基于SCoT分子标记的48份石斛种质遗传多样性分析

doi:10.16178/j.issn.0528-9017.20230814

摘要/Abstract

摘要：

为研究不同品种石斛的遗传多样性和亲缘关系,该研究收集48份不同品种的石斛叶片,提取基因组DNA作为实验材料,利用随机选取的2个DNA样品为模板,从36条引物中筛选出具有扩增条带清晰、重复性好的15条目标起始密码子多态性(start codon targeted polymorphism,SCoT)标记引物,进行所有样品的SCoT-PCR扩增反应和遗传多样性研究。结果显示,挑选得出的15条引物每条引物可检测到12~20个等位位点,共扩增获得224条清晰条带,其中217条为多态性条带,多态性比率达到96.88%。聚类分析结果图中显示,48份供试石斛在遗传相似系数在0.55~0.78,在遗传系数0.659水平上可以将供试石斛分为5个类群。15条SCoT引物对48种石斛材料进行有效区分,并将其分为5个组。48种石斛材料的遗传背景存在显著差异,为石斛资源保护提供理论指导。

关键词: 石斛, SCoT分子标记, 遗传多样性

Abstract:

To investigate the genetic diversity and relatedness of different species of Dendrobium,the genomic DNA of 48 Dendrobium leaves of different varieties was collected, and 15 SCoT primers with rich polymorphism and good repeatability were screened from 36 primers by using 2 randomly selected DNA samples, and then the collected genomic DNA of 48 Dendrobium were amplified by SCoT-PCR and the genetic diversity was analyzed. The results showed that a total of 224 clear bands were amplified from the 15 primers selected, of which 217 were polymorphic bands, and the polymorphism ratio (PPR) was 96.88%. Each primer can detect 12 to 20 allelic sites. The results of cluster analysis showed that the genetic similarity coefficient of 48 samples of Dendrobium was between 0.55 and 0.78. The tested Dendrobium can be divided into 5 groups at the level of genetic coefficient 0.659. Fifteen SCoT primers were used to effectively distinguish 48 Dendrobium materials and divide them into 5 groups. There are significant differences in the genetic background of 48 types of Dendrobium materials, providing theoretical guidance for the protection of Dendrobium resources.

Key words: Dendrobium, SCoT markers, genetic diversity

中图分类号:

S567.9

庞敏霞, 姚国祥, 金波, 戚勇, 谢雄杰. 基于SCoT分子标记的48份石斛种质遗传多样性分析[J]. 浙江农业科学, 2024, 65(9): 2093-2099.

PANG Minxia, YAO Guoxiang, JIN Bo, QI Yong, XIE Xiongjie. Genetic diversity analysis of 48 accessions of Dendrobium germplasms based on SCoT markers[J]. Journal of Zhejiang Agricultural Sciences, 2024, 65(9): 2093-2099.

图/表 11

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序号	拉丁学名	种名	序号	拉丁学名	种名
1	Dendrobium officinale Kimura et Migo	铁皮石斛	25	Dendrobium lituiflorum Lindl.	喇叭唇石斛
2	Dendrobium pendulum Roxb.	肿节石斛	26	Dendrobium heterocarpum Lindl.	尖刀唇石斛
3	Dendrobium nobile Lindl.	金钗石斛	27	Dendrobium lindleyi Stendel	聚石斛
4	Dendrobium wardianum Warner	大苞鞘石斛	28	Dendrobium ochreatum.	黑喉石斛
5	Dendrobium moschatum (Buch.-Ham.)Sw.	杓唇石斛	29	Dendrobium minutiflorum S.C.Chen et Z.H.Tsi	勐海石斛
6	Dendrobium findlayanum Par. et Rchb.f.	棒节石斛	30	Dendrobium Signatum Rchb.f.	黄喉石斛
7	Dendrobium aphyllum (Roxb.)C. E. Fischer	兜唇石斛	31	Dendrobium aurantiacum Rchb.f.	线叶石斛
8	Dendrobium tosaense Makino	黄石斛	32	Dendrobium acinaciforme Roxb.	剑叶石斛
9	Dendrobium fimbriatum Hook.	流苏石斛	33	Dendrobium hancockii Rolfe	细叶石斛
10	Dendrobium chrysanthum Lindl	束花石斛	34	Dendrobium henryi Schltr.	疏花石斛
11	Dendrobium parishii Rchb. f.	紫瓣石斛	35	Dendrobium hookerianum Lindl.	金耳石斛
12	Dendrobium brymerianum Rchb.f.	长苏石斛	36	Dendrobium exile Schltr.	景洪石斛
13	Dendrobium chrysotoxum Lindl.	鼓槌石斛	37	Ddrobiurm bellatulum Rolfe	矮石斛
14	Dendrobium densiflorum Lindl.	密花石斛	38	Dendrobium mivakei Schltr.	红花石斛
15	Dendrobium thyrsiflorum Rchb. f.	球花石斛	39	Dendrobium pseudotenellum Guillaum.	针叶石斛
16	Dendrobium cariniferum Rchb. f.	翅萼石斛	40	Dendrobium wilsonii Rolfe	广东石斛
17	Dendrobium infundibulum Lindl.	高山石斛	41	Dendrobium equitans Kraenzl.	燕石斛
18	Dendrobium williamsonii Day et Rchb. f.	黑毛石斛	42	Dendrobium hercoglossum Rchb.f.	重唇石斛
19	Dendrobium strongylanthum Rchb. f.	梳唇石斛	43	Dendrobium chameleon Ames	长爪石斛
20	Dendrobium secundum.	红牙刷石斛	44	Dendrobium christyanum Rchb.f.	喉红石斛
21	Dendrobium denneanum Kerr.	叠鞘石斛	45	Dendrobium furcatopedicellatum Hayata	双花石斛
22	Dendrobium pulchellum	蜻蜓石斛	46	Dendrobium kingianum Bidwill ex Lindl.	澳洲鸽石斛
23	Dendrobium Jinawianum Rchb.f.	矩唇石斛	47	Dendrobium nobile Lindl	虎牙石斛
24	Dendrobium capillipes Rchb. f.	短棒石斛	48	Dendrobium polyanthum Wall. ex Lindl.	黄贝壳石斛

序号	拉丁学名	种名	序号	拉丁学名	种名
1	Dendrobium officinale Kimura et Migo	铁皮石斛	25	Dendrobium lituiflorum Lindl.	喇叭唇石斛
2	Dendrobium pendulum Roxb.	肿节石斛	26	Dendrobium heterocarpum Lindl.	尖刀唇石斛
3	Dendrobium nobile Lindl.	金钗石斛	27	Dendrobium lindleyi Stendel	聚石斛
4	Dendrobium wardianum Warner	大苞鞘石斛	28	Dendrobium ochreatum.	黑喉石斛
5	Dendrobium moschatum (Buch.-Ham.)Sw.	杓唇石斛	29	Dendrobium minutiflorum S.C.Chen et Z.H.Tsi	勐海石斛
6	Dendrobium findlayanum Par. et Rchb.f.	棒节石斛	30	Dendrobium Signatum Rchb.f.	黄喉石斛
7	Dendrobium aphyllum (Roxb.)C. E. Fischer	兜唇石斛	31	Dendrobium aurantiacum Rchb.f.	线叶石斛
8	Dendrobium tosaense Makino	黄石斛	32	Dendrobium acinaciforme Roxb.	剑叶石斛
9	Dendrobium fimbriatum Hook.	流苏石斛	33	Dendrobium hancockii Rolfe	细叶石斛
10	Dendrobium chrysanthum Lindl	束花石斛	34	Dendrobium henryi Schltr.	疏花石斛
11	Dendrobium parishii Rchb. f.	紫瓣石斛	35	Dendrobium hookerianum Lindl.	金耳石斛
12	Dendrobium brymerianum Rchb.f.	长苏石斛	36	Dendrobium exile Schltr.	景洪石斛
13	Dendrobium chrysotoxum Lindl.	鼓槌石斛	37	Ddrobiurm bellatulum Rolfe	矮石斛
14	Dendrobium densiflorum Lindl.	密花石斛	38	Dendrobium mivakei Schltr.	红花石斛
15	Dendrobium thyrsiflorum Rchb. f.	球花石斛	39	Dendrobium pseudotenellum Guillaum.	针叶石斛
16	Dendrobium cariniferum Rchb. f.	翅萼石斛	40	Dendrobium wilsonii Rolfe	广东石斛
17	Dendrobium infundibulum Lindl.	高山石斛	41	Dendrobium equitans Kraenzl.	燕石斛
18	Dendrobium williamsonii Day et Rchb. f.	黑毛石斛	42	Dendrobium hercoglossum Rchb.f.	重唇石斛
19	Dendrobium strongylanthum Rchb. f.	梳唇石斛	43	Dendrobium chameleon Ames	长爪石斛
20	Dendrobium secundum.	红牙刷石斛	44	Dendrobium christyanum Rchb.f.	喉红石斛
21	Dendrobium denneanum Kerr.	叠鞘石斛	45	Dendrobium furcatopedicellatum Hayata	双花石斛
22	Dendrobium pulchellum	蜻蜓石斛	46	Dendrobium kingianum Bidwill ex Lindl.	澳洲鸽石斛
23	Dendrobium Jinawianum Rchb.f.	矩唇石斛	47	Dendrobium nobile Lindl	虎牙石斛
24	Dendrobium capillipes Rchb. f.	短棒石斛	48	Dendrobium polyanthum Wall. ex Lindl.	黄贝壳石斛

引物编号	引物序列(5'—3')	引物编号	引物序列(5'—3')
SCoT8	CAACAATGGCTACCACGT	SCoT23	CACCATGGCTACCACCAG
SCoT9	CAACAATGGCTACCAGCA	SCoT24	CACCATGGCTACCACCAT
SCoT10	CAACAATGGCTACCAGCC	SCoT26	ACCATGGCTACCACCGTC
SCoT14	ACGACATGGCGACCACGC	SCoT27	ACCATGGCTACCACCGTG
SCoT16	ACCATGGCTACCACCGAC	SCoT28	CCATGGCTACCACCGCCA
SCoT18	ACCATGGCTACCACCGCC	SCoT29	CCATGGCTACCACCGGCC
SCoT20	ACCATGGCTACCACCGCG	SCoT30	CCATGGCTACCACCGGCG
SCoT21	ACGACATGGCGACCCACA

引物编号	引物序列(5'—3')	引物编号	引物序列(5'—3')
SCoT8	CAACAATGGCTACCACGT	SCoT23	CACCATGGCTACCACCAG
SCoT9	CAACAATGGCTACCAGCA	SCoT24	CACCATGGCTACCACCAT
SCoT10	CAACAATGGCTACCAGCC	SCoT26	ACCATGGCTACCACCGTC
SCoT14	ACGACATGGCGACCACGC	SCoT27	ACCATGGCTACCACCGTG
SCoT16	ACCATGGCTACCACCGAC	SCoT28	CCATGGCTACCACCGCCA
SCoT18	ACCATGGCTACCACCGCC	SCoT29	CCATGGCTACCACCGGCC
SCoT20	ACCATGGCTACCACCGCG	SCoT30	CCATGGCTACCACCGGCG
SCoT21	ACGACATGGCGACCCACA

引物编号	条带总数/条	多态性条带/条	多态性比率/%
SCoT8	13	13	100.00
SCoT9	16	14	87.50
SCoT10	20	20	100.00
SCoT14	16	16	100.00
SCoT16	12	12	100.00
SCoT18	16	16	100.00
SCoT20	14	14	100.00
SCoT21	17	15	88.24
SCoT23	14	14	100.00
SCoT24	16	15	100.00
SCoT26	12	12	100.00
SCoT27	13	13	100.00
SCoT28	15	14	93.33
SCoT29	17	17	100.00
SCoT30	12	12	100.00
平均值	14.9	14.5	96.88