普通油茶与小果油茶的叶绿体基因分子鉴定标记

Molecular identification of chloroplast gene markers of Camellia oleifera and C.meiocarpa

[目的]开发能够准确、便捷、快速鉴定普通油茶与小果油茶的叶绿体基因分子标记,为油茶遗传资源的挖掘与利用提供参考。[方法]基于高通量测序技术组装了普通油茶和小果油茶的叶绿体全基因组,通过比较基因组学技术寻找两者叶绿体全基因组的差异,并根据其差异来开发可用于鉴定普通油茶与小果油茶的分子标记。为验证标记的有效性,分别选取具有代表性的12个普通油茶居群和6个小果油茶居群的个体,提取叶绿体全基因组,进行PCR扩增和凝胶电泳检验。[结果]组装的普通油茶与小果油茶叶绿体基因组序列全长分别为156 977、156 539 bp,两者叶绿体全基因组具有高度的保守性,但小果油茶在atpH基因与atpI基因间的非编码区存在长度为452 bp的大片段缺失。所开发引物的PCR扩增结果表明,所有普通油茶样本均可扩增出609 bp的条带,而小果油茶扩增出227 bp的条带,条带差异显著且稳定。[结论]使用所开发的分子标记能够稳定、便捷、精确地鉴定出普通油茶与小果油茶,但是使用时应先确定所鉴定的物种为普通油茶和小果油茶中的一种。

【Objective】The development of chloroplast gene molecular markers for accurate,convenient and rapid identification of Camellia oleifera and C.meiocarpa will provide scientific and technological support for the efficient exploitation and utilization of C.oleifera genetic resources.【Method】The chloroplast genomes of C.oleifera and C.meiocarpa were assembled using high-throughput sequencing.Finding the difference of chloroplast genome between C.oleifera and C.meiocarpa by comparative genomics.According to the differences,the molecular markers for the identification of C.oleifera and C.meiocarpa were developed.In order to verify the validity of molecular markers,12 representative populations of C.oleifera and 6 populations of C.meiocarpa were selected,and the whole chloroplast genome were extracted for PCR amplification and gel electrophoresis.【Result】The chloroplast genome size of C.oleifera and C.meiocarpa were 156977 bp and 156539 bp respectively.And both chloroplast genomes were highly conserved.But at the atpH-atpI intergenic region,a 452 bp long fragment deletion was found in the chloroplast genome of C.meiocarpa being compared to that of C.oleifera.The PCR amplification results showed that bands of all C.oleifera samples about 609 bp,while bands of all C.meiocarpa samples about 227 bp.And the bands were significantly different and stable.【Conclusion】The molecular markers developed in this study can identify C.oleifera and C.meiocarpa stably,conveniently and accurately.However,it should be noted that the identified species is one of the C.oleifera and C.meiocarpa.