油棕种壳厚度控制基因SHELL的SNP分子标记开发

石 鹏<sup>1; 2</sup>; 夏 薇<sup>3</sup>; 肖 勇<sup>1; 2</sup>; 王 永<sup>1; 2</sup>; 曹红星<sup>1; 2</sup>; 李东霞<sup>1; 2</sup>; 雷新涛<sup>1; 2*</sup>

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*油棕种壳厚度控制基因SHELL的SNP分子标记开发*
石鹏^{1, 2}, 夏薇³, 肖勇^{1, 2}, 王永^{1, 2}, 曹红星^{1, 2}, 李东霞^{1, 2}, 雷新涛^{1, 2*}
1. 中国热带农业科学院椰子研究所, 海南文昌 571339;2. 海南省热带油料作物生物学重点实验室, 海南文昌 571339;3. 海南大学, 海口 570228

摘要:

油棕属棕榈科多年生木本油料作物,果实含油量高达50%,且单位面积产油量高,享有“世界油王”美誉。油棕果实由外果皮、中果皮、内果皮(种壳)、种子四个部分组成,产油部分主要是中果皮和种子,其中种壳厚度是影响果实含油量的重要因素。SHELL基因控制种壳厚度,是一类MADS-box同源基因,SHELL基因在厚壳种和无壳种中的变异主要是第一个外显子上的两个SNP位点。该研究根据两个SNP位点进行特异标记开发,根据已知的油棕SHELL基因的序列,设计了4对SNP引物。4对SNP引物以2个SNP位点设计,每个SNP位点设计2对SNP标记,并均在引物3'端第二位引入强错配碱基。以2份薄壳种油棕材料和2份厚壳种油棕材料DNA为模板,扩增筛选油棕SHELL基因SNP引物。通过PCR扩增发现,设计的SHELL基因特异SNP标记EgSh(N)-f/EgSh(SNP)-2r能够鉴别油棕厚壳种和薄壳种。再用24株油棕树进行特异性验证,发现该标记能较准确地判断油棕的厚薄壳。该研究结果表明SNP标记EgSh(N)-f/EgSh(SNP)-2r可用来进行油棕种质资源早期分子鉴定,为高产油棕品种选育提供了技术支撑。

关键词: 油棕, 种壳厚度, SHELL基因, 错配碱基, SNP标记

DOI：10.11931/guihaia.gxzw201702002

分类号:Q949.9, S59

文章编号:1000-3142(2018)02-0195-07

Fund project:海南省自然科学基金(20163145); 中国热带农业科学院基本科研业务费专项资金(17CXTD-13); 海南省重点研发计划(ZDYF2016214); 国家农业部“948”项目(2016-X40)[Supported by Natural Science Foundation of Hainan Province(20163145); Central Public-Interest Scientific Institution Foundamental Research Fund for Chinese Academy of Tropical Agricultural Sciences(17CXTD-13); Key Research and Development Program of Hainan Province(ZDYF2016214); “948” Program from Ministry of Agriculture of China(2016-X40)]。

SNP markers development of SHELL controlling shell thickness in oil palm(Elaesis guineensis)

SHI Peng^{1, 2}, XIA Wei³, XIAO Yong^{1, 2}, WANG Yong^{1, 2}, CAO Hongxing^{1, 2}, LI Dongxia^{1, 2}, LEI Xintao^{1, 2*}

1. Coconut Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wenchang 571339, Hainan, China;2. Hainan Key Biological Laboratory of Tropical Oil Crops, Wenchang 571339, Hainan, China;3. Hainan University, Haikou 570228, China

Abstract:

Oil palm(Elaeis guineensis )belongs to palmae perennial woody oil crop, known as “oil king of the world”, and its fruit oil content is up to 50%. Fruit of oil palm consists of exocarp, mesocarp, endocarp(shell)and seed, mesocap and seed is the resource of oil, and shell thicknees play an important role in fruit oil content. SHELL gene controls the shell thickness, which is a kind of MADS-box homologous gene. Moreover, variation of SHELL between dura and pisifera is mainly two SNP loci in the first exon. Specific markers were developed according to two SNP loci, in order to evaluate germplasm resources of oil palm early. Four SNP markers were designed according to SHELL gene sequence. Four pairs of SNP primers were designed with two SNP loci, which existed strong mismatch base in the second position from 3'-end respectively. Four pairs of SNP primers were amplified in two tenera palms and two dura palms to screen effective primers. PCR result showed that SNP marker EgSh(N)-f/EgSh(SNP)-2r was able to identify dura and tenera palms effectively. Verification experiment using 24 plants revealed that this marker could accurately determine the shell thickness of oil palm. In this study, results showed that SNP marker EgSh(N)-f/EgSh(SNP)-2r might be used for early molecular identification, which would provide technical support for high yield breeding of oil palm.

Key words: oil palm, shell thickness, SHELL gene, mismatch bases, SNP markers