华东地区地方鸡品种mtDNA控制区遗传多样性

doi:10.17520/biods.2017012

摘要/Abstract

摘要：

为了探明我国华东地区地方鸡品种的遗传多样性和群体遗传结构, 追溯其母系起源和进化过程, 利用PCR技术扩增了11个地方鸡品种的线粒体DNA (mtDNA)控制区(D-loop)序列, 并结合NCBI数据库中已发表的红色原鸡(Gallus gallus) D-loop区全序列, 分析了它们的遗传多样性与亲缘关系, 构建了11个品种与红色原鸡系统发生邻接树。结果表明：11个地方品种mtDNA D-loop区全长为1,231或1,232 bp, 其中1,231 bp的序列有196条, 1,232 bp的序列有123条, 经过比对发现, 两者在859 bp处存在单碱基缺失。11个地方品种319个个体共计检测到变异位点37个, 总体单倍型多样度核苷酸多样度和平均核苷酸差异分别为0.901 ± 0.009、0.00573 ± 0.000001和6.833。按照鸡mtDNA单倍型分类通用标准, 共包含35种单倍型, 可以分为A、B、C和E共4个分支(单倍型群), 分别包括11、10、9和5个单倍型。中介网络图中11个鸡品种也很明显地分成了4个支系, 分别含有100、118、47和54条序列。系统发育树分为4个大枝, 海南亚种(G. gallus jabouillei)自成一枝; C单倍型群与4个亚种的红色原鸡聚为一枝; E单倍型群与2个亚种红色原鸡聚为一枝; A和B单倍型群只与滇南亚种(G .gallus spadiceus)聚为一枝。11个品种中, 除了狼山和丝羽乌骨鸡2个标准化品种外, 都有很高的遗传多样性, 可开发选择潜力很大。没有发现线粒体品种特异性DNA序列。华东地区地方品种至少有4个母系起源, 部分品种可能受到了欧美高产品系的渗入。

关键词: 系统发育, 红色原鸡, 控制区, 地方品种

Abstract

The objective of this study was to determine the origin and evolution of chickens in Eastern China by assessing the genetic diversities and structures of 11 local chicken breeds. The complete D-loop region of mitochondrial DNA (mtDNA) of 319 chickens from 11 local breeds were sequenced and analyzed together with published data for the red junglefowl. These sequences were then used in a neighbor-joining method to construct the phylogenetic tree of these breeds and the red junglefowl. The D-loop regions of the local breeds were characterized by 1,231 and 1,232 bp; the 1,231 bp haplotype had 196 sequences, while the 1,232 bp haplotype had 123 sequences, with a base C deficiency from the 859 bp site in the 1,231 bp haplotype. A total of 37 mutation sites were detected in the 319 individuals. The average haplotype diversity, nucleotide diversity, and nucleotide difference were 0.901 ± 0.009, 0.005 73 ± 0.000001 and 6.833, respectively. A total of 35 haplotypes were identified which belonged to four previously published clades, i.e. Clades A, B, C, and E, which contained 11, 10, 9, and 5 haplotypes, respectively. Median-joining network profiles of the haplotype indicated the 11 local breeds were divided into four maternal clades containing 100, 118, 47, and 54 sequences. The cluster of five subspecies of the red junglefowl and 11 local breeds were divided into four distinct groups. Gallus gallus jabouillei was grouped alone. Haplotype C and four subspecies of junglefowl were clustered in one group. Haplotype E and two subspecies of junglefowl were clustered in another group. Haplotypes A, B, and Gallus gallus spadiceus were clustered in another group. Chickens from eastern China revealed abundant genetic diversity in the mtDNA D-loop region, except for the Langshan and Silky breeds. No breed-specific matrilineal clade was observed. We conclude that chicken breeds from eastern China likely share four common maternal lineages, and that some chicken populations may have been mixed with exotic lineage chickens.

Key words: phylogeny, red junglefowl, D-loop region, local breed

贾晓旭, 唐修君, 樊艳凤, 陆俊贤, 黄胜海, 葛庆联, 高玉时, 韩威 (2017) 华东地区地方鸡品种mtDNA控制区遗传多样性. 生物多样性, 25, 540-548. DOI: 10.17520/biods.2017012.

Xiaoxu Jia, Xiujun Tang, Yanfeng Fan, Junxian Lu, Shenghai Huang, Qinglian Ge, Yushi Gao, Wei Han (2017) Genetic diversity of local chicken breeds in East China based on mitochondrial DNA D-loop region. Biodiversity Science, 25, 540-548. DOI: 10.17520/biods.2017012.

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链接本文: https://www.biodiversity-science.net/CN/10.17520/biods.2017012

https://www.biodiversity-science.net/CN/Y2017/V25/I5/540

图/表 7

参考文献 26

[1]	Bao WB, Chen GH, Wu XS, Xu Q, Wu SL, Shu JT, Weigend S (2007) Genetic diversity of red junglefowl in China (Gallus gallus spadiceus) and red junglefowl (Gallus gallus gallus) in Thailand. Hereditas (Beijing) , 29, 587-592. (in Chinese with English abstract)
	[包文斌, 陈国宏, 吴信生, 徐琪, 吴圣龙, 束婧婷, Steffen Weigend (2007) 中国红原鸡和泰国红原鸡遗传多样性分析. 遗传, 29, 587-592.]
[2]	Bao WB, Shu JT, Wang CB, Zhang HX, Weigend S, Chen GH (2008) Investigation on genetic diversity and systematic evolution in Chinese domestic fowls and red jungle fowls by analyzing the mtDNA control region. Acta Veterinaria et Zootechnica Sinica, 39, 1449-1459. (in Chinese with English abstract)
	[包文斌, 束婧婷, 王存波, 张红霞, Steffen Weigend, 陈国宏 (2008) 中国家鸡和红色原鸡mtDNA控制区遗传多态性及系统进化分析. 畜牧兽医学报, 39, 1449-1459.]
[3]	Chang CS, Chen CF, Berthouly-Salazar C, Chazara O, Lee YP, Chang CM, Chang KH, Bed’Hom B, Tixier-Boichard M (2012) A global analysis of molecular markers and phenotypic traits in local chicken breeds in Taiwan. Animal Genetics, 43, 172-182.
[4]	Chen KW, Li HF, Wang JY, Tang QP, Shen JC, Zhang SJ (2006) Study on genetic diversity of 27 indigenous chicken breeds or strains in East China. Acta Veterinaria et Zootechnica Sinica, 37, 7-11. (in Chinese with English abstract)
	[陈宽维, 李慧芳, 王金玉, 汤青萍, 沈见成, 章双杰 (2006) 华东27个地方鸡品种(品系)的遗传变异. 畜牧兽医学报, 37, 7-11.]
[5]	China National Commission of Animal Genetic Resources(2011) Animal Genetic Resources in China: Poultry. China Agriculture Press, Beijing. (in Chinese)
	[国家畜禽遗传资源委员会(2011) 中国畜禽遗传资源志:家禽志. 中国农业出版社, 北京.]
[6]	Fumihito A, Miyake T, Sumi S, Takada M, Ohno S, Kondo N (1994) One subspecies of the red junglefowl (Gallus gallus gallus) sufficies as the matriarchic ancestor of all domestic breeds. Proceedings of the National Academy of Sciences, USA, 91, 12505-12509.
[7]	Gao YS, Tang XJ, Tu YJ, Lu JX, Xue MY, Shi ZH, Zhang XY (2011) Studies on the DNA barcoding of fifteen chicken breeds by mtDNA COI gene. Scientia Agricultura Sinica, 44, 587-594. (in Chinese with English abstract)
	[高玉时, 唐修君, 屠云洁, 陆俊贤, 薛茂云, 施祖灏, 张小燕 (2011) 基于线粒体COI基因15个鸡种的DNA编码研究. 中国农业科学, 44, 587-594.]
[8]	Gao YS, Jia XX, Tang XJ, Tang MJ, Fan YF, Lu JX, Gu R, Ge QL, Su YJ (2015) The genetic diversity and origin analysis of Anyi Tile-like chickens (Gallus gallus domestiaus) based on mitochondrial DNA D-loop sequence. Journal of Agricultural Biotechnology, 23, 940-944. (in Chinese with English abstract)
	[高玉时, 贾晓旭, 唐修君, 唐梦君, 樊艳凤, 陆俊贤, 顾荣, 葛庆联, 苏一军 (2015) 基于线粒体基因组D-loop区全序列分析安义瓦灰鸡遗传多样性及其起源进化关系. 农业生物技术学报, 23, 940-944.]
[9]	Guan FR (2001) A review of Dongxiang blue egg chicken. Livestock and Poultry Industry, (7) , 22-23.
	(in Chinese) [官发荣 (2001) 东乡黑羽绿壳蛋鸡综述. 畜禽业, (7) , 22-23.]
[10]	Huang XH, Chen JB, He DL, Zhang XQ, Zhong FS (2016) DNA barcoding of indigenous chickens in China: a reevaluation. Scientia Agricultura Sinica, 49, 2622-2633. (in Chinese with English abstract)
	[黄勋和, 陈洁波, 何丹林, 张细权, 钟福生 (2016) DNA条形码技术鉴定中国地方鸡品种的重新评估. 中国农业科学, 49, 2622-2633.]
[11]	Jia XX, Tang XJ, Lu JX, Fan YF, Chen DW, Tang MJ, Gu R, Gao YS (2016) The investigation of genetic diversity and evolution of Daweishan Mini chicken based on the complete mitochondrial (mt)DNA D-loop region sequence. Mitochondrial DNA, 27, 3001-3004.
[12]	Kanginakudru S, Metta M, Jakati RD, Nagaraju J (2008) Genetic evidence from Indian red jungle fowl corroborates multiple domestication of modern day chicken. BMC Evolutionary Biology, 8, 174.
[13]	Liao Y, Mo G, Sun J, Wei F, Liao DJ (2016) Genetic diversity of Guangxi chicken breeds assessed with microsatellites and the mitochondrial DNA D-loop region. Molecular Biology Reports, 43, 1-11.
[14]	Liu YP, Wu GS, Yao YG, Miao YW, Luikart G, Baig M, Beja-Pereira A, Ding ZL, Palanichamy MG, Zhang YP (2006) Multiple maternal origins of chickens: out of the Asian jungles. Molecular Phylogenetics & Evolution, 38, 12-19.
[15]	Lu JX, Jia XX, Tang XJ, Fan YF, Tang MJ, Gao YS, Su YJ (2016) Genetic diversity of two local Yunnan chicken breeds and their relationships with red junglefowl. Journal of Zhejiang University (Agriculture and Life Sciences), 42, 385-390. (in Chinese with English abstract)
	[陆俊贤, 贾晓旭, 唐修君, 樊艳凤, 唐梦君, 高玉时, 苏一军 (2016) 2个云南原始鸡种遗传多样性及其与红色原鸡的亲缘关系. 浙江大学学报(农业与生命科学版), 42, 385-390.]
[16]	Miao YW, Peng MS, Wu GS, Ouyang YN, Yang ZY, Yu N, Liang JP, Piangchou G, Beja-Pereira A, Mitra B, Palanichamy MG, Baig M, Chaudhuri TK, Shen YY, Kong QP, Murphy RW, Yao YG, Zhang YP (2013) Chicken domestication: an updated perspective based on mitochondrial genomes. Heredity, 110, 277-282.
[17]	Mwacharo JM, Bjørnstad G, Mobegi V, Nomura K, Hanada H, Amano T (2011) Mitochondrial DNA reveals multiple introductions of domestic chicken in East Africa. Molecular Phylogenetics & Evolution, 58, 374-382.
[18]	Osman SAM, Yonezawa T, Nishibori M (2016) Origin and genetic diversity of Egyptian native chickens based on complete sequence of mitochondrial DNA D-loop region. Poultry Science, 95, 1248-1256.
[19]	Rozas J, Sanchezelbarrio JC, Messeguer X, Rozas R (2003) DnaSP, DNA polymorphism analyses by the coalescent and other methods. Bioinformatics, 19, 2496-2497.
[20]	Shi NN, Fan L, Yao YG, Peng MS, Zhang YP (2014) Mitochondrial genomes of domestic animals need scrutiny. Molecular Ecology, 23, 5393-5397.
[21]	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.
[22]	Thompsom JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The Clustal-X Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 25, 4876-4882.
[23]	Xiang H, Gao JQ, Yu BQ, Zhou H, Cai DW, Zhang YW, Chen XY, Wang XI, Hofreiter M, Zhao XB (2014) Early Holocene chicken domestication in northern China. Proceedings of the National Academy of Sciences, USA, 111, 17564-17569.
[24]	Yang N (2002) Poultry Production. China Agriculture Press， Beijing. (in Chinese)
	[杨宁 (2002) 家禽生产学. 中国农业出版社, 北京.]
[25]	Zhu WQ, Li HF, Wang JY, Shu JT, Zhu CH, Song WT, Song C, Ji GG, Liu HX (2014) Molecular genetic diversity and maternal origin of Chinese black-bone chicken breeds. Genetics & Molecular Research, 13, 3275-3282.
[26]	Zhuang GY, Gong YB (2015) Investigation and utilization of Langya chicken. Poultry Science, (9), 45-50. (in Chinese)
	[庄桂玉, 龚玉波 (2015) 琅琊鸡品种资源调查与保种利用. 家禽科学, (9), 45-50.]

品种及代号 Breed and code	原产地 Origin	现有种群规模及采样 Population (sample size)	经济用途 Economic use
安义瓦灰 Anyi Gray (AY)	江西安义 Anyi, Jiangxi	600,000 (30)	肉蛋兼用 E & M
白耳黄 Baier Yellow (BE)	江西广丰 Guangfeng, Jiangxi	20,000,000 (30)	蛋用 E
崇仁麻 Chongren Partridge (CP)	江西崇仁 Chongren, Jiangxi	52,000,000 (30)	肉蛋兼用 E & M
东乡绿壳 Dongxiang Blue-eggshell (DX)	江西东乡 Dongxiang, Jiangxi	200,000 (30)	肉蛋兼用 E & M
丝羽乌骨 Silkies (SL)	江西泰和 Taihe, Jiangxi	16,500,000 (30)	肉药兼用 Med & M
金湖乌凤 Jinhu Black-bone (JH)	福建泰宁 Taining, Fujian	160,000 (29)	肉药兼用 Med & M
仙居 Xianju (XJ)	浙江仙居 Xianju, Zhejiang	1,484,000 (30)	蛋用 E
琅琊 Langya (LY)	山东日照 Rizhao, Shandong	110,000 (30)	肉蛋兼用 E & M
寿光 Shouguang (SG)	山东寿光 Shouguang, Shandong	201,000 (30)	肉蛋兼用 E & M
狼山 Langshan (LS)	江苏如东 Rudong, Jiangsu	10,000 (20)	肉蛋兼用 E & M
鹿苑 Luyuan (LU)	江苏张家港 Zhangjiagang, Jiangsu	20,000 (30)	肉蛋兼用 E & M

品种及代号 Breed and code	原产地 Origin	现有种群规模及采样 Population (sample size)	经济用途 Economic use
安义瓦灰 Anyi Gray (AY)	江西安义 Anyi, Jiangxi	600,000 (30)	肉蛋兼用 E & M
白耳黄 Baier Yellow (BE)	江西广丰 Guangfeng, Jiangxi	20,000,000 (30)	蛋用 E
崇仁麻 Chongren Partridge (CP)	江西崇仁 Chongren, Jiangxi	52,000,000 (30)	肉蛋兼用 E & M
东乡绿壳 Dongxiang Blue-eggshell (DX)	江西东乡 Dongxiang, Jiangxi	200,000 (30)	肉蛋兼用 E & M
丝羽乌骨 Silkies (SL)	江西泰和 Taihe, Jiangxi	16,500,000 (30)	肉药兼用 Med & M
金湖乌凤 Jinhu Black-bone (JH)	福建泰宁 Taining, Fujian	160,000 (29)	肉药兼用 Med & M
仙居 Xianju (XJ)	浙江仙居 Xianju, Zhejiang	1,484,000 (30)	蛋用 E
琅琊 Langya (LY)	山东日照 Rizhao, Shandong	110,000 (30)	肉蛋兼用 E & M
寿光 Shouguang (SG)	山东寿光 Shouguang, Shandong	201,000 (30)	肉蛋兼用 E & M
狼山 Langshan (LS)	江苏如东 Rudong, Jiangsu	10,000 (20)	肉蛋兼用 E & M
鹿苑 Luyuan (LU)	江苏张家港 Zhangjiagang, Jiangsu	20,000 (30)	肉蛋兼用 E & M

单倍型 Haplotype	变异位点 Variable sites	单倍型在品种的分布(频率) Breeds (frequency)	总计 Total
	0000000000 0000000000 0000000000 0000111 1112222222 2222222222 3333333333 4677222 3690111122 3444445678 1134566679 4819112 3797027925 9012366101 0502813726 6612455
NC007235	TTTACATCAC ACAGTTTCTA CTCAAACTAT CGGGCAA
A1	.C...G...T .....C.... .C........ .....G.	BE8, DX13, SL21, XJ8, LU17, LY2	69
A2	.C.G.G...T .....C.... .C........ .....G.	SG8	8
A3	.C...G...T .....C.... .C...T.... .....G.	LU6	6
A4	.C...G...T .....C...G .C.......C .....GG	CP2	2
A5	.C...G...T .....C.... .C.......C .....G.	SG5	5
A6	.C...G...T G....C.... .C........ .....G.	JH1	1
A7	.CC..G...T .....C.... .C......G. .....G.	LY1	1
A8	.C...G...T .....CC... .C........ .....G.	SL2	2
A9	.C...G...T .....C.... .C........ .A...G.	AY3	3
A10	.C..TG...T .....C.... .C........ .A...G.	JH2	2
A11	.C.......T .....C.... .C........ .....G.	LY1	1
B1	.......... .......... .......... .......	XJ1, AY7, CP10, DX4, LY2, SG9, LS20	53
B2	C......... .......... .......... .......	CP1	1
B3	.......... .......... ..T....... .......	SL7	7
B4	....T..... .......... .......... .......	LY2	2
B5	.......T.. .......T.. .......... .......	LY1	1
B6	.......... .......... .......... ...A...	XJ1, AY5, BE1, SG1, LY1	9
B7	.......... ......C... .......... ...A...	BE4	4
B8	.......... .......... .......... .A.A...	JH11, XJ2, BE1	14
B9	.......... G......... ....G..... .A.A...	BE15	15
B10	.......... .......... T......... .A.....	CP12	12
C1	.....G.... ...ACCC..G TC.G..TC.. .....G.	XJ18, AY6, DX2, JH1, SG2	29
C2	.....G.... ...ACCCT.G TC.G..TC.. .....G.	LY3	3
C3	.....G.... .T.ACCC..G TC.G..TC.. .....G.	LY1	1
C4	.....G.... .T.ACC...G TC.G..TC.. ..A..G.	LU1	1
C5	.....G.... ...ACCCT.G TC....TC.. .....G.	AY2, LY1	3
C6	.....G.... ...ACCCTCG TC....TC.. .....G.	CP1	1
C7	.....G.... ...ACCC... TC.G..TC.. .....G.	BE1	1
C8	.........T ..GACCC..G TC....TC.. .....G.	AY7	7
C9	.......... ...ACCC..G TC....TC.. .....G.	LY1	1
E1	.....GC... ....CCCT.. TC........ T...TG.	JH13, CP3, LY6, LU6	28
E2	.....GC.G. ....CCCT.. TCT....... T...TG.	CP1	1
E3	.....GC... ....CCCT.. TC.G...... TA..TG.	LY8	8
E4	..C..GC... ....CC.T.. TC........ T...TG.	DX11	11
E5	..C..GC... ....CCCT.. TC........ T...TG.	JH1, SG5	6

单倍型 Haplotype	变异位点 Variable sites	单倍型在品种的分布(频率) Breeds (frequency)	总计 Total
	0000000000 0000000000 0000000000 0000111 1112222222 2222222222 3333333333 4677222 3690111122 3444445678 1134566679 4819112 3797027925 9012366101 0502813726 6612455
NC007235	TTTACATCAC ACAGTTTCTA CTCAAACTAT CGGGCAA
A1	.C...G...T .....C.... .C........ .....G.	BE8, DX13, SL21, XJ8, LU17, LY2	69
A2	.C.G.G...T .....C.... .C........ .....G.	SG8	8
A3	.C...G...T .....C.... .C...T.... .....G.	LU6	6
A4	.C...G...T .....C...G .C.......C .....GG	CP2	2
A5	.C...G...T .....C.... .C.......C .....G.	SG5	5
A6	.C...G...T G....C.... .C........ .....G.	JH1	1
A7	.CC..G...T .....C.... .C......G. .....G.	LY1	1
A8	.C...G...T .....CC... .C........ .....G.	SL2	2
A9	.C...G...T .....C.... .C........ .A...G.	AY3	3
A10	.C..TG...T .....C.... .C........ .A...G.	JH2	2
A11	.C.......T .....C.... .C........ .....G.	LY1	1
B1	.......... .......... .......... .......	XJ1, AY7, CP10, DX4, LY2, SG9, LS20	53
B2	C......... .......... .......... .......	CP1	1
B3	.......... .......... ..T....... .......	SL7	7
B4	....T..... .......... .......... .......	LY2	2
B5	.......T.. .......T.. .......... .......	LY1	1
B6	.......... .......... .......... ...A...	XJ1, AY5, BE1, SG1, LY1	9
B7	.......... ......C... .......... ...A...	BE4	4
B8	.......... .......... .......... .A.A...	JH11, XJ2, BE1	14
B9	.......... G......... ....G..... .A.A...	BE15	15
B10	.......... .......... T......... .A.....	CP12	12
C1	.....G.... ...ACCC..G TC.G..TC.. .....G.	XJ18, AY6, DX2, JH1, SG2	29
C2	.....G.... ...ACCCT.G TC.G..TC.. .....G.	LY3	3
C3	.....G.... .T.ACCC..G TC.G..TC.. .....G.	LY1	1
C4	.....G.... .T.ACC...G TC.G..TC.. ..A..G.	LU1	1
C5	.....G.... ...ACCCT.G TC....TC.. .....G.	AY2, LY1	3
C6	.....G.... ...ACCCTCG TC....TC.. .....G.	CP1	1
C7	.....G.... ...ACCC... TC.G..TC.. .....G.	BE1	1
C8	.........T ..GACCC..G TC....TC.. .....G.	AY7	7
C9	.......... ...ACCC..G TC....TC.. .....G.	LY1	1
E1	.....GC... ....CCCT.. TC........ T...TG.	JH13, CP3, LY6, LU6	28
E2	.....GC.G. ....CCCT.. TCT....... T...TG.	CP1	1
E3	.....GC... ....CCCT.. TC.G...... TA..TG.	LY8	8
E4	..C..GC... ....CC.T.. TC........ T...TG.	DX11	11
E5	..C..GC... ....CCCT.. TC........ T...TG.	JH1, SG5	6

品种名 Breed	样本量 sample size	单倍型数 No. of haplotypes	单倍型群(对应个体数) Haplogroup (number of individuals observed)	单倍型多样度 Haplotype diversity (SD)	核苷酸多样度 Nucleotide diversity (SD)	平均核苷酸差异 Average number of nucleotide differences
安义瓦灰 AY	30	6(A = 1; B = 2; C = 3)	A(3); B(12); C(15)	0.837(0.027)	0.00613(0.00140)	7.540
白耳黄 BE	30	6(A = 1; B = 4; C = 1)	A(8); B(21); C(1)	0.680(0.066)	0.00424(0.00133)	5.218
崇仁麻 CP	30	7(A = 1; B = 3; C = 1; E = 2)	A(2); B(23); C(1); E(4)	0.736(0.054)	0.00425(0.00178)	5.232
东乡绿壳 DX	30	4(A = 1; B = 1; C = 1; E = 1)	A(13); B(4); C(2); E(11)	0.678(0.049)	0.00457(0.00138)	5.444
金湖乌凤 JH	29	6(A = 2; B = 1; C = 1; E = 2)	A(3); B(11); C(1); E(14)	0.670(0.057)	0.00615(0.00174)	7.571
丝羽乌骨 SL	30	3(A = 2; B = 1)	A(23); B(7)	0.467(0.087)	0.00221(0.00075)	2.720
仙居 XJ	30	5(A = 1; B = 3; C = 1)	A(8); B(4); C(18)	0.582(0.079)	0.00492(0.00127)	6.053
琅琊 LY	30	13(A = 3; B = 4; C = 4; E = 2)	A(4); B(6); C(6); E(14)	0.887(0.038)	0.00647(0.00184)	7.968
寿光 SG	30	6(A = 2; B = 2; C = 1; E = 1)	A(13); B(10); C(2); E(5)	0.805(0.035)	0.00553(0.00165)	6.805
狼山 LS	20	1(B = 1)	B(20)	0	0	0.000
鹿苑 LU	30	3(A = 2; E = 1)	A(25); E(5)	0.549(0.081)	0.00237(0.00088)	2.917
总计 All	319	35(A = 11; B = 10; C = 9; E = 5)	(A = 100; B = 118; C = 47; E = 54)	0.901(0.009)	0.00573(0.000001)	6.833