Genetic diversity and genetic structure of Tilia miqueliana population

doi:10.12302/j.issn.1000-2006.202110049

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2023, Vol. 47 ›› Issue (1): 145-153.doi: 10.12302/j.issn.1000-2006.202110049

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Genetic diversity and genetic structure of Tilia miqueliana population

WANG Huanli(), YAN Lingjun, HUANG Xi, WANG Zhongwei, TANG Shijie()

Institute of Botany Jiangsu Province and Chinese Academy of Science,Jiangsu Key Laboratory for the Research and Utilization of Plant Resources, Nanjing 210014, China

Received:2021-10-25 Accepted:2021-11-29 Online:2023-01-30 Published:2023-02-01
Contact: TANG Shijie E-mail:wanghuanli@cnbg.net;tangshijie69@aliyun.com

Abstract

Abstract:

【Objective】 Tilia miqueliana is an important native tree species in Jiangsu Province, and its wild germplasm resources are particularly scarce. Exploration of the genetic diversity and the genetic structure of T. miqueliana in the field population provides the scientific basis for its resource protection, variety breeding and genetic improvement.【Method】 Fifteen polymorphic EST-SSR primers were used to analyze the genetic diversity and population structure of 93 individuals from five natural populations of T. miqueliana located on Baohua Mountain (Jiangsu Province, P1), Niushou Mountain (Jiangsu Province, P2), Huangcangyu Nature Reserve (Anhui Province, P3), Shu Mountain (Anhui Province, P4) and Tiantai Mountain (Zhejiang Province, P5). 【Result】 (1) A total of 96 alleles (A) were detected using 15 EST-SSR primer pairs. The average allelic richness at a locus was 6.4. The total number of tetraploid genotypic richness (G) and tetraploid specific genotypes (G_i) were 441 and 251, respectively. The average of the specific genotype ratio (R₁) and the germplasm identification rate (R₂) were 45.73% and 17.99%, respectively. (2) In the five populations, the mean number of alleles per locus (A_loc) and the tetraploid genotypic richness (G_loc) were 5.50 ± 2.43 and 9.41 ± 4.29, respectively. The mean observed heterozygosity (H_o) and the mean expected heterozygosity (H_e) were 0.61 ± 1.43 and 0.62 ± 0.14, respectively. Based on the values for G_loc and H_e, the P1 and P3 populations were identified as having high genetic diversity. (3) The genetic differentiation coefficient (G_st) was 0.030, indicating a low genetic differentiation among the five populations. Analysis of molecular variance (AMOVA) showed that the variation in genetic diversity predominantly came from within the population (96%). (4) Cluster analysis and genetic structure analysis by structure showed that the five tested populations could be divided into two groups (Group1. P1, P2 and P5; Group2. P3 and P4). The Mantel test results showed that there was no significant correlation between the genetic differentiation and the geographic distance. 【Conclusion】 High levels of genetic diversity were found within the five populations of T. miqueliana, among which Baohua Mountain population and Huangcangyu Nature Reserve population showed considerably higher genetic diversity than that the other populations showed. Combined with the results from the clustering and genetic structure analysis, we can infer that population expansion may have been centered on these two populations and migrated to other regions through human activities. There was no pronounced genetic differentiation among the different geographic populations, which may be due to the long life of individuals, the lack of natural regeneration of populations, and seed dispersal among populations through human activities. Therefore, we propose to establish isolation areas, identify priority populations for protection, increase plant outcrossing, and use artificial breeding and germplasm regeneration to protect the wild populations.

Key words: Tilia miqueliana, SSR marker, genetic structure, genetic diversity

CLC Number:

S722

WANG Huanli, YAN Lingjun, HUANG Xi, WANG Zhongwei, TANG Shijie. Genetic diversity and genetic structure of Tilia miqueliana population[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2023, 47(1): 145-153.

Figures/Tables 10

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种群编号 population No.	采集地点 location of collection	东经/ (°) longitude (E)	北纬/ (°) latitude (N)	个体数 sample size
P1	江苏宝华山 Baohua Mountain, Jiangsu	119.08	32.13	15
P2	江苏牛首山 Niushou Mountain, Jiangsu	118.75	31.90	16
P3	安徽皇藏峪 Huangcangyu, Anhui	117.07	34.02	33
P4	安徽蜀山 Shu Mountain, Anhui	117.18	31.84	17
P5	浙江天台山 Tiantai Mountain, Zhejiang	121.03	29.16	12

编号 No.	引物序列(5'→3') primer squence	产物长度/bp length of products	退火温度/℃ annealing tempreature	荧光标记 flourescent label
B1045	F: ACGGCTCCCTTATCACACTG R: TGCCATCTCACGTAGCTTTG	280	56.5	5'-FAM
B2405	F: TCTGCAAAGGAACTCTTCATCA R: TGGATTGCTAGTTGTCAAGCAT	218	54.0	5'-HEX
B2410	F: TATAGATGCCCGTTCCAAGC R: CATCAAGACAAGGGCAATGA	195	52.5	5'-HEX
B2550	F: ATTGTGCTGTGGTGGCATTA R: TACCCCTGCATGTTTTCCAT	226	53.0	5'-HEX
B2915	F: CGACCTGGTTCTCTGGTTTC R: CGACCAGGTGCATATAGCCT	268	54.5	5'-FAM
B485	F: TGGATGGTGATGGTAGGGTT R: GTGTGTCAACTGGCAGCAAT	278	54.0	5'-FAM
B75	F: GGAGAGAGAAAGGCAGAGCA R: TAAGGTGGTGATGACGTTGG	189	54.5	5'-TAMRA
B90	F: AGAGCAGAGCCGTTGGATAA R: GGAGAGAGAAAGGCAGAGCA	193	54.0	5'-FAM
C1050	F: ACCTGGAGGCTCTGGACTT R: ACACAGAGGTTCGGAAAATCG	259	54.0	5'-FAM
C2155	F: CGGTGTGTCGACATTGTTTC R: CTCTGGAAGAAGGGAACACG	219	53.5	5'-HEX
C2700	F: GCCAGCCATGTTTTGTTTCT R: CGTTCAATGGCCTCTTCATT	181	58.0	5'-FAM
C3235	F: GTTAGGAGCAATGATGCCGT R: CCACAGTCGCATCTTCTTCA	162	57.0	5'-FAM
E5	F: TTGCTGTTTCATTCTGCTGG R: AGTTTCGAATAGCGCCTTGA	178	53.0	5'-TAMRA
Tc11	F:AGCTATGAAAGAACTATCAAGAGAAAG R: CCCCAAGACATTGCAGTAGAAC	146	55.0	5'-TAMRA
Tc8	F: CGAAGAAACTGTCAAAACAACG R: AGCTGGGTTTTAGAGGATAGGG	160	58.0	5'-FAM

引物名称 primer	A	A_i	G	G_i	R₁/%	R₂/%	C_PIC	I
C1050	4	2	13	7	53.85	7.53	0.660 2	1.456 7
C2155	4	3	13	2	15.38	2.15	0.814 0	2.015 6
C2700	3	2	13	3	23.08	3.23	0.801 2	1.968 8
C3235	12	3	62	42	67.74	45.16	0.978 8	3.997 7
E05	4	2	15	4	26.67	4.30	0.829 5	2.189 8
Tc8	3	2	8	2	25.00	2.15	0.740 9	1.557 4
Tc11	6	2	33	13	39.39	13.98	0.943 7	3.162 9
B2250	6	2	21	8	38.10	8.60	0.888 4	2.569 1
B1045	4	2	18	5	27.78	5.38	0.888 4	2.472 9
B485	10	3	65	50	76.92	53.76	0.969 4	3.941 7
B2405	7	3	49	30	61.22	32.26	0.968 4	3.667 9
B75	9	3	43	28	65.12	30.11	0.914 1	3.185 3
B291 5	6	2	29	13	44.83	13.98	0.894 7	2.789 1
B2410	9	2	31	19	61.29	20.43	0.820 9	2.562 6
B90	9	3	42	25	59.52	26.88	0.952 7	3.387 8
总计total	96	—	441	251	—	—	—	—
均值mean	6.4	2.3	30.3	17	45.73	17.99	0.871 0	2.728 4

群体 population	N	A_l	A_loc	G_loc	H_o	H_e
P1	15	5.73±2.52	2.35±0.43	9.60±3.85	0.64±0.15	0.65±0.11
P2	16	5.27±2.43	2.34±0.38	8.33±4.66	0.63±0.16	0.63±0.15
P3	33	5.87±2.67	2.25±0.43	13.20±6.54	0.60±0.14	0.62±0.14
P4	17	5.20±2.54	2.23±0.44	9.20±3.14	0.59±0.15	0.61±0.14
P5	12	5.00±2.00	2.28±0.51	6.73±3.28	0.59±0.20	0.58±0.19
均值mean	18.6	5.50±2.43	2.29±0.44	9.41±4.29	0.61±1.43	0.62±0.14

引物名称 primer name	H_t	H_s	D_st	G_st
C1050	0.433	0.385	0.048	0.110
C2155	0.628	0.621	0.007	0.012
C2700	0.579	0.578	0.001	0.002
C3235	0.778	0.769	0.009	0.011
E5	0.597	0.567	0.030	0.051
Tc8	0.521	0.515	0.006	0.011
Tc11	0.686	0.669	0.017	0.024
B2250	0.514	0.491	0.023	0.045
B1045	0.681	0.647	0.034	0.050
B485	0.841	0.811	0.030	0.036
B2405	0.788	0.772	0.016	0.020
B75	0.756	0.746	0.010	0.013
B2915	0.540	0.520	0.020	0.036
B2410	0.425	0.434	0.011	0.026
B90	0.740	0.739	0.001	0.002
均值mean	0.634	0.616	0.018	0.030

Genetic diversity and genetic structure of Tilia miqueliana population

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变异来源 source of variation	自由度 df	平方和 sum of square	变异组成 variance component	变异百分比/% percentage of variation	P_hi_PT
群体间 among populations	4	91.206	0.557	4	0.042
群体内 within populations	88	1 130.515	12.847	96
总变异 total	92	1 221.720	13.404	100