亚洲杂草稻红色果皮分子鉴定及其Rc-bHLH序列多态性分析  

Molecular Identification of Red Pericarp and Variation Analysis of Rc-b HLH Sequence in Asian Weedy Rice

作　　者：张树林[1,2] 李娟[1] 董陈文华 熊海波[1] 张利东[1] 朱骞[1] 张小玲[1] 伍腾飞[1] 李东宣[1] 陈丽娟[1] 

机构地区：[1]云南农业大学稻作研究所,昆明650201 [2]安阳工学院生物与食品工程学院,安阳455000

出　　处：《分子植物育种》2015年第1期51-60,共10页Molecular Plant Breeding

基　　金：国家自然科学基金项目(30860057;31260257);云南省水稻产业技术体系(A3006516);云南省应用基础研究(2013FD018)共同资助

摘　　要：多数杂草稻的果皮是红色,该特征可用于稻田中杂草稻的早期的初步鉴定,也可用于水稻果皮Rc/rc基因的起源进化研究。本研究利用红色果皮Rc基因第六外显子的In Del标记RID14对来自亚洲不同稻作区的199份杂草稻（O.sativa f.spontanea）、24份栽培稻（O.sativa）和9份野生稻（O.rufipogon）进行基因型鉴定并与表型比对;同时对9份代表性杂草稻Rc-b HLH位点的DNA序列进行测序,并将其与下载的29份栽培稻（籼,粳）和14份野生稻的对应序列进行比对分析。研究结果表明：（1）亚洲杂草稻的果皮颜色的表型多样,多数（84.4%）为红色或褐色（由浅至深）、少数（15.6%）为白色,其基因型与表型鉴定结果高度一致;杂草稻红色果皮Rc基因第六外显子均为无14 bp缺失的类型,仅有一份中国东北杂草稻除外（为14 bp缺失型）,揭示用RID14标记可以早期快速的鉴定稻田中红色果皮杂草稻;（2）测试的9份杂草稻其Rc-b HLH位点的DNA序列均较为保守,其中出现4个突变位点;（3）在Rc-b HLH部分区域（1-115 bp）,红色果皮的籼型栽培稻、杂草稻和野生稻的DNA序列相同,但红色果皮粳型栽培稻与白色果皮籼型栽培稻和杂草稻在106 bp处存在一个A/G替换;（4）通过对比野生稻和栽培稻,杂草稻在Rc-b HLH位点核苷酸多态性多态性最高,但是野生稻在该位点Tajima＇D检验值呈显著,说明Rc-b HLH位点在野生稻中有选择性;通过聚类分析发现分为2个类群,类群1由白色果皮的杂草稻、籼型栽培稻和红/白果皮的粳型栽培稻组成;类群2由红色果皮的栽培稻（籼型）、杂草稻和野生稻组成。该结果暗示：在粳稻区,白色果皮的亚洲杂草稻可能由栽培稻退化返祖而形成的;在籼稻区,红色果皮的亚洲杂草稻可能经由栽培稻及野生稻杂交（如籼籼交或栽野交）演化而来。Many of weedy rice accessions （O. sativa f. spontanea） have a feature of red pericarp, which can be used to identify the weedy rice in the rice field at the early stage for its control as well as for understanding the evolution or origin of Rc/rc gene in rice （O. sativa）. We analyzed the genotypes of 199 weedy rice accessions, 24 cultivated rice varieties, and 9 wild rice accessions from different rice cropping areas of Asia using InDel molecular markers RID14 in 6 exon ofRc/rc gene to compare with their phenotypes; DNA sequences at Rc-bHLH locus of 9 weedy rice accessions were isolated through multiple clone method and then comprehensively analyzed with locus sequence from 29 cultivated rice （indica and japonica） and 14 wild rice accessions that downloaded fromthe NCBI database. The result showed that： （1） Pericarp color of Asian weedy rice has the various phenotypes with 84.4% as a red or brown （from light to dark） and 15.6% as white, which is highly consistent with the results from genotype identification; genotypes of the red pericarp weedy rice accessions did not have 14 bp deletion in the 6 exon ofRc/rc gene except an accession from Northeast in China; （2） 9 DNA sequences of weedy rice accessions at the Rc-bHLH locus were highly conserved except 4 mutation sites; （3） Red pericarp of weed rice, indica cultivars and wild rice had the same sequence from 1 bp to 115 bp at the Rc-bHLH locus, but there was A/G substitution at 106 bp between red pericarp japonica cultivars and white pericarp indica cultivars and white pericarp weedy rice accessions; （4） Asian weedy rice has the high genetic diversity at the Rc-bHLH locus by comparing with wild and cultivated rice; but the statistical Tajima＇D significance was found in wild rice population, which indicated the Rc-bHLH locus in wild rice should be selected; all entries were divided into two groups by cluster analysis, the first group included white pericarp weedy rice, white pericarp indica cultivars and japonica cultivars with wh

关 键 词：杂草稻 栽培稻 野生稻 红色果皮Rc/rc基因 起源 演化 

分 类 号：S511[农业科学—作物学]