将200个c-DNA克隆分类并划分为14个功能类别。根据CombiMatrix技术和4x2K Custom Array TM ，使用630个序列（200个SSH库，48个 Kalanchoe 属，382个其他物种的数据库基因）作为微阵列分析的探针。 sup>被合成。在长时间的光周期（允许条件）下，在芽发射前（5至50毫米），在发育的7个阶段从叶片边缘提取RNA。每个样品准备三份重复。从双链cDNA合成反义RNA（a RNA），并掺入氨基-alil-UTP并与Alexa FluorReg偶联。 647.根据CombiMatrix协议杂交微阵列。使用CombiMatrix Microarray Imager软件提取数据，然后将其导出到Microsoft Excel中以计算平均值，中值和标准差。计算人的相关性并标准化数据。去除背景后，探针减少至484个。使用“倍数变化”方法（FC = 2）比较样品基因表达的不同水平。微阵列统计的显着性分析（SAM方法）产生263个重要的调制基因，错误发现率（FDR）为5％。

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