Cover Image

Outcrossing Rates and Gene Flow in Natural Population of the Endangered Endemic Aquatic Lycophyte Isoetes yunguiensis as Revealed by ISSR Markers

Mengxue MA, Xiang DONG, Kuo LIAO, Fei HAO, Rui QIN, Hong LIU

Abstract


In this study, mating system, genetic diversity, and genetic structure of the endangered endemic aquatic Isoetes yunguiensis in China was investigated using ISSR markers. The results of ISSR analyses showed that the estimate of multilocus outcrossing rate (tm) was high at species level (tm = 0.955), indicating that diploid I. yunguiensis is a predominant outcrossing species. Nine selected ISSR primers used in the study amplified 66 reproducible bands, 41 of which were polymorphic among 37 individuals. High level of genetic diversity was detected at the species level (PPB = 62.12%), whereas, relatively low genetic diversity existed within populations (PPB = 39.39%). Analysis of molecular variance (AMOVA) revealed that 31.99% of the genetic variation was attributable to differences between populations and the rest (68.01%) to variability within populationsof I. yunguiensis. Value of Fst (0.320) indicated that genetic differentiation between populations also was significant. These results showed that I. yunguiensis predominantly favors crossing, and has a high level of genetic diversity and highly significant genetic variation between and within populations. Gene flow (Nm) among populations is equal to 1.177. High outcrossing rates may be responsible for the high levels of genetic diversity observed in the I. yunguiensis population. To maintain the current level of genetic diversity for this species, we recommend increasing in situ conservation sites.

 

*********

In press - Online First. Article has been peer reviewed, accepted for publication and published online without pagination. It will receive pagination when the issue will be ready for publishing as a complete number (Volume 47, Issue 1, 2019). The article is searchable and citable by Digital Object Identifier (DOI). DOI number will become active after the article will be included in the complete issue.


Keywords


endangered; genetic structure; ISSR; Isoetes yunguiensis; outcrossing rate

Full Text:

PDF

References


Cánovas JL, Jiménez JF, Mota JF, Gómez PS (2015). Genetic diversity of Viola cazorlensis Gand., an endemic species of Mediterranean dolomitic habitats: implications for conservation. Systematics and Biodiversity 13:571-580.

Carrasco B, Ávila P, Perez-Diaz J, Muñoz P, Garcia R, Lavandero B, Zurita-Silva A, Retamales JB, Caligari PDS (2009). Genetic structure of highland papayas (Vasconcellea pubescens (Lenné et C. Koch) Badillo) cultivated along a geographic gradient in Chile as revealed by Inter Simple Sequence Repeats (ISSR). Genetic Resources and Crop Evolution 56:331-337.

Chen JM, Gituru WR, Liu X, Wang QF (2007). Genetic diversity in Isoetes yunguiensis, a rare and endangered endemic fern in China. Frontiers of Biology in China 2:46-49.

Chen YY, Ye QG, Li ZZ (2004). Genetic structure of Xiuning population of Isoetes sinensis, a critically endangered species in China. Chinese Biodiversity 12:564-571.

Chen JM, Liu X, Wang QF. (2005a). Genetic diversity in Isoetes yunguiensis, a rare and endangered endemic fern in china. Wuhan University Journal(Natural Science Edition) 51:767-770.

Chen JM, Liu X, Wang JY, Gituru WR, Wang QF (2005b). Genetic variation within the endangered quillwort Isoëtes hypsophila (Isoëtaceae) in China as evidenced by ISSR analysis. Aquatic Botany 82:89-98.

Chen JM, Wang QF (2006). Genetic diversity in the rare and endangered fern Isoetes orientalis. Journal of Wuhan Botanical Research 24:569-573.

Chen GQ, Huang HW, Crawford DJ, Pan BR, Ge XJ (2009). Mating system and genetic diversity of a rare desert legume Ammopiptanthus nanus (Leguminosae). Journal of Systematic and Evolution 47:57-66.

Chen YY, Liao Li, Li W, Li ZZ (2010a). Genetic diversity and population structure of the endangered alpine quillwort Isoetes hypsophila Hand.-Mazz. revealed by AFLP markers. Plant Systematics and Evolution 290:127-139.

Chen YY, Han QX, Cheng Y, Li ZZ, Li W (2010b). Genetic variation and clonal diversity of the endangered aquatic fern Ceratopteris pteridoides as revealed by AFLP analysis. Biochemical Systematics and Ecology 38:1129-1136.

Cheng JY, Jiang DZ, Cheng H, Zhou XH, Fang YY, Zhang XH, Xiao X, Deng XZ, Li LL (2018). Determination of Camellia oleifera Abel. germplasm resources of genetic diversity in China using ISSR markers. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 46:165-172.

Dong ZR, Chen JM, Wang QF (2006). RAPD analysis for mating system in Caldesia parnassifolia. Journal of Wuhan Botanical Research 24:167-170.

Dong YH, Chen JM, Gituru WR, Wang QF (2007). Gene flow in populations of the endangered aquatic fern Ceratopteris pteridoides in China as revealed by ISSR. Aquatic Botany 87:69-74.

Dong YH, Gituru WR, Wang QF (2010). Genetic variation and gene flow in the endangered aquatic fern Ceratopteris pteridoides in China, and conservation implication. Annales Botanici Fennici 47:34-44.

Dong X, Liao K, Hao F, Liu H,Qin R (2018a). Mating system and genetic variability of the endangered endemic aquatic lycophyte, Isoetes yunguiensis, in China determined using AFLP markers. American Fern Journal 108(2):47-61.

Dong X, Liu H, Gao W, Qin R, Gichira AW, Wang M, Liao K (2018b). Estimation of mating system in the endangered aquatic fern Ceratopteris pteridoides in China based on AFLP molecular marker and selfing test: implications for conservation. Notulae Botanicae Horti Agrobotanici Cluj-Napoca 46:688-699.

Doyle JJ, Doyle JL (1987). A rapid DNA isolation method for small quantities of fresh tissues. Phytochemical Bulletin 19:11-15.

Esselman EJ, Li JQ, Crawford D, Winduss JL, Wolfe AD (1999). Clonal diversity in the rare Calamagrostis porteri ssp. insperata (Poaceae): comparative results for allozymes and random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. Molecular Ecology 8:443-451.

Excoffier L, Lischer H (2015). Arlequin ver. 3.5.2: An integrated software package for population genetics data analysis. Institute of Ecology and Evolution, University of Berne, Switzerland. http://cmpg.unibe.ch/software/arlequin3.

Fernando DD, Discenza JJ, Bouchard JR, Leopold DJ (2015). Genetic analysis of the threatened American hart’s-tongue fern (Asplenium scolopendrium var. americanum [Fernald] Kartesz and Gandhi): Insights into its mating system and implications for conservation. Biochemical Systematics and Ecology 62:25-35.

Gaiotto FA, Bramucci M, Grattapaglia D (1997). Estimation of outcrossing rate in a breeding population of Eucalyptus urophylla with dominant RAPD and AFLP markers. Theoretical and Applied Genetics 95:842-849.

Han YC, Teng CZ, Wahiti GR, Zhou MQ, Hu ZL, Song YC (2009). Mating system and genetic diversity in natural populations of Nelumbo nucifera (Nelumbonaceae) detected by ISSR markers. Plant Systematics and Evolution 277:13-20.

Hickok LG, Warne TR, Fribourg RS (1995). The biology of the fern Ceratopteris and its use as a model system. International Journal of Plant Sciences 156:332-345.

Hoot SB, Taylor WC (2001). The utility of nuclear ITS, a LEAFY homolog intron, and chloroplast atpB-rbcL spacer region data in phylogenetic analyses and species delimitation in Isoetes. American Fern Journal 91:166-177.

Holsinger KE (2000). Reproductive systems and evolution in vascular plants. Proceedings of the National Academy of Sciences of the United States of America 97:7037-7042.

Kim C, Shin H, Choi HK (2009). Genetic diversity and population structure of diploid and polyploidy species of Isoetes in East Asia based on AFLP marker. International Journal of Plant Sciences 170: 496-504.

Korpelainen H (1995). Mating system and distribution of enzyme genetic variation in bracken (Pteridium aquilinum). Canadian Journal of Botany 73:1611-1617.

Kang M, Ye Q, Huang H (2005). Genetic consequence of restricted habitat and population decline in endangered Isoetes sinensis (Isoetaceae). Annals of Botany 96(7):1265-1274.

Li JM, Jin ZX (2008). Genetic structure of endangered Emmenopterys henryi Oliv. Based on ISSR polymorphism and implications for its conservation. Genetica 133: 227-234.

Li XD, Tan GJ, Li XX, Liu BD (2015). Comparative study about development of microgametopyte of two endangered endemic quillwort species in China. Acta Botanica Boreali-Occidentalia Sinica 35:1142-1147.

Liu H, Wang QF, Taylor WC (2005). Isoetes orientalis (Isoetaceae), a new hexaploid quillwort from China. Novon 15:164-167.

Liu X, Gituru WR, Wang QF (2004). Distribution of basic diploid and polyploid species of Isoetes in East Asia. Journal of Biogeography 31:1239-1250.

Nei M, Li WH (1979). Mathematical model for studying genetic variation in terms of restriction endonucleases. Proceedings of the National Academy of Sciences of the United States of America 76:5269-5273.

Orive ME, Asmussen MA (2000). The effect of pollen and seed migration on nuclear-dicytoplasmic systems. II. A new method for estimating plant gene flow from joint nuclear-cytoplasmic data. Genetics 155:833-854.

Pang XA, Liu X, Liu H, Wu C, Wang JY, Yang SX, Wang QF (2003). The geographic distribution and habitat of the Isoetes plants in China. Biodiversity Science 11:288-294.

Peredo EL, Méndez-Couz M, Revilla MA, Fernández H (2013). Mating system in Blechnum spicant and Dryopteris affinis ssp. affinis correlates with genetic variability. American Fern Journal 103:27-39.

Pometti CL, Bessega CF, Vilardi JC, Saidman BO (2013). Comparison of mating system parameters and genetic structure in three natural scenarios of Acacia visco (Leguminosae, Mimosoideae). Plant Systematics and Evolution 299:761-771.

Primack RB (1993). Essentials of conservation biology. Sinauer Associates, Inc, Sunderland, MA.

Ritland K, Jain S (1981). A model for the estimation of outcrossing rate and gene frequencies using n independent loci. Heredity 47: 35-52.

Ritland K (1990). A series of FORTRAN computer programs for estimating plant mating systems. Journal of Heredity 81:235-237.

Ritland K (2009). Multilocus mating system program MLTR. Version 3.4. University of British Columbia, Canada. http://genetics.forestry.ubc.ca/ritland/programs.html.

Rohlf FJ (1998). NTSYSpc: Numerical taxonomy and multivariate analysis system, version 2.02. Exeter Software, Setauket, New York.

Saki S, Bagheri H, Deljou A, Zeinalabedini M (2016). Evaluation of genetic diversity amongst Descurainia sophia L. genotypes by inter-simple sequence repeat (ISSR) marker. Physiology and Molecular Biology of Plants 22:97-105.

Small R, Hickey RJ (1997). Levels and patterns of genetic variation in Isoetes karstenii with observations on I. palmeri. American Fern Journal 87:104-115.

Soltis PS, Soltis DE (1990). Genetic variation within and among populations of ferns. American Fern Journal 80:161-172.

Suwarni E, Omondi SF, Dancik BP, Khasa DP (2018). Estimation of pollen contamination and mating system in Pinus merkusii seedling seed orchard using allozyme markers. Journal of Tropical Forest Science 30:95-105.

Wang QF, Liu X, Taylor WC, He ZR (2002). Isoetes yunguiensis (Isoetaceae), a new basic diploid quillwort from China. Novon 12:587-591.

Wright S (1978). Variability within and among natural populations. University of Chicago Press, Chicago.

Wu ZH, Qin RC (1991). Fern Families and Genera of China. Science Press, Beijing.

Yang H, Chen YY, Xu YX, Li ZZ (2011). Gene flow dynamics of ex-situ conservation populations in two endangered Isoetes species: genetic implications for reintroduction, conservation and management. Plant Science Journal 20:319-330.

Yeh FC, Yang R, Boyle TJ (2000). PopGene32: Microsoft Windows-based Freeware for Population Genetic Analysis (version 1.32). Molecular Biology and Biotechnology Centre, University of Alberta, Alberta, Canada.

Yu YF (1999). A milestone of wild plant conservation in China. Plants 5:3-11.

Zietkiewicz E, Rafalski A, Labuda D (1994). Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20:176-183.

Zhao HB, Zhou LH, Liu HH, Bao ZY (2014). Genetic effects of different mating modes in Sinocalycanthus chinensis (Cheng et S.Y. Chang) Cheng et S.Y. Chang, an endangered species endemic to Zhejiang Province, China. Biochemical Systematics and Ecology 54:8-14.




DOI: http://dx.doi.org/10.15835/nbha47111264

June 1, 2018: Notulae Botanicae Horti Agrobotanici Cluj-Napoca in Scopus – Elsevier CiteScore 2017=0.78, Horticulture; Agronomy and Crop Science; Plant Science


 
http://not-bot-horti-agrobo.blogspot.com/
https://www.facebook.com/NotBotHA
https://twitter.com/NotBotHA