基于TrnL-F与ISSR的凹脉金花茶与3种近缘种的分子鉴别及其遗传多样性

陈莹; 曾菁菁; 吕祉龙; 仇明月; 郭蓓琳; 姚丽敏; 闫淑君

doi:10.19303/j.issn.1008-0384.2022.010.008

基于TrnL-F与ISSR的凹脉金花茶与3种近缘种的分子鉴别及其遗传多样性

doi: 10.19303/j.issn.1008-0384.2022.010.008

福建农林大学风景园林与艺术学院，福建　福州　350100

基金项目: 福建省自然科学基金项目（2021J01132）；福建农林大学学科专业建设项目（YSYL-bdpy-4）；福建农林大学科技创新专项基金（CXZX2020052A）；福建农林大学创新训练项目（202110389165）

详细信息

作者简介:
陈莹（1981−），女，博士，副教授，主要从事观赏药用植物和园林生态相关研究（E-mail：25220528@qq.com）

通讯作者:
闫淑君（1975−），女，博士，教授，主要从事城市植被生态相关研究（E-mail：fjyansj@tom.com）

中图分类号: S 682.39
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出版历程
- 收稿日期: 2022-04-30
- 修回日期: 2022-07-14
- 网络出版日期: 2022-11-29
- 刊出日期: 2022-10-30

TrnL-F and ISSR Primer-based Molecular Identification and Genetic Diversity Determination on Camellia impressinervis and Three Closely Related Species

College of Landscape Architecture and Art, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350100, China

摘要

摘要: 目的凹脉金花茶是一种具有较高价值的观赏与药用两用植物，与其同属中的金花茶、显脉金花茶、东兴金花茶外形相似。通过分子指纹技术分析这4个近源种之间的遗传多样性，可为凹脉金花茶的鉴定及其种质资源保护和利用提供理论指导。方法利用金花茶、凹脉金花茶、显脉金花茶、东兴金花茶的19个样品评估TrnL-F条形码在这4种山茶中鉴定凹脉金花茶的潜力，并利用简单序列间重复（ISSR）引物检测4个山茶组种的遗传多样性。结果基于TrnL-F序列构建的进化树与检测到的单核苷酸多态性位点的差异相一致，所有凹脉金花茶样品单独聚为一类。ISSR分析显示，4个凹脉金花茶植物样本单独聚为一类，与其他3个山茶组物种的相似指数为0.57。结论 TrnL-F可有效鉴别凹脉金花茶与3种近缘种。本研究揭示了这4个近缘种的分子亲缘关系，从分子水平证实显凹脉金花茶具有异于其他3种山茶的遗传特性。
- 金花茶 /
- TrnL-F /
- 分子鉴定 /
- 遗传多样性 /
- ISSR
Abstract: Objective Molecular identification and genetic diversity of Camellia impressinervis were studied to improve the preservation and utilization of the endangered species. Method A highly valuable ornamental and medicinal, but extremely endangered as classified by the International Union for the Conservation of Nature, C. impressinervis was compared with the closely related C. nitidissima, C. euphlebia, and C. tunghinensis. Using 19 specimens of the 4 species, validity of applying trnL-F barcode to confidently identify them was evaluated, and their genetic diversity examined based on the inter-simple sequence repeat (ISSR) primers. Result TrnL-F could accurately differentiate C. impressinervis from those closely related camellia cultivars and clustered all in a same group. The ISSR analysis grouped the 4 C. impressinervis specimens together and showed a similarity index of 0.57 with the other 3 camellia species. Conclusion The genetic fingerprint in terms of trnL-F barcode and ISSR primers successfully identified the close relationship of C. impressinervis with, but distinguishable from, C. nitidissima, C. euphlebia, and C. tunghinensis.
- Camellia /
- trnL-F /
- molecular identification /
- genetic diversity /
- ISSR

HTML全文

图 1 TrnL-F系列代表性位点

Figure 1. Representative loci of trnL-F series

下载: 全尺寸图片幻灯片

图 2 不同金花茶TrnL-F序列的系统发育树

Figure 2. Phylogenetic tree of C. sinensis trnL-F sequences

下载: 全尺寸图片幻灯片

图 3 引物UBC846聚合酶链反应扩增

M：Marker；1、8；9、10、12、14为金花茶；2、6；11、18为显脉金花茶，3、4、7、15为凹脉金花茶，5、13、16、17、19为东兴金花茶。

Figure 3. PCR amplification with primer UBC846

M: Marker; 1,8,9,10,12,14 are C . nitidissima; 2, 6, 11,18 are C.euphlebia; 3,4,7,15 are C. impressinervis; 5,13,16,17,19 are C.tunghinensis.

下载: 全尺寸图片幻灯片

图 4 4种金花茶种质资源的分类鉴定及亲缘关系

Figure 4. Classification, identification, and genetic relationship of 4 camellia germplasms

下载: 全尺寸图片幻灯片

表 1 ISSR扩增引物

Table 1. Amplified ISSR primers

引物 Primer	引物序列（3′-5′） Primer sequence（3′-5′）	退火温度 Annealing temperature/℃
UBC811	GAGAGAGAGAGAGAGAC	51.2
UBC815	CTCTCTCTCTCTCTCTG	50.5
UBC825	ACACACACACACACACT	50.5
UBC834	AGAGAGAGAGAGAGAGYT	49
UBC835	AGAGAGAGAGAGAGAGYC	56
UBC836	AGAGAGAGAGAGAGAGYA	52
UBC840	GAGAGAGAGAGAGAGAYT	49
UBC841	GAGAGAGAGAGAGAGAYC	52
UBC844	CTCTCTCTCTCTCTCTRC	52
UBC845	CTCTCTCTCTCTCTCTRG	50.5
UBC846	CACACACACACACACART	51.2
UBC848	CACACACACACACACARG	55
UBC851	GTGTGTGTGTGTGTGTYG	52
UBC856	ACACACACACACACACYA	52

下载: 导出CSV

表 2 ISSR引物扩增产物的多态性

Table 2. Polymorphisms of ISSR primers amplified products

引物 Primer	扩增条带总数 Total number of amplified bands/条	多态性条带数 Polymorphic strips/条	多态性条带占比 Percentage of polymorphic bands/%
UBC811	20	20	100.0
UBC815	12	12	100.0
UBC825	18	16	88.9
UBC834	13	13	100.0
UBC835	15	15	100.0
UBC836	16	13	81.3
UBC840	12	12	100.0
UBC841	12	10	83.3
UBC844	15	15	100.0
UBC845	12	12	100.0
UBC846	16	16	100.0
UBC848	3	3	100.0
UBC851	9	8	88.9
UBC856	9	8	88.9
总数 Total	182	173	95.1

下载: 导出CSV

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