Using the SSR fluorescent labling to establish SSR fingerprint for 131 cultivars in Bougainvillea (Nyctaginaceae)

doi:10.3969/j.issn.1000-2006.201812045

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2019, Vol. 43 ›› Issue (6): 60-66.doi: 10.3969/j.issn.1000-2006.201812045

Using the SSR fluorescent labling to establish SSR fingerprint for 131 cultivars in Bougainvillea (Nyctaginaceae)

SUN Lina¹^,²(), ZHOU Qun³, HUANG Meiping², ZHOU Changpin¹, XU Shisong⁴, YANG Shuting², DU Ling², GAN Siming¹^,²^,^*(), TANG Qing²^,^*()

1. Key Laboratory of State Forestry Administration on Tropical Forestry Research, Research Institute of Tropical Forestry, Chinese Academy of Forestry, Guangzhou 510520, China
2. Guangxi Key Laboratory of Superior Timber Trees Resource Cultivation & Key Laboratory of Central South Fast-growing Timber Cultivation of Forestry Ministry of China, Guangxi Forestry Research Institute, Nanning 530002, China
3. Xiamen Botanical Garden, Xiamen 361003, China
4. Tropical Crops Genetic Resources Institute of Chinese Academy of Tropical Agricultural Sciences, Danzhou 571737, China

Received:2018-12-26 Revised:2019-06-24 Online:2019-11-30 Published:2019-11-30
Contact: GAN Siming,TANG Qing E-mail:sunlina8958170@126.com;Siming_Gan@126.com;335221247@qq.com

Abstract

Abstract:

【Objective】 Find a reliable molecular marker to identifyBougainvillea cultivars that are difficult to distinguish, due to cultivar richness and cultivar homonymy and synonymy. 【Method】 A total of 15 simple sequence repeats (SSR) markers were used to amplify DNA samples from 131 Bougainvillea cultivars. SSR amplicons were detected by a capillary electrophoresis method. Cultivar polymorphism and genetic distance were calculated; they were used for clustering analysis and fingerprint construction of all the 131 cultivars.【Result】A total of 85 different alleles were collectively generated by the 15 SSR loci, averaging 5.60 alleles per locus. Shannon’s information index, observed heterozygosity value, and expected heterozygosity value ranged from 0.38 to 1.53, 0.11 to 0.94 and 0.16 to 0.73, with an average of 1.04, 0.50 and 0.57 per locus, respectively. The polymorphism information content ranged from 0.15 to 0.69, with an average of 0.51 per locus. The genetic distance between cultivars ranged from 0.00 to 0.65, with an average of 0.33. Clustering analysis showed that most of the cultivars within a species tended to fall in the same cluster, but some cultivars of the same, whereas certain cultivars from different species were grouped in the same cluster. Several pairs of cultivars were either homonyms or synonyms. Only 11 SSR loci were shown to be effective for identifying the 131 cultivars; thus, the molecular fingerprints of all these cultivars were constructed based on allelic combinations of the 11 markers. 【Conclusion】Further research should be conducted to verify either the Chinese or Latin names of some suspicious cultivars. SSR-based fingerprints ofBougainvillea cultivars constitute a reliable technique for cultivar identification, registration, and intellectual property protection as well as for cultivar clarification in production practices.

Key words: Bougainvillea, cultivar classitication, SSR, fingerprints, genetic relationship

CLC Number:

SUN Lina, ZHOU Qun, HUANG Meiping, ZHOU Changpin, XU Shisong, YANG Shuting, DU Ling, GAN Siming, TANG Qing. Using the SSR fluorescent labling to establish SSR fingerprint for 131 cultivars in Bougainvillea (Nyctaginaceae)[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2019, 43(6): 60-66.

Figures/Tables 3

Table 1

The 131 Bougainvillea cultivars tested in this study "

序号 No.	品种 cultivar	序号 No.	品种 cultivar	序号 No.	品种 cultivar
1	‘青叶紫’(‘小叶浅紫’) (B. glabra ‘Elizabeth Angus’)	45	‘斑叶小叶紫’(B. × buttiana ‘Sanderiana Variegata’)	89	‘粉蝶’(B. sp. ‘Feathery Fantasy Double Colour’)
2	‘粉粧’(B. glabra ‘Eva’)	46	‘宝花橙’(B. × buttiana ‘Louise Wathen’)	90	‘红蝶’(B. sp. ‘Ratana Pink’)
3	‘黄金叶’(B. glabra ‘Golden Lady’)	47	‘彩虹黄’(B. × buttiana ‘Yellow Wonder’)	91	‘红莲花’(B. sp. ‘Red Lotus’)
4	‘茄色’(B. glabra ‘Merial Fitzpartick’)	48	‘橙冰’(B. × buttiana ‘Louise Wathen Variegata’)	92	‘黄金大奖’(B. sp. ‘MonSam’)
5	‘塔紫’(B. glabra ‘Pink Pixie’)	49	‘红粉佳人’(B. × buttiana ‘Alick Lancaste’)	93	‘绿叶玉宝’(B. sp. ‘Ladybird’)
6	‘水红’(B. glabra ‘Rosa’)	50	‘加州黄金’(B. × buttiana ‘California Gold’)	94	‘蒙娜丽莎黄’(B. sp. ‘Mona Lisa Yellow’)
7	‘光叶斑叶紫花’(B. glabra ‘Variegata’)	51	‘金边叶重粉’(B. × buttiana ‘Los Banos Variegata’)	95	‘孟加拉红’(B. sp. ‘Royal Bengal Red’)
8	‘橙红’(B. spectabilis ‘Auratus’)	52	‘金发女郎’(B. × buttiana ‘Blondie’)	96	‘热带花’(B. sp. ‘Tropical Bouquet’)
9	‘胭脂红’(B. spectabilis ‘China Beauty’)	53	‘蓝月亮’(B. × buttiana ‘Blue Moon’)	97	‘塔黄’(B. sp. ‘Pixie Yellow’)
10	‘大花深红’(B. spectabilis ‘Crimonlake’)	54	‘绿叶橙’(B. × buttiana ‘Afterglow’)	98	‘蜥蜴橙’(B. sp. ‘Lguana Variegata’)
11	‘大花深紫’(B. spectabilis ‘Senjakala’)	55	‘绿叶柠檬黄’(B. × buttiana ‘Golden Glow’)	99	‘小粉雀’(B. sp. ‘Firecracker Purple’)
12	‘樱花’(B. × spectoglabra ‘Imperial Delight’)	56	‘水红’(B. × buttiana ‘Miss Manila’)	100	‘小金雀’(B. sp. ‘Firecracker Yellow’)
13	‘金心’(B. × spectoglabra ‘Thimma’)	57	‘香港丽人’(B. × buttiana ‘Hong Kong Beauty’)	101	‘银边叶浅紫’(B. sp. ‘Pixie Orange’)
14	‘皱叶深红’(B. × buttiana ‘Barbara Karst’)	58	‘小花大红’(B. × buttiana ‘Mrs. Butt’)	102	‘暗斑叶加州黄金’(B. sp.)
15	‘暗斑叶橙红’(B. × buttiana ‘Lateritia’)	59	‘猩红皇后’(B. × buttiana ‘San Diego Red’)	103	‘冰斑美人’(B. sp.)
16	‘柠檬黄’(B. × buttiana ‘Mrs McLean’)	60	‘怡锦’(B. × buttiana ‘Cherry Blossom’)	104	‘波依斯玫瑰’(B. sp.)
17	‘重瓣大红’(B. × buttiana ‘Pretoria’)	61	‘印度画报’(B. × buttiana ‘Chitra’)	105	‘大蜜叶精灵’(B. sp.)
18	‘斑叶塔紫’(B. × buttiana ‘Temple Fire’)	62	‘重斑粉’(B. × buttiana ‘Los Banos Beauty’)	106	‘红宝石’(B. sp.)
19	‘福摩斯’(B. glabra ‘Formosa’)	63	‘重瓣橙’(B. × buttiana ‘Roseville’s Delight’)	107	‘红粉夫人’(B. sp.)
20	‘安格斯’(B. glabra ‘Elizabeth Angus’)	64	‘重瓣红’(B. × buttiana ‘Mahara’)	108	‘红九月’(B. sp.)
21	‘暗斑叶白’(B. glabra ‘Bambino Beesnees’)	65	‘重瓣黄’(B. × buttiana ‘Aussie Gold’)	109	‘红叶大红’(B. sp.)
22	‘白雪公主’(B. glabra ‘Alba’)	66	‘重瓣枣红’(B. × buttiana )	110	‘金边小叶紫’(B. sp.)
23	‘斑叶怡锦’(B. glabra ‘Mrs. Eusenia Raja Singhe’)	67	‘重瓣白’(B. × buttiana )	111	‘金龟紫’(B. sp.)
24	‘光叶紫’(B. glabra ‘Sanderiana’)	68	‘帕塔’(B. peruviana ‘Partha’)	112	‘金叶浅粉’(B. sp.)
25	‘金边安格斯’(B. glabra ‘Elizabeth Angus Variegata’)	69	‘斑叶印度橙粉’(B. peruviana ‘Partha Variegata’)	113	‘拉菲泰’(B. sp.)
26	‘金边叶浅紫’(B. glabra ‘Mrs. Eva Variegata Mauve II’)	70	‘红心樱花’(B. peruviana ‘Makris’)	114	‘绿叶浅黄’(B. sp.)
27	‘金叶白’(B. glabra ‘JinYu’)	71	‘金双色’(B. peruviana ‘Thimma’)	115	‘浅色樱花’(B. sp.)
28	‘哭泣的美人’(B. glabra ‘Variegata’)	72	‘口红’(B. peruviana ‘Begum Sikander’)	116	‘仨斑叶紫’(B. sp.)
29	‘柳叶白’(B. glabra ‘Shweta’)	73	‘绿叶宫粉’(B. peruviana ‘Mrs. H.C. Buck’)	117	‘洒金叶橙’(B. sp.)
30	‘绿叶浅紫’(B. glabra ‘Mrs. Eva’)	74	‘蒙娜丽莎’(B. peruviana ‘Mona Lisa’)	118	‘沙瓣叶艳红’(B. sp.)
31	‘塔红’(B. glabra ‘Pink Pixie’)	75	‘洋红公主’(B. peruviana ‘Mrs. H.C. Buck’)	119	‘沙瓣叶紫’(B. sp.)
32	‘小叶紫’(B. glabra ‘Royal Purple’)	76	‘印度橙粉’(B. peruviana ‘Partha’)	120	‘莎莎’(B. sp.)
33	‘新加坡白’(B. glabra ‘Singapore White’)	77	‘樱花’(B. peruviana ‘Imperial Delight’)	121	‘索菲亚’(B. sp.)
34	‘新加坡粉’(B. glabra ‘Singapore Beauty’)	78	‘火焰’(B. spectabilis ‘Flame’)	122	‘顽皮’(B. sp.)
35	‘雪紫’(B. glabra ‘Snow Purple’)	79	‘毛叶’(B. spectabilis ‘Splendens’)	123	‘小花粉紫’(绿叶) (B. sp.)
36	‘牙买加白’(B. glabra ‘Jamaica White’)	80	‘玫红’(B. spectabilis ‘B.J.F. Bailey’)	124	‘小叶小红粉’(B. sp.)
37	‘胭脂红’(B. glabra ‘Zinia Barat’)	81	‘热火桑巴’(B. spectabilis ‘Flame’)	125	‘异叶红’(B. sp.)
38	‘银边叶白’(B. glabra ‘Mrs. Eva Variegata White’)	82	‘小红雀’(B. spectabilis ‘Tomato Red’)	126	‘银河’(B. sp.)
39	‘银河’(B. glabra ‘Galaxy’)	83	‘婴儿玫瑰’(B. spectabilis ‘Baby Rose’)	127	‘银沙丽人’(B. sp.)
40	‘紫蝶’(B. glabra ‘Mrs. Eva Purple Ratana’)	84	‘砖红’(B. spectabilis ‘Lateritia’)	128	‘杂斑叶大红’(B. sp.)
41	‘紫丁香美女’(B. glabra ‘Lilac Beauty’)	85	‘暗斑叶樱花’(B. sp. ‘Blushing Beauty’)	129	‘杂斑叶水红’(B. sp.)
42	‘暗斑叶橙’(B. × buttiana ‘Jinda Orange’)	86	‘斑叶塔橙’(B. sp. ‘Pixie Orange Variegata’)	130	‘紫叶布丁’(B. sp.)
43	‘暗斑叶红’(B. × buttiana ‘Juanita Hatten’)	87	‘橙灯笼’(B. sp. ‘Chinese Lantern’)	131	‘总统紫’(B. sp.)
44	‘巴特夫人’(B. × buttiana ‘Mrs. Butt’)	88	‘橙蝶’(B. sp. ‘Ratana Orange’)

Table 1

Table 2

Fig.1

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EST-SSR引物 EST-SSR primer	N_a	N_e	I	H_o	H_e	PIC
BOUeSSR01	8.00	3.36	1.42	0.91	0.70	0.65
BOUeSSR02	3.00	1.98	0.76	0.15	0.49	0.39
BOUeSSR03	5.00	2.37	1.01	0.37	0.58	0.49
BOUeSSR04	9.00	3.63	1.53	0.49	0.73	0.69
BOUeSSR05	7.00	3.34	1.39	0.44	0.70	0.66
BOUeSSR06	6.00	2.50	1.07	0.84	0.60	0.53
BOUeSSR08	4.00	3.08	1.16	0.94	0.68	0.61
BOUeSSR09	4.00	2.33	0.96	0.78	0.57	0.48
BOUeSSR10	4.00	1.83	0.78	0.16	0.45	0.39
BOUeSSR11	7.00	2.28	1.12	0.58	0.56	0.51
BOUeSSR12	4.00	1.90	0.70	0.14	0.47	0.37
BOUeSSR13	5.00	1.19	0.38	0.11	0.16	0.15
BOUeSSR14	4.00	2.91	1.10	0.84	0.66	0.58
BOUeSSR15	6.00	2.59	1.15	0.60	0.61	0.56
BOUeSSR16	9.00	2.58	1.15	0.20	0.61	0.54
均值mean	5.60	2.52	1.04	0.50	0.57	0.51

Using the SSR fluorescent labling to establish SSR fingerprint for 131 cultivars in Bougainvillea (Nyctaginaceae)

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