花椰菜种子遗传纯度检测与品种鉴定的分子标记分析
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摘要
花椰菜(Brasica oleracea var.botrytis L.)是世界上重要的蔬菜作物之一,由于其营养价值高,适应性广,种植面积大,在我国蔬菜产业占有重要的地位。花椰菜是异花授粉作物,具有显著的杂种优势,目前市场上广泛利用的均为杂交一代的种子。杂种优势的充分表现与种子遗传纯度有着密切的联系,如果种子纯度太低就会给生产带来很大的影响。因而,建立准确、高效、快速地鉴定花椰菜品种与检测种子遗传纯度的方法是目前蔬菜生产中的一个重要课题。
本研究利用三种分子标记体系RAPD、ISSR和SSR对上海及周边地区广泛种植的银冠花椰菜种子遗传纯度进行检测。以银冠花椰菜F_1及父母本为材料,筛选多态性引物,结果发现两个RAPD引物(NAURP23和NAURP56),两个ISSR引物(NAUISR43和NAUISR75)及两对SSR引物(NAUSSR2和NAUSSRl4)能够同时在F_1代中产生父母本特异条带,这些特异条带可以用来进行检测杂交种种子遗传纯度。检测结果表明,210个单株中发现了3株为假杂种,分别为4号、178号、184号单株。六个引物扩增的结果都显示这三个单株扩增的图谱与其母本的扩增图谱很相似,都未产生父本特异标记,推测其为母本自交产生的假杂种。这一结果与田间形态学鉴定结果高度一致。因而,银冠花椰菜种子遗传纯度为98.57%。本研究表明,RAPD、ISSR、SSR这三种标记体系都能够有效地鉴定花椰菜种子遗传纯度,而且综合利用三种标记的检测结果比单一使用其中一种标记检测的结果更加准确可靠。
同时,本研究利用RAPD、ISSR、SRAP三种分子标记技术对全国广泛种植的30个花椰菜品种及1个青花菜、1个甘蓝品种进行了指纹图谱的分子标记分析,分别构建了三种分子标记数字化核酸指纹图谱。分析结果显示:32个品种间存在着较高的多态性,综合这两个引物的分析结果可以完全区分开32个供试品种。利用三种分子标记构建的指纹图谱进行了聚类分析,结果均显示甘蓝、青花菜与花椰菜品种间的亲缘关系较远。花椰菜不同品种间的亲缘关系与其分布的地理位置及其品种特性有很大的关系。研究结果表明RAPD、ISSR、SRAP三种分子标记技术均能有效地应用于花椰菜作物的品种鉴定及遗传多样性分析领域,为新品种的选育及新品种的保护提供有力的理论依据和可靠的技术保障。
Cauliflower(Brasica oleracea var.botrytis L.),one of the most popular vegetable crops,has high nutrition value and large production area,play an important role in the vegetable industry of China.It is a typical cross-pollinated crop exhibited prominent heterosis of quality,yield and disease resistance in F_1 hybrid progeny.Cauliflower F_1 hybrid seeds have been wildly used in the production.If the seed genetic purity was very low,it could have bad effect on the production.Therefore,it is necessary that quality control assessment of cauliflower hybrid seed is carried out before seed being released in the market.
In this study,cauliflower hybrid 'yin'guan' and its parents were studied for identification with three PCR based molecular markers,RAPD,ISSR and SSR.Two RAPD primers(NAURP23 and NAURP56),two ISSR primers(NAUISR43 and NAUISR75) and two SSR primers(NAUSSR2 and NAUSSR14),produced male and female parent-specific markers simultaneously in the hybrid,were found to be useful in determining the hybrid seed genetic purity.The results indicated that three of 210 'yin'guan' hybrid individuals appeared to be false hybrids(Nos.4,178,184).Three putative false hybrids confirmed with all six primers,exhibited similar band patterns to be female parent,suggesting that they could be derived from selfing of the female parent.It was high accordance with the result from field grow-out trails.Therefore,the genetic purity of hybrid 'yin'guan' seed lots was 98.57%.The present study showed that all three molecular marker systems were useful tools for testing the cauliflower hybrid seed genetic purity,and using the combination of RAPD,ISSR and SSR markers were more reliable and efficient than using the three markers in isolation.
In this study,three molecular marker systems RAPD,ISSR and SRAP,were employed to identify cultivar and the relationship of 29 cauliflower cultivars and one broccoli cultivar and one cabbage cultivar,and construct the fingerprint of these cultivars,respectively.The result showed that 32 cultivars existed higher polymorphism.32 cultivars studied were identified with two primers.Cluster analysis and relatively genetic similarity coefficient indicated that the relationships of cabbage with cauliflower and broccoli and cauliflower were larger.The genetic relationship of these cauliflower cultivars was tightly associated with its origin and its characters.The study showed RAPD,ISSR and SRAP marker were useful of identify cauliflower cultivars and analysis genetic diversity,and this could be useful for the new cultivar breeding and the new cultivar protection.
本研究利用三种分子标记体系RAPD、ISSR和SSR对上海及周边地区广泛种植的银冠花椰菜种子遗传纯度进行检测。以银冠花椰菜F_1及父母本为材料,筛选多态性引物,结果发现两个RAPD引物(NAURP23和NAURP56),两个ISSR引物(NAUISR43和NAUISR75)及两对SSR引物(NAUSSR2和NAUSSRl4)能够同时在F_1代中产生父母本特异条带,这些特异条带可以用来进行检测杂交种种子遗传纯度。检测结果表明,210个单株中发现了3株为假杂种,分别为4号、178号、184号单株。六个引物扩增的结果都显示这三个单株扩增的图谱与其母本的扩增图谱很相似,都未产生父本特异标记,推测其为母本自交产生的假杂种。这一结果与田间形态学鉴定结果高度一致。因而,银冠花椰菜种子遗传纯度为98.57%。本研究表明,RAPD、ISSR、SSR这三种标记体系都能够有效地鉴定花椰菜种子遗传纯度,而且综合利用三种标记的检测结果比单一使用其中一种标记检测的结果更加准确可靠。
同时,本研究利用RAPD、ISSR、SRAP三种分子标记技术对全国广泛种植的30个花椰菜品种及1个青花菜、1个甘蓝品种进行了指纹图谱的分子标记分析,分别构建了三种分子标记数字化核酸指纹图谱。分析结果显示:32个品种间存在着较高的多态性,综合这两个引物的分析结果可以完全区分开32个供试品种。利用三种分子标记构建的指纹图谱进行了聚类分析,结果均显示甘蓝、青花菜与花椰菜品种间的亲缘关系较远。花椰菜不同品种间的亲缘关系与其分布的地理位置及其品种特性有很大的关系。研究结果表明RAPD、ISSR、SRAP三种分子标记技术均能有效地应用于花椰菜作物的品种鉴定及遗传多样性分析领域,为新品种的选育及新品种的保护提供有力的理论依据和可靠的技术保障。
Cauliflower(Brasica oleracea var.botrytis L.),one of the most popular vegetable crops,has high nutrition value and large production area,play an important role in the vegetable industry of China.It is a typical cross-pollinated crop exhibited prominent heterosis of quality,yield and disease resistance in F_1 hybrid progeny.Cauliflower F_1 hybrid seeds have been wildly used in the production.If the seed genetic purity was very low,it could have bad effect on the production.Therefore,it is necessary that quality control assessment of cauliflower hybrid seed is carried out before seed being released in the market.
In this study,cauliflower hybrid 'yin'guan' and its parents were studied for identification with three PCR based molecular markers,RAPD,ISSR and SSR.Two RAPD primers(NAURP23 and NAURP56),two ISSR primers(NAUISR43 and NAUISR75) and two SSR primers(NAUSSR2 and NAUSSR14),produced male and female parent-specific markers simultaneously in the hybrid,were found to be useful in determining the hybrid seed genetic purity.The results indicated that three of 210 'yin'guan' hybrid individuals appeared to be false hybrids(Nos.4,178,184).Three putative false hybrids confirmed with all six primers,exhibited similar band patterns to be female parent,suggesting that they could be derived from selfing of the female parent.It was high accordance with the result from field grow-out trails.Therefore,the genetic purity of hybrid 'yin'guan' seed lots was 98.57%.The present study showed that all three molecular marker systems were useful tools for testing the cauliflower hybrid seed genetic purity,and using the combination of RAPD,ISSR and SSR markers were more reliable and efficient than using the three markers in isolation.
In this study,three molecular marker systems RAPD,ISSR and SRAP,were employed to identify cultivar and the relationship of 29 cauliflower cultivars and one broccoli cultivar and one cabbage cultivar,and construct the fingerprint of these cultivars,respectively.The result showed that 32 cultivars existed higher polymorphism.32 cultivars studied were identified with two primers.Cluster analysis and relatively genetic similarity coefficient indicated that the relationships of cabbage with cauliflower and broccoli and cauliflower were larger.The genetic relationship of these cauliflower cultivars was tightly associated with its origin and its characters.The study showed RAPD,ISSR and SRAP marker were useful of identify cauliflower cultivars and analysis genetic diversity,and this could be useful for the new cultivar breeding and the new cultivar protection.
引文
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