| 利用ISSR与SRAP分子标记分析金线莲种质资源遗传多样性 |
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| 引用本文： | 黄锦春,万思琦,陈扬,李丽红,张自力,朱建军,吴梅,邢丙聪,邵清松,陆晨飞.利用ISSR与SRAP分子标记分析金线莲种质资源遗传多样性[J].浙江农林大学学报,2023,40(1):22-29. |
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| 作者姓名： | 黄锦春 万思琦 陈扬 李丽红 张自力 朱建军 吴梅 邢丙聪 邵清松 陆晨飞 |
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| 作者单位： | 1.浙江农林大学 浙江省特色中药资源保护与创新利用重点实验室，浙江 杭州 3113002.温州科技职业学院 温州市园艺植物育种重点实验室，浙江 温州 3250063.金华市农业科学研究院，浙江 金华 321000 |
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| 基金项目： | 国家自然科学基金资助项目(82173916)；国家级大学生创新创业训练计划项目(202210341058)；浙江农林大学学生科研训练项目(S202210341017) |
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| 摘 要： | 目的 研究浙江与福建等地引种、杂交与野生的金线莲Anoectochilus roxburghii样品间遗传多样性和亲缘关系，为金线莲种质资源鉴定及优良新品种(系)选育提供科学依据。 方法 以48份新鲜金线莲叶片为材料，分别采用简单重复序列扩增多态性(ISSR)与序列相关扩增多态性(SRAP)分子标记技术，各挑选11条(对)多态性好、扩增条带清晰的引物。经琼脂糖凝胶电泳成像后，统计其扩增条带数，运用NTSYS-PC 2.1和POPGENE 32软件进行非加权组平均法(UPGMA)聚类分析。 结果 ISSR分子标记技术共扩增出86条条带，其中多态性条带84条，多态位点百分率为97.67%；SRAP分子标记技术共扩增出88条条带，其中多态性条带86条，多态位点百分率为97.73%，ISSR和SRAP标记均表现出较高的多态性。不同样本间的遗传距离和遗传一致度表明:浙江省与福建省的金线莲种质混杂严重，而遗传多样性结果显示:福建省的金线莲种群遗传多样性更高。此外，基于ISSR和SRAP标记的UPGMA聚类结果显示:48份不同种源的金线莲依亲缘关系的远近可分为四大类，聚类的划分受到一定地域性的影响，但各地域的金线莲品种在这四大类中均互有混杂。 结论 金线莲在物种水平上遗传多样性较高，在各地区、各品种(系)间遗传交流频繁，ISSR与SRAP分子标记技术可以从分子水平上揭示浙江与福建等地金线莲的遗传多样性，且结合ISSR标记与SRAP标记的分析结果要优于单一的分子标记结果。图4表3参24
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| 关 键 词： | 金线莲 遗传多样性 分子标记 种质资源 ISSR SRAP |
| 收稿时间： | 2022-07-19 |
Genetic diversity of Anoectochilus roxburghii based on ISSR and SRAP molecular markers |
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| HUANG Jinchun,WAN Siqi,CHEN Yang,LI Lihong,ZHANG Zili,ZHU Jianjun,WU Mei,XING Bingcong,SHAO Qingsong,LU Chenfei.Genetic diversity of Anoectochilus roxburghii based on ISSR and SRAP molecular markers[J].Journal of Zhejiang A&F University,2023,40(1):22-29. |
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| Authors: | HUANG Jinchun WAN Siqi CHEN Yang LI Lihong ZHANG Zili ZHU Jianjun WU Mei XING Bingcong SHAO Qingsong LU Chenfei |
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| Affiliation: | 1.Zhejiang Provincial Key Laboratory of Resources Protection and Innovation of Traditional Chinese Medicine, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China2.Wenzhou Key Laboratory of Horticultural Plant Breeding, Wenzhou Vocational College of Science & Technology, Wenzhou 325006, Zhejiang, China3.Agricultural Sciences Academy of Jinhua, Jinhua 321000, Zhejiang, China |
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| Abstract: | Objective The objective is to study the genetic diversity and genetic relationship among introduced, crossbred and wild samples of Anoectochilus roxburghii in Zhejiang and Fujian Provinces, so as to provide a scientific reference for the identification of germplasm resources of A. roxburghii and the breeding of new cultivars (lines). Method 48 samples of A. roxburghii fresh leaves were used as materials, and 11 pairs of primers with good polymorphism and clear amplification bands were selected by ISSR and SRAP, respectively. After agarose gel electrophoresis imaging, the number of amplified bands was counted, and UPGMA clustering analysis was performed using NTSYS-PC 2.1 and POPGENE 32 software. Result A total of 86 bands were amplified by ISSR, including 84 polymorphic bands (PPB = 97.67%). A total of 88 bands and 86 polymorphic bands were amplified by SRAP, and the PPB was 97.73%. Both ISSR and SRAP markers showed high polymorphism. The genetic distance and genetic consistency between different samples indicated that the germplasm of A. roxburghii in Zhejiang and Fujian was seriously mixed, while the genetic diversity in Fujian was higher. In addition, the UPGMA clustering results based on ISSR+SRAP markers showed that the 48 samples from different provenances could be divided into 4 major groups according to the distance of their genetic relationship. The classification of clusters was affected by certain regions, but the cultivars in each region were mixed in the 4 categories. Conclusion The genetic diversity of A. roxburghii is relatively high at the species level, and genetic exchanges among regions and cultivars are frequent. The genetic diversity of A. roxburghii in Zhejiang and Fujian can be revealed at the molecular level by ISSR and SRAP molecular markers, and the analysis results of ISSR and SRAP markers combined are better than those of single molecular markers. Ch, 4 fig. 3 tab. 24 ref.] |
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