金耳SSR分子标记开发及指纹编码构建

doi:10.16420/j.issn.0513-353x.2024-0767

摘要/Abstract

摘要：

以7个金耳菌株及转录组测序获得的序列为试材，对金耳转录组序列上的SSR类型及分布进行了研究，设计了100对引物开展试验验证，总计检索到6种SSR类型的4 268个位点，其中主要以三核苷酸重复类型（P3）和六核苷酸重复类型（P6）为主，占总SSR的64.20%。由试验验证获得7对SSR引物，并开展分析。UPGMA聚类结果显示，7个金耳菌株主要分为3个类群。同时，基于7对SSR引物进行金耳的指纹编码构建，获得了7个金耳菌株的14位数字分子指纹数据库编码。以期为金耳的种质资源鉴定、品种选育、分子标记辅助育种等研究提供参考依据。

关键词: 金耳, 转录组, SSR, 标记开发

Abstract:

Based on experimental validation and bioinformatics，SSR analysis result were exploited from the transcriptiome of Naematelia aurantialba and seven strains. In the meantime，100 SSR were selected for primer design and experimental verification. A total of 4 268 sequences of 6 SSR types were retrieved. The main type of trinucleotide（P3）and hexanucleotide（P6），accounting for 64.20% of the total SSR number. Seven pairs of primers were obtained by verification test and analyzed. UPGMA clustering results showed that the seven N. aurantialba strains were mainly divided into three groups. At the same time，the DNA fingerprint coding of seven pairs of primers was constructed and the 14-digit molecular fingerprint database codes of seven N. aurantialba strains were obtained. In order to provide a theoretical basis for the germplasm resources identification，variety selection and molecular marker-assisted breeding of N. aurantialba.

Key words: Naematelia aurantialba, transcriptiome, SSR, mark development

李雪松, 华蓉, 孙达锋, 岳万松, 张俊波, 李建英, 刘绍雄. 金耳SSR分子标记开发及指纹编码构建[J]. 园艺学报, 2025, 52(7): 1780-1788.

LI Xuesong, HUA Rong, SUN Dafeng, YUE Wansong, ZHANG Junbo, LI Jianying, and LIU Shaoxiong. Development of SSR Markers Based on Transcriptiome of Naematelia aurantialba[J]. Acta Horticulturae Sinica, 2025, 52(7): 1780-1788.

图/表 6

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标记名称 Markers	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer	SSR 类型 SSR type	SSR 基元 SSR motif
JESSR003	CATCCCTATGCAATGATTCCACG	TAGAACAAGACTAGAACCGGGAC	P2	（TC）8
JESSR010	GAAGGACGTACAAGGCCGTG	GTGACTAGATTTGATGCTGGCAG	P6	（CCCAGC）4
JESSR032	GCTCTGTTTACGAGGGGATAGG	CCGAATCTGACACCATCATCTCT	P3	（TTG）6
JESSR075	GAGTCTTGAGCGGAGGAGTAC	GACGTACCATGCTGATGCTGAT	P6	（GATGCC）4
JESSR090	TAAAGCACTTCAATTGCGTCTCG	CTAGAGGAGGCCAAGAGGAAGAA	P6	（CATGTG）4
JESSR098	CCTATACCTTAGCCTGAGACAGC	GAGGATCACCATGTCCGTCTC	P3	（CGT）6
JESSR100	CTTGGGGAGGTGGAGAGTATAGG	ACTGTTCTTCGCTTGACCAAATT	P6	（CTGGCA）6

标记名称 Markers	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer	SSR 类型 SSR type	SSR 基元 SSR motif
JESSR003	CATCCCTATGCAATGATTCCACG	TAGAACAAGACTAGAACCGGGAC	P2	（TC）8
JESSR010	GAAGGACGTACAAGGCCGTG	GTGACTAGATTTGATGCTGGCAG	P6	（CCCAGC）4
JESSR032	GCTCTGTTTACGAGGGGATAGG	CCGAATCTGACACCATCATCTCT	P3	（TTG）6
JESSR075	GAGTCTTGAGCGGAGGAGTAC	GACGTACCATGCTGATGCTGAT	P6	（GATGCC）4
JESSR090	TAAAGCACTTCAATTGCGTCTCG	CTAGAGGAGGCCAAGAGGAAGAA	P6	（CATGTG）4
JESSR098	CCTATACCTTAGCCTGAGACAGC	GAGGATCACCATGTCCGTCTC	P3	（CGT）6
JESSR100	CTTGGGGAGGTGGAGAGTATAGG	ACTGTTCTTCGCTTGACCAAATT	P6	（CTGGCA）6

标记名称Markers	荧光标记Fluorescence labeling	扩增片段数Fragment number	片段大小/bp Fragment size
JESSR003	FAM	3	172，174，182
JESSR010	FAM	4	166，172，178，184
JESSR032	HEX	3	161，164，167
JESSR075	ROX	3	159．161，179
JESSR090	TAMRA	3	162，168，174
JESSR098	TAMRA	3	129，131，135
JESSR100	TAMRA	3	136，138，142

标记名称Markers	荧光标记Fluorescence labeling	扩增片段数Fragment number	片段大小/bp Fragment size
JESSR003	FAM	3	172，174，182
JESSR010	FAM	4	166，172，178，184
JESSR032	HEX	3	161，164，167
JESSR075	ROX	3	159．161，179
JESSR090	TAMRA	3	162，168，174
JESSR098	TAMRA	3	129，131，135
JESSR100	TAMRA	3	136，138，142

引物Primer	赋值Assignment
引物Primer	0	1	2	3	4
JESSR003	-	173	174	182	-
JESSR010	-	166	172	178	184
JESSR032	-	161	164	167	-
JESSR075	-	159	161	179	-
JESSR090	-	162	168	174	-
JESSR098	-	129	131	135	-
JESSR100	-	136	138	142	-