Biodiversity Science ›› 2013, Vol. 21 ›› Issue (5): 601-609.doi: 10.3724/SP.J.1003.2013.09098

• Orginal Article • Previous Article     Next Article

Genetic diversity and genetic structure of the rare and endangered species, Primula ranunculoides

Deyun Wang1, Jie Peng1, Yajing Chen1, Guosheng Lü1, Xiaoping Zhang1, 2, Jianwen Shao1, 3, *()   

  1. 1 College of Life Science, Anhui Normal University, Wuhu, Anhui 241000
    2 Key Laboratory of Biotic Environment and Ecological Safety in Anhui Province, Anhui Normal University, Wuhu, Anhui 241000
    3 Key Laboratory of Conservation and Employment of Biological Resources of Anhui, Wuhu, Anhui 241000
  • Received:2013-04-18 Accepted:2013-07-01 Online:2013-10-08
  • Shao Jianwen E-mail:shaojw@mail.ahnu.edu.cn

The rare and endangered species Primula ranunculoides is endemic to China and is of reasonably high potential horticultural value. The genetic diversity and structure of the species was examined by surveying variation at nine microsatellite loci across 222 individuals sampled from seven natural populations. The results indicated that there was relatively low genetic diversity present within populations (He = 0.330, Ho = 0.286) and high genetic differentiation among populations. Levels of population genetic diversity and effective population size were both correlated with plant density, rather than population census size or population area. Gene flow between populations was low (Nm = 0.730) and an AMOVA analysis showed that 48.08% of the total genetic diversity was attributable to among populations and the rest (51.92%) to variation within populations. Bayesian assignment and principal coordinate analyses supported clustering of the seven populations into four groups, which were correlated with topographical features. This suggested that Lianghu Plain (Dongtinghu Plain and Jianghan Plain) and the farming area of Xiushui and Wuning counties were major barriers to gene flow, causing high divergence between different mountain populations. Based on the genetic structure of P. ranunculoides, four management units for conservation purposes are proposed, i.e. the Jiulingshan, Mufushan, Qizimeishan and Yinlu units.

Key words: Primula ranunculoides, genetic diversity, genetic structure, microsatellites, conservation proposal

Table 1

Locality and information of studied populations of Primula ranunculoides"

种群
Population
地点
Locality
生境
Habitat
地理位置
Geographical location
海拔
Altitude
(m)
面积
Area (m2)
花朵类型
Flower type
种群大小
Population size
杨家坪A YJPA 江西省修水县杨家坪
Yangjiaping, Xiushui County, Jiangxi
林下溪边或路边
Brook-side or roadside under forests
28°47'45"N, 114°43'56"E 300-375 720 两型花
Distyle
~2,000
杨家坪B YJPB 江西省修水县杨家坪
Yangjiaping, Xiushui County, Jiangxi
林下溪边或路边
Brook-side or roadside, under forest
28°47'04"N, 114°44'13"E 350-360 1,060 两型花
Distyle
~6,000
毛竹山
MZS
江西省修水县毛竹山
Maozhushan, Xiushui County, Jiangxi
林下溪边或路边
Brook-side or roadside, under forest
28°50'31"N, 114°52'00"E 390-410 1,650 两型花
Distyle
~1,500
三界A
SJA
湖北省通山县三界
Sanjie, Tongshan County, Hubei
林下溪边或路边
Brook-side or roadside, under forest
29°24'16"N, 114°27'15"E 255-285 545 两型花
Distyle
~1,000
三界B
SJB
湖北省通山县三界
Sanjie, Tongshan County, Hubei
林下溪边或路边
Brook-side or roadside, under forest
29°22'57"N, 114°27'08"E 265-280 385 两型花
Distyle
~700
七姊妹山
JPT
湖北省宣恩县七姊妹山
Qizimei Mount, Xuanen County, Hubei
林下溪边
Brook-side, under forest
30°02'09"N, 109°43'57"E 1,310-1,365 1,200 同型花
Monostyle
~600
银炉
YL
江西省武宁县银炉
Yinlu, Wuning County, Jiangxi
溪边石壁
Rock near brook-side
28°59'31''N, 114°49'39''E 420 50 两型花
Distyle
~200

Fig. 1

Geographic location of the studied populations and the first two boundaries (a to b) detected using BARRIER. Population codes see Table 1."

Table 2

The sequences of microsatellite loci and primers in this study"

位点
Locus
序列
Repeat motif
引物
Primer (5'-3')
退火温度(℃)
Annealing temperature
荧光标记
Fluorescent label
等位基因数
No. of alleles
Pm1 (TC)10(CT)3 F:ATCTTTGAGGTCCTTTTA 50 FAM 7
R: ATCGCCCAATGGAGTGAA
Pm2 (AG)14 F: CGCCTACAGTGTTTGGGA 55 FAM 2
R: CTATCTCACCTGCGTTCT
Pm7 (AG)3GG(AG)8 F: TTGTTCACCGACGCATAC 54 HEX 18
R: TTACACGCACCAAATCAT
Pm9 (TTTC)2(TC)6 F: AGACTCACGAGGAATACG 50 HEX 6
R: AGAAAAGGAGGAGACAAA
Pm12 (TC)10(CT)3 F: TAAAACTCCTGGAGGGGTAC 51 TAMRA 10
R: ATCGCCCAATGGAGTGAA
Pm13 (GATAGG)2GAT(AG)7 F: GAGGACAGGCACCACAGA 54 TAMRA 6
R: TCCCCAACTTCATGCTCTT
Pm14 (AG)3(GA)12 F: TCGCCCAATGGAGTGAAC 50 TAMRA 9
R: TCTTTGAGGTCCTTTTAT
Pm16 (GAGGGA)3(GA)3 F: AACCACTCGTCGTCCTAA 51 FAM 5
R: CGATAGATTGCCTTACCC
Pm17 (GA)9 F: TAAATCAAGGTAGCAACT 51 HEX 9
R: TACCTACCATTACTCCC

Table 3

Genetic characteristics of Primula ranunculoides populations"

种群
Population
取样数
Sample size
有效种群大小
Effective population size
期望杂合度(He)
Expected heterozygosity
观察杂合度(Ho)
Observed heterozygosity
稀有等位基因
Rare alleles
近交系数(Fis)
Inbreeding coefficient
YJPA 32 166 0.430 0.424 2 0.014 ns
YJPB 31 231 0.412 0.399 4 0.032**
MZS 30 105 0.297 0.155 2 0.484***
SJA 33 169 0.348 0.330 4 0.055***
SJB 31 149 0.281 0.230 1 0.184***
JPT 33 127 0.203 0.132 2 0.350***
YL 32 153 0.340 0.328 2 0.037**
平均 Mean 31.7 157.1 0.330 0.286 2.4

Table 4

Pairwise population differentiation (below diagonal) and genetic distances (above diagonal) among Primula ranunculoides populations"

YJPA YJPB MZS SJA SJB JPT YL
YJPA - 0.091 0.208 0.420 0.528 0.875 0.702
YJPB 0.096 - 0.167 0.311 0.363 0.793 0.588
MZS 0.256 0.222 - 0.336 0.378 0.827 0.742
SJA 0.301 0.262 0.399 - 0.030 0.921 0.473
SJB 0.385 0.332 0.476 0.047 - 0.929 0.467
JPT 0.510 0.526 0.645 0.569 0.599 - 0.318
YL 0.445 0.424 0.536 0.392 0.428 0.367 -

Table 5

Analysis of molecular variance (AMOVA) within/among Primula ranunculoides populations"

变异来源
Source
自由度
d.f.
方差和
Sum of squares
变异组分
Variance components
占总变异比例%
Percentage of variation
显著性检验
Significance tests
种群间 Among populations 6 225.635 0.6596 48.08 P<0.001
种群内 Within populations 215 310.851 0.7124 51.92 P<0.001
总和 Total 222 536.486 1.3720 100%

Fig. 2

The result of Principal coordinates (PCO) analysis of Primula ranunculoides populations. The first and second axis extracted 40.0% and 26.9% of the total genetic variance, respectively. Population codes see Table 1."

Fig. 3

The results of genetic structure analyses by STRUCTURE soft. (a) Plot of mean posterior probability lnP(D) values of each K; (b) The corresponding △K statistics calculated according to Evanno et al. (2005); (c) Histogram of the structure analysis for the model with K = 2-4. Population codes see Table 1."

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