基于SSR分子标记的枣品种鉴定及DNA分子身份证构建

Jujube variety identification based on SSR molecular markers and construction of DNA molecular identity card

【目的】利用荧光SSR分子标记对北京枣主栽品种的亲缘关系及遗传多样性分析并构建枣品种DNA分子身份证,为枣新品种选育、生产利用奠定基础。【方法】收集103个枣品种材料,筛选SSR荧光标记引物,采用多重荧光毛细管电泳检测技术分析扩增条带分子量和等位基因,获得相应的扩增带型,从而对每个枣种质材料进行标记并构建供试样品的DNA分子身份证。【结果】基于SSR分子标记技术,筛选出19对SSR引物用于103个枣品种遗传多样性分析,并筛选了7对核心基因组合构建DNA分子身份证。结果表明:19对引物对103个枣品种样品经SSR-PCR扩增后共检测到了156个等位标记点,每对引物平均8.211个,有效等位基因(Ne)变化范围为1.263~12.568,平均为3.467;Shannon’s信息指数(I)分布范围0.547~2.664,平均为1.351;多态信息含量PIC值变幅在0.32~0.917,平均值为0.603,观测杂合度(Ho)和期望杂合度(He)范围分别为0.208~0.920和0.133~0.990,Ho和He的平均值为0.639和0.637,显示出有较高的杂合度;遗传相似系数为0.85~0.98,且在相似系数为0.85处时被分为两大类。【结论】北京地区京枣31和京枣311、马牙枣和牙枣等栽培品种的遗传背景较狭窄,遗传相似度较高,不同品系间基因交流较少;京枣60、朗家园枣、北京蜂蜜罐、北京泡泡枣、京枣28等枣的亲缘关系较远,遗传多样性丰富。本研究结果为枣品种鉴定和培育新品种提供了重要依据。

【Objective】The genetic relationship and genetic diversity of the main cultivars of Beijing jujube were analyzed by fluorescent SSR molecular markers,and the DNA molecular identity card of jujube varieties was constructed,which laid a foundation for the breeding,production and utilization of new jujube varieties.【Method】A total of 103 jujube germplasm materials were collected,and the SSR fluorescent marker primers were screened.The molecular weight and alleles of the amplified bands were analyzed by multiple fluorescence capillary electrophoresis,and the corresponding amplified bands were obtained.Each jujube germplasm material was marked and the DNA molecular identity of the test samples was constructed.【Result】Based on SSR molecular marker technology,19 pairs of SSR primers were screened for genetic diversity analysis of 103 jujube varieties,and 7 pairs of core gene combinations were screened to construct DNA molecular identity cards.The results showed that a total of 156 alleles were detected in 103 jujube cultivars by 19 pairs of primers after SSR-PCR amplification,with an average of 8.211 alleles per pair of primers.The effective alleles (Ne) ranged from 1.263 to 12.568,with an average of 3.467.Shannon’s information index (I) ranged from 0.547 to 2.664,with an average of 1.351.The polymorphism information content (PIC) ranged from 0.32 to 0.917,with an average value of 0.603.The observed heterozygosity (Ho) and expected heterozygosity (He) ranged from 0.208 to 0.920 and 0.133 to 0.990,respectively.The average values of Ho and He were 0.639 and 0.637,showing a high heterozygosity.The genetic similarity coefficient was 0.85-0.98,and it was divided into two categories when the similarity coefficient was 0.85.【Conclusion】In summary,the genetic background of Jingzao 31,Jingzao 311,Mayazao and Yazao in Beijing is relatively narrow,the genetic similarity is high,and the gene exchange between different strains is less.The genetic relationship of Jingzao 60,Langjiayuanzao,Beijing Fengmiguan,Beijing paopaozao,Jingzao 28 and other jujubes is far away,and the genetic diversity is rich.The results provided an important basis for the identification of jujube varieties and the cultivation of new varieties.