基于SSR标记的丛生竹杂种鉴定、遗传分析和指纹图谱构建

doi:10.12302/j.issn.1000-2006.202012046

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南京林业大学学报（自然科学版） ›› 2021, Vol. 45 ›› Issue (5): 10-18.doi: 10.12302/j.issn.1000-2006.202012046

所属专题：专题报道

基于SSR标记的丛生竹杂种鉴定、遗传分析和指纹图谱构建

袁金玲¹, 马婧瑕², 钟远标¹, 岳晋军¹

¹ 中国林业科学研究院亚热带林业研究所, 浙江杭州 311400
² 安吉竹子博览园有限公司,浙江湖州 313300

收稿日期:2020-12-28 接受日期:2021-03-10 出版日期:2021-09-30 发布日期:2021-09-30
基金资助:
浙江省农业新品种选育重大专项(2021C02070-4)

SSR-based hybrid identification, genetic analyses and fingerprint development of hybridization progenies from sympodial bamboo (Bambusoideae, Poaceae)

YUAN Jinling¹, MA Jingxia², ZHONG Yuanbiao¹, YUE Jinjun¹

¹ Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Hangzhou 311400, China
² Anji Bamboo Exposition Garden Co., Ltd., Huzhou 313300, China

Received:2020-12-28 Accepted:2021-03-10 Online:2021-09-30 Published:2021-09-30

摘要/Abstract

摘要：

【目的】 准确鉴定丛生竹杂交种,分析杂种及其亲本的遗传关系,开发可以利用的指纹图谱。【方法】 以孝顺竹(Bambusa multiplex)×麻竹(Dendrocalamus latiflorus)、孝顺竹(B. multiplex)×粉单竹(B. chungii)两个杂交群体为材料,从已公布的竹子SSR引物中随机选取30对引物进行筛选、检测和分析。【结果】 发现6对SSR引物(P8、P9、P18、P19、P20、P27)适于孝顺竹×麻竹组合的杂种鉴定和遗传分析,鉴定结果表明34个子代全部为真实杂种,杂种真实率为100%;另外6对SSR引物(P2、P9、P10、P11、P20、P28)则适于孝顺竹×粉单竹组合的杂种鉴定和遗传分析,鉴定结果表明59个子代中有54个真实杂种,杂种真实率为91.5%。孝顺竹×麻竹的杂种遗传了较多的母本位点,而孝顺竹×粉单竹的杂种则遗传了较多的父本位点。利用SSR引物扩增出的位点分别构建了两个杂交群体的指纹图谱,为品种保护提供了保障。【结论】 SSR引物能有效地鉴定丛生竹杂交种的真伪,父母本的遗传位点在不同杂交组合的子代中遗传概率不同,杂交群体指纹图谱的构建可为品种保护提供保障。

关键词: 丛生竹, 杂交子代, SSR, 杂种鉴定, 遗传分析, 指纹图谱

Abstract:

【Objective】 The study aims to accurately verify sympodial bamboo hybrids, clarify genetic relationships between hybrids and their parents, and develop applicable fingerprints. 【Method】 We screened, tested and analyzed 30 pairs of randomly selected published bamboo simple sequence repeat (SSR) primers using hybrids derived from Bambusa multiplex × Dendrocalamus latiflorus and B. multiplex × B. chungii crosses. 【Result】 According to the results, six SSR primer pairs (P8, P9, P18, P19, P20 and P27) were suitable for the hybrid identification and genetic analysis of the B. multiplex × D. latiflorus cross, which revealed that all 34 progenies were true hybrids (100%). Six SSR primer pairs (P2, P9, P10, P11, P20 and P28) were suitable for the hybrid identification and genetic analysis of the B. multiplex × B. chungii cross, which revealed that 54 of the 59 progenies were true hybrids (91.5%). Hybrids of the B. multiplex × D. latiflorus cross inherited more maternal alleles, while hybrids of the B. multiplex × B. chungii cross inherited more paternal alleles. The alleles detected using SSR primers were used to construct hybrid fingerprints of the two crosses for variety protection. 【Conclusion】 The SSR primers could be used to effectively verify sympodial bamboo hybrids as the progeny inherited alleles from different parents at varying probabilities in different crosses. In addition, the development of hybrid fingerprints could facilitate variety protection.

Key words: sympodial bamboo, hybridization progenies, simple sequence repeat (SSR), hybrid identification, genetic analyses, fingerprint

中图分类号:

S722

袁金玲,马婧瑕,钟远标,等. 基于SSR标记的丛生竹杂种鉴定、遗传分析和指纹图谱构建[J]. 南京林业大学学报（自然科学版）, 2021, 45(5): 10-18.

YUAN Jinling, MA Jingxia, ZHONG Yuanbiao, YUE Jinjun. SSR-based hybrid identification, genetic analyses and fingerprint development of hybridization progenies from sympodial bamboo (Bambusoideae, Poaceae)[J].Journal of Nanjing Forestry University (Natural Science Edition), 2021, 45(5): 10-18.DOI: 10.12302/j.issn.1000-2006.202012046.

图/表 8

表1

图1

图2

图3

Table 2

图4

图5

Table 3

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No. 序号	marker 引物名称	primer sequence (5'→ 3') 引物序列		No. 序号	marker 引物名称	primer sequence (5'→ 3') 引物序列
No. 序号	marker 引物名称	F-primer正向	R-primer反向	No. 序号	marker 引物名称	F-primer 正向	R-primer 反向
P1	PBM031	CGCCGAGTTCCCTATTATTATTT	AGCACAGCCTCCGTGATTG	P16	DOM01	TGGAGGCATTAAGCACAG	TTGACACTCGTGGTAGGC
P2	PBM032	TTTCCCAAATAAAACCTCACC	GTCCATTTAGGGTTCCACTGA	P17	DOM02	GTTCAGCCCCTTACCTCC	TGATCAATTCAGATTCCACTT
P3	PBM033	CTGACTGTGCGTGCGTCTC	CTTGGTCTCGCTCATCTCCTC	P18	DOM03	AAGGAGTTCATCTCCCGACA	CACCACGCAAACCAAGCT
P4	PBM034	TCGGCTCGGCGTGATGGAT	ATCGGCATCCGCGACTGCC	P19	DOM04	GGAGTTCATCTCCCGACA	TCTTTAAGGCAAACAACG
P5	PBM035	ACCGTGATGACTACCGCCGCGACC	TGCTGCCTCCACCCCTCCGTCC	P20	DOM05	TTGGAGGAGACTGAATGA	CCTTTGCGAAAATACACG
P6	PBM036	CACATGGACCGCCTCATCC	GCAACAAAACGAGAACCAGAC	P21	DOM06	TCTAGCATGGACGGTCAG	TCTCGCATAAGCCACATC
P7	PBM037	TGCAAGCCTGCTATACGTTT	GAAGTGGGAGTACATACTTCCCA	P22	DOM07	GGGCTGTCTGGCTTGTTA	TGAAAGGATAGGTTAGGTTGC
P8	PBM038	GGTCGGCTCATTTTGTAGTGT	CAACCTTCAGGCAATAGATTACAT	P23	DOM08	CCTGAAGGTTGACTGGGATA	TCCTCCGCCTTAGTGTCT
P9	PBM039	CATCCTCAGTTTCTCACCG	CAGCTTCACCAACTTGTGG	P24	DOM09	CACCACGCAAACCAAGCT	GGAGTTCATCTCCCGACA
P10	PBM040	GAATCATCTGGGAAGAAGAAGGA	TGCATTGCATTTGGCTTAGTAGT	P25	DOM10	TGCTTCATTATCTGCTGGTT	TGCACGTCGCAAGTTTCA
P11	PBM041	TGGTGTTGCCTGTGACCTTAC	CCCACCTCCACCTCTACTACG	P26	DOM11	GCCCGACTTTCTTTCATCTCCC	CGCATTTCGCTACGTTCTTCAT
P12	PBM042	TCCTTTACGGCTTTACCCC	GCCCCAGCTTAGTACACCAC	P27	DOM12	CCGGAGCGATCTCAAGGA	CCAAGAGTCTGGGAGTAGGG
P13	PBM043	CTCACCGCCCCACCTCGCA	CGGCTGCTGATGCGGAGGA	P28	DOM13	CCTGACCGAAGCAGACCG	GCCGAGATATCCGTTGCC
P14	PBM044	AAGGCCCACGTTGCCAGAC	GTTCCCGTTGATGCCCCAC	P29	DOM14	GTGCCTTCCCAGTTTGTC	CCTTTGCTATTGCCTTGC
P15	PBM045	TGAGCGAGGTAGTTTCATTTTAGTTA	CCTACGACGAGTAGATTGCGAGT	P30	DOM15	ATCCTAGTGCGCCTCGTC	TTGGTGGGCTCCCTTTGT

primer 引物	B1	B2	B3	B4	B5	B6	B7	B8	B9	B10	B11	B12	B13	B14	B15	B16	B17	B18	B19	B20	B21	B22	B23	B24	B25	B26	B27	B28	B29	B30	B31	B32	B33	B34
	1	1	1	0	1	0	0	1	0	1	0	0	1	1	0	0	0	0	0	0	1	1	0	1	0	1	1	1	0	1	0	0	0	1
P8	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
	0	0	0	1	0	1	1	0	1	0	1	1	0	0	1	1	1	1	1	1	0	0	1	0	1	0	0	0	1	0	1	1	1	0
	0	0	0	0	0	0	1	1	0	1	0	1	0	0	0	0	0	0	0	0	1	0	0	0	0	1	0	1	1	1	0	1	1	0
	0	0	0	0	0	0	1	1	0	1	0	1	0	0	0	0	0	0	0	0	1	0	0	0	0	1	0	1	1	1	0	1	1	0
	1	1	1	1	1	0	0	0	1	0	1	1	1	0	1	1	0	1	1	1	1	1	0	1	1	1	0	1	0	0	1	0	0	1
P20	1	1	1	1	1	0	0	0	1	0	1	1	1	0	1	1	0	1	1	1	1	1	0	1	1	1	0	1	0	0	1	0	0	1
	0	0	0	0	0	1	1	1	0	1	0	0	0	1	0	0	1	0	0	0	0	0	1	0	0	0	1	0	1	1	0	1	1	0
	1	1	1	1	1	1	1	1	1	1	1	0	1	1	1	1	1	1	1	1	0	1	1	1	1	0	1	0	1	1	1	1	1	1
	1	1	1	1	1	1	0	0	1	0	1	0	1	1	1	1	1	1	1	1	0	1	1	1	1	0	1	0	0	0	1	0	0	1
	1	0	0	1	1	0	1	0	1	0	1	0	0	1	1	0	0	1	0	1	0	1	0	1	0	1	0	1	1	0	1	1	1	1
P27	0	1	1	0	0	1	0	1	0	1	0	1	1	0	0	1	1	0	1	0	1	0	1	0	1	0	1	0	0	1	0	0	0	0
	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1

primer 引物	D1	D2	D3	D4	D5	D6	D7	D8	D9	D10	D11	D12	D13	D14	D15	D16	D17	D18
P2	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
	1	0	1	0	0	0	1	0	1	0	1	0	0	1	1	0	1	1
	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
P9	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
	1	1	1	0	1	1	1	1	0	1	1	1	1	1	1	0	1	1
	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
P11	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	0	1	1
	1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	1	0	0
	0	1	0	1	1	0	1	0	0	1	1	1	0	1	1	0	0	1
	0	0	0	0	0	0	0	1	1	0	0	0	0	0	0	0	1	0
	1	0	1	0	0	1	0	1	1	0	0	0	1	0	0	1	1	0
primer 引物	D19	D20	D22	D23	D24	D25	D26	D27	D28	D29	D30	D31	D32	D33	D34	D35	D36	D37
P2	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
	0	0	0	0	1	1	0	1	1	0	1	1	0	1	1	0	0	1
	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
P9	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
	0	0	0	0	1	0	0	1	0	0	0	0	0	1	1	1	1	0
	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
P11	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
	0	1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	0	0	0	0	1	1	0	1	1	1	0	1	1	0	0	0	1	1
	0	0	0	0	0	0	1	0	0	0	0	0	0	0	0	0	0	0
	1	0	1	1	0	0	1	0	0	0	1	0	0	1	1	1	0	0
primer 引物	D38	D39	D40	D41	D42	D43	D44	D46	D47	D49	D50	D51	D52	D53	D54	D55	D56	D59
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	1	0	0	1	0	0	1	0	0	0	1	0	1	0	1	0	1	0
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	0	0	1	0	0	1	1	1	1	0	0	1	1	0	1	1	0	1
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P11	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1
	0	0	0	0	0	0	0	0	0	0	0	0	0	1	0	0	0	0
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	0	1	0	1	0	1	1	0	1	0	1	1	0	0	0	1	1	1

基于SSR标记的丛生竹杂种鉴定、遗传分析和指纹图谱构建

SSR-based hybrid identification, genetic analyses and fingerprint development of hybridization progenies from sympodial bamboo (Bambusoideae, Poaceae)

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