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林业科学 ›› 2015, Vol. 51 ›› Issue (1): 157-164.doi: 10.11707/j.1001-7488.20150119

• 研究简报 • 上一篇    下一篇

基于SRAP标记的油橄榄品种遗传多样性分析

占明明1, 杨毅2, 程子彰1, 苏光灿3, 胡伟3, 陈华萍1, 黄乾明1   

  1. 1. 四川农业大学理学院 雅安 625014;
    2. 四川农业大学生命科学学院 雅安 625014;
    3. 四川省凉山州中泽新技术开发有限责任公司 西昌 615000
  • 收稿日期:2014-04-01 修回日期:2014-10-22 出版日期:2015-01-25 发布日期:2015-01-23
  • 通讯作者: 黄乾明
  • 基金资助:

    四川省科技厅科技支撑计划项目(12ZC2220).

Genetic Diversity of Olive Varieties Based on SRAP Markers

Zhan Mingming1, Yang Yi2, Cheng Zizhang1, Su Guangcan3, Hu Wei3, Chen Huaping1, Huang Qianming1   

  1. 1. College of Science, Sichuan Agricultural University Ya'an 625014;
    2. College of Life Sciences, Sichuan Agricultural University Ya'an 625014;
    3. Sichuan Liangshan New Technology Development Co., Ltd. Xichang 615000
  • Received:2014-04-01 Revised:2014-10-22 Online:2015-01-25 Published:2015-01-23

摘要:

[目的]油橄榄作为重要的油料经济作物,其健康价值越来越受到人们的重视,相关产业在我国已初具规模.引种品种混乱和遗传关系不明确影响了我国油橄榄事业的进一步发展,因此我国油橄榄品种的遗传变异有待评估.[方法]采用SRAP技术对32个来自四川西昌市的油橄榄品种进行遗传多样性分析,其中5个油橄榄品种为国内育种品种,27个油橄榄品种为国外引种品种.利用改良的CTAB法提取得到32个油橄榄品种的基因组DNA.[结果]25对SRAP引物共扩增出293条多态性条带,平均每对引物扩增出11.72条多态性条带,平均多态性比率为90.75%;引物组合M5E5扩增出的多态性条带最多(37条);期望杂合度(He)为0.804~0.958(平均0.896),多态性信息含量(PIC)为0.773~0.955(平均0.884),一致性概率(PI)为0.004~0.067(平均0.024).筛选得到的25对SRAP引物扩增条带多态性好,具有更强的鉴别能力.基于UPGMA法的聚类分析结果显示,所有品种分为3大类,品种间的遗传相似性为0.59~0.89(平均0.74),遗传相似性最高的2个品种(‘希腊3号’和‘配多灵’)为0.89.国内育种品种分布于不同的类群中, ‘中山24’为通过‘阿斯’实生选优而来的,二者在形态学和遗传学方面均有较高的相似性,进一步证实了国内品种育种途径以及与国外品种的亲缘关系. ‘哥朗米扎’与‘未知米扎’聚在一类,‘希腊3号’与‘配多灵’聚在一类,且遗传相似性系数最高.主坐标分析(PCA)显示,所有油橄榄品种可分为3个群体,累计贡献率为20.4%.UPGMA分析中的第Ⅱ类的所有品种都聚在PCA分析的Group Ⅰ内,两者分析结果一致.[结论]综合不同油橄榄品种形态学(果质量、含油率、叶片形状等)和遗传学数据(育种背景、遗传相似性系数等)分析,部分引种品种的分类并未按照其地理起源来划分,而是由其遗传学和形态学数据共同来决定的.由以上结果可知,SRAP标记技术用于分析油橄榄遗传多样性简便、可靠.油橄榄遗传多样性比较丰富,我国引种油橄榄品种存在同种异名、命名混乱、受外界影响存在高度的遗传变异等情况.今后要加大运用SRAP和其他一些新型分子标记对油橄榄进行遗传多样性分析和种质鉴定,开辟基因定位和功能鉴定新途径,加大遗传图谱构建的密度和遗传距离;将传统育种方法与分子标记相结合,缩短育种年限,提高育种效率.

关键词: 油橄榄, SRAP, 遗传多样性

Abstract:

[Objective]As an important economic crop, the value of olive(Olea europaea) for health has been paid more and more attention. And the olive industry in China has reached a certain scale. Mislabeling and synonyms in introduced varieties have affected the development of China's olive industry. So there is a need to assess genetic variation of olive varieties in China. [Method]SRAP technology was applied to conduct genetic diversity analysis for 32 olive varieties from Xichang, Sichuan Province. 5 olive varieties among them were bred in China and 27 varieties were introduced. Genomic DNA was extracted for 32 olive varieties by the improved CTAB method. [Result]A total of 293 (90.75%) polymorphic bands were amplified by 25 pairs of primers and 11.72 polymorphic bands in average were amplified from each pair of primers. The most polymorphic bands (37) were amplified by primers M5E5. The expected heterozygoisty (He) varied from 0.804 to 0.958 (mean, 0.896), while the values of polymorphic information content(PIC) varied from 0.773 to 0.955 (mean, 0.884). The probability of identity (PI) varied from 0.004 to 0.067 (mean, 0.024). As the clustering analysis indicated, all varieties could be clustered into 3 groups and the genetic similarity (GS) varied from 0.59 to 0.89 (mean, 0.74). The highest genetic similarity between two varieties (‘Greece 3#’ and ‘Pendolino’) was 0.89. Varieties bred in China were distributed in 3 groups. ‘Zhongshan 24’ was developed from seedling selection of ‘Ascolana Tenera’, which displayed high similarity in both morphology and genetics. This further proved the relations between varieties bred in China and introduced foreign varieties. ‘Mixaj I Dukat’-‘Mixaj’ and ‘Greece 3#’-‘Pendolino’ were clustered together with high genetic similarity. The PCA analysis indicated that all varieties could be clustered into 3 groups explaining 20.8% of the total variation. Varieties of the group Ⅰ of PCA analysis were all contained cluster Ⅱ of UPGMA, indicating consistence between the two ways of analysis. [Conclusion]According to the morphology of different olive varieties (fruit mass, oil content, leaf shape etc.) and genetic (breeding background and genetic similarity etc.) data, the cluster of some introduced varieties were not in consistence with their geographical origins, probably due to both genetic and morphological data. SRAPs can be applied to olive genetic diversity analysis more simply and reliably. The introduced olive varieties displayed a genetic diversity, and there were confusions of variety names and high genetic variation due to environment effects. In the future, SRAP markers and other new molecular markers can be used to further analyze genetic diversity and germplasm identification. Moreover, new technologies of gene mapping and identification of gene functions can be also used to construct high-density genetic linkage maps. Traditional breeding and molecular markers should be integrated to accelerate the process of breeding.

Key words: Olea europaea, SRAP, genetic diversity

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