基于ISSR分子标记的白鹤芋遗传多样性分析

郭甜馨; MuhammadMoaaz Ali; 颜喜玺; 李雪; 简丽观; 彭金彬; 陈歆妍; 赵锦河; 彭俊贤; 陈发兴

doi:10.19303/j.issn.1008-0384.2022.005.011

基于ISSR分子标记的白鹤芋遗传多样性分析

doi: 10.19303/j.issn.1008-0384.2022.005.011

1.
福建农林大学园艺学院，福建　福州　350002
2.
泉州市泉美生物科技有限公司，福建　泉州　362000
3.
泉州市洛江区林业局技术推广站，福建　泉州　362000
4.
泉州市洛江区市政公用事业管理中心，福建　泉州　362000
5.
安溪县林业局，福建　泉州　 362400
6.
泉州市洛江区自然资源局，福建　泉州　362000

基金项目: 现代设施农业福建省高等学校工程研究中心开放研究基金项目（G2- KF001)

详细信息

作者简介:
郭甜馨（2001−），女，研究方向：花卉与观赏园艺（E-mail：tianxinguo2022@163.com）

通讯作者:
陈发兴（1967−），男，博士，研究员，研究方向：园艺植物生理与分子生物学（E-mail： fxchen@fafu.edu.cn）

中图分类号: S 682.36
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- 文章访问数: 451
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- 被引次数: 0
出版历程
- 收稿日期: 2022-03-04
- 修回日期: 2022-05-26
- 网络出版日期: 2022-06-20
- 刊出日期: 2022-05-28

Genetic diversity analysis based on ISSR molecular markers in Spathiphyllum

1.
College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China
2.
Quanmei Biotechnology Company of Quanzhou City, Quanzhou, Fujian　362000, China
3.
Luojiang District Forestry Bureau Technology Extension Station of Quanzhou , Quanzhou Fujian　362000, China
4.
Municipal Public Utilities Management Center, Quanzhou Luojiang District , Quanzhou, Fujiian　362000, China
5.
Forestry Bureau of Anxi County, Quanzhou, Fujian　362400, China
6.
Luojiang District Natural Resources Bureau of Quanzhou City, Quanzhou, Fujian　362000, China

摘要

摘要: 目的探讨白鹤芋种质资源的亲缘关系及遗传多样性，为其开发、利用与保护提供理论依据。方法利用ISSR分子标记技术对14份栽培品种、7份杂种品系共21份白鹤芋种质资源的亲缘关系及遗传多样性进行分析。结果采用9条扩增清晰、多态性较好的ISSR引物对21份种质进行PCR扩增，共扩增出133个位点，其中多态性位点有120个，多态性比率（PPB）为90.23%；NTsys聚类分析表明14份栽培品种在相似系数为0.6692处可分为3组，7份杂交品系在相似系数为0.6700处可分为2组；应用TBtools相关性、主成分分析，在第一主坐标上14份栽培品种被分为2组，聚类分析结果准确。结论 ISSR分子标记技术可有效揭示白鹤芋种质资源的遗传多样性和品种间的亲缘关系；ISSR聚类结果与白鹤芋的种质来源、佛焰苞花序性状等吻合度较高；主成分分析印证聚类结果的同时，更直观显示出种质间的亲缘关系。
- 白鹤芋 /
- 种质资源 /
- ISSR标记 /
- 遗传多样性
Abstract: Objective In order to provide theoretical basis for the development, utilization and protection of Spathiphyllum germplasm resources, the genetic diversity and relationship of germplasm resources were discussed. Method ISSR markers were used to analyze the genetic relationship and genetic diversity of 21 Spathiphyllum germplasm resources from 14 cultivars and 7 hybrid strains. Result Nine clearly amplified and highly polymorphic ISSR primers were used for PCR amplification of 21 germplasm. A total of 133 loci were amplified, including 120 polymorphic loci, and the polymorphism ratio (PPB) was 90.23%. According to NTsys cluster analysis, the 14 cultivars could be divided into three groups at the similarity coefficient of 0.6692. The 7 hybrid strains could be divided into two groups at the similarity coefficient of 0.6700. Using TBtools correlation and principal component analysis, 14 cultivars were divided into two groups on the first principal coordinate, which proved the accuracy of cluster analysis results. Conclusion ISSR molecular markers could effectively reveal the genetic diversity and the genetic relationship between cultivars; the genetic diversity of the spathiphyllum is abundant. ISSR clustering results were in good agreement with the germplasm origin and the character of spathes; principal component analysis confirmed the clustering results and showed the genetic relationship between varieties more directly.
- Spathiphyllum /
- germplasm resources /
- ISSR marker /
- genetic diversity

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图 1 供试白鹤芋的佛焰苞

Figure 1. Spathes of the different cultivars of Spathiphyllum

下载: 全尺寸图片幻灯片

图 2 引物UBC895、899对21份白鹤芋种质的扩增电泳图谱

上为UBC899；下为UBC900。

Figure 2. Electrophoretic patterns of 21 Spathiphyllum germplasm samples amplified with primer UBC895 and UBC899

the upper is UBC895；the below is UBC899.

下载: 全尺寸图片幻灯片

图 3 14个白鹤芋栽培品种ISSR扩增条带的UPGMA聚类结果

Figure 3. UPGMA clustering results of ISSR amplification bands of 14 cultivars of Spathiphyllum

下载: 全尺寸图片幻灯片

图 4 14个白鹤芋栽培品种ISSR扩增条带的相关性分析、主成分分析

Figure 4. Correlation analysis and principal component analysis results of ISSR amplification bands of 14 cultivars

下载: 全尺寸图片幻灯片

图 5 7个杂交品系与亲本间ISSR扩增条带的UPGMA聚类分析

Figure 5. UPGMA clustering of ISSR amplification bands between 7 hybrid strains and their parents

下载: 全尺寸图片幻灯片

图 6 Z7 双苞片变异

Figure 6. Z7 double bract variation

下载: 全尺寸图片幻灯片

表 1 供试白鹤芋信息

Table 1. Spathiphyllum information of participants in the trial

序号 Number	代号 Code number	名称　　　 Name	来源　　 Source	类型 Types	植株紧凑性 Plant compactness	肉质花序颜色 Fleshy inflorescence color	肉质花序凸起 Convex appearance of fleshy inflorescence
1	SPAT01	娇小白掌 Spathiphyllum 'Petite'	中国广东 Guangdong，China	栽培种 Cultivar	高 High	浅黄 Slight yellow	明显 Clear
2	SPAT10	超级较小白掌 Spathiphyllum 'Super Petite'	美国 America	栽培种 Cultivar	中等 Middle	乳黄 Yellow	明显 Clear
3	SPAT24	小巨人 Spathiphyllum 'Dwarf Sensation'	中国广东 Guangdong，China	栽培种 Cultivar	中等 Middle	浅黄 Slight yellow	明显 Clear
4	SPAT29	科丽丝 Spathiphyllum 'Chris'	中国福建 Fujian，China	栽培种 Cultivar	高 High	乳黄 Yellow	极明显 Very clear
5	SPAT30	美酒 Spathiphyllum ‘Mojo'	中国福建 Fujian，China	栽培种 Cultivar	高 High	浅黄 Slight yellow	极明显 Very clear
6	SPAT33	克劳迪娅 Spathiphyllum 'Claudia'	美国 America	栽培种 Cultivar	高 High	浅黄 Slight yellow	极明显 Very clear
7	SPAT34	维客 Spathiphyllum ‘Vickily'	中国福建 Fujian，China	栽培种 Cultivar	高 High	乳黄 Yellow	极明显 Very clear
8	SPAT37	—	澳洲 Australia	栽培种 Cultivar	中等 Middle	浅黄 Slight yellow	明显 Clear
9	SPAT38	—	中国福建 Fujian，China	栽培种 Cultivar	高 High	浅黄 Slight yellow	不明显 Not clear
10	SPAT50	阳光之梦 Sunshine dream	SPAT12×SPAT03杂交种 Hybrid of SPAT12×SPAT03	杂交种 Hybrid	高 High	米白White	极明显 Very clear
11	SPAT51	阳光之恋 Sunshine love	SPAT12×SPAT14杂交种 Hybrid of SPAT12×SPAT14	杂交种 Hybrid	高 High	米白 White	极明显 Very clear
12	SPAT52	阳光之星 Sunshine star	SPAT18×SPAT12杂交种 Hybrid of SPAT18×SPAT12	杂交种 Hybrid	高 High	米白 White	明显 Clear
13	SPAT53	阳光明珠 Sunshine pearl	SPAT10×SPAT12杂交种 Hybrid of SPAT10×SPAT12	杂交种 Hybrid	中等 Middle	米白 White	明显 Clear
14	SPAT70	—	SPAT36×SPAT51杂交种 Hybrid of SPAT36×SPAT51	杂交种 Hybrid	中等 Middle	米白 White	不明显 Not clear
15	Z1	—	SPAT33×SPAT51杂交种 Hybrid of SPAT33×SPAT51	杂交品系 Hybrid strain	中等 Middle	浅黄 Slight yellow	极明显 Very clear
16	Z2	—	SPAT33×SPAT51杂交种 Hybrid of SPAT33×SPAT51	杂交品系 Hybrid strain	高 High	米白/绿色 White/Green	明显 Clear
17	Z3	—	SPAT33×SPAT51杂交种 Hybrid of SPAT33×SPAT51	杂交品系 Hybrid strain	高 High	米白 White	明显 Clear
18	Z4	—	SPAT33×SPAT51杂交种 Hybrid of SPAT33×SPAT51	杂交品系 Hybrid strain	中等 Middle	米白/绿色 White/Green	不明显 Not clear
19	Z5	—	SPAT33×SPAT51杂交种 Hybrid of SPAT33×SPAT51	杂交品系 Hybrid strain	高 High	绿色 Green	明显 Clear
20	Z6	—	SPAT33×SPAT51杂交种 Hybrid of SPAT33×SPAT51	杂交品系 Hybrid strain	高 High	绿色 Green	不明显 Not clear
21	Z7	—	SPAT33×SPAT51杂交种 Hybrid of SPAT33×SPAT51	杂交品系 Hybrid strain	高 High	绿色 Green	极明显 Very clear

下载: 导出CSV

表 2 筛选出的9条引物信息

Table 2. Information of nine selected primers

引物名称 Primer name	序列（5′—3′） Sequence (5′—3′)	T_m值 Melting temperature/℃	最佳退火温度 Annealing temperature/℃
UBC-866	CTCCTCCTCCTCCTCCTC	59.5	62.5
UBC-873	GACAGACAGACAGACA	49.2	44.9
UBC-876	GATAGATAGACAGACA	44.0	40.8
UBC-879	CTTCACTTCACTTCA	43.0	38.0
UBC-881	GGGTGGGGTGGGGTG	59.4	58.2
UBC-892	TAGATCTGATATCTGAATTCCC	52.1	47.1
UBC-895	AGAGTTGGTAGCTCTTGATC	53.4	49.1
UBC-899	CATGGTGTTGGTCATTGTTCCA	55.8	52.7
UBC-900	ACTTCCCCACAGGTTAACACA	55.6	52.5

下载: 导出CSV

表 3 9条引物多态性信息

Table 3. Polymorphism information of 9 primers

引物编号 Primer number	条带数 Number of binds/条	多态性条带数 Number of polymorphic bands/条	多态条带百分率 Percentage of polymorphic bands/%	多态信息含量 Polymorphism information content
UBC-866	11	9	81.82	0.835
UBC-873	15	13	86.67	0.905
UBC-876	8	7	87.50	0.827
UBC-879	15	12	80.00	0.908
UBC-881	14	13	92.86	0.893
UBC-892	16	16	100.00	0.902
UBC-895	16	14	87.50	0.908
UBC-899	18	16	88.89	0.909
UBC-900	20	20	100.00	0.938
总计 Total	133	120	90.23	—

下载: 导出CSV

参考文献(24)

[1]	于丽萍. 花叶俱美的室内植物: 白掌 [J]. 绿化与生活, 2002(2):28. YU L P. Spathiphyllum, an indoor plant with beautiful flowers and leaves [J]. Greening and Life, 2002(2): 28.(in Chinese)
[2]	戴桦均, 徐维杰, 廖飞雄. 不同白鹤芋种/品种开花特性和形态的观察[C]//中国观赏园艺研究进展, 2012: 148 − 153.
[3]	陈汉鑫, 万学锋, 康月惠, 等. 白掌新品种的引进与栽培管理 [J]. 亚热带植物科学, 2012, 41(2):62−64. doi: 10.3969/j.issn.1009-7791.2012.02.016 CHEN H X, WAN X F, KANG Y H, et al. The introduction of five varieties of Spathiphyllum [J]. Subtropical Plant Science, 2012, 41(2): 62−64.(in Chinese) doi: 10.3969/j.issn.1009-7791.2012.02.016
[4]	来伊楠, 陈波, 卢山. 天南星科室内观赏植物对苯的净化研究 [J]. 浙江理工大学学报, 2015, 33(3):280−284. LAI Y N, CHEN B, LU S. Study on purification of benzene in indoor air by Araceae ornamental plants [J]. Journal of Zhejiang Sci-Tech University, 2015, 33(3): 280−284.(in Chinese)
[5]	MAYO S J, BOGNER J, BOYCE P C. The genera of Araceae[M]. Royal Botanic Gardens Kew, UK, 1997: 110.
[6]	HENNY R J, CHEN J J. Cultivar development of ornamental foliage plants[M]//Plant Breeding Reviews. Oxford, UK: John Wiley & Sons, Inc. , 2010: 245-290.
[7]	NORMAN R J D J, CHEN J. Progress in ornamental aroid breeding research [J]. Annals of the Missouri Botanical Garden, 2004, 91(3): 465−473.
[8]	ZIETKIEWICZ E, RAFALSKI A, LABUDA D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification [J]. Genomics, 1994, 20(2): 176−183. doi: 10.1006/geno.1994.1151
[9]	钱韦, 葛颂, 洪德元. 采用RAPD和ISSR标记探讨中国疣粒野生稻的遗传多样性 [J]. 植物学报, 2000, 42(7):741−750. QIAN W, GE S, HONG D Y. Assessment of genetic variation of Oryza granulata detected by RAPDs and ISSRs [J]. Acta Botanica Sinica, 2000, 42(7): 741−750.(in Chinese)
[10]	马国华, 朱西儒, 张云开, 等. 液体振荡培养高速繁殖白鹤芋丛芽 [J]. 园艺学报, 1993, 20(3):307−308. doi: 10.3321/j.issn:0513-353X.1993.03.013 MA G H, ZHU X R, ZHANG Y K, et al. High speed propagation of Spathiphyllum shoots in liquid shake culture [J]. Acta Horticulturae Sinica, 1993, 20(3): 307−308.(in Chinese) doi: 10.3321/j.issn:0513-353X.1993.03.013
[11]	朱根发. 白鹤芋属观赏植物的组织培养和快速繁殖技术研究 [J]. 中国农学通报, 2003, 19(3):75−76,88. doi: 10.3969/j.issn.1000-6850.2003.03.023 ZHU G F. Tissue culture and propagation of Spathiphyllum [J]. Chinese Agricultural Science Bulletin, 2003, 19(3): 75−76,88.(in Chinese) doi: 10.3969/j.issn.1000-6850.2003.03.023
[12]	李银华. 白鹤芋规模化生产技术 [J]. 北方园艺, 2012(18):104−105. LI Y H. Large-scale production technologies of Spathiphyllum kochii [J]. Northern Horticulture, 2012(18): 104−105.(in Chinese)
[13]	詹启成, 蒋雄辉, 王奎, 等. 不同浓度赤霉素对甜芝白掌催花效果的影响 [J]. 北方园艺, 2013(8):83−84. ZHAN Q C, JIANG X H, WANG K, et al. Influence of different concentrations of gibberellin acid on flower stimulation of Spathiphyllum ‘sweet chico’ [J]. Northern Horticulture, 2013(8): 83−84.(in Chinese)
[14]	刘小飞, 廖飞雄, 李冬梅, 等. 基于SRAP标记的白鹤芋属资源鉴定与亲缘关系分析 [J]. 广东农业科学, 2017, 44(11):26−31. LIU X F, LIAO F X, LI D M, et al. Genetic relationship analysis and cultivar identification of Spathiphyllum using SRAP marker [J]. Guangdong Agricultural Sciences, 2017, 44(11): 26−31.(in Chinese)
[15]	刘小飞, 孙映波, 黄丽丽, 等. 白掌SSR引物筛选及12份种质资源亲缘关系分析[J/OL]. 分子植物育种, 2021, 1(16). https://kns.cnki.net/kcms/detail/46.1068. S. 20201224.0858.002.html. LIU X F, SUN Y B, HUANG L L, et al. Screening of SSR primers and genetic relationship analysis of 12 germplasm resources of Spathiphyllum[J/OL]. Molecular Plant Breeding, 2021, 1(16). https://kns.cnki.net/kcms/detail/46.1068.S.20201224.0858.002.html.
[16]	李金璐, 王硕, 于婧, 等. 一种改良的植物DNA提取方法 [J]. 植物学报, 2013, 48(1):72−78. doi: 10.3724/SP.J.1259.2013.00072 LI J L, WANG S, YU J, et al. A modified CTAB protocol for plant DNA extraction [J]. Chinese Bulletin of Botany, 2013, 48(1): 72−78.(in Chinese) doi: 10.3724/SP.J.1259.2013.00072
[17]	黄敏, 黄旭萍, 李斌奇, 等. 枫香ISSR-PCR反应体系的优化及引物筛选[J/OL]. 分子植物育种, 2019, 1(12). https://kns.cnki.net/kcms/detail/46.1068.s.20210429.1428.010.html. HUANG M, HUANG X P, LI B Q, et al. Optimization of Liquidambar formosana ISSR-PCR reaction system and primer selection[J/OL]. Molecular Plant Breeding, 2019, 1(12). https://kns.cnki.net/kcms/detail/46.1068.s.20210429.1428.010.html.
[18]	张晨光. 太白山独叶草的小尺度分布成因及其克隆结构、遗传多样性研究[D]. 西安: 西北大学, 2018. ZHANG C G. The study on the distribution at small-scale, clonal structure and genetic diversity of Kingdonia uniflora on Taibai Mountain[D]. Xi'an: Northwest University, 2018. (in Chinese)
[19]	丁刘慧子, 邳植, 吴则东. 甜菜品种SSR指纹图谱的构建及遗传多样性分析 [J]. 作物杂志, 2021(5):72−78. DING L H Z, PI Z, WU Z D. Construction of SSR fingerprint and analysis of genetic diversity of sugar beet varieties [J]. Crops, 2021(5): 72−78.(in Chinese)
[20]	何予卿, 张宇, 孙梅, 等. 利用ISSR分子标记研究栽培稻和野生稻亲缘关系 [J]. 农业生物技术学报, 2001, 9(2):123−127. doi: 10.3969/j.issn.1674-7968.2001.02.005 HE Y Q, ZHANG Y, SUN M, et al. Studies on the relationship between cultivated and wild rice using ISSR markers [J]. Journal of Agricultural Biotechnology, 2001, 9(2): 123−127.(in Chinese) doi: 10.3969/j.issn.1674-7968.2001.02.005
[21]	岑彩娴. 白掌品种的观赏特性评价和杂交技术研究[D]. 广州: 华南农业大学, 2016. CEN C X. Ornamental characteristics evaluation and hybridization technique's study of Spathiphyllum floribundum[D]. Guangzhou: South China Agricultural University, 2016. (in Chinese)
[22]	刘小飞, 廖飞雄, 詹启成. 白掌新品种银心梭的选育 [J]. 热带农业科学, 2021, 41(11):50−53. LIU X F, LIAO F X, ZHAN Q C. Breeding and selection of a new Spathiphyllum variety ‘Yinxinsuo’ [J]. Chinese Journal of Tropical Agriculture, 2021, 41(11): 50−53.(in Chinese)
[23]	刘金梅, 刘小飞, 于波, 等. 不同白掌组合授粉结实和种子特性研究 [J]. 热带作物学报, 2015, 36(7):1213−1219. doi: 10.3969/j.issn.1000-2561.2015.07.005 LIU J M, LIU X F, YU B, et al. Fruit-setting and seed characteristics of Spathiphyllum with different pollination combinations [J]. Chinese Journal of Tropical Crops, 2015, 36(7): 1213−1219.(in Chinese) doi: 10.3969/j.issn.1000-2561.2015.07.005
[24]	周辉明, 林辉锋, 莫智龙, 等. 四倍体白掌新品种‘绿萌’的选育 [J]. 热带作物学报, 2022, 43(5):964−970. ZHOU H M, LIN H F, MO Z L, et al. Breeding a new tetraploid Spathiphyllum floribundum ‘lyumeng' variety [J]. Chinese Journal of Tropical Crops, 2022, 43(5): 964−970.(in Chinese)