EST-PCR markers representing watermelon fruit genes are polymorphic among watermelon heirloom cultivars sharing a narrow genetic base
EST-PCR markers representing watermelon fruit genes are polymorphic among watermelon heirloom cultivars sharing a narrow genetic base
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DOI：
10.1017/s1479262108014366
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发表时间：
2009-04-01
期刊：
影响因子：
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通讯作者：
Davis, Angela
中科院分区：
文献类型：
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作者：
Levi, Amnon;Wechter, Patrick;Davis, Angela
到目前为止，只有几个序列标记位点（STS）标记与控制西瓜果实品质的基因。西瓜（Citrullus lanatus var. lanatus）构建。cDNA克隆的序列分析产生了4700个在西瓜果实中表达的非冗余EST（EST unigenes）（http：www.ncbi.nlm.nih.gov，http://www.icugi.org）。其中100个EST单基因[包括40个含有简单重复序列（SSR）基序的EST单基因（EST-SSR）和60个常规EST单基因（不含SSR基序）]用于设计PCR实验的引物对。用25个西瓜传家宝品种和13个西瓜属的美国植物介绍（US Pis）的基因组DNA在PCR实验中测试EST引物对，包括四个C。毛状变种5个C lanatus var. citroides Pis和四种西瓜属（Citrullus colocynthis Pis）。40对EST-SSR引物和60对EST引物在西瓜和西瓜品种间分别扩增出108和142个EST-PCR标记。在108个与EST-SSR相关的EST-PCR标记中，103个存在于西瓜中，64个存在于栽培品种中。在这64个标记中，有45个（70.3%）在品种间具有多态性。在与常规EST相关的142个EST-PCR标记中，134个存在于西瓜中，108个存在于栽培品种中。在这108个标记中，有86个（79.6%）在品种间具有多态性。本研究结果表明，西瓜品种果实表达基因编码区存在多态性。利用上述数据共获得131个西瓜果实基因多态性EST-PCR标记（45个与EST-SSR相关，86个与常规ESTs相关）。这些分子标记可用于西瓜品种和育种系的DNA指纹图谱、亲缘关系的评估和西瓜遗传图谱的构建。
To date, there are only a few sequenced-tagged site (STS) markers associated with genes controlling fruit quality in watermelon. A normalized cDNA library for watermelon fruit (Citrullus lanatus var. lanatus) was constructed. Sequence analysis of the cDNA clones resulted in the development of 4700 non-redundant ESTs (EST unigenes) expressed in watermelon fruit (http://www.ncbi.nlm.nih.gov, http://www.icugi.org). One hundred of these EST unigenes [including 40 EST unigenes that contain simple sequence repeat (SSR) motives (EST-SSRs) and 60 Customary EST unigenes (not containing SSR motives)] were used for designing primer pairs for PCR experiments. The EST primer pairs were tested in PCR experiments with genomic DNA of 25 watermelon heirloom cultivars and 13 United States Plant Introductions (US Pis) of Citrullus sp., including four C. lanatus var. lanatus, five C lanatus var. citroides Pis and four Citrullus colocynthis Pis. The 40 EST-SSR and 60 EST primer pairs produced 108 and 142 EST-PCR markers, respectively, among the Citrullus Pis and watermelon cultivars, A large number of the EST-PCR markers were polymorphic between the Citrullus Pis and the watermelon cultivars, but significantly less polymorphic among the cultivars. Of the 108 EST-PCR markers associated with EST-SSRs, 103 exist in the Citrullus Pis and 64 in the cultivars. Of these 64 markers, 45 (70.3%) were polymorphic among the cultivars. Of the 142 EST-PCR markers associated with customary ESTs, 134 exist in the Citrullus Pis and 108 in the cultivars. Of these 108 markers, 86 (79.6%) were polymorphic among cultivars. The results in this study indicate that polymorphism exists in coding regions of genes expressed in fruits of watermelon cultivars. In total, 131 polymorphic EST-PCR markers (45 associated with EST-SSRs and 86 associated with customary ESTs) related to watermelon fruit genes were generated using the above data. These markers should be useful for DNA fingerprinting of cultivars and breeding lines, for assessing genetic relationships and for genetic mapping of watermelon.