In tra-sp ecific G enetic R ela tionsh ip am ong 20 Cu ltiva rs fromCamellia japon ica Ba sed on ISSR M olecu lar M arkers

NI Sui; LI Ji-yuan; WANG Qiang

Volume 22 Issue 5

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In tra-sp ecific G enetic R ela tionsh ip am ong 20 Cu ltiva rs fromCamellia japon ica Ba sed on ISSR M olecu lar M arkers

1.
R esearch Institute of Sub trop icalForestry, CAF, Fuyang 311400, Zhejiang, Ch in a
2.
Facu lty of Life Science and Biotechno logy ofNingboUn ivers ity, Ningbo 315211, Zhe jiang, Ch ina
3.
Ningbo Ins titu te ofTechnology, Zh ejiangUn ivers ity, N ingbo 315100, Zh ejiang, Ch ina

Received Date: 2008-09-11

Abstract

Camellia japonica has various ecologica,l horticultural and economical uses. An approach was establishedusing ISSR ( Inter-simple sequence repeat) molecu lar makers to c lassify and analyze the intra-specific geneticrelationship among twenty cultivars represented from C. japonica, collected in Ch ina and abroad. Twenty-onecoup les of primer sequences which cou ld amplify c learly and reproducible e lectrophoresis bandswere screened fromsixty couples of primers. Using these primers, 153 discern ib le DNA fragment were generated with 146 ( 95. 4% )being polymorphic, ind icating pronounced genetic variation at the cu ltivars leve;l the band in fingerprintingwereconverted into / 00 or / 10, where / 10 and / 00 ind icated the presence and absence of a band, respectively. Ne ipsd iversity ind ices (H ) ranged from 0. 40 to 0. 48 and Shannon. s information index ( I) ranged from 0. 57 to 0. 67.The gene differentiation coeffic ien ts ranged from 0. 5 to 0. 7 andNm ranged from 0. 2 to 0. 5. The genetic similaritycoefficients (H s) among the tested cu ltivars ranged from 0. 50 to 0. 74, averaging 0. 63. The resu lts showed that thegenetic differences among the twenty cultivars were relatively little. Comb ing the results of flowermorphology andUPGMA cluster analysis, we cou ld divide the twenty cultivars into two b ig groups. Therewere four cu ltivars in group?, which all grows in Zhejiang Province and has red flowers. There were sixteen cu ltivars in groupò, intraspec ific genetic relationsh ip comp lexity. In add ition, the results also showed that ISSRmolecu larmakerwas suitable to ana lyze the intra-specific genetic diversities and genetic relationships ofC. japonica.
- Camellia japonica,
- ISSR molecu larmaker,
- intra-specific genetic re lationships,
- c luster analysis

References

[1]	张宏达, 任善湘. 中国植物志: 第49 卷第3 分册[M]. 北京: 科学出版社, 1998
[2]	高继银, 陈绍山, 徐碧玉. 世界名贵茶花[M]. 杭州: 浙江科学技术出版社, 1998
[3]	闵天禄. 世界山茶属的研究[M]. 昆明: 云南科技出版社, 2000
[4]	邹天才. 小黄花茶和长柱红山茶种质资源利用的研究[J]. 贵州科学, 2000, 18 (3): 209- 215
[5]	Zh ou Jian lin, J iYucheng, JiangYanbo, et al. D evelopment of s imp le sequ ence repeats (SSR) markers of ram ie and comparison of SSRand inter-SSR marker systems [J]. Progess in N atural Scien ce,2005, 15(2): 137- 142
[6]	Cu lleyTM, Sb ita S J, W ick A. Popu lat ion genet ic effects of u rbanhab itat fragmen tat ion in the perennial herb Viola pubescens (V iolaceae) us ing ISSR markers [J]. Annals of Botany, 2007, 100 (1):91- 100
[7]	PowellW, Morgan teM, Andr.C, et al. The comparison ofRFLP,RAPD, AFLP and SSR (m icrosatell ite) markers for germp lasm analysis[J]. Molecu lar Breed ing, 1996, -: 225- 238
[8]	Du ran Y, Frat in i R, Garcia P. An in tersubspecific genet ic map ofLens[J]. Th eoretical andApp liedG enetics, 2004, 108 (7): 1265-1273
[9]	Lai Jou ann, Y angWeichen, X iao Juying. A n assessment of genet icrelationsh ip s in cu ltivated tea clones and nat ivew ild tea inT aiwan using RAPD and ISSR mark ers[J]. Botan ical Bu llet in of Academ iaSin ica, 2001, 42: 93 - 100
[10]	FernandezM E, F igu eiras A M, Ben ito C. The use of ISSR andPARD mark ers for d etect ingDNA polymorph ism, genotype ident ification and genet ic d iversity among b arley cu lt ivarsw ith known orig in[J]. Theoretical andA pp lied Genet ics, 2002, 104: 845 - 851
[11]	姚明哲, 陈亮, 王新超, 等. 我国茶树无性系品种遗传多样性和亲缘关系的ISSR 分析[J]. 作物学报, 2007, 33 (4):598- 604
[12]	邓辉, 黄晓燕, 乙引. ISSR-PCR 技术在植物种间和种下关系中的应用[J]. 种子, 2006, 25(4): 55- 57
[13]	Cho S, Kumar J, Shu ltz JL, et al. Mapp ing genes for doub le podd ing and oth er morpholog ical traits in ch ickpea [J]. Euphytica,2002, 128 (2): 285 - 292
[14]	Mondal T K. A ssessmen t of genetic d iversity of tea (Camelliasinensis (L.) O. Kuntze) by in ter-simp le sequence repeat polymerase ch ain reaction[J]. Euphyt ica, 2002, 128: 307- 315
[15]	Zh aoWeigao, MiaoXu exia, Zhang Bo, et a l. Con struction of fingerp rint ing and genetic d iversity ofmulberry cu ltivars in Ch ina byISSR markers[J]. A cta G enetica Sin ica, 2006, 33(9): 851- 860
[16]	X iaoT iaojiang, Parks C R. Molecu lar analysis of genu sCamellia[M]. American Camellia Y earbook, 2002: 52 - 58
[17]	邓白罗, 谭晓风, 漆龙霖, 等. 山茶属红山茶组植物的RAPD分析及分类研究[J]. 林业科学, 2006, 42 (5): 36- 42
[18]	申屠文月, 陈析丰, 马伯军. 3 个山茶花变异品种的形态鉴定和RAPD分析[J]. 浙江师范大学学报, 2006, 29 (3): 317- 321
[19]	宾晓芸, 唐绍清. 金花茶遗传多样性的ISSR分析[J]. 武汉植物学研究, 2005, 23 (1): 20- 26
[20]	Doyle J J, Doyle J L. A rap id DNA isolat ion procedu re for sma llquan tit ies of fresh leaf t issue [J]. Phytochem Bu l,l 1987, 19:11- 15

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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In tra-sp ecific G enetic R ela tionsh ip am ong 20 Cu ltiva rs fromCamellia japon ica Ba sed on ISSR M olecu lar M arkers

1. R esearch Institute of Sub trop icalForestry, CAF, Fuyang 311400, Zhejiang, Ch in a

2. Facu lty of Life Science and Biotechno logy ofNingboUn ivers ity, Ningbo 315211, Zhe jiang, Ch ina

3. Ningbo Ins titu te ofTechnology, Zh ejiangUn ivers ity, N ingbo 315100, Zh ejiang, Ch ina

Received Date: 2008-09-11

Abstract: Camellia japonica has various ecologica,l horticultural and economical uses. An approach was establishedusing ISSR ( Inter-simple sequence repeat) molecu lar makers to c lassify and analyze the intra-specific geneticrelationship among twenty cultivars represented from C. japonica, collected in Ch ina and abroad. Twenty-onecoup les of primer sequences which cou ld amplify c learly and reproducible e lectrophoresis bandswere screened fromsixty couples of primers. Using these primers, 153 discern ib le DNA fragment were generated with 146 ( 95. 4% )being polymorphic, ind icating pronounced genetic variation at the cu ltivars leve;l the band in fingerprintingwereconverted into / 00 or / 10, where / 10 and / 00 ind icated the presence and absence of a band, respectively. Ne ipsd iversity ind ices (H ) ranged from 0. 40 to 0. 48 and Shannon. s information index ( I) ranged from 0. 57 to 0. 67.The gene differentiation coeffic ien ts ranged from 0. 5 to 0. 7 andNm ranged from 0. 2 to 0. 5. The genetic similaritycoefficients (H s) among the tested cu ltivars ranged from 0. 50 to 0. 74, averaging 0. 63. The resu lts showed that thegenetic differences among the twenty cultivars were relatively little. Comb ing the results of flowermorphology andUPGMA cluster analysis, we cou ld divide the twenty cultivars into two b ig groups. Therewere four cu ltivars in group?, which all grows in Zhejiang Province and has red flowers. There were sixteen cu ltivars in groupò, intraspec ific genetic relationsh ip comp lexity. In add ition, the results also showed that ISSRmolecu larmakerwas suitable to ana lyze the intra-specific genetic diversities and genetic relationships ofC. japonica.

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Reference (20)

[1]	张宏达, 任善湘. 中国植物志: 第49 卷第3 分册[M]. 北京: 科学出版社, 1998
[2]	高继银, 陈绍山, 徐碧玉. 世界名贵茶花[M]. 杭州: 浙江科学技术出版社, 1998
[3]	闵天禄. 世界山茶属的研究[M]. 昆明: 云南科技出版社, 2000
[4]	邹天才. 小黄花茶和长柱红山茶种质资源利用的研究[J]. 贵州科学, 2000, 18 (3): 209- 215
[5]	Zh ou Jian lin, J iYucheng, JiangYanbo, et al. D evelopment of s imp le sequ ence repeats (SSR) markers of ram ie and comparison of SSRand inter-SSR marker systems [J]. Progess in N atural Scien ce,2005, 15(2): 137- 142
[6]	Cu lleyTM, Sb ita S J, W ick A. Popu lat ion genet ic effects of u rbanhab itat fragmen tat ion in the perennial herb Viola pubescens (V iolaceae) us ing ISSR markers [J]. Annals of Botany, 2007, 100 (1):91- 100
[7]	PowellW, Morgan teM, Andr.C, et al. The comparison ofRFLP,RAPD, AFLP and SSR (m icrosatell ite) markers for germp lasm analysis[J]. Molecu lar Breed ing, 1996, -: 225- 238
[8]	Du ran Y, Frat in i R, Garcia P. An in tersubspecific genet ic map ofLens[J]. Th eoretical andApp liedG enetics, 2004, 108 (7): 1265-1273
[9]	Lai Jou ann, Y angWeichen, X iao Juying. A n assessment of genet icrelationsh ip s in cu ltivated tea clones and nat ivew ild tea inT aiwan using RAPD and ISSR mark ers[J]. Botan ical Bu llet in of Academ iaSin ica, 2001, 42: 93 - 100
[10]	FernandezM E, F igu eiras A M, Ben ito C. The use of ISSR andPARD mark ers for d etect ingDNA polymorph ism, genotype ident ification and genet ic d iversity among b arley cu lt ivarsw ith known orig in[J]. Theoretical andA pp lied Genet ics, 2002, 104: 845 - 851
[11]	姚明哲, 陈亮, 王新超, 等. 我国茶树无性系品种遗传多样性和亲缘关系的ISSR 分析[J]. 作物学报, 2007, 33 (4):598- 604
[12]	邓辉, 黄晓燕, 乙引. ISSR-PCR 技术在植物种间和种下关系中的应用[J]. 种子, 2006, 25(4): 55- 57
[13]	Cho S, Kumar J, Shu ltz JL, et al. Mapp ing genes for doub le podd ing and oth er morpholog ical traits in ch ickpea [J]. Euphytica,2002, 128 (2): 285 - 292
[14]	Mondal T K. A ssessmen t of genetic d iversity of tea (Camelliasinensis (L.) O. Kuntze) by in ter-simp le sequence repeat polymerase ch ain reaction[J]. Euphyt ica, 2002, 128: 307- 315
[15]	Zh aoWeigao, MiaoXu exia, Zhang Bo, et a l. Con struction of fingerp rint ing and genetic d iversity ofmulberry cu ltivars in Ch ina byISSR markers[J]. A cta G enetica Sin ica, 2006, 33(9): 851- 860
[16]	X iaoT iaojiang, Parks C R. Molecu lar analysis of genu sCamellia[M]. American Camellia Y earbook, 2002: 52 - 58
[17]	邓白罗, 谭晓风, 漆龙霖, 等. 山茶属红山茶组植物的RAPD分析及分类研究[J]. 林业科学, 2006, 42 (5): 36- 42
[18]	申屠文月, 陈析丰, 马伯军. 3 个山茶花变异品种的形态鉴定和RAPD分析[J]. 浙江师范大学学报, 2006, 29 (3): 317- 321
[19]	宾晓芸, 唐绍清. 金花茶遗传多样性的ISSR分析[J]. 武汉植物学研究, 2005, 23 (1): 20- 26
[20]	Doyle J J, Doyle J L. A rap id DNA isolat ion procedu re for sma llquan tit ies of fresh leaf t issue [J]. Phytochem Bu l,l 1987, 19:11- 15

In tra-sp ecific G enetic R ela tionsh ip am ong 20 Cu ltiva rs fromCamellia japon ica Ba sed on ISSR M olecu lar M arkers

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通讯作者: 陈斌, bchen63@163.com

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In tra-sp ecific G enetic R ela tionsh ip am ong 20 Cu ltiva rs fromCamellia japon ica Ba sed on ISSR M olecu lar M arkers

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