安徽三大水系入侵物种克氏原螯虾的种群遗传格局

doi:10.3724/SP.J.1003.2010.398

摘要/Abstract

摘要：

克氏原螯虾(Procambarus clarkii)是长江中下游地区较为常见的外来物种, 目前已经扩散到安徽境内的主要水系。我们采集了安徽境内长江、淮河和新安江三大流域的4个地区9个地点的182个样本, 利用9对微卫星分子标记, 通过PCR扩增和微卫星分型, 分析了其种群遗传多样性水平和遗传结构, 构建了地方种群的遗传距离格局。结果表明本研究区域克氏原螯虾种群遗传多样性处于较高水平, 总的期望杂合度(H_E)和观测杂合度(H_O)分别为0.78和0.36。望江种群遗传多样性最高(期望杂合度为0.76), 肥西种群遗传多样性最低(期望杂合度为0.56)。克氏原螯虾种群显示了极强的杂合子缺失, 人为捕捞可能是造成其种群私有等位基因数目较多和杂合子缺失的主要原因。地方种群分化系数(F_st)与地理距离呈显著正相关(r= 0.33, P< 0.05), 望江种群和城东湖种群存在明显的跳跃性扩散(jump dispersal)。因此, 水系间的交流是种群扩散的主要途径, 人为贸易促使克氏原螯虾种群在不同地区的交流, 对其种群扩散也有一定的作用。

关键词: Procambarus clarkii, 微卫星, 遗传结构, 生物入侵, 物种扩散

Abstract

Procambarus clarkii is an alien invasive crayfish species that occurs in the major river basins along the middle and lower reaches of the Yangtze River. To understand genetic diversity and population structure of the species in the region, we collected 182 individuals from nine sites in four regions in the Yangtze, Huaihe, and Xin'anjiang River basins in Anhui Province. Using nine pairs of microsatellite markers, we studied the genetic differentiation, genetic structure and genetic clustering pattern of these P. clarkiipopulations. The expected heterozygosity (H_E) was 0.78 while the observed heterozygosity (H_O) was 0.36. Of the nine populations, the highest expected heterozygosity (0.76) was in the Wangjiang population, and the lowest (0.56) was for the Feixi population. Using a Bottleneck Test, we found a significant signal of heterozygote deficiency, indicating a recent decline in these populations. Our studies also showed that there were high levels of gene flow among populations, and a significant correlation found between genetic distance and geographic distance (r= 0.33, P< 0.05). We detected evidence of a significant jump dispersal event between the Xin’anjiang and Chengdonghu populations, thus indicating that human-mediated dispersal has played an important role in the present genetic structure ofProcambarus clarkiipopulation.

Key words: Procambarus clarkii, microsatellite DNA, genetic structure, biological invasion, species dispersal

曹玲亮, 周立志, 张保卫 (2010) 安徽三大水系入侵物种克氏原螯虾的种群遗传格局. 生物多样性, 18, 398-407. DOI: 10.3724/SP.J.1003.2010.398.

Lingliang Cao, Lizhi Zhou, Baowei Zhang (2010) Genetic patterns of an invasive Procambarus clarkii population in the three river basins of Anhui Province. Biodiversity Science, 18, 398-407. DOI: 10.3724/SP.J.1003.2010.398.

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://www.biodiversity-science.net/CN/10.3724/SP.J.1003.2010.398

https://www.biodiversity-science.net/CN/Y2010/V18/I4/398

图/表 8

参考文献 64

[1]	Barbaresi S, Fani R, Gherardi F, Mengoni A, Souty-Grosset C (2003) Genetic variability of European populations of an invasive American crayfish: preliminary results. Biological Invasions, 5,269-274.
[2]	Barbaresi S, Gherardi F, Mengoni A, Souty-Grosset C (2007) Genetics and invasion biology in fresh waters:a pilot study of Procambarus clarkii in Europe. In: Biological Invaders in Inland Waters: Profile, Distribution and Threats (ed. Gherardi F), pp.381-400. Springer, Dordrecht.
[3]	Belfiore NM, May B (2000) Variable microsatellite loci in red swamp crayfish, Procambarus clarkii, and their characterization in other crayfish taxa. Molecular Ecology, 9,2231-2234. URL PMID
[4]	Berg DJ, Garton DW, Macisaac HJ, Panov VE, Telesh IV (2002) Changes in genetic structure of North American Bythotrephes populations following invasion from Lake Ladoga, Russia. Freshwater Biology, 47,275-282.
[5]	Castric V, Bernatchez L, Belkhir K, Bonhomme F (2002) Heterozygote deficiencies in small lacustrine populations of brook charr Salvelinus fontinalis Mitchill (Pices, Salmonidae): a test of alternative hypotheses. Heredity, 89,27-35. URL PMID
[6]	Chen YF (陈毅峰), Yan YZ (严云志) (2005) Evolutionary biology of invasions. Acta Hydrobiologica Sinica (水生生物学报), 29,220-224. (in Chinese)
[7]	Chu D (褚栋), Zhang YJ (张友军), Wan FH (万方浩) (2007) Application of molecular marker techniques in invasion ecology. Chinese Journal of Applied Ecology (应用生态学报), 18,1383-1387. (in Chinese with English abstract)
[8]	DeAssis JB, DeLaat DM, Peixoto MGCD, Bergmann JAG, Fonseca CG, Carvalho MRS (2009) Genetic diversity and population structure in Brazilian Mangalarga Marchador horses. Genetics and Molecular Research, 8,1519-1524. DOI URL PMID
[9]	Facon B, Pointier JP, Glaubrecht M, Poux C, Jarne P, David P (2003) A molecular phylogeography approach to biological invasions of the New World by parthenogenetic Thiarid snails. Molecular Ecology, 12,3027-3039. URL PMID
[10]	Friar EA, Ladoux TA, Roalson EH, Robichaux RH (2000) Microsatellite analysis of a population crash and bottleneck in the Mauna Kea silverword, Argyroxiphium sandwicense ssp. sandwicense (Asteraceae), and its implications for reintroduction. Molecular Ecology, 9,2027-2034. DOI URL PMID
[11]	Gaffney PM, Scott TM, Koehn RK, Diehl WJ (1990) Interrelationships of heterozygosity, growth rate heterozygote deficiencies in the coot clam, Mutinia lateralis. Genetics and Molecular Biology, 124,687-699.
[12]	Geiger W, Alcorlo P, Baltanas A, Montes C (2005) Impact of an introduced Crustacean on the trophic webs of Mediterranean wetlands. Biological Invasions, 7,49-73.
[13]	Gherardi F (2006) Crayfish invading Europe: the case study of Procambarus clarkii. Marine and Freshwater Behaviour and Physiology, 39,175-191.
[14]	Gherardi F, Silvia B (2000) Invasive crayfish: activity patterns of Procambarus clarkii in the rice fields of the Lower Guadalquivir (Spain). Archiv für Hydrobiologie, 150,153-168. (in Spanish with English abstract).
[15]	Gregorius HR (1980) The probability of losing an allele when diploid genotype are sampled. Biometrics, 36,632-652.
[16]	Guo XM (郭晓鸣), Zhu SQ (朱松泉) (1997) A preliminary study on the larval development of the crayfish Procambarus clarkii. Acta Zoologica Sinica (动物学报), 43,372-381. (in Chinese with English abstract)
[17]	Hassan M, Bonhomme F (2005) No reduction in neutral variability of mitochondrial and nuclear genes for a Lessepsian migrant, Upeneus moluccensis. Journal of Fish Biology, 66,865-870.
[18]	Hedrick PW (1984) Population Biology: The Evolution and Ecology of Populations. Jones and Bartlett Publishers, Boston.
[19]	Herborg LM, Weetman D, Oosterhout C, Hänfling B (2007) Genetic population structure and contemporary dispersal patterns of a recent European invader, the Chinese mitten crab, Eriocheir sinensis. Molecular Ecology, 16,231-242. URL PMID
[20]	Hernández L, Maeda-Martínez AM, Ruiz-Campos G, Rodríguez-Almaraz G, Alonzo-Rojo F, Sainz JC (2007) Geographic expansion of the invasive red crayfish Procambarus clarkii (Girard, 1852) (Crustacea: Decapoda) in Mexico. Biological Invasions, 10,977-984.
[21]	Huner (1988) Procambarus in North America and elsewhere. In: Freshwater Crayfish: Biology, Management and Exploitation (eds Holdich DM, Lowery RS), pp.239-261. Timber Press, London.
[22]	Jensen JL, Bohonak AJ, Kelley ST (2005) Isolation by distance, web service. BMC Genetics, 6,13. URL PMID
[23]	Kerby JL, Riley SPD, Kats LB, Wilson P (2005) Barriers and flow as limiting factors in the spread of an invasive crayfish (Procambarus clarkii) in southern California streams. Biological Conservation, 126,402-409. DOI URL
[24]	Kitamoto N, Honjo M, Ueno S, Takenaka A, Tsumura Y, Washitani I, Ohsawa R (2005) Spatial genetic structure among and within populations of Primula sieboldii growing beside separate streams. Molecular Ecology, 14,149-157. DOI URL PMID
[25]	Kolbe JJ, Glor RE, Schettino LR, Lara AC, Larson A, Losos JB (2004) Genetic variation increases during the biological invasion of a Cuban lizard. Nature, 431,177-181. URL PMID
[26]	Kolbe JJ, Larson A, Losos JB, Queiroz K (2008) Admixture determines genetic diversity and population differentiation in the biological invasion of a lizard species. Biology Letters, 4,434-437. URL PMID
[27]	Kumar S, Nei M, Dudley J, Tamura K (2008) MEGA: a biologist-centric software for evolutionary analysis of DNA and protein sequences. Briefings in Bioinformatics, 9,299-306. URL PMID
[28]	Lee CE (2002) Evolutionary genetics of invasive species. Trends in Ecology and Evolution, 17,386-391.
[29]	Li CC (1995) Population Genetics. The University of Chicago Press, Chicago and London.
[30]	Li LP (李浪平), Lü JL (吕建林), Gong SY (龚世园), Shu XY (舒新亚), Gong LJ (龚珞军) (2006) Studies on the morphology of Procambarus clarkii. Reservoir Fisheries (水利渔业), 26(3),40-42. (in Chinese)
[31]	Li SC (李顺才), Xu XY (徐兴友), Du LQ (杜利强), Yin XL (尹秀玲), Meng XD (孟宪东), Xie JY (解敬瑶) (2005) Investigation on and analysis of alien invasions in Chinese farming industry. Chinese Agricultural Science Bulletin (中国农学通报), 21(6),156-159. (in Chinese with English abstract)
[32]	Li ZY (李振宇), Xie Y (解焱) (2002) Invasive Alien Species in China (中国外来入侵种). China Forestry Publishing House, Beijing. (in Chinese)
[33]	Manel S, Schwartz MK, Gordon L, Pierre T (2003) Landscape genetics: combining landscape ecology and population genetics. Trends in Ecology and Evolution, 18,189-197. DOI URL
[34]	Manning AD, Lindenmayer DB, Nix HA (2004) Continua and umwelt: novel perspectives on viewing landscapes. Oikos, 104,621-628.
[35]	Maria JC, Rebelo R (2007) Colonization of freshwater habitats by an introduced crayfish, Procambarus clarkii, in Southwest Iberian Peninsula. Hydrobiologia, 575,191-201.
[36]	Morrison LJ, Tweedie A, Black A, Pinchbeck GL, Christley RM, Schoenefeld A, Hertz-Fowler C, MacLeod A, Turner CM, Tait A (2009) Discovery of mating in the major African livestock pathogen Trypanosoma congolense. PLoS ONE, 4,e5564. URL PMID
[37]	Park SDE (2001) Trypanotolerance in West African Cattle and the Population Genetic Effects of Selection. PhD dissertation of University of Dublin, Dublin.
[38]	Parker IM, Simberloff D, Lonsdale WM, Goodell K, Wonham M, Kareiva PM, Williamson MH, Holle BV, Moyle PB, Byers JE, Goldwasser L (1999) Impact: toward a framework for understanding the ecological effects of invaders. Biological Invasions, 1,3-19.
[39]	Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics, 155,945-959. URL PMID
[40]	Raymond M, Rousset F (1995) GENEPOP (version 1.2): population genetics software for exact tests and ecumenicism. Journal of Heredity, 86,248-249.
[41]	Rousset F (2008) GENEPOP' 007: a complete re-implemen- tation of the GENEPOP software for Windows and Linux. Molecular Ecology Resources, 8,103-106. DOI URL PMID
[42]	Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4,406-425. URL PMID
[43]	Sakai AK, Allendorf FW, Holt JS, Lodge DM, Molofsky J, With KA, Baughman S, Cabin RJ, Cohen JE, Ellstrand NC, McCauley DE, O'Neil P, Parker IM, Thompson JN, Weller SG (2001) The population biology of invasive species. Annual Review of Ecology and Systematics, 32,305-332.
[44]	Sambrook J, Russell DW (2001) Molecular Cloning: A Laboratory Manual, 3rd edn. Cold Spring Harbor Laboratory Press, New York.
[45]	Serrano M, Calvo JH, Martínez M, Marcos-Carcavilla A, Cuevas J, González C, Jurado JJ, de Tejada PD (2009) Microsatellite based genetic diversity and population structure of the endangered Spanish Guadarrama goat breed. BMC Genetics, 10,61. URL PMID
[46]	Shi GR (史刚荣), Ma CC (马成仓) (2006) Biological characteristics of alien plants successful invasion. Chinese Journal of Applied Ecology (应用生态学报), 17,727-732. (in Chinese with English abstract)
[47]	Simberloff D (2005) The politics of assessing risk for biological invasions: the USA as a case study. Trends in Ecology and Evolution, 20,216-222. URL PMID
[48]	Sjogren P, Wyoni PI (1994) Conservation genetics and detection of rare alleles in finite populations. Conservation Biology, 8,267-270.
[49]	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. URL PMID
[50]	Stepien CA, Taylor CD, Dabrowska KA (2002) Genetic variability and phylogeographical patterns of a nonindigenous species invasion: a comparison of exotic vs. native zebra and quagga mussel populations. Journal of Evolutionary Biology, 15,314-328.
[51]	Suarez AV, Holway DA, Case TJ (2001) Patterns of spread in biological invasions dominated by long-distance jump dispersal: insights from Argentine ants. Proceedings of the National Academy of Sciences, USA, 98,1095-1100.
[52]	Tiunov AV, Cindy H, Andrew H, Tamara VP (2006) Invasion patterns of Lumbricidae into the previously earthworm-free areas of northeastern Europe and the western Great Lakes region of North America. Biological Invasions, 8,1223-1234.
[53]	Tsutsui ND, Suarez AV, Holway DA, Case TJ (2001) Relationships among native and introduced populations of the Argentine ant, Linepithema humile and the source of introduced populations. Molecular Ecology, 10,2151-2161. URL PMID
[54]	Valles-Jimenez R, Cruz P, Perez-Enriquez R (2005) Population genetic structure of pacific white shrimp (Litopenaeus vannamei) from Mexico to Panama microsatellite DNA variation. Marine Biotechnology, 6,475-484. URL PMID
[55]	Villanelli F, Gherardi F (1998) Breeding in the crayfish, Austropotamobius pallipes: mating patterns, mate choice and intermale competition. Freshwater Biology, 40,305-315. DOI URL
[56]	Wang CZ (王长忠), Li Z (李忠), Liang HW (梁宏伟), Hu GF (呼光富), Wu QC (吴勤超), Zou GW (邹桂伟), Luo XZ (罗相忠) (2009) Genetic diversity in four Procambarus clarkii populations in the lower reaches of the Yangtze River. Biodiversity Science (生物多样性), 17,518-523. (in Chinese with English abstract)
[57]	Wu ZJ (武正军), Cai FJ (蔡凤金), Jia YF (贾运锋), Lu JX (鲁建鑫), Jiang YF (蒋勇福), Huang CM (黄乘明) (2008) Predation impact of Procambarus clarkii on Rana limnocharis tadpoles in Guilin area. Biodiversity Science (生物多样性), 16,150-155. (in Chinese with English abstract)
[58]	Xie Y (解焱), Wang S (汪松) (2001) Review on invasive species in China. In: Protecting the Biodiversity of China (II) (保护我国的生物多样性) (eds Schei P, Xie Y (解焱), Wang S (汪松)), pp.91-96. China Environmental Science Press, Beijing. (in Chinese)
[59]	Xu RM (徐汝梅), Ye WH (叶万辉) (2003) Biological Invasion: Theory and Practice (生物入侵: 理论与实践). Science Press, Beijing. (in Chinese)
[60]	Yan LN (闫路娜), Zhang DX (张德兴) (2004) Effects of sample size on various genetic diversity measures in population genetic study with microsatelite DNA markers. Acta Zoologica Sinica (动物学报), 50,279-290. (in Chinese with English abstract)
[61]	Yasuda N, Nagai S, Hamaguchi M, Okaji K, Gérard K, Nadaoka K (2009) Gene flow of Acanthaster planci (L.) in relation to ocean currents revealed by microsatellite analysis. Molecular Ecology, 18,1574-1590. DOI URL PMID
[62]	Yue GH, Li JL, Bai ZY, Wang CM, Feng F (2010) Genetic diversity and population structure of the invasive alien red swamp crayfish. Biological Invasions, 12,2697-2706.
[63]	Zalewski A, Piertney SB, Zalewska H, Lambin X (2009) Landscape barriers reduce gene flow in an invasive carnivore: geographical and local genetic structure of American mink in Scotland. Molecular Ecology, 18,1601-1615. DOI URL PMID
[64]	Zhu ZY, Yue GH (2008) Eleven polymorphic microsatellites isolated from red swamp crayfish, Procambarus clarkii. Molecular Ecology Resources, 8,796-798. URL PMID

地方种群 Local populations	编号 Code	采集点 Sampling site	样本量 Sample size	期望杂合度 H_E	观测杂合度 H_O	平均等位基因数 MNA	私有等位基因数 Np
淮河 Huaihe River
城东湖 Chengdong Lake	Cd	霍邱黄泊渡 Huangbodu, Huoqiu	24	0.72	0.31	6.62	5
江淮之间 Between Yangtze River and Huaihe River
巢湖 Chaohu Lake	Ch	义城 Yicheng	7	0.65	0.24	4.89	5
巢湖 Chaohu Lake	Ch	长临河口 Changlin River Mouths	3	0.65	0.24	4.89	0
肥西 Feixi	Fx	柿树岗 Stream in Shishugang	12	0.56	0.39	4.44	3
合肥 Hefei	Hf	董铺水库 Dongpu Reservoir	6	0.57	0.33	4.75	2
合肥 Hefei	Hf	三十头支流 Stream in Sanshitou	9	0.57	0.33	4.75	1
长江 Yangtze River
菜子湖 Caizi Lake	Czh	王湾咀 Wangwanzui	7	0.67	0.47	9.89	4
		杨湾 Yangwan	3				0
		肖店 Xiaodian	21				6
		双店 Shuangdian	12				3
升金湖 Shengjin Lake	Sj	升金湖 Shenjin Lake	13	0.71	0.56	6.33	5
东至 Dongzhi	Dz	香口乡 Stream in Xiangkou	21	0.74	0.33	7.55	9
望江 Wangjiang	Wj	华阳河 Huayang River	20	0.76	0.17	7.44	18
新安江 Xin'anjiang River
新安江 Xin'anjiang River	Xa	屯光镇 Tunguang	24	0.64	0.39	6.00	3
总计 Total			182	0.67	0.35	-	-

地方种群 Local populations	编号 Code	采集点 Sampling site	样本量 Sample size	期望杂合度 H_E	观测杂合度 H_O	平均等位基因数 MNA	私有等位基因数 Np
淮河 Huaihe River
城东湖 Chengdong Lake	Cd	霍邱黄泊渡 Huangbodu, Huoqiu	24	0.72	0.31	6.62	5
江淮之间 Between Yangtze River and Huaihe River
巢湖 Chaohu Lake	Ch	义城 Yicheng	7	0.65	0.24	4.89	5
巢湖 Chaohu Lake	Ch	长临河口 Changlin River Mouths	3	0.65	0.24	4.89	0
肥西 Feixi	Fx	柿树岗 Stream in Shishugang	12	0.56	0.39	4.44	3
合肥 Hefei	Hf	董铺水库 Dongpu Reservoir	6	0.57	0.33	4.75	2
合肥 Hefei	Hf	三十头支流 Stream in Sanshitou	9	0.57	0.33	4.75	1
长江 Yangtze River
菜子湖 Caizi Lake	Czh	王湾咀 Wangwanzui	7	0.67	0.47	9.89	4
		杨湾 Yangwan	3				0
		肖店 Xiaodian	21				6
		双店 Shuangdian	12				3
升金湖 Shengjin Lake	Sj	升金湖 Shenjin Lake	13	0.71	0.56	6.33	5
东至 Dongzhi	Dz	香口乡 Stream in Xiangkou	21	0.74	0.33	7.55	9
望江 Wangjiang	Wj	华阳河 Huayang River	20	0.76	0.17	7.44	18
新安江 Xin'anjiang River
新安江 Xin'anjiang River	Xa	屯光镇 Tunguang	24	0.64	0.39	6.00	3
总计 Total			182	0.67	0.35	-	-

位点 Loci	重复单元Repeat unit	引物序列 Primer sequence(5'-3')	退火温度 Annealing temperature (°C)	长度 Size range (bp)	登录号 Accession no.	N_a	H_E	H_O	PIC	F_is	F_it	F_st
PclG33	(GT)₂₁	F: FAM-TTCGAGGCGTTGCTGATTGTAAGT R: CAAGGAAGCGTATAGCCGGAGTCT	56	120-180	AF290936	23	0.83	0.46	0.81	0.50	0.56	0.110
PCL29	(GA)₂₇	F: HEX-CACTCAAGCCTGCCCTCACTC R: GTCTCTTCCTCCCCCATTCTCAC	56	170-214	EF564126	14	0.87	0.44	0.85	0.55	0.55	0.108
PCL47	(TG)₁₃	F: TAMRA-ACTCTGCCCATTGTTTCTCGG R: AGCCCTTGGACCCCGCCTATC	56	324-344	EF564128	10	0.79	0.82	0.75	0.57	0.62	0.110
PCL33	(TG)₃₀	F: FAM-ACTCCTGTCCCATTTCACTAC R: ACAACTAACTGCAACTCATTCTA	56	132-158	EF564127	24	0.81	0.43	0.78	0.49	0.54	0.100
PCL28	(CA)₂₉	F: HEX-CCTACCAGAGAACCCAAAACAGAA R: GTCAGCCTCCACCACATCACTT	56	229-239	EF564125	23	0.82	0.35	0.80	0.50	0.56	0.111
PCL17	(TC)₁₇	F: TAMRA-TTCACTACCGCCCACAGGATG R: TAGGCACCGCACTTATTAGACCAG	56	370-422	EF564122	18	0.71	0.10	0.69	0.45	0.51	0.111
PclG48	(CA)₁₂	F: FAM-CTGTTGGTGATTTCCGTCAATTTT R: AGATTCAACGCTGTGTTCCTGATC	56	146-190	AF290941	22	0.81	0.24	0.79	0.47	0.53	0.108
PCL02	(TG)₃₀	F: HEX-GAAGACGGGACACCACGAG R: ATCAAATCAAACGAAGCAAGAAAG	56	245-271	EF564119	24	0.86	0.23	0.84	0.46	0.52	0.113
PCL50	(GA)₁₂	F: TAMRA-AAGCGCTGAAATGCACAAACAAGA R: CAAGCCCCGAGGTCAAAGGTC	56	424-525	EF564129	19	0.53	0.17	0.51	0.48	0.53	0.105

位点 Loci	重复单元Repeat unit	引物序列 Primer sequence(5'-3')	退火温度 Annealing temperature (°C)	长度 Size range (bp)	登录号 Accession no.	N_a	H_E	H_O	PIC	F_is	F_it	F_st
PclG33	(GT)₂₁	F: FAM-TTCGAGGCGTTGCTGATTGTAAGT R: CAAGGAAGCGTATAGCCGGAGTCT	56	120-180	AF290936	23	0.83	0.46	0.81	0.50	0.56	0.110
PCL29	(GA)₂₇	F: HEX-CACTCAAGCCTGCCCTCACTC R: GTCTCTTCCTCCCCCATTCTCAC	56	170-214	EF564126	14	0.87	0.44	0.85	0.55	0.55	0.108
PCL47	(TG)₁₃	F: TAMRA-ACTCTGCCCATTGTTTCTCGG R: AGCCCTTGGACCCCGCCTATC	56	324-344	EF564128	10	0.79	0.82	0.75	0.57	0.62	0.110
PCL33	(TG)₃₀	F: FAM-ACTCCTGTCCCATTTCACTAC R: ACAACTAACTGCAACTCATTCTA	56	132-158	EF564127	24	0.81	0.43	0.78	0.49	0.54	0.100
PCL28	(CA)₂₉	F: HEX-CCTACCAGAGAACCCAAAACAGAA R: GTCAGCCTCCACCACATCACTT	56	229-239	EF564125	23	0.82	0.35	0.80	0.50	0.56	0.111
PCL17	(TC)₁₇	F: TAMRA-TTCACTACCGCCCACAGGATG R: TAGGCACCGCACTTATTAGACCAG	56	370-422	EF564122	18	0.71	0.10	0.69	0.45	0.51	0.111
PclG48	(CA)₁₂	F: FAM-CTGTTGGTGATTTCCGTCAATTTT R: AGATTCAACGCTGTGTTCCTGATC	56	146-190	AF290941	22	0.81	0.24	0.79	0.47	0.53	0.108
PCL02	(TG)₃₀	F: HEX-GAAGACGGGACACCACGAG R: ATCAAATCAAACGAAGCAAGAAAG	56	245-271	EF564119	24	0.86	0.23	0.84	0.46	0.52	0.113
PCL50	(GA)₁₂	F: TAMRA-AAGCGCTGAAATGCACAAACAAGA R: CAAGCCCCGAGGTCAAAGGTC	56	424-525	EF564129	19	0.53	0.17	0.51	0.48	0.53	0.105

	Ch	Czh	Fx	Sj	Dz	Wj	Hf	Xa	Cd
Ch		4.00	1.56	4.08	2.59	1.63	2.86	1.52	3.13
Czh	0.06		1.67	9.49	2.52	1.62	5.75	1.38	1.92
Fx	0.14	0.13		2.89	1.23	2.00	1.71	0.86	1.27
Sj	0.06	0.03	0.08		2.72	2.58	3.97	1.27	2.03
Dz	0.09	0.09	0.17	0.08		2.66	1.54	2.72	2.26
Wj	0.13	0.13	0.11	0.09	0.09		1.80	1.92	2.41
Hf	0.08	0.04	0.13	0.06	0.14	0.12		1.11	1.57
Xa	0.14	0.15	0.23	0.16	0.08	0.12	0.18		1.69
Cd	0.07	0.12	0.16	0.11	0.10	0.09	0.14	0.13