7 ISSR 13 Flemingia macrophylla Leguminosae Flemingia Roxb DNA 11 0.% 260 ~ 1 00 m Beckman DU 00 ng μl PCR 07 UBC 02 ISSR 2 11 μl 60 1

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1 Guihaia Jul http / /journal.gxzw.gxib.cn http / / DOI.1131/guihaia.gxzw ISSR J WANG GJ XIAO WX ANG SX et al. Analysis of genetic diversity of Flemingia macrophylla resources from South Yunnan by ISSR J. Guihaia ISSR * ISSR 7 Flemingia macrophylla PPL 4.5% Ne Nei s He Shannon s Ho PPL = 43.44% Ne = He = Ho = 0.24 Nei s Gst % 3.75% MY QB NE ISSR Q4.7 A Analysis of genetic diversity of Flemingia macrophylla resources from South Yunnan by ISSR WANG Gui-Juan * XIAO Wen-Xiang ANG Shou-Xian Xishuangbanna ropical Botanical Garden Chinese Academy of Sciences Mengla China Abstract he genetic diversity of 135 individuals collected from 7 wild populations of Flemingia macrophylla was estimated using ISSR method. An high level of genetic diversity was detected in wild populations of F. macrophylla from South Yunnan Province. At species level the percentage of polymorphic loci PPL = 4.5% effective number of alleles Ne = Nei s gene diversity He = and Shannon s information index Ho = at population level PPL = 43.44% Ne = Nei s gene diversity He = and Shannon s in formation index Ho = he level of genetic differentiation 3. 75% among populations is lower than that within populations 60.25%. Based on the results we suggest that the protected areas of F. macrophylla should be build in MY QB and NE because of the high level of genetic diversity. Key words Flemingia macrophylla genetic diversity ISSR Supported by Innovation Project Pilot Funding of Xishuangbana ropical Botanical Garden Chinese Academy of Sciences wanggj@ xtbg.org.cn *

2 7 ISSR 13 Flemingia macrophylla Leguminosae Flemingia Roxb DNA 11 0.% 260 ~ 1 00 m Beckman DU 00 ng μl PCR 07 UBC 02 ISSR 2 11 μl PCR Bio-Rad PCR ISSR Inter-Simple Sequence Repeat ISSR μl 1 Buffer 2.0 mmol L -1 MgCl μmol μl -1 dnps 0.5 μmol μl U aq 07 Wang et al 12 Liu et al 13 CAB DNA DNA OYOBO 25 ng DNA PCR 11 ISSR min s ~ s s 72 7 min 1.4 PCR 1% 0 V 30 min Marker 0 bp DNA lader UVP 0 11 ISSR 1.5 DNA ~ bp 0 /1 POPGENE 1 Yeh et al PPL Na Ne Nei s 173 He Shannon Ho Ht Hs Gst Nei s 17 D I Nei s NSYS-pc2.1

3 14 36 able 1 1 Locality habitat altitude geographical location and sample size of Flemingia macrophylla populations from Yunnan Population code Locality and habitat Sample size Altitude m Geographical location LC 17 he secondary forest in Lancang Puer City ' N 57' E NE he field in Liming Ninger Puer City ' N 0 5' E QB he field in Qiubei Wenshan City ' N 4 36' E XM he secondary forest in Xiangming Mengla Jinghong City ' N 1 ' E MY he secondary forest in Yiwu Mengla Jinghong City ' N 1 3' E YY he secondary forest between Yangwu and Shiping Yuxi City ' N 2 ' E GS he swidden land in Nanlin Gasa Jinghong City ' N 0 30' E ISSR able 2 ISSR primers used for generating ISSR markers from 135 individuals of 7 Flemingia populations Primer 5'-3' Sequence 5'-3' Annealing temperature No. of bands scored No. of polymorphic bands 0 AG C GA 13 C 14 C A 16 CA 1 25 CA G 2 AC G 35 A AG YC 1 56 AC Y Fig. 1 Locations of the 7 sampled Flemingia macrophylla AC populations YA Population codes are the same as those in able 1 Note Y = C. UPGMA Rohlf 00 Nm = Gst /Gst 4.5% Nei s He Shannon Ho Na ~ Ne ~ ISSR 7 PPL 25.77% ~ 62.% 43.44% 6 ~ 11 MY 2 PPL YY Nei s He

4 7 ISSR Ht = Hs = able 3 Genetic diversity of Flemingia populations Dst = Ht-Hs Nei s Nei's Shannon Gst = % Population PPL 60.25% Na Ne He Ho % LC NE QB XM MY 1.62 YY GS Mean At species level Note Na. Observed number of alleles Ne. Effective number of alleles He. Nei s gene diversity Ho. Shannon s information index PPL. Percentage of polymorphic loci. 4 Nei's 17 able 4 Nei s 17 analysis of gene diversity in Flemingia populations ISSR Item Nm Nei s Ht Gst Hs He Mean Standard deviation He = Nybom & Bartish 04 Nybom 00 Note Ht. total gene diversity Hs. gene diversity within populations Gst. coefficient of gene differentiation Nm. gene flow. Nm = Gst / Gst Nm = Gst /Gst Nei s I ~ D 0.0 ~ UPGMA GS YY GS LC He ~ Shannon Ho ~ MY > Hamrick et al 11 Hamrick & Godt 1 QB > NE > XM > PPL = 4. 5% Na = GS > LC > YY He = Gst = MY PPL He PPL = 62.% Ne = Gst He = Ho = PPL = N e = He = 0.04 Ho = MY 2.2 PPL 62.% He

5 able 5 7 Nei's 17 I D Nei s 17 genetic identity and genetic distance of 7 Flemingia populations LC NE QB XM MY YY GS LC **** NE **** QB **** XM **** MY **** YY **** GS **** Nei s Note Nei s genetic identity above diagonal and genetic distance below diagonal. 2 Nei s UPGMA Fig. 2 UPGMA dendrogram for 7 Flemingia populations based on Nei s genetic identity GUAN YH MA J PENG JM et al 0. Investigation on resources of Flemingia Roxb. ex Ait. et Ait. f. in Xishuangbanna J. Chin Med Mat J GUAN ZB ZHANG LX GAO WW 11. Study on germination XM PPL characteristics of Flemingia macrophylla seeds J. Chin Agric Sci Bull % He J HAMRICK JL GOD MJW MURAWSKI DA et al 11. Correlations between species traits and allozyme diversity implications 3.2 for conservation biology M / /FALK DA HOLSINGER % KE. Genetics and conservation of rare plants. New York Oxford University Press % HAMRICK JL GOD MJW 1. Allozyme diversity in plant species C / / BROWN AHD CLEGG M KAHLER AL et al. Plant population genetics breeding and genetic resources C. Sunderland Massachusetts USA Sinauer LI BQ SONG QS 0. Chemical constituents in roots of Nm Flemingia macrophylla J. Chin radit Herb Drugs < J LI CS ZHANG LX ZHAO JL et al 11. Comparison of 2 7 isoflavones content of Moghania macrophylla from different areas

6 7 ISSR 17 of Yunnan Province J. Mod Chin Med and relationship of global faba bean Vicia faba L germplasm revealed by ISSR markers J. heor Appl Genet J LI L QING MJ ZHANG LX et al 0. Advance in the study on the chemcal composition and biological activity of Flemingia Roxb. ex Ait. et Ait. f. J. Drugs Clin WEI YZ 11. Preliminary study on the China Flemingia genus J. Guihaia J XIANG ZY SONG SQ WANG GJ et al 07. Genetic diversity of 0. Jatropha curcas Euphorbiaceae collected from southern Yunnan detected J LIU JF SHI SQ CHANG E et al 13. Genetic diversity of the critically endangered huja sutchuenensis revealed by ISSR markers and the implications for conservation J. Intern J Mol Sci LV FJ YANG CY ANG QR et al 11. Study of various economic benefit Pueraria of Wallichii and Flemingia macrophylla by inter-simple sequence repeat ISSR J. Acta Bot Yunnan ISSR J YEH FC YANG RC BOYLE et al 17. POPGENE the user friendly shareware for population genetic analysis M. Edmonton University of Alberta. J. For Invent Plann ZHANG LX PENG JM MA J 07. Research progress of Flemingia 11. J. J. Chin Med Mat NYBOM H BARISH IV 00. Effects of life history traits and 07. J ZHANG ZL ZHANG LX SONG MF et al 11. Analysis of sampling strategies on genetic diversity estimates obtained with genetic relationship and quality assessment on plants in RAPD markers in plants J. Persp Plant Ecol Evol Syst NYBOM H 04. Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants J. Mol Ecol ROHLF FJ 00. NSYS-pc numerical taxonomy and multivariate analysis system version 2.1 M. New York Exeter Software. WANG HF ZONG XX GUAN JP et al 12. Genetic diversity Flemingia Roxb. ex Ait. et Ait. f. J. Chin rad Herb Drugs J ZHAO Q 02. Flemingia macrophylla-excellent forage germplasm resource in tropical area of Yunnan J. Sichuan Grassl J 櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍櫍 72 Continue from page 72 ZHOU CM ZHAO P NI GY et al 11. Whole -tree water use bution to transpiration J. Chin J Appl Ecol characteristics of Schima superba in wet and dry seasons based on sap flow and soil-leaf water potential gradient analysis J. Chin J J. Ecol ZHOU CM HUANG YQ GU DX et al 15. he radial variation of J xylem sap. flow deusity in trunk of Eucalyptus urophylla E. grandis J. Chin J Ecol nighttime stem sap flow of Schima superba in Guangzhou region of 15. ZHOU CM ZHAO P NI GY et al 12. Water recharge through Guangdong Province South China affecting factors and contri- J

AFLP % 1. 1 DNA 5 RAPD DNA 0. 1g % ~ ph 6 10 DNA 9 40μl TE SSR STS 20ng /L

AFLP % 1. 1 DNA 5 RAPD DNA 0. 1g % ~ ph 6 10 DNA 9 40μl TE SSR STS 20ng /L 2012 13 2 201-206 Journal of Plant Genetic Resources / 071001 41 SSR 21 SSR 41 52 2 ~ 4 2. 5 Shannon 0. 21 ~ 1. 40 0. 73 5. 54 3 7 9 13. 16% ~ 186. 49% 5. 48 6 11 SSR Genetic Diversity of Peanut Arachis