利用SSR构建薄壳山核桃主要品种的分子身份证

张成才,王亚萍,王开良,常君,任华东,姚小华

植物遗传资源学报. 2022, 23(6): 1835-1846

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植物遗传资源学报 ›› 2022, Vol. 23 ›› Issue (6) : 1835-1846. DOI: 10.13430/j.cnki.jpgr.20220611001
论文

利用SSR构建薄壳山核桃主要品种的分子身份证

  • 张成才, 王亚萍, 王开良, 常君, 任华东, 姚小华
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Construction of Molecular IDs for Major Cultivars in Pecan (Carya illinoinensis) Using SSR Markers

  • Zhang Cheng-cai, Wang Ya-ping, Wang Kai-liang, Chang Jun, Ren Hua-dong, Yao Xiao-hua
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摘要

薄壳山核桃是著名的干果和木本油料树种,当前存在“同名异物、同物异名”现象影响产业发展。本研究在薄壳山核桃基因组中筛选到80个简单重复序列(SSR),利用8个品种的基因组DNA,对80对SSR引物进行筛选,初筛到23对目标条带清晰、检测结果稳定的引物,发现它们分布于13条染色体上。进一步在‘波尼’(‘Pawnee’)、‘马罕’(‘Mahan’)、‘斯图亚特’(‘Stuart’)等36个常用品种中开展基因分型,共检测到70个等位位点;其中18对引物具有遗传多态性,检测到的等位基因数介于2~8,多态性信息含量(PIC)介于0.03 ~ 0.72。聚类分析发现,各个样本按照遗传背景聚类,不同品种之间遗传距离介于0.026~0.6359,同一品种的不同来源个体之间遗传距离介于0 ~ 0.0127。按PIC值从大到小的顺序,依次增加引物的数量,直至能够区分每个品种,发现最少可使用Ciz91、Ciz85、Ciz81、Ciz140和Ciz107等5个标记实现所有品种的有效区分,进而将这组引物确定为核心引物组。最后,使用核心引物组分别构建每个品种特异的分子指纹图谱、分子身份证条形码和二维码。本研究,利用SSR标记构建品种特异的分子指纹图谱和分子身份证,为薄壳山核桃品种鉴别、种苗纯度检测、品种追溯等提供理论基础。

Abstract

Pecan [Carya illinoinensis (Wangenh.) K. Koch] is an important woody oil plant and a famous nut tree species, which has been widely cultivated in China. Adequate identification of different cultivars is showing the top priority in marketing. This study aimed to develop SSR markers which would be used for analyzing the pecan genetic diversity and constructing the molecular fingerprint and IDs. A total of 80 primer pairs were used for analyzing the polymorphisms in eight cultivars. Twenty-three (28.75%) SSR primer pairs from 13 chromosomes could amplify the target fragments. These primers were further used for genotyping in 45 samples of 36 pecan cultivars, such as ‘Pawnee’, ‘Mahan’, ‘Stuart’, ‘Kanza’, and ‘Shawnee’, and produced a total of 70 alleles in all samples. Eighteen primer pairs were detected with polymorphisms in different cultivars, showing the number of alleles (NA) ranged from 2 to 8, and the polymorphism information content (PIC) values from 0.03 to 0.72. The genetic distance between different cultivars ranged from 0.026 to 0.6359, and the genetic distance between samples from different sources of the same cultivar ranged from 0 to 0.0127. A dendrogram generated by the UPGMA method suggested three groups, of which group I contained 32 cultivars and most of them have the genetic backgrounds of ‘Schley’, ‘Success’, and ‘Major. Group II and Group III contained three and one cultivars, respectively. By use of (at least) five SSR primer pairs, including Ciz91, Ciz85, Ciz81, Ciz140, and Ciz107, all the cultivars were classified. These primers were selected as core SSR markers to construct molecular fingerprints and IDs. The molecular IDs were illustrated as bar codes and QR codes. Collectively, this study provided markers applicable for the cultivar identification and traceability in pecan, which has implication in the progress of pecan cultivation and production.

关键词

薄壳山核桃;品种鉴定;分子标记;SSR;指纹图谱;分子身份证

Key words

Carya illinoinensis;cultivar identification;molecular marker;SSR;DNA fingerprints;molecular ID.

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张成才,王亚萍,王开良,常君,任华东,姚小华. 利用SSR构建薄壳山核桃主要品种的分子身份证. 植物遗传资源学报. 2022, 23(6): 1835-1846 https://doi.org/10.13430/j.cnki.jpgr.20220611001
Zhang Cheng-cai,Wang Ya-ping,Wang Kai-liang,Chang Jun,Ren Hua-dong and Yao Xiao-hua. Construction of Molecular IDs for Major Cultivars in Pecan (Carya illinoinensis) Using SSR Markers. Journal of Plant Genetic Resources. 2022, 23(6): 1835-1846 https://doi.org/10.13430/j.cnki.jpgr.20220611001

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