江苏农业学报
江囌農業學報
강소농업학보
JIANGSU JOURNAL OF AGRICULTURAL SCIENCES
2014年
5期
997-1002
,共6页
戚维聪%程计华%黄邦全%李坦%林峰
慼維聰%程計華%黃邦全%李坦%林峰
척유총%정계화%황방전%리탄%림봉
海甘蓝%RNA-Seq%EST-SSR%分子标记
海甘藍%RNA-Seq%EST-SSR%分子標記
해감람%RNA-Seq%EST-SSR%분자표기
Crambe abyssinica%RNA-Seq%EST-SSR%molecular marker
利用油料作物海甘蓝(十字花科)发育时期种子的RNA-Seq测序数据组装获得186778条cDNA重叠群( Contigs)序列,通过MISA 和Primer 3程序设计了6639个EST-SSR分子标记。在这些标记中,除了单核苷酸重复(45%)外,三核苷酸重复的 SSR 是最常见的碱基重复类型(29%),其次是双核苷酸型(10%)、五核苷酸型(7%)、六核苷酸型(5%)和四核苷酸型(2%)型。采用电子定位的方法将1206个EST-SSR标记定位到近缘种白菜( Brassica napa)的基因组上。依据引物在白菜基因组中的分布,挑选了20条EST-SSR引物在海甘蓝中进行PCR验证,其结果显示所有引物均能够扩增出符合预期大小的PCR片段。这些新开发的EST-SSR引物可以作为功能标记应用于海甘蓝的分类鉴定、遗传图谱构建、种质资源鉴定以及分子标记辅助育种工作中。
利用油料作物海甘藍(十字花科)髮育時期種子的RNA-Seq測序數據組裝穫得186778條cDNA重疊群( Contigs)序列,通過MISA 和Primer 3程序設計瞭6639箇EST-SSR分子標記。在這些標記中,除瞭單覈苷痠重複(45%)外,三覈苷痠重複的 SSR 是最常見的堿基重複類型(29%),其次是雙覈苷痠型(10%)、五覈苷痠型(7%)、六覈苷痠型(5%)和四覈苷痠型(2%)型。採用電子定位的方法將1206箇EST-SSR標記定位到近緣種白菜( Brassica napa)的基因組上。依據引物在白菜基因組中的分佈,挑選瞭20條EST-SSR引物在海甘藍中進行PCR驗證,其結果顯示所有引物均能夠擴增齣符閤預期大小的PCR片段。這些新開髮的EST-SSR引物可以作為功能標記應用于海甘藍的分類鑒定、遺傳圖譜構建、種質資源鑒定以及分子標記輔助育種工作中。
이용유료작물해감람(십자화과)발육시기충자적RNA-Seq측서수거조장획득186778조cDNA중첩군( Contigs)서렬,통과MISA 화Primer 3정서설계료6639개EST-SSR분자표기。재저사표기중,제료단핵감산중복(45%)외,삼핵감산중복적 SSR 시최상견적감기중복류형(29%),기차시쌍핵감산형(10%)、오핵감산형(7%)、륙핵감산형(5%)화사핵감산형(2%)형。채용전자정위적방법장1206개EST-SSR표기정위도근연충백채( Brassica napa)적기인조상。의거인물재백채기인조중적분포,도선료20조EST-SSR인물재해감람중진행PCR험증,기결과현시소유인물균능구확증출부합예기대소적PCR편단。저사신개발적EST-SSR인물가이작위공능표기응용우해감람적분류감정、유전도보구건、충질자원감정이급분자표기보조육충공작중。
Crambe abyssinica of Brassicaceae family is a new crop and cultivated for its erucic concentration as high as 60%. Although as a high-value commercial crop, the germplasm collection and breeding achievements of C. abyssinica have lagged behind other crops due to its limited molecular markers. In this study, 6 639 EST-SSR primers according to 186 778 cDNA contigs derived from 86 224 256 RNA-Seq tags of the developing seeds were designed with the aid of MISA and Primer 3 programs. In addition to mononucleotide repeats ( 45%) , trinucleotides SSR is the dominant repeat type (29%) followed by 10% dinucleotide, 7% pentanucleotide, 5% hexanucleotide, and 2% tetranucleotide. The 1 206 EST-SSR markers were in silico mapped onto the Brassica napa genome. Based on their distribution on Brassic napa ge-nome, 20 primers were selected and validated in Crambe abyssinica. All the primers successfully amplified the expected PCR band. The newly developed EST-SSR primers, so-called functional markers, will be powerful resource for further applications such as taxonomy, linkage map con-struction and molecular assisted breeding.