Yancai Shi, Xiao Wei, Jiqing Wei, Yongtao Li, Shengfeng Chai, Jianming Tang


Heteroplexis microcephala Y. L. Chen. is an endemic and endangered species found only in karst limestone regions in the Yangshuo County of the Guangxi Zhuang Autonomous Region in China: it is a habitat representative of species in the Heteroplexis genus. To provide basic genetic information for its conservation, in this study we evaluated the genetic variation and differentiation among six wild populations of H. microcephala by random amplified polymorphic DNA markers (RAPD). The leaves of 141 individuals were sampled. Based on 12 primers, 113 DNA fragments were generated. Genetic diversity was low at the population level (Nei’s gene diversity (h)=0.0579; Shannon information index (I)=0.0924; percentage of polymorphic bands (PPB)=23.30%), but relatively high at the species level (h=0.1701; I=0.2551; PPB=46.34%). The coefficient of genetic differentiation based on Nei’s genetic diversity analysis (0.6661) was high, indicating that there was significant genetic differentiation among populations, which was confirmed by AMOVA analysis exhibiting population differentiation among populations of 68.77%. Low gene flow among populations (0.2507) may result from several factors, such as a harsh pollination environment, population isolation and low seed dispersal distance. Limited gene flow and self-compatibility are the primary reasons for the high genetic differentiation observed for this species. We propose the collection of seeds from more populations with fewer individuals and core populations for ex situ conservation and suggest methods to increase seed germination rates.


Keywords: endangered species; random amplified polymorphic DNA; genetic variability; population differentiation; conservation


Received: September 21, 2015; Revised: October 12, 2015; Accepted: October 20, 2015; Published online: June 16, 2016

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Haining T, Yan L. Qin HN, Liu Y. [A checklist of vascular plants of Guangxi]. Beijing: Science Press; 2010. p. 625. Chinese.

Fu LG. Chinese plant red book. Beijing: Science Press; 1992. p. 654. Chinese.

Futuyma DJ. The evolution of evolutionary ecology. Isr J Ecol Evol. 2013;59(4):172-80.

Jeong JH, Kim EH, Guo WH, Yoo KO, Jo DG, Kim ZS. Genetic diversity and structure of the endangered species Megaleranthis saniculifolia in Korea as revealed by allozyme and ISSR markers. Plant Syst Evol. 2010;289(1):67-76.

Furches MS, Wallace LE, Helenurm K. High genetic divergence characterizes populations of the endemic plant Lithophragma maximum (Saxifragaceae) on San Clement Island. Conserv Genet. 2009;10(1):115-26.

Fan XN, Lin S, Zhu CG, Hu JF, Hu JF, Liu Y, Chen XG, Chen NH, Wang WJ, Shi JG. Terpenoids of Heteroplexis micocephala and their bioactivities (in Chinese). China J Chinese Materia Medica. 2010;35(3):315-22.

Zhang Gl, Guo X. The karyotype analysis of Anemoclema glaucifolium and Heteroplexis microcephala both endemic to China (in Chinese). Acta Bot Yunnanica. 2002;24(6):765-8.

Wang CM, Huan SX, Wang Y. A preliminary study on ecological and biologcal characteristics of Heteroplexis microcephala-A protected and endemic species from Guangxi (in Chinese). Guihaia. 1994;12(4):356-8.

Qiu YX, Hong DY, Fu CX, Cameron KM. Genetic variation in the endangered and endemic species Changium smyrnioides (Apiaceae). Biochem Syst Ecol. 2004;32(6):583-96.

Badfar-Chaleshtori S, Shiran B, Kohgard M, Mommeni H. Hafizi A, Khodambashi M, Mirakhorli N, Sorkheh K. Assessment of genetic diversity and structure of Imperial Crown (Fritillaria imperialis L.) populations in the Zagros region of Iran using AFLP, ISSR and RAPD markers and implications for its conservation. Biochem Syst Ecol. 2012;42:35-48.

Pineda-Martos R, Velasco L, Perez-Vich B. Identification, characterisation and discriminatory power of microsatellite markers in the parasitic weed Orobanche cumana. Weed Res. 2014;54(2):120-32.

Nybom H. Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants. Mol Ecol. 2004;13(5):1143-55.

Excoffier L, Smouse PE, Quattro JM. Analysis of molecular variance inferred from metric distances among DNA haplotypes: Application to human mitochondrial DNA restriction data. Genetics. 1992;131(2):479-91.

Huff DR, Peakall R, Smouse PE. RAPD variation within and among natural populations of outcrossing buffalograss [Buchloe dactyloides (Nutt.) Engelm.]. Theo Appl Genet. 1993;86(8):927-34.

Diaz V, Muniz LM, Ferrer E. Random amplified polymorphic DNA and amplified fragment length polymorphism assessment of genetic variation in Nicaraguan populations of Pinus oocarpa. Mol Ecol. 2001;10:2593-603.

del Hoyo A, Lopez-Pujol J, Chung MY, de la Vega BL. Population genetics and conservation of the extremely narrow Pyrenean palaeoendemic Glandora oleifolia (Boraginaceae). Plant Ecol Divers. 2012;5(4):501-11.

Bruetting C, Hensen I, Wesche K. Ex situ cultivation affects genetic structure and diversity in arable plants. Plant Biol. 2013;15(3):505-13.

Sharma SK, Bhowmik SSD, Kumaria S, Tandon P, Rao SR. Low genetic diversity as revealed by SPAR methods possibly leads to extinction of two critically-endangered and endemic species of Mantisia. Biol Plant. 2012;56(2):292-300.

Chai XY, Chen SL, Xu W. Using inter-simple sequence repeat markers to analyze the genetic structure of natural Pteroceltis tatarinowii populations and implications for species conservation. Plant Syst Evol. 2010;285(1):65-73.

Zhou TH, Wu KX, Qian ZQ, Zhao GF, Liu ZL, Li S. Genetic diversity of the threatened Chinese endemic plant, Sinowilsonia henryi Hemsi. (Hamamelidaceae), revealed by inter-simple sequence repeat (ISSR) markers. Biochem Syst Ecol. 2014;56:171-7.

Silva L, Elias RB, Moura M, Meimberg H, Dias E. Genetic variability and differentiation among populations of the Azorean endemic Gymnosperm Juniperus brevifolia: baseline information for a conservation and restoration perspective. Biochem Genet. 2011;49(11):715-34.

Yang Q, Fu Y, Wang YQ, Wang Y, Zhang WH, Li XY, Reng YQ, Zhang J. Genetic diversity and differentiation in the critically endangered orchid (Amitostigma hemipilioides): implications for conservation. Plant Syst Evol. 2014;300(5):871-9.

Ge XJ, Sun M. Reproductive biology and genetic diversity of a cryptoviviparous mangrove Aegiceras corniculatum (Myrsinaceae) using allozyme and inter-simple sequence repeat (ISSR) analysis. Mol Ecol. 1999;8(12):2061-9.

Lacerda DR, Acedo MDP, Lemos Filho JP, Lovato MB. Genetic diversity and structure of natural populations of Plathymenia reticulate (Mimosoideae), a tropical tree from the Brazilian Cerrado. Mol Ecol. 2001;10(5):1143-52.

Dostalek T, Munzbergova Z, Plackova I. Genetic diversity and its effect on fitness in an endangered plant species, Dracocephalum austriacum L. Conserv Genet. 2010;11(3):773-83.

Sozen E, Ozaydin B. A preliminary study on the genetic diversity of the critically endangered Centaurea nivea (Asteraceae). Ann. Bot. Fenn. 2009;46(6):541-8.

Nguyen HTT, Choi KS, Park S. Genetic diversity and differentiation of a narrowly distributed and endemic species, Aster spathulifolius Maxim (Asteraceae), revealed with inter simple sequence repeat markers. J Korean Soc Appl Biol Chem. 2013;56(3):255-62.

Luan SS, Chiang TY, Gong X. High genetic diversity vs. low genetic differentiation in Nouelia insignis (Asteraceae), a narrowly distributed and endemic species in China, revealed by ISSR fingerprinting. Ann Bot. 2006;98(3):583-9.

Rossetto M, Weaver PK, Dixon KW. Use of RAPD analysis in devising conservation strategies for the rare and endangered Grevillea scapigera (Proteaceae). Mol Ecol. 1995;4(3):321-9.

Schaal BA, Hayworth DA, Olsen KM, Rauscher JT, Smith WA. Phylogeographic studies in plants: problems and prospects. Mol Ecol. 1998;7(4):65-474.

Rajeb C, Messaoud C, Chograni H, Bejaoui A, Boulila A, Rejeb MN, Boussaid M. Genetic diversity in Tunisian Crataegus azarolus L. var. aronia L. populations assessed using RAPD markers. Ann For Sci. 2010;67(5):512.

Chen DX, Li LY, Zhang X, Wang Y, Zhang Z. Genetic diversity in wild Dipsacus chinensis populations from China based on ISSR markers. Genet Mol Res. 2013;12(2):1205-13.

Harish, Gupta AK, Phulwaria M, Rai MK, Shekhawat NS. Conservation genetics of endangered medicinal plant Commiphora wightii in Indian Thar Desert. Gene. 2014;535(2):266-72.


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