中国甘薯（Ipomoea batatas）品种资源遗传多样性及分子鉴定研究
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摘要
甘薯(Ipomoea batatas(L.)Lam.)属于番薯属(Ipomoea L.)，旋花科(Convolvulaceae)，是蔓生性多年生或一年生草本。甘薯原产南美洲，现在广泛分布于世界热带、亚热带和温带地区。甘薯利用根、茎进行无性繁殖，四季可种，不择土壤，高产稳产，适应性广，再生能力强，根、茎、叶均能食用，营养丰富。甘薯是重要的粮食作物，在我国粮食作物中居第四位。但甘薯在遗传上的高度杂合性和种间、种内广泛存在的杂交不亲和性，严重制约了甘薯的生产和发展，培育高产、优质、抗逆性强、无病毒、耐储藏的优良甘薯品种，仍然是十分迫切的事。
本研究采用随机扩增多态性DNA(Random Amplified Polymorphic DNA，简称RAPD)分子标记技术，首次对采自山东农业科学院和徐州甘薯研究中心的50个国内外甘薯品种进行遗传多样性和分子鉴定研究，分析这些品种之间的亲缘关系，进行种质资源分析和品种鉴定，旨在为引种鉴定、选育和杂交育种提供一定的理论依据。
本文从386个随机引物中筛选出24个扩增强、重复性好、带型清晰、不同品种间多态性高的引物进行扩增。结果表明：24个有效引物共产生157条DNA片段，大小分布在100bp-3500bp之间，其中91条具有遗传多态性，约占总数的58％，平均每个引物扩增的DNA带数为6.54条。将任一扩增带看作一个性状，按带的有无列出二元数据矩阵，利用NTSYS-pc软件计算出材料间的Jaccard遗传相似性系数。50个甘薯品种基因型的遗传相似性分析表明，其相似性系数分布在0.496-0.930之间，这表明这些品种基因型之间的遗传多态性较为丰富。通过非加权算术平均聚类(UPGMA)的方法，绘制出这些甘薯品种基因型之间的遗传关系树状图。根据相似性系数和聚类结果，在相似性系数0.70处，这些甘薯品种明显地聚为4类：A类、B类、C类、D类。在相似性系数0.75处，这些品种可分为14个组。各组内的相似性系数都大于0.73，表明组内亲缘关系较近，但也存在一定的遗传差异。
A类分为A1、A2两个组，B类分为B1、B2两个组，C类包括C1、C2、C3、C4四个组，D类包括D1、D2、D3、D4、D5、D6六个组。A1组包括：1 470042、26南放、11胜南，A2组包括：36烟薯6号。B1组包括：21济薯2号、32鲁薯2号、34华北52-45、37青农8号、47徐236，B2组包括：28南京92、35济薯5号、33洛红3号、45许薯4号。C1组包括：4 460188、19栗子香、23徐941、40苏薯1号，C2组包括：17徐1-2、27一窝红、30新种花、31浙薯1号、43宁远30日早、49宛薯596，C3组包括：5 460103、6 460027、14济南红、25南京51、39禺北白、50陕薯1号，C4组包括：15烟薯8号、29安薯07、44胜利百号、48丰收黄、42徐61。D1组包括：2 460363、24美国红、18徐薯18，
DZ组包括：9台农 9号、12板栗黄、22辽薯 224，D3：组包括：3 460021、8
460097、20南瑞菩、7 460379．10苏薯2号、46高系14，D4组包括：16郑
颖红、41 安薯1号，DS组只包括38烟薯2号，D6组只包括13香薯。
从供试的这些品种来看，国外引进品种属于几个不同的组。这表明来源不同
的品种之间亲缘关系较远，遗传差异较大。从所有的供试材料来看，各个材料之
间的遗传距离都大于零值，又都能聚在一起，这表明这些材料之间有相同的遗传
背景，但相互间又存在一定的差异。
通过随机扩增，部分品种产生了特异带，部分品种不具有某些特异带。据此，
可以对其进行品种鉴定。有的品种用一个引物进行RAPD扩增难以区别，必须采用
几个引物。从实验结果来看，利用多个引物根据 gi条多态带在 50个甘薯品种间
的表现情况是可以将这些品种分开的。
Ipomoea batatas (L.) Lam., belonging to Ipomoea of Convolvulaceae, is a perennial or annual trailing herb. It initially originated from South America, and now it is widely distributed in tropical and subtropical and template regions of the world. Asexul reproduction can be carried out by root and stem. It can be planted in the four seasons and it doesn't choose soil. It has high and stable yields, wide adaptability and strong regeneration. It is rich in nutrition and its roots, stems and leaves can be edible, so it is an important crop and it is in the 4th place of crop in China. But high heterozygosity on heredity and interspecific and intraspecific hybridization incompatibility, which wildly exsisted, gravely limit its production and development. It is still urgent to cultivate fine sweetpotato cultivars of high yield, high quality, strong adversity, nonvirus and easy to store.
In this paper, the technique of RAPD molecular markers was firstly used to study genetic diversity and molecular identification of 50 native and abroad sweetpotato cultivars from Shandong Agricultural Acadamy of Science and Xuzhou Sweetpotato Research Center, and to analyse their genetic relationship and germplasm resourses in order to offer theoretical foundation for identification, selection, and hybridization breeding.
In order to select a set of suitable primers, 3 cultivars were initially surveyed with 386 10-base primers.24 were chosen for they showed strong, reproducible amplification and distinct polymorphisms from 386 10-base arbitrary primers, and a total of 157 DNA fragments were amplified from 100bp-3500bp, among which 91 were polymorphic, and which accounted for 58%. The average number of DNA band produced by each primer was 6.54. Any amplified band was reguarded as a character, and the binary data matrix was set out according as the bands existed or not. The Jaccard coefficient was worked out by using NTSYS-pc software. The result of genetic similarity analysis for 50 sweetpotato cultivar genotypes showed that Jaccard coefficient ranged from 0.496 to 0.930, suggesting that there was a close genetic relationship among them and a rich genetic polymorphism among the sweetpotato cultivars due to the Jaccard coefficient. A DNA molecular dendrogram was established for 50 sweetpotato cultivars genotypes based on UPGMA (unweighed pair-group method with arithmetic mean) cluster analysis of the 157 DNA bands amplified by 24 primers. According to the coefficient and the cluster result, when the coefficient is 0.70, these cultivar genotypes were divided into 4 groups: Group A, Group B, Group C, Group D. when the coefficient is
0.75, these cultivar genotypes were divided into 14 sections. The coefficient in each section all surpasses 0.75, which suggests that the relationship among sections was fairly near, but some genetic difference exists.
Group A was divided into section Al and A2. Group B was divided into section Bl and B2. Group C was divided into section Cl, C2, C3, and C4. Group D was divided into section Dl, D2, D3, D4, D5, and D6. Al included 3 genotypes, viz., 470042, Nanfang, Shengnan. A2 only included 1 genotypes, viz., No.6 Yanshu. Bl included 5 genotypes, viz., No.2 Jishu, No.2 Lushu, Huabei 52-45, No.8 Qingnong, Xu 236. B2 included 4 genotypes, viz., Nanjing 92, No.5 Jishu, No.3 Luohong, No.4 Xushu. Cl included 4 genotypes, viz., 460188, Lizixiang, Xu 941, NO.l sushu. C2 included 6 genotypes, viz., Xu 1-2, Yiwohong, Xinzhonghua, No.l Zheshu, Ningyuansanshirizao, Wanshu 596. C3 included 6 genotypes, viz., 460103, 460027, Ji'nanhong, Nanjing 51, Yubeibai, No.l Shanshu. C4 included 5 genotypes, viz., No.8 Yanshu, Anshu 07, Shenglibaihao, Fengshouhuang, Xu 61. Dl included 3 genotypes, viz., 460363, Meiguohong, Xushu 18. D2 included 3 genotypes, viz., No.9 Tainong, Banlihuang, Liaoshu 224. D3 included 6 genotypes, viz., 4600^1, 460097, Nanruishao, 46039, No.2 Sushu, Gaoxi 14. D4 included 2 genotypes, viz.,Zaengyinghong, No.l Anshu,. D5 included 1 genotypes, viz., NO.2 Yanshu. D4 included 1 genotypes, viz., Xiangshu.
本研究采用随机扩增多态性DNA(Random Amplified Polymorphic DNA，简称RAPD)分子标记技术，首次对采自山东农业科学院和徐州甘薯研究中心的50个国内外甘薯品种进行遗传多样性和分子鉴定研究，分析这些品种之间的亲缘关系，进行种质资源分析和品种鉴定，旨在为引种鉴定、选育和杂交育种提供一定的理论依据。
本文从386个随机引物中筛选出24个扩增强、重复性好、带型清晰、不同品种间多态性高的引物进行扩增。结果表明：24个有效引物共产生157条DNA片段，大小分布在100bp-3500bp之间，其中91条具有遗传多态性，约占总数的58％，平均每个引物扩增的DNA带数为6.54条。将任一扩增带看作一个性状，按带的有无列出二元数据矩阵，利用NTSYS-pc软件计算出材料间的Jaccard遗传相似性系数。50个甘薯品种基因型的遗传相似性分析表明，其相似性系数分布在0.496-0.930之间，这表明这些品种基因型之间的遗传多态性较为丰富。通过非加权算术平均聚类(UPGMA)的方法，绘制出这些甘薯品种基因型之间的遗传关系树状图。根据相似性系数和聚类结果，在相似性系数0.70处，这些甘薯品种明显地聚为4类：A类、B类、C类、D类。在相似性系数0.75处，这些品种可分为14个组。各组内的相似性系数都大于0.73，表明组内亲缘关系较近，但也存在一定的遗传差异。
A类分为A1、A2两个组，B类分为B1、B2两个组，C类包括C1、C2、C3、C4四个组，D类包括D1、D2、D3、D4、D5、D6六个组。A1组包括：1 470042、26南放、11胜南，A2组包括：36烟薯6号。B1组包括：21济薯2号、32鲁薯2号、34华北52-45、37青农8号、47徐236，B2组包括：28南京92、35济薯5号、33洛红3号、45许薯4号。C1组包括：4 460188、19栗子香、23徐941、40苏薯1号，C2组包括：17徐1-2、27一窝红、30新种花、31浙薯1号、43宁远30日早、49宛薯596，C3组包括：5 460103、6 460027、14济南红、25南京51、39禺北白、50陕薯1号，C4组包括：15烟薯8号、29安薯07、44胜利百号、48丰收黄、42徐61。D1组包括：2 460363、24美国红、18徐薯18，
DZ组包括：9台农 9号、12板栗黄、22辽薯 224，D3：组包括：3 460021、8
460097、20南瑞菩、7 460379．10苏薯2号、46高系14，D4组包括：16郑
颖红、41 安薯1号，DS组只包括38烟薯2号，D6组只包括13香薯。
从供试的这些品种来看，国外引进品种属于几个不同的组。这表明来源不同
的品种之间亲缘关系较远，遗传差异较大。从所有的供试材料来看，各个材料之
间的遗传距离都大于零值，又都能聚在一起，这表明这些材料之间有相同的遗传
背景，但相互间又存在一定的差异。
通过随机扩增，部分品种产生了特异带，部分品种不具有某些特异带。据此，
可以对其进行品种鉴定。有的品种用一个引物进行RAPD扩增难以区别，必须采用
几个引物。从实验结果来看，利用多个引物根据 gi条多态带在 50个甘薯品种间
的表现情况是可以将这些品种分开的。
Ipomoea batatas (L.) Lam., belonging to Ipomoea of Convolvulaceae, is a perennial or annual trailing herb. It initially originated from South America, and now it is widely distributed in tropical and subtropical and template regions of the world. Asexul reproduction can be carried out by root and stem. It can be planted in the four seasons and it doesn't choose soil. It has high and stable yields, wide adaptability and strong regeneration. It is rich in nutrition and its roots, stems and leaves can be edible, so it is an important crop and it is in the 4th place of crop in China. But high heterozygosity on heredity and interspecific and intraspecific hybridization incompatibility, which wildly exsisted, gravely limit its production and development. It is still urgent to cultivate fine sweetpotato cultivars of high yield, high quality, strong adversity, nonvirus and easy to store.
In this paper, the technique of RAPD molecular markers was firstly used to study genetic diversity and molecular identification of 50 native and abroad sweetpotato cultivars from Shandong Agricultural Acadamy of Science and Xuzhou Sweetpotato Research Center, and to analyse their genetic relationship and germplasm resourses in order to offer theoretical foundation for identification, selection, and hybridization breeding.
In order to select a set of suitable primers, 3 cultivars were initially surveyed with 386 10-base primers.24 were chosen for they showed strong, reproducible amplification and distinct polymorphisms from 386 10-base arbitrary primers, and a total of 157 DNA fragments were amplified from 100bp-3500bp, among which 91 were polymorphic, and which accounted for 58%. The average number of DNA band produced by each primer was 6.54. Any amplified band was reguarded as a character, and the binary data matrix was set out according as the bands existed or not. The Jaccard coefficient was worked out by using NTSYS-pc software. The result of genetic similarity analysis for 50 sweetpotato cultivar genotypes showed that Jaccard coefficient ranged from 0.496 to 0.930, suggesting that there was a close genetic relationship among them and a rich genetic polymorphism among the sweetpotato cultivars due to the Jaccard coefficient. A DNA molecular dendrogram was established for 50 sweetpotato cultivars genotypes based on UPGMA (unweighed pair-group method with arithmetic mean) cluster analysis of the 157 DNA bands amplified by 24 primers. According to the coefficient and the cluster result, when the coefficient is 0.70, these cultivar genotypes were divided into 4 groups: Group A, Group B, Group C, Group D. when the coefficient is
0.75, these cultivar genotypes were divided into 14 sections. The coefficient in each section all surpasses 0.75, which suggests that the relationship among sections was fairly near, but some genetic difference exists.
Group A was divided into section Al and A2. Group B was divided into section Bl and B2. Group C was divided into section Cl, C2, C3, and C4. Group D was divided into section Dl, D2, D3, D4, D5, and D6. Al included 3 genotypes, viz., 470042, Nanfang, Shengnan. A2 only included 1 genotypes, viz., No.6 Yanshu. Bl included 5 genotypes, viz., No.2 Jishu, No.2 Lushu, Huabei 52-45, No.8 Qingnong, Xu 236. B2 included 4 genotypes, viz., Nanjing 92, No.5 Jishu, No.3 Luohong, No.4 Xushu. Cl included 4 genotypes, viz., 460188, Lizixiang, Xu 941, NO.l sushu. C2 included 6 genotypes, viz., Xu 1-2, Yiwohong, Xinzhonghua, No.l Zheshu, Ningyuansanshirizao, Wanshu 596. C3 included 6 genotypes, viz., 460103, 460027, Ji'nanhong, Nanjing 51, Yubeibai, No.l Shanshu. C4 included 5 genotypes, viz., No.8 Yanshu, Anshu 07, Shenglibaihao, Fengshouhuang, Xu 61. Dl included 3 genotypes, viz., 460363, Meiguohong, Xushu 18. D2 included 3 genotypes, viz., No.9 Tainong, Banlihuang, Liaoshu 224. D3 included 6 genotypes, viz., 4600^1, 460097, Nanruishao, 46039, No.2 Sushu, Gaoxi 14. D4 included 2 genotypes, viz.,Zaengyinghong, No.l Anshu,. D5 included 1 genotypes, viz., NO.2 Yanshu. D4 included 1 genotypes, viz., Xiangshu.
引文
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