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2025年8月20日 星期三
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大白菜硫代葡萄糖甙合成基因SSR标记的开发
巩红影1,罗双霞2,顾爱侠2,王彦华2,赵建军1,申书兴1
1. 河北农业大学园艺学院
2.
Development of SSR Markers Linked to Glucosinolate Synthesis Genes in Chinese cabbage(Brassica rapa ssp. pekinensis)
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摘要 硫代葡萄糖甙(glucosinolates, GS)是一种含氮、硫的重要植物次生代谢产物,是十字花科蔬菜风味的主要来源。本研究根据拟南芥(Arabidopsis thaliana)和大白菜(Brassica rapa ssp. pekinensis)基因组序列信息,比对GS生物合成相关基因的序列信息,发现大白菜中与GS含量相关的候选基因102个,分布于10条染色体上。拟南芥GS合成基因在大白菜中的同源基因个数分别为:零拷贝基因8个、单拷贝基因13个、 2拷贝基因17个、2个以上多拷贝基因14个。通过FASTPCR6.0软件分析目标基因上下游序列各5 kb,共开发出237个简单重复序列(SSR)位点,16个基因没有发现SSR位点,72个基因存在1~4个SSR位点,5个基因存在5个SSR位点,4个基因存在6个SSR位点,4个基因存在7个SSR位点,1个基因存在8个SSR位点。所开发的SSR位点中,共发现4种重复类型:单核苷酸重复最多,共122个,占SSR总数的51.4%;其次是二核苷酸重复类型,共48个,占SSR总数的20.3%;三核苷酸重复类型最少,共7个,占SSR总数的3.0%,另外60个SSR无明显的重复类型。具有SSR位点的86个基因中,77个基因可以设计SSR特异引物,其中49对特异引物在供试的75份大白菜高代自交系中得到有效扩增。其中16对引物具有多态性,占全部引物的20.8%; 18对引物无多态性,占全部引物的23.4%;15对引物杂带多,占全部引物的19.5%;另外28对引物无扩增产物,占全部引物的36.4%。最终11对获得了清晰多态性扩增产物,共检测出26个等位基因。本研究为分子标记辅助选择培育高有益GS成分的大白菜新品种提供了基础资料,进而加快大白菜营养品质育种的进程。
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巩红影
罗双霞
顾爱侠
王彦华
赵建军
申书兴
关键词 大白菜硫代葡萄糖甙(GS)简单重复序列(SSR)分子标记    
Abstract:Glucosinolates(GS), as one of important plant secondary metabolites containing nitrogen and sulfur, is one major flavor source of cruciferous vegetables. In this study we developed SSRs linked to GS content in Chinese cabbage(Brassica rapa ssp. pekinensis), aiming to exploit the variation in gene specific SSR loci or alleles crossed over Chinese cabbage accessions. According to the genomic sequence information of Arabidopsis thaliana and Chinese Cabbage, gene sequences related to GS biosynthesis were alignment and analysis, finding 102 candidate genes associated with GS content distributed in 10 chromosomes. Compared with Arabidopsis, the homologous genes for GS biosynthensis were detected in Chinese cabbage, with 8 zero copy, 13 single copies, 17 double copies, 14 more than 2 copies. By analyzing sequences of candidate genes and their up/down franking 5 kb nucleotides with software FASTPCR 6.0, 237 SSRs were developed in 102 genes, 16 of them had no SSRs, 72 of them had 1~4 SSRs, 5 of them had 5 SSRs, 4 of them had 6 SSRs, 4 of them had 7 SSRs, and 1 of them had 8 SSRs. Within all detected SSRs, four replicated types were found. Most of types were single nucleotide replication, with number of 122 and 51.4% of total SSRs; forty eight replication types of double nucleotide were found, with 20.3% of total SSRs; the most less types were three nucleotide replications, with only 7 SSRs and with 3.0%; the rest 60 replicated types had no clear replications. Seventy seven SSR specific primers linked to GS synthesis genes could be designed from 86 genes with SSR locus. Forty nine SSR primers could amplify available and clear bands in 75 inbred lines of Chinese cabbage, 16 of them (20.8%) were polymorphic, 18 of them (23.4%) had no polymorphism, 15 of them (19.5%) had more bands, 28 of them (36.4%) had no amplification products. Finally 11 pairs of primer were amplified in all accessions and produced 26 SSR alleles. The results can offer basic materials used for molecular marker assistant in breeding new varieties with high valuable GS compositions, and accelerate the process of breeding in nutritional quality in Chinese cabbage.
Key wordsChinese cabbage    Glucosinolates(GS)    Simple sequence repeat(SSR)    Molecular marker
收稿日期: 2014-03-24     
通讯作者: 赵建军   
引用本文:   
巩红影1,罗双霞2,顾爱侠2,王彦华2,赵建军1,申书兴1. 大白菜硫代葡萄糖甙合成基因SSR标记的开发[J]. , 2014, 22(10): 1232-1241.
, , , , ,. Development of SSR Markers Linked to Glucosinolate Synthesis Genes in Chinese cabbage(Brassica rapa ssp. pekinensis). , 2014, 22(10): 1232-1241.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2014/V22/I10/1232
 
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