Genetic diversity of the habitat-forming red alga Gracilaria vermiculophylla along Chinese coasts

doi:10.17520/biods.2016038

Abstract

Abstract:

As a habitat-forming species, the red alga Gracilaria vermiculophylla has been noted to play an essential role in shaping coastal marine communities and maintaining intertidal ecosystems along Chinese coasts. However, few studies have been conducted that analyze the genetic structure of the population and the distribution patterns of genetic variation. In this study, we compiled partial mitochondrial DNA cytochrome c oxidase subunit I gene (cox1) of 19 G. vermiculophylla populations (461 individuals) and performed phylogenetic analysis and population genetic surveys. A total of 461 mt-DNA cox1 sequences were obtained and edited into 641 bp, which contained 21 polymorphic sites and yielded 15 haplotypes. Phylogenetic inferences, haplotype networking, and principal coordinate analysis consistently indicated that G. vermiculophylla populations diverged into two groups: the northern group is comprised of specimens from the Yellow-Bohai Sea and the Shengsi Islands, while the southern group is comprised of specimens from six locations south to Xiamen, Fujian Province. Analysis of the molecular variance and Nei’s genetic distances indicated low genetic differentiation between populations in each group while group-level genetic divergence is comparable to the degree of subspecies differentiation.

http://jtp.cnki.net/bilingual/detail/html/SWDY201607006

Key words: Gracilaria vermiculophylla, mitochondrial cox1, phylogenetics, genetic diversity, population genetic differentiation

Ruoyu Liu, Zhongmin Sun, Jianting Yao, Zimin Hu, Delin Duan. Genetic diversity of the habitat-forming red alga Gracilaria vermiculophylla along Chinese coasts[J]. Biodiv Sci, 2016, 24(7): 781-790.

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URL: https://www.biodiversity-science.net/EN/10.17520/biods.2016038

https://www.biodiversity-science.net/EN/Y2016/V24/I7/781

Figures/Tables 7

References 36

[1]	Bandelt HJ, Forster P, Röhl A (1999) Median-joining networks for inferring intraspecific phylogenies. Molecular Biology and Evolution, 16, 37-48.
[2]	Bellorin AM, Olivera MC, Olivera EC (2004) Gracilaria vermiculophylla: a western Pacific species of Gracilariaceae (Rhodophyta) first recorded from the eastern Pacific. Phycological Research, 52, 69-79.
[3]	Boo GH, Kim KM, Nelson WA, Rafael RR, Yoon KJ, Boo SM (2014) Taxonomy and distribution of selected species of the agarophyte genus Gelidium (Gelidiales, Rhodophyta). Journal of Applied Phycology, 26, 1243-1251.
[4]	Cheang CC, Chu KH, Put AO (2010) Phylogeography of the marine macroalga Sargassum hemiphyllum (Phaeophyceae, Heterokontophyta) in northwestern Pacific. Molecular Ecology, 19, 2933-2948.
[5]	Chen Z, Zhang J, Fu HF, Xu ZZ, Deng KZ, Zhang JY (2012) On the validity of the species Phenacoccus solenopsis based on morphological and mitochondrial COI data, with the description of a new body color variety. Biodiversity Science, 20, 443-450. (in Chinese with English abstract)
	[陈哲, 张姜, 傅杭飞, 许争争, 邓坤正, 张加勇 (2012) 基于形态特征和线粒体COI基因探讨扶桑绵粉蚧物种的有效性并记述一体色变异型扶桑绵粉蚧. 生物多样性, 20, 443-450.]
[6]	Chu D, Liu GX, Fu HB, Xu W (2009) Phylogenetic analysis of mt-COI reveals the cryptic lineages in Phenacoccus solenopsis complex (Hemiptera: Pseudococcidae). Acta Entomologica Sinica, 52, 1261-1265.
[7]	Geraldino PJL, Yang EC, Kim MS, Boo SM (2009) Systematics of Hypnea asiatica sp. nov. (Hypneaceae, Rhodophyta) based on morphology and nrDNA SSU, plastid rbcL, and mitochondrial cox1. Taxon, 58, 606-616.
[8]	Guillemin ML, Faugeron S, Destombe C, Viard F, Correa JA, Valero M (2008) Genetic variation in wild and cultivated populations of the haploid-diploid red alga Gracilaria chilenesis: how farming practices favor asexual reproduction and heterozygosity. Evolution, 62, 1500-1519.
[9]	Guiry MD, Guiry GM (2016) AlgaeBase. World-wide electronic publication, National University of Ireland, Galway.. (accessed on 2016-04-12.
[10]	Gulbransen DJ, McGlathery KJ, Marklund M, Norris JN, Gurgel CFD (2012) Gracilaria vermiculophylla (Rhodophyta, Gracilariales) in the Virginia coastal bays, USA: cox1 analysis reveals high genetic richness of an introduced macroalga. Journal of Phycology, 48, 1278-1283.
[11]	Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95-98.
[12]	Hebert PD, Cywinska A, Ball SL, deWaard JR (2003) Biological identifications through DNA barcodes. Proceedings of the Royal Society B: Biological Sciences, 270, 313-321.
[13]	Hu ZM, Zhang J, Lopez-Bautista J, Duan DL (2013) Asymmetric genetic exchange in the brown seaweed Sargassum fusiforme (Phaeophyceae) driven by oceanic currents. Marine Biology, 160, 1407-1414.
[14]	Hu ZM, Juan LB (2014) Adaptation mechanisms and ecological consequences of seaweed invasions: a review case of agarophyte Gracilaria vermiculophylla. Biological Invasions, 16, 967-976.
[15]	Kim SY, Weinberger F, Boo SM (2010) Genetic data hint at a common donor region for invasive Atlantic and Pacific populations of Gracilaria vermiculophylla (Gracilariales, Rhodophyta). Journal of Phycology, 46, 1346-1349.
[16]	Librado P, Roas J (2009) DNAsp v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformations, 25, 1451-1452.
[17]	Li JJ, Zhang J, Hu ZM, Duan DL (2013) Population genetics and demographic history of red seaweed, Palmaria palmata, from the Canada-northwest Atlantic. Biodiversity Science, 21, 306-314. (in Chinese with English abstract)
	[李晶晶, 张杰, 胡自民, 段德麟 (2013) 加拿大西北大西洋地区掌形藻的种群遗传结构与动态变化. 生物多样性, 21, 306-314.]
[18]	Peakall R, Smouse PE (2012) GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research—an update. Bioinformatics, 28, 2537-2539.
[19]	Peason EA, Murray SN (1997) Patterns of reproduction, genetic diversity and genetic differentiation in California populations of the geniculate coralline alga Lithothrix aspergillum (Rhodophyta). Journal of Phycology, 33, 753-763.
[20]	Posada D, Crandall KA (1998) Modeltest: testing the model of DNA substitution. Bioinformatics, 14, 817-818.
[21]	Riosmena RR, Talavera SA, Acosta VB, Gardner SC (2009) Heavy metals dynamics in seaweeds and seagrasses in Bahía Magdalena, BCS, México. Journal of Applied Phycology, 22, 283-291.
[22]	Robba L, Russell SJ, Barker GL, Brodie J (2006) Assessing the use of the mitochondrial Cox1 marker for use in DNA barcoding of red algae (Rhodophyta). American Journal of Botany, 93, 1101-1108.
[23]	Rueness J (2005) Life history and molecular sequences of Gracilaria vermiculophylla (Gracilariales, Rhodophyta), a new introduction to European waters. Phycologia, 44, 120-128.
[24]	Saunders GW (2005) Applying DNA barcoding to red macroalgae: a preliminary appraisal holds promise for future applications. Philosophical Transactions of the Royal Society of London Series B: Biological Sciences, 360, 1879-1888.
[25]	Saunders GW (2009) Routine DNA barcoding of Canadian Gracilariales (Rhodophyta) reveals the invasive species Gracilaria vermiculophylla in British Columbia. Molecular Ecology Resources, 9(Suppl. 1), 140-150.
[26]	Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution, 30, 2725-2729.
[27]	Terada R, Yamamoto H (2002) Review of Gracilaria vermiculophylla and other species in Japan and Asia. Taxonomy of Economic Seaweeds, 8, 215-224.
[28]	Thomsen MS, Stæhr PA, Nejrup L, Schiel DR (2013) Effects of the invasive macroalgae Gracilaria vermiculophylla on two co-occurring foundation species and associated invertebrates. Aquatic Invasions, 8, 133-145.
[29]	Tseng CK, Chen JF (1959) Reproductive of Gracilaria vermiculophylla and cultivation seedings inlaboratory. 90, 202-203. (in Chinese)
	[曾呈奎, 陈椒芬 (1959) 真江蓠的繁殖习性和幼苗的室内培育. 中国科学院海洋研究所调查研究报告, 90, 202-203.]
[30]	Tseng CK (2008) Seaweeds in Yellow Sea and Bohai Sea of China. Science Press, Beijing. (in Chinese)
	[曾呈奎 (2008) 中国黄渤海海藻. 科学出版社, 北京.]
[31]	Wang PX (1999) Response of western Pacific marginal seas to glacial cycles: paleoceanographic and sedimentological features. Marine Geology, 156, 5-39.
[32]	Xia BM, Chang CF (1999) Flora Algarum Marinarum Sinicarum Tomus II, Rhodophyta no. V Ahnfeltiales, Gigartinales, Rhodymeniales. Science Press, Beijing. (in Chinese)
	[夏邦美, 张峻甫 (1999) 中国海藻志第二卷红藻门(第五分册): 伊谷藻目, 杉藻目, 红皮藻目. 科学出版社, 北京.]
[33]	Yang EC, Kim MS, Geraldino PJL, Sahoo D, Shin JA, Boo SM (2008) Mitochondrial cox1 and plastid rbcL genes of Gracilaria vermiculophylla (Gracilariaceae, Rhodophyta). Journal of Applied Phycology, 20, 161-168.
[34]	Zhao XB, Pang SJ, Shan TF, Liu F (2013) Applications of three DNA barcodes in assorting intertidal red macroalgal flora in Qingdao, China. Journal of Ocean University of China, 12, 139-145.
[35]	Zheng WJ, Zhu SH, Shen XQ, Liu BQ, Pan ZC, Ye YF (2009) Genetic differentiation of Tegillarca granosa based on mitochondrial COI gene sequences. Zoological Research, 30, 17-23. (in Chinese with English abstract)
	[郑文娟, 朱世华, 沈锡权, 刘必谦, 潘志崇, 叶央芳 (2009) 基于线粒体COI基因序列探讨泥蚶的遗传分化. 动物学研究, 30, 17-23.]
[36]	Zhong CH, Huang RF, Lin Q, Zheng YY, Li LB, Liu B (2014) Molecular identification of a green type of Gracilaria vermiculophylla. Journal of Applied Oceanography, 33, 183-189. (in Chinese with English abstract)
	[钟晨辉, 黄瑞芳, 林琪, 郑雅友, 李雷斌, 刘波 (2014) 一种绿色真江蓠的分子鉴定. 应用海洋学报, 33, 183-189.]

代号 Code	地理位置 Location	经纬度 Longitude/ Latitude	N	Np	Nh	Hn(n)	Hd	π (× 10^-2)	采样人 Collectors
南方群体 Region South
YT	北海银滩 Yingtang beach, Beihai	109.34 ºE, 21.45 ºN	24	7	4	H0(21), H2, H3, H4	0.239	0.143	刘若愚, 胡自民 Ruoyu Liu, Zimin Hu
JH	北海金海岸 Jinhaian, Beihai	109.11 ºE, 21.49 ºN	33	1	2	H0(32), H2	0.061	0.009	刘若愚, 胡自民 Ruoyu Liu, Zimin Hu
TC	湛江特呈岛 Techeng island, Zhanshan,	110.44 ºE, 21.15 ºN	30	0	1	H0(30)	0.000	0.000	孙忠民, 姚建亭 Zhongmin Sun, Jianting Yao
NA	汕头南澳岛 Nanao island, Shantou	117.04 ºE, 23.45 ºN	23	1	2	H0(19), H1(4)	0.300	0.047	孙忠民, 姚建亭 Zhongmin Sun, Jianting Yao
ZP	漳州漳浦六鳌 Zhangpu liuao, Zhanzhou,	117.77 ºE, 23.92 ºN	35	0	1	H0(35)	0.000	0.000	孙忠民, 黄超华 Zhongmin Sun, Chaohua Huang
XL	厦门杏林湾 Xingling bay, Xiamen	118.08 ºE, 24.60 ºN	5	1	2	H0(4), H1(1)	0.400	0.062	钟晨辉等 Chenhui Zhong et al
小计 Subtotal			150	8	5	H0(141), H1(5), H2(2), H3, H4	0.116	0.027
北方群体 Region North
SS	舟山嵊泗岛 Shengsi island, Zhoushan	122.44 ºE, 30.68 ºN	19	0	1	H6(19)	0.000	0.000	胡自民 Zimin Hu
YY	青岛一浴 No. 1 beach, Qingdao	120.34 ºE, 36.05 ºN	21	2	3	H5(19), H7, H10	0.186	0.030	刘若愚 Ruoyu Liu
EY	青岛二浴 No. 2 beach, Qingdao	120.34 ºE, 36.05 ºN	21	0	1	H5(21)	0.000	0.000	刘若愚 Ruoyu Liu
SY	青岛三浴 No. 3 beach	120.36 ºE, 36.05 ºN	39	0	1	H5(39)	0.000	0.000	刘若愚 Ruoyu Liu
YH	青岛银海国际 Yinhai, Qingdao	120.42 ºE, 36.06 ºN	36	2	3	H5(26), H8(9), H13	0.427	0.069	刘若愚 Ruoyu Liu
LR	青岛石老人 Shilaoren sea, Qingdao	120.49 ºE, 36.09 ºN	36	2	3	H5(32), H9(3), H12	0.208	0.033	刘若愚 Ruoyu Liu
SD	威海石岛 Stone island, Weihai	122.41 ºE, 36.91 ºN	13	0	1	H5(13)	0.000	0.000	李晶晶, 张杰 Jingjing Li, Jie Zhang
DC	威海东楮岛 Dongchu island, Weihai	122.56 ºE, 37.04 ºN	32	1	2	H5(16), H14(16)	0.516	0.081	刘若愚, 胡自民Ruoyu Liu, Zimin Hu
JM	威海鸡鸣岛 Jiming island, Weihai	122.48 ºE, 37.45 ºN	7	0	1	H5(7)	0.000	0.000	李晶晶, 张杰 Jingjing Li, Jie Zhang
CD	烟台长岛 Long island, Yantai	120.72 ºE, 37.94 ºN	16	0	1	H5(16)	0.000	0.000	胡自民 Zimin Hu
HN	大连黄泥川 Huangninchuan, Dalian	121.56 ºE, 38.82 ºN	35	1	2	H5(34), H11	0.057	0.009	刘若愚, 胡自民 Ruoyu Liu, Zimin Hu
HS	大连黑石礁 Heshijiao, Dalian	121.56 ºE, 38.87 ºN	8	0	1	H5(8)	0.000	0.000	刘若愚, 胡自民 Ruoyu Liu, Zimin Hu
ZZ	大连獐子岛 Zhangzi island, Dalian	122.74 ºE, 39.04 ºN	28	0	1	H5(28)	0.000	0.000	姚建亭, 孙忠民 Ruoyu Liu, Zimin Hu
小计 Subtotal			311	9	10	H5(259), H6(19), H7, H8(9), H9(3) H10, H11, H12, H13, H14(16)	0.300	0.050

代号 Code	地理位置 Location	经纬度 Longitude/ Latitude	N	Np	Nh	Hn(n)	Hd	π (× 10^-2)	采样人 Collectors
南方群体 Region South
YT	北海银滩 Yingtang beach, Beihai	109.34 ºE, 21.45 ºN	24	7	4	H0(21), H2, H3, H4	0.239	0.143	刘若愚, 胡自民 Ruoyu Liu, Zimin Hu
JH	北海金海岸 Jinhaian, Beihai	109.11 ºE, 21.49 ºN	33	1	2	H0(32), H2	0.061	0.009	刘若愚, 胡自民 Ruoyu Liu, Zimin Hu
TC	湛江特呈岛 Techeng island, Zhanshan,	110.44 ºE, 21.15 ºN	30	0	1	H0(30)	0.000	0.000	孙忠民, 姚建亭 Zhongmin Sun, Jianting Yao
NA	汕头南澳岛 Nanao island, Shantou	117.04 ºE, 23.45 ºN	23	1	2	H0(19), H1(4)	0.300	0.047	孙忠民, 姚建亭 Zhongmin Sun, Jianting Yao
ZP	漳州漳浦六鳌 Zhangpu liuao, Zhanzhou,	117.77 ºE, 23.92 ºN	35	0	1	H0(35)	0.000	0.000	孙忠民, 黄超华 Zhongmin Sun, Chaohua Huang
XL	厦门杏林湾 Xingling bay, Xiamen	118.08 ºE, 24.60 ºN	5	1	2	H0(4), H1(1)	0.400	0.062	钟晨辉等 Chenhui Zhong et al
小计 Subtotal			150	8	5	H0(141), H1(5), H2(2), H3, H4	0.116	0.027
北方群体 Region North
SS	舟山嵊泗岛 Shengsi island, Zhoushan	122.44 ºE, 30.68 ºN	19	0	1	H6(19)	0.000	0.000	胡自民 Zimin Hu
YY	青岛一浴 No. 1 beach, Qingdao	120.34 ºE, 36.05 ºN	21	2	3	H5(19), H7, H10	0.186	0.030	刘若愚 Ruoyu Liu
EY	青岛二浴 No. 2 beach, Qingdao	120.34 ºE, 36.05 ºN	21	0	1	H5(21)	0.000	0.000	刘若愚 Ruoyu Liu
SY	青岛三浴 No. 3 beach	120.36 ºE, 36.05 ºN	39	0	1	H5(39)	0.000	0.000	刘若愚 Ruoyu Liu
YH	青岛银海国际 Yinhai, Qingdao	120.42 ºE, 36.06 ºN	36	2	3	H5(26), H8(9), H13	0.427	0.069	刘若愚 Ruoyu Liu
LR	青岛石老人 Shilaoren sea, Qingdao	120.49 ºE, 36.09 ºN	36	2	3	H5(32), H9(3), H12	0.208	0.033	刘若愚 Ruoyu Liu
SD	威海石岛 Stone island, Weihai	122.41 ºE, 36.91 ºN	13	0	1	H5(13)	0.000	0.000	李晶晶, 张杰 Jingjing Li, Jie Zhang
DC	威海东楮岛 Dongchu island, Weihai	122.56 ºE, 37.04 ºN	32	1	2	H5(16), H14(16)	0.516	0.081	刘若愚, 胡自民Ruoyu Liu, Zimin Hu
JM	威海鸡鸣岛 Jiming island, Weihai	122.48 ºE, 37.45 ºN	7	0	1	H5(7)	0.000	0.000	李晶晶, 张杰 Jingjing Li, Jie Zhang
CD	烟台长岛 Long island, Yantai	120.72 ºE, 37.94 ºN	16	0	1	H5(16)	0.000	0.000	胡自民 Zimin Hu
HN	大连黄泥川 Huangninchuan, Dalian	121.56 ºE, 38.82 ºN	35	1	2	H5(34), H11	0.057	0.009	刘若愚, 胡自民 Ruoyu Liu, Zimin Hu
HS	大连黑石礁 Heshijiao, Dalian	121.56 ºE, 38.87 ºN	8	0	1	H5(8)	0.000	0.000	刘若愚, 胡自民 Ruoyu Liu, Zimin Hu
ZZ	大连獐子岛 Zhangzi island, Dalian	122.74 ºE, 39.04 ºN	28	0	1	H5(28)	0.000	0.000	姚建亭, 孙忠民 Ruoyu Liu, Zimin Hu
小计 Subtotal			311	9	10	H5(259), H6(19), H7, H8(9), H9(3) H10, H11, H12, H13, H14(16)	0.300	0.050

单倍型 Haplotype	单倍型数量 Number	核苷酸多态位点 Polymorphic sites																					GenBank accession number
单倍型 Haplotype	单倍型数量 Number	4	25	52	64	65	112	151	250	346	352	370	406	433	460	517	558	562	593	604	605	634	GenBank accession number
H0	141	T	G	A	A	C	G	A	A	G	T	T	T	A	G	G	C	T	G	G	G	A	KU136384
H1	5	T	A	A	A	C	G	A	A	G	T	T	T	A	G	G	C	T	G	G	G	A	KF789527
H2	2	T	G	A	A	C	G	A	A	G	T	T	T	A	G	G	C	T	G	G	G	G	KU136385
H3	1	T	G	T	A	C	T	T	A	G	T	T	T	A	G	G	C	T	T	G	A	A	KU136387
H4	1	A	G	A	A	C	G	A	A	G	T	T	T	A	G	G	C	T	G	G	G	A	KU136386
H5	259	T	G	A	A	C	G	A	A	C	T	T	T	A	G	A	C	C	G	T	G	A	KU136374
H6	19	T	G	A	A	C	G	A	A	C	G	T	T	A	G	A	C	C	G	T	G	A	KU136375
H7	16	T	G	A	G	C	G	A	A	C	T	T	T	A	G	A	C	C	G	T	G	A	KU136383
H8	9	T	G	A	A	T	G	A	A	C	T	T	T	A	G	A	C	C	G	T	G	A	KU136377
H9	3	T	G	A	A	C	G	A	A	C	T	A	T	A	G	A	C	C	G	T	G	A	KU136378
H10	1	T	G	A	A	C	G	A	A	C	T	T	C	A	G	A	C	C	G	T	G	A	KU136379
H11	1	T	G	A	A	C	G	A	A	C	T	T	T	A	G	A	A	C	G	T	G	A	KU136380
H12	1	T	G	A	A	C	G	A	A	C	T	T	T	A	A	A	C	C	G	T	G	A	KU136381
H13	1	T	G	A	A	C	G	A	T	C	T	T	T	A	G	A	C	C	G	T	G	A	KU136382
H14	1	T	G	A	A	C	G	A	A	C	T	T	T	T	G	A	C	C	G	T	G	A	KU136376

单倍型 Haplotype	单倍型数量 Number	核苷酸多态位点 Polymorphic sites																					GenBank accession number
单倍型 Haplotype	单倍型数量 Number	4	25	52	64	65	112	151	250	346	352	370	406	433	460	517	558	562	593	604	605	634	GenBank accession number
H0	141	T	G	A	A	C	G	A	A	G	T	T	T	A	G	G	C	T	G	G	G	A	KU136384
H1	5	T	A	A	A	C	G	A	A	G	T	T	T	A	G	G	C	T	G	G	G	A	KF789527
H2	2	T	G	A	A	C	G	A	A	G	T	T	T	A	G	G	C	T	G	G	G	G	KU136385
H3	1	T	G	T	A	C	T	T	A	G	T	T	T	A	G	G	C	T	T	G	A	A	KU136387
H4	1	A	G	A	A	C	G	A	A	G	T	T	T	A	G	G	C	T	G	G	G	A	KU136386
H5	259	T	G	A	A	C	G	A	A	C	T	T	T	A	G	A	C	C	G	T	G	A	KU136374
H6	19	T	G	A	A	C	G	A	A	C	G	T	T	A	G	A	C	C	G	T	G	A	KU136375
H7	16	T	G	A	G	C	G	A	A	C	T	T	T	A	G	A	C	C	G	T	G	A	KU136383
H8	9	T	G	A	A	T	G	A	A	C	T	T	T	A	G	A	C	C	G	T	G	A	KU136377
H9	3	T	G	A	A	C	G	A	A	C	T	A	T	A	G	A	C	C	G	T	G	A	KU136378
H10	1	T	G	A	A	C	G	A	A	C	T	T	C	A	G	A	C	C	G	T	G	A	KU136379
H11	1	T	G	A	A	C	G	A	A	C	T	T	T	A	G	A	A	C	G	T	G	A	KU136380
H12	1	T	G	A	A	C	G	A	A	C	T	T	T	A	A	A	C	C	G	T	G	A	KU136381
H13	1	T	G	A	A	C	G	A	T	C	T	T	T	A	G	A	C	C	G	T	G	A	KU136382
H14	1	T	G	A	A	C	G	A	A	C	T	T	T	T	G	A	C	C	G	T	G	A	KU136376

	YT	JH	TC	NA	ZP	XL	SS	YY	EY	SY	YH	LR	SD	DC	JM	CD	HN	HS	ZZ
YT	0.000
JH	0.000	0.000
TC	0.000	0.000	0.000
NA	0.002	0.001	0.001	0.000
ZP	0.000	0.000	0.000	0.001	0.000
XL	0.003	0.002	0.002	0.000	0.002	0.000
SS	0.277	0.272	0.272	0.276	0.272	0.277	0.000
YY	0.217	0.213	0.213	0.216	0.213	0.217	0.049	0.000
EY	0.215	0.212	0.211	0.215	0.211	0.215	0.049	0.000	0.000
SY	0.215	0.212	0.211	0.215	0.211	0.215	0.049	0.000	0.000	0.000
YH	0.222	0.218	0.217	0.221	0.217	0.222	0.052	0.003	0.003	0.003	0.000
LR	0.217	0.213	0.213	0.216	0.213	0.217	0.049	0.001	0.000	0.000	0.003	0.000
SD	0.215	0.212	0.211	0.215	0.211	0.215	0.049	0.000	0.000	0.000	0.003	0.000	0.000
DC	0.233	0.230	0.229	0.233	0.229	0.234	0.062	0.012	0.012	0.012	0.015	0.013	0.012	0.000
JM	0.215	0.212	0.211	0.215	0.211	0.215	0.049	0.000	0.000	0.000	0.003	0.000	0.000	0.012	0.000
CD	0.215	0.212	0.211	0.215	0.211	0.215	0.049	0.000	0.000	0.000	0.003	0.000	0.000	0.012	0.000	0.000
HN	0.216	0.212	0.212	0.215	0.212	0.216	0.049	0.000	0.000	0.000	0.003	0.000	0.000	0.012	0.000	0.000	0.000
HS	0.215	0.212	0.211	0.215	0.211	0.215	0.049	0.000	0.000	0.000	0.003	0.000	0.000	0.012	0.000	0.000	0.000	0.000
ZZ	0.215	0.212	0.211	0.215	0.211	0.215	0.049	0.000	0.000	0.000	0.003	0.000	0.000	0.012	0.000	0.000	0.000	0.000	0.000