生物多样性 ›› 2013, Vol. 21 ›› Issue (6): 723-731.doi: 10.3724/SP.J.1003.2013.09117

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伯乐树种群遗传多样性及遗传结构

徐刚标1, *(), 梁艳1, 蒋燚2, 刘雄盛1, 胡尚力1, 肖玉菲1, 郝博搏1   

  1. 1 中南林业科技大学林木遗传育种实验室, 长沙 410004
    2 广西林业科学研究院, 南宁 530001
  • 收稿日期:2013-05-13 接受日期:2013-09-10 出版日期:2013-11-20
  • 通讯作者: 徐刚标 E-mail:gangbiaoxu@163.com
  • 基金项目:
    国家林业公益性科研专项经费项目(201104033)

Genetic diversity and population structure of Bretschneidera sinensis, an endangered species

Gangbiao Xu1, *(), Yan Liang1, Yan Jiang2, Xiongsheng Liu1, Shangli Hu1, Yufei Xiao1, Bobo Hao1   

  1. 1 Laboratory of Forest Genetics, Central South University of Forestry and Technology, Changsha 410004
    2 Guangxi Forestry Research Institute, Nanning 530001
  • Received:2013-05-13 Accepted:2013-09-10 Online:2013-11-20
  • Contact: Xu Gangbiao E-mail:gangbiaoxu@163.com

了解种内遗传变异信息是制定种群遗传多样性保护策略的前提。伯乐树(Bretschneidera sinensis)是古老的单科属孑遗植物, 被列为国家一级保护植物。为了揭示伯乐树天然种群的遗传多样性和遗传结构, 采用7条ISSR引物分析了采自湖南、江西、广东、广西和贵州5个省区15个天然种群的219株个体的样本。结果显示, 伯乐树遗传多样性水平较高, 物种和种群水平上的多态位点百分率(PPB)分别为74.42%和38.06%, Shannon’s表型多样性指数(I)分别为0.3630和0.2081, Nei’s基因多样性指数(He)分别为0.2397和0.1405。种群间遗传分化显著, 基于表型多样性指数和分子方差分析揭示的伯乐树天然种群间遗传分化系数分别为FST = 0.4267、GST = 0.2973。UPGMA聚类表明, 参试的15个天然种群可分为2大组群; Mantel检测发现, 种群间遗传距离与其地理距离存在显著相关性(r = 0.3096, P = 0.008)。基于上述研究结果, 我们认为, 伯乐树濒危原因不是种群遗传进化潜力小, 而是由于生境破坏严重, 以及自身繁殖能力低、适应性差、竞争力弱等生物学特征导致的。建议优先保护遗传多样性较为丰富的阳明山、莽山、乳阳、八面山种群, 并对种群近交衰退开展相应的监测工作。

关键词: Bretschneidera sinensis, ISSR标记, 种群遗传结构, 濒危机制

Amounts and distribution of intraspecific genetic variation provide benchmarks for developing conservation strategies. Bretschneidera sinensis is a monotypic relic species listed in the First Grade of the List of Wild Plants Under State Protection (First Batch) in China. We examined the genetic diversity and genetic structure of 219 B. sinensis individuals sampled from 15 natural populations distributed in Hunan, Jiangxi, Guangdong, Guangxi, and Guizhou using inter-simple sequence repeat (ISSR) markers generated by seven ISSR primers. The percentage of polymorphic bands (PPB) at the species and population level was 74.42% and 38.06%, respectively. Shannon’s index (I) of phenotypic diversity at the species and population level was 0.3630 and 0.2081, respectively, and Nei’s genetic diversity (He) at the species and population level was 0.2397 and 0.1405, respectively. These results indicate that B. sinensis contains relatively high levels of genetic diversity. Analysis of molecular variance (AMOVA) and estimates of the coefficient of genetic differentiation based on phenotypic diversity index also indicated high levels of population subdivision (GST = 0.2973; FST = 0.4267) in the species. Analysis of the ISSR data using UPGMA further revealed that populations were genetically clustered into two groups, while a Mantel test showed that genetic divergence was significantly correlated with geographical distance among populations (Mantel test; r = 0.3096, P = 0.008). We conclude from our results that B. sinensis is not endangered due to low evolutionary potential stemming from low genetic diversity, but by habitat destruction coupled with a low reproductive capacity, poor adaptability and weak competitiveness. The Mt. Yangming, Mt. Mangshan, Ruyang, and Mt. Bamianshan populations of the species with higher genetic diversity should be given priority for conservation, and inbreeding depression monitoring should be conducted.

Key words: Bretschneidera sinensis, ISSR markers, population genetic structure, endangered mechanism

表1

伯乐树种群采样信息及样本量"

种群
Population
位置
Location
经纬度
Latitude and longitude
海拔
Altitude (m)
株数
Sample size
YMS 湖南阳明山省级自然保护区
Yangmingshan Provincial Nature Reserve, Hunan
26º07′ N, 111º50′ E 1,390 22
MS 湖南莽山国家级自然保护区
Mangshan National Nature Reserve, Hunan
24º58′ N, 112º51′ E 1,230 23
SHS 湖南舜皇山国家级自然保护区
Shunhuangshan National Nature Reserve, Hunan
26º35′ N, 110º00′ E 950 22
BMS 湖南八面山国家级自然保护区
Bamianshan National Nature Reserve, Hunan
26º10′ N, 113º39′ E 1,020 15
HS 湖南黄桑国家级自然保护区
Huangsang National Nature Reserve, Hunan
26º38′ N, 110º55′ E 1,180 16
LP 贵州黎平国家森林公园
Liping National Forest Park, Guizhou
25º42′ N, 109º13′ E 500 14
DY 广西大瑶山国家级自然保护区
Dayaoshan National Nature Reserve, Guangxi
24º14′ N, 110º12′ E 920 11
MRS 广西猫儿山国家级自然保护区
Maoershan National Nature Reserve, Guangxi
24º25′ N, 111º29′ E 800 10
HP 广西花坪国家级自然保护区
Huaping National Nature Reserve, Guangxi
25º18′ N, 110º53′ E 1,070 5
TX 广东田心省级自然保护区
Tianxin Provincial Nature Reserve, Guangdong
25°01′ N, 112°30′ E 970 15
DDS 广东南岭国家级自然保护区大东山管理处
Dadongshan Administration Bureau of the Nanling National Nature Reserve, Guangdong
24°38′ N, 113°12′ E 950 16
RY 广东南岭国家级自然保护区乳阳管理处
Rurang Administration Bureau of the Nanling National Nature Reserve, Guangdong
24°45′ N, 112°53′ E 1,000 21
JGS 江西井冈山国家级自然保护区
Jinggangshan National Nature Reserve, Jiangxi
26°26′ N, 114°15′ E 450 14
NKS 广东南昆山省级自然保护区
Nankunshan Provincial Nature Reserve, Guangdong
23°40′ N, 113°50′ E 600 10
JLS 江西九连山国家级自然保护区
Jiulianshan National Nature Reserve, Jiangxi
24°33′ N, 114°28′ E 580 5

表2

ISSR-PCR引物及扩增结果"

引物
Primer code
序列(5′ to 3′)
Sequence (5′ to 3′)
退火温度
Annealing temperature (℃)
总条带数
Scored bands
多态条带数
No. of polymorphic bands
多态条带百分比
% of polymorphic bands
UBC824 (TC)8G 51 11 6 54.54
UBC825 (AC)8T 52 13 12 92.30
UBC834 (AG)8YT 53 12 11 91.66
UBC836 (AG)8YA 52 11 8 72.72
UBC843 (CT)8RA 53 8 4 50.00
UBC844 (CT)8RC 52 16 12 75.00
UBC872 (GATA)4 40 15 11 73.33
平均 Mean 12 9 72.80
物种 Species level 86 64 74.42

表3

伯乐树15个种群的遗传多样性参数的估算"

种群
Population
Shannon表型多样性指数 Shannon's phenotypic diversity index I PPB (%) He
UBC824 UBC825 UBC834 UBC836 UBC843 UBC844 UBC872
阳明山 YMS 0.1411 0.5048 0.3045 0.3181 0.2545 0.3374 0.2520 0.3079 53.49 0.2057
莽山 MS 0.0955 0.5595 0.2976 0.2004 0.0700 0.3008 0.1728 0.2566 45.35 0.1742
舜皇山 SHS 0.0843 0.4068 0.3353 0.1152 0.2191 0.2371 0.2288 0.2382 43.02 0.1608
八面山 BMS 0.1678 0.2706 0.3439 0.2200 0.1838 0.3210 0.2119 0.2522 46.51 0.1698
黄桑 HS 0.2377 0.2266 0.2901 0.1870 0.1258 0.1903 0.2378 0.2176 38.37 0.1486
黎平 LP 0.1241 0.2029 0.3086 0.2841 0.1512 0.2671 0.0860 0.2047 37.21 0.1373
大瑶山 DY 0.1699 0.1481 0.0908 0.1573 0.0522 0.1815 0.1051 0.1339 26.74 0.0881
猫儿山 MRS 0.0625 0.1420 0.0919 0.1948 0.0920 0.1922 0.0940 0.1279 26.74 0.0834
花坪 HP 0.1110 0.1518 0.0562 0.0792 0.1527 0.0955 0.0581 0.0972 18.60 0.0644
田心 TX 0.1752 0.2798 0.2685 0.2125 0.1202 0.3412 0.1671 0.2331 41.86 0.1578
大东山 DDS 0.2163 0.2111 0.2097 0.2226 0.1111 0.3094 0.1819 0.2170 39.53 0.1466
乳阳 RY 0.2650 0.2446 0.2693 0.2859 0.1311 0.2660 0.1468 0.2323 45.35 0.1538
井冈山 JGS 0.2127 0.1014 0.2787 0.2018 0.0539 0.2452 0.1651 0.1867 36.05 0.1255
南昆山 NKS 0.0856 0.3194 0.1156 0.2362 0.1973 0.3532 0.2950 0.2411 41.86 0.1641
九连山 JLS 0.0600 0.2047 0.1377 0.3304 0.1644 0.1567 0.2223 0.1833 30.23 0.1269
平均 Mean 0.1472 0.2650 0.2266 0.2134 0.1386 0.2522 0.1750 0.2081 38.06 0.1405
物种 Species level 0.2622 0.4607 0.3942 0.3190 0.2279 0.4369 0.3528 0.3630 74.42 0.2397

图1

伯乐树15个种群219株个体的遗传结构(Q值为个体归属各组群的比例。种群代号见表1)"

表4

伯乐树天然种群的分子方差分析"

变异来源
Source of variation
自由度
df
均方和
Sum of squares
方差组分
Variance components
变异百分比
% of variation
遗传分化系数(GST)
Genetic differentiation index
P
种群间 Among populations 14 685.9610 2.9145 29.73 0.2973 <0.001
种群内 Within populations 204 1,405.0889 6.8877 70.27 <0.001
总和 Total 218 2,091.0501 9.8022

表5

伯乐树天然种群间遗传分化系数(ΦST)估算值(对角线下方)与地理距离(km, 对角线上方)的相关性"

种群
Population
黎平
LP
阳明山YMS 莽山
MS
舜皇山SHS 田心
TX
大东山DDS 八面山BMS 大瑶山DY 井冈山JGS 猫儿山MRS 黄桑
HS
乳阳
RY
花坪
HP
南昆山NKS 九连山JLS
黎平 LP 292 397 194 416 443 480 240 507 127 106 342 138 554 574
阳明山 YMS 0.3157 152 100 126 210 198 289 228 160 189 170 200 331 312
莽山 MS 0.2752 0.2443 240 55 47 155 291 223 275 325 40 267 180 170
舜皇山 SHS 0.3674 0.2600 0.3593 204 289 283 267 308 92 95 250 150 413 398
田心 TX 0.3787 0.2611 0.2713 0.3231 75 167 270 235 236 300 75 245 195 204
大东山 DDS 0.3793 0.2683 0.2219 0.3751 0.3922 162 321 228 322 364 29 315 124 135
八面山 BMS 0.3439 0.1488 0.2457 0.3004 0.1349 0.1030 435 66 341 370 154 378 261 165
大瑶山 DY 0.4976 0.3949 0.3849 0.4525 0.2516 0.2201 0.2881 483 194 274 312 135 377 450
井冈山 JGS 0.4056 0.3065 0.2841 0.4081 0.1065 0.1145 0.1686 0.2216 380 401 212 422 319 215
猫儿山 MRS 0.4460 0.3387 0.3608 0.4204 0.2200 0.2251 0.2681 0.0563 0.1429 79 284 56 422 425
黄桑 HS 0.4253 0.2805 0.3683 0.2019 0.3109 0.3125 0.2562 0.4188 0.3995 0.3762 356 119 488 473
乳阳 RY 0.3940 0.3149 0.3098 0.3833 0.0716 0.0922 0.2160 0.2572 0.1176 0.2027 0.3423 291 175 116
花坪 HP 0.4857 0.3426 0.3938 0.4311 0.2046 0.2309 0.2798 0.1641 0.1762 0.0741 0.3836 0.1680 419 440
南昆山 NKS 0.3319 0.2463 0.2092 0.3185 0.1825 0.1652 0.1899 0.3955 0.3146 0.3802 0.3385 0.2765 0.3885 136
九连山 JLS 0.3441 0.2508 0.2588 0.3268 0.1940 0.2088 0.2469 0.4163 0.3212 0.3721 0.3661 0.2954 0.4030 0.0391

图2

基于Nei’s遗传距离的15个伯乐树种群的UPGMA聚类(种群代号见表1)"

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