濒危植物毛柄小勾儿茶片断化居群的遗传多样性

doi:10.17521/cjpe.2006.0022

摘要/Abstract

摘要：

采用扩增片段长度多态性(AFLP)标记对我国特有的濒危植物毛柄小勾儿茶(Berchemiella wilsonii var. pubipetiolata)现存于浙江和安徽的4个片断化居群中的89株个体进行了遗传多样性和遗传结构的研究。结果表明,与其它木本濒危植物相比,毛柄小勾儿茶具有与它们相当的遗传多样性,8对选择扩增引物共扩增出122条清晰的条带,居群的平均多态位点百分率为P_p=26.4%,其中马家河居群最高(29.5%)而湍口居群最低(23.8%),居群的平均基因多样度为H_ep=0.162 8(0.140 5~0.172 4);而在物种水平上的遗传多样性为P_s=36.9%, H_es=0.202 4。居群间的遗传分化系数F_ST=0.193 9,表明居群间有显著的遗传分化,进一步利用AMOVA软件对遗传变异进行等级剖分发现:24.88%的遗传变异存在于地理宗间(浙江地理宗和安徽地理宗),14.71%的遗传变异存在于居群间,60.42%存在于居群内。该研究结果表明,由于人为干扰引起的生境片断化和居群减小导致了毛柄小勾儿茶居群的遗传多样性丧失和遗传分化,并对毛柄小勾儿茶的生存造成潜在威胁。该文还就保育策略进行了讨论。

关键词: 毛柄小勾儿茶, 生境片断化, AFLP, 保育遗传, 濒危物种

Abstract:

Berchemiella wilsonii var. pubipetiolata (Rhamnaceae) is an endangered tree species with a narrow distribution in fragmented habitat patches in Tianmu Mountain, Zhejiang Province and Dabie Mountain, Anhui Province, east of China. The habitat has been severely disturbed because of agricultural practices by local residents. Over the past several decades, there has been a decrease in the population number and size. Information on genetic variation and structure is critical for developing successful conservation strategies for this species. The amplified fragment length polymorphisms (AFLPs) were used to evaluate the genetic diversity and population structure of four extant populations of B. wilsonii var. pubipetiolata. A total of 122 unambiguous bands were generated using eight primer combinations of which 45 (36.9%) were polymorphic across all individuals. In comparison with other endangered woody plants, B. wilsonii var. pubipetiolata has similar genetic diversity at the population level (P_p=26.4%, H_ep=0.162 8) and species level (P_s=36.9%, H_es=0.202 4). A relatively high value of F_ST (0.193 9) indicated significant genetic differentiation among populations. Analysis of molecular variance (AMOVA) further revealed that 24.88% of the AFLP variation resided between geographic domes (Zhejiang and Anhui Province), 14.71% among populations and 60.42% within populations. The results of this study suggest that habitat fragmentation and reduction of population size caused by anthropogenic activities have caused genetic erosion and population differentiation in B. wilsonii var. pubipetiolata, which poses a potential threat to the long-term survivability of this species. Genetic information obtained in the present study provides useful baseline data for formulating conservation strategies. Conservation management should include both in situ conservation and ex situ conservation programs and need to be well designed. An ex situ conservation strategy for extensive sampling of all extant populations to maximize genomic representation of the species is recommended.

Key words: Berchemiella wilsonii var. pubipetiolata, Habitat fragmentation, AFLP, Conservation genetics, Endangered species

许凤华, 康明, 黄宏文, 江明喜. 濒危植物毛柄小勾儿茶片断化居群的遗传多样性. 植物生态学报, 2006, 30(1): 157-164. DOI: 10.17521/cjpe.2006.0022

XU Feng-Hua, KANG Ming, HUANG Hong-Wen, JIANG Ming-Xi. GENETIC DIVERSITY IN FRAGMENTED POPULATIONS OF BERCHEMIELLA WILSONII VAR. PUBIPETIOLATA, AN ENDANGERED PLANT ENDEMIC TO EASTERN CHINA. Chinese Journal of Plant Ecology, 2006, 30(1): 157-164. DOI: 10.17521/cjpe.2006.0022

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https://www.plant-ecology.com/CN/Y2006/V30/I1/157

图/表 5

参考文献 41

[1]	Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Stuhl K (1987). Current Protocols in Molecular Biology. John Wiley & Sons, New York.
[2]	Brown AHD (1978). Isozymes, plant population genetic structure and genetic conservation. Theoretical and Applied Genetics, 52,145-157. URL PMID
[3]	Buza L, Young A, Thrall P (2000). Genetic erosion, inbreeding and reduced fitness in fragmented populations of the endangered tetraploid pea Swainsona recta. Biological Conservation, 93,177-186.
[4]	Chung MG (1999). Allozyme diversity in the endangered shrub Abeliophyllum distichum (Oleaceae): a monotypic Korean genus. International Journal of Plant Science, 160,553-559.
[5]	Coart E, Vekemans X, Smulders MJM, Wagner I, van Huylenbroeck J, van Bockstaele E, Roldn-Ruiz I (2003). Genetic variation in the endangered wild apple ( Malus sylvestris (L.) Mill.) in Belgium as revealed by amplified fragment length polymorphism and microsatellite markers. Molecular Ecology, 12,845-857. DOI URL PMID
[6]	Dolan RW, Yahr R, Menges ES, Halfhill MD (1999). Conservation implications of genetic variation in three rare species endemic to Florida rosemary scrub. American Journal of Botany, 86,1556-1562. URL PMID
[7]	Drummond RSM, Keeling DJ, Richardson TE, Gardner RC, Wright SD (2000). Genetic analysisand conservation of 31 surviving individuals of a rare New Zealand tree,Metrosideros bartlettii (Myrtaceae). Molecular Ecology, 9,1149-1157. URL PMID
[8]	Excoffier L, Smouse PE, Quattro JM (1992). Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data. Genetics, 131,479-491. URL PMID
[9]	Excoffier L (1995). AMOVA 1.55 (Analysis of Molecular Variance). Genetics and Biometry Laboratory, University of Geneva, Suitzerland.
[10]	Falk DA, Holsinger KE (1991). Genetics and Conservation of Rare Plants. Oxford University Press, Oxford.
[11]	Frankham R, Ballou JD, Briscoe DA (2002). Introduction to Conservation Genetics. Cambridge University Press, UK.
[12]	Ge S (葛颂) (1998). The review and prospect of researches in genetic structure of plant populations. In: Li CS (李承森) ed. Progress in plant scienceⅠ (植物科学进展(第一卷)). Higher Education Press, Beijing,1-15. (in Chinese)
[13]	Juan A, Crespo MB, Cowan RS, Lexer C, Fay MF (2004). Patterns of variability and gene flow in Medicago citrina, an endangered endemic of islands in the western Mediterranean, as revealed by amplified fragment length polymorphism (AFLP). Molecular Ecology, 13,2679-2690. DOI URL PMID
[14]	Jorgensen SM, Hamrick JL (1997). Biogeography and population genetics of whitebark pine,Pinus albicaulis. Canadian Journal of Forestry Research, 27,1574-1585.
[15]	Kahmen S, Poschlod P (2000). Population size, plant performance, and genetic variation in the rare plant Arnica montana L. in the Rhøn, Germany. Basic and Applied Ecology, 1,43-51.
[16]	Kwon JA, Morden CM (2002). Population genetic structure of two rare tree species ( Colubrina oppositifolia and Alphitonia ponderosa, Rhamnaceae) from Hawaiian dry and mesic forests using random amplified polymorphic DNA markers. Molecular Ecology, 11,991-1001. URL PMID
[17]	Lacy RC (1987). Loss of genetic diversity from managed populations: interacting effects of drift, mutation, immigration, selection, and population subdivision. Conservation Biology, 1,143-158.
[18]	Lande R (1988). Genetics and demography in biological conservation. Science, 241,1455-1460.
[19]	Llorens TM, Ayre DJ, Whelan RJ (2004). Evidence for ancient genetic subdivision among recently fragmented populations of the endangered shrub Grevillea caleyi (Proteaceae). Heredity, 92,519-526. DOI URL PMID
[20]	Martinez-Palacios A, Eguiarte LE, Furnier GR (1999). Genetic diversity of the endangered endemic Agave victoriae-regineae (Agavaceae) in the Chihuahuan desert. American Journal of Botany, 86,1093-1098. URL PMID
[21]	Miller MP (1997). AMOVA-PREP, a Program for the Preparation of AMOVA Input Files for Use with WINAMOUA. Department of Biological Sciences, Northern Arizona University, Flagstaff, AZ.
[22]	Mueller UG, Wolfenbarger LL (1999). AFLP genotyping and fingerprinting. Trends in Ecology and Evolution, 14,389-394. DOI URL PMID
[23]	Nei M (1978). Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics, 89,583-590. URL PMID
[24]	Primack RB (1993). Essentials of Conservation Biology. Sinauer Associates, Sunderland, USA.
[25]	Qian H (钱宏) (1988). A study on the cenus Berchemiella Nakai (Rhamnaceae) endemic to eastern Asia. Bulletin of Botanical Research (植物研究), 8,119-126. (in Chinese with English abstract)
[26]	Rogers SO, Bendich AJ (1985). Extraction of DNA from milligram amount of fresh, herbarium and mummified plant tissues. Plant Molecular Biology, 5,69-76. DOI URL PMID
[27]	Rohlf FJ (1994). NTSYS-pc, Numerical Taxonomy and Multivariate Analysis System* Version 2.0. Exeter Software*, Setauket, New York.
[28]	Schmidt K, Jensen K (2000). Genetic structure and AFLP variation of remnant populations in the rare plant Pedicularis palustris (Scrophulariaceae) and its relation to population size and reproductive components. American Journal of Botany, 87,678-689. URL PMID
[29]	Shea KL, Furnier GR (2002). Genetic variation and population structure in central and isolated populations of balsam fir, Abies balsamea (Pinaceae). American Journal of Botany, 89,783-791.
[30]	Sork VL, Nason J, Campbell DR, Fernandez JF (1999). Landscape approaches to historical and contemporary gene flow in plants. Trends in Ecology and Evolution, 14,219-224. DOI URL PMID
[31]	Tomimatsu H, Ohara M (2003). Genetic diversity and local population structure of fragmented populations of Trillium camschatcense (Trilliaceae). Biological Conservation, 109,249-258.
[32]	Travis SE, Maschinski J, Keim P (1996). An analysis of genetic variation in Astragalus cremnophylax var. cremnophylax, a critically endangered plant, using AFLP markers. Molecular Ecology, 5,735-745. URL PMID
[33]	Vasseur L (2001). Allozymic diversity of Allium tricoccum (Ait.) Solander var. burdickii Hanes in isolated populations of Nova Scotia (Canada). Plant Systematics and Evolution, 228,71-79.
[34]	Vekemans X, Beauwens T, Lernaire M, Roldan-Ruiz I (2002). Data from amplified fragment length polymorphism (AFLP) makers show indication of size homoplasy and relationship between degree of homoplasy and fragment size. Molecular Ecology, 11,139-151. URL PMID
[35]	Vos P, Hogers R, Bleeker M, Reijans M, Van de Lee T, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M, Zebeau M (1995). AFLP: a new technique for DNA fingerprinting. Nucleic Acids Research, 23,4407-4414. URL PMID
[36]	Vucetich JA, Peterson RO, Waite TA (2000). Is one migrant per generation sufficient for the genetic management of fluctuating populations? Animal Conservation, 3,261-266.
[37]	Wang Y (王燕), Tang SQ (唐绍清), Li XK (李先琨) (2004). The genetic diversity of the endangered plant Abies yuanbaoshanensis. Biodiversity Science (生物多样性), 12,269-273. (in Chinese with English abstract)
[38]	Winfield MO, Arnold GM, Cooper F, Leray M, White J, Karp A, Edwards KJ (1998). A study of genetic diversity in Populus nigra subsp. betulifolia in the upper Severn area in the UK using AFLP markers. Molecular Ecology, 7,3-10.
[39]	Wright S (1969). Evolution and Genetics of Populations Vol.2.The Theory of Gene Frequencies. University of Chicago Press, Chicago.
[40]	Young A, Boyle T, Brown T (1996). The population genetic consequences of habitat fragmentation in plants. Trends in Ecology and Evolution, 11,413-418. URL PMID
[41]	Zhivotovsky LA (1999). Estimating population structure in diploids with multilocus DNA markers. Molecular Ecology, 8,907-913. URL PMID

居群 Population	取样数(N) No. of sampling	多态位点百分率(P) Polymorphism(%)	基因多样度(H_e) Gene diversity	居群特有带 Population specific bands
湍口Tuankou	23	23.8	0.140 5 (0.015 12)	0
马啸Maxiao	13	26.2	0.171 4 (0.014 87)	0
马家河Majiahe	31	29.5	0.166 8 (0.015 38)	2
万佛山Wanfoshan	22	26.2	0.172 4 (0.014 66)	1
居群平均Population	22.25	26.4	0.162 8 (0.007 52)
物种水平Species	89	36.9	0.202 4

居群 Population	取样数(N) No. of sampling	多态位点百分率(P) Polymorphism(%)	基因多样度(H_e) Gene diversity	居群特有带 Population specific bands
湍口Tuankou	23	23.8	0.140 5 (0.015 12)	0
马啸Maxiao	13	26.2	0.171 4 (0.014 87)	0
马家河Majiahe	31	29.5	0.166 8 (0.015 38)	2
万佛山Wanfoshan	22	26.2	0.172 4 (0.014 66)	1
居群平均Population	22.25	26.4	0.162 8 (0.007 52)
物种水平Species	89	36.9	0.202 4

引物组合 Primer combination	引物序列 Primer sequence (5'-3')	总位点数 Total bands	多态位点数 Polymorphic bands	多态位点百分率 Polymorphism(%)
EAAC/MCGT	GACTGCGTACCAATTCAAC	16	8	50.0
	GATGAGTCCTGAGTAACGT
EACT/MCAT	GACTGCGTACCAATTCACT	15	7	46.7
	GATGAGTCCTGAGTAACAT
EATC/MCAA	GACTGCGTACCAATTCATC	13	3	23.1
	GATGAGTCCTGAGTAACAA
EAAG/MCGA	GACTGCGTACCAATTCAAG	20	9	45.0
	GATGAGTCCTGAGTAACGA
EAAG/MCGT	GACTGCGTACCAATTCAAG	17	4	23.5
	GATGAGTCCTGAGTAACGT
EAAG/MCAT	GACTGCGTACCAATTCAAG	15	6	40.0
	GATGAGTCCTGAGTAACAT
EACA/MCAA	GACTGCGTACCAATTCACA	12	7	58.3
	GATGAGTCCTGAGTAACAA
EACA/MCAT	GACTGCGTACCAATTCACA	14	1	7.1
	GATGAGTCCTGAGTAACAT
总计Total		122	45	36.9

引物组合 Primer combination	引物序列 Primer sequence (5'-3')	总位点数 Total bands	多态位点数 Polymorphic bands	多态位点百分率 Polymorphism(%)
EAAC/MCGT	GACTGCGTACCAATTCAAC	16	8	50.0
	GATGAGTCCTGAGTAACGT
EACT/MCAT	GACTGCGTACCAATTCACT	15	7	46.7
	GATGAGTCCTGAGTAACAT
EATC/MCAA	GACTGCGTACCAATTCATC	13	3	23.1
	GATGAGTCCTGAGTAACAA
EAAG/MCGA	GACTGCGTACCAATTCAAG	20	9	45.0
	GATGAGTCCTGAGTAACGA
EAAG/MCGT	GACTGCGTACCAATTCAAG	17	4	23.5
	GATGAGTCCTGAGTAACGT
EAAG/MCAT	GACTGCGTACCAATTCAAG	15	6	40.0
	GATGAGTCCTGAGTAACAT
EACA/MCAA	GACTGCGTACCAATTCACA	12	7	58.3
	GATGAGTCCTGAGTAACAA
EACA/MCAT	GACTGCGTACCAATTCACA	14	1	7.1
	GATGAGTCCTGAGTAACAT
总计Total		122	45	36.9

居群 Population	湍口 Tuankou	马啸 Maxiao	马家河 Majiahe	万佛山 Wanfoshan
湍口Tuankou	-	1.982	0.588	0.643
马啸Maxiao	0.112 0	-	1.341	0.912
马家河Majiehe	0.298 5	0.157 1	-	3.247
万佛山Wanfoshan	0.280 0	0.215 1	0.071 5	-