爱玉子性别与品系的分子鉴别研究
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摘要
爱玉子(Ficus awkeotsang Makino)为桑科榕属植物,雌雄异株,为我国特有。其瘦果制成的爱玉冻是种高贵的天然食品,人工栽培具有广阔的市场前景。在苗期及营养期很难从形态特征上来鉴别爱玉子的雌、雄以及品系,因而找到一种简单、迅速、准确的鉴别方法,是目前生产上亟待解决的难题。本实验采用SRAP分子标记技术,研究爱玉子性别鉴别、爱玉子优良品系鉴别及其遗传多样性,为爱玉子栽培和种质资源改良提供理论依据。
通过对CTAB法进行改良,建立了适合爱玉子DNA提取的方法,获得了浓度和纯度都较高的基因组DNA。利用128个SRAP引物组合对爱玉子雌、雄DNA池进行扩增,从1228个扩增条带中找到一个雌性特异的标记Me5Em14-230,可用于爱玉子的早期性别鉴别。对福建省主栽的爱玉子品系(24个雌性品系,16个雄性品系)进行品系鉴别研究,结果表明:Me1Em8引物组合能够区分全部雄性品系,Me6Em6和Me3Em2双引物组合可以区分全部雌性品系;并在此基础上建立了雌、雄爱玉子各品系独特的数字化核酸指纹。进行Nei's遗传距离的计算,并以UPGMA法聚类,结果表明,雌、雄爱玉子各品系之间存在着较为复杂的亲缘关系,具有较丰富的遗传多样性。
Ficus awkeotsang Makino is a species of Moraceae in Ficus, and it is a dioecism and a unique woody vine growing in China. Its achenes can be utilized to produce jelly curd, which is an excellent food in the market. So, its artificial cultivation has broad market prospects. But it is hard to discriminate Ficus awkeotsang Makino's sex and strain from its morphologic characteristics when they are not in blossom. A simple, rapid and accurate method of identification should be developed and applied in practical production. Sex and strains identification and the genetic diversity of Ficus awkeotsang Makino were studied in this work using SRAP(Sequence-related amplified polymorphism) molecular marker. The result could provide a theoretical basis for the cultivation and the improvement of germplasm resources in Ficus awkeotsang Makino.
The genomic DNA of Ficus awkeotsang Makino with high concentration and purity could be obtained with an improved CTAB method, which was suitable for molecular analysis. 128 SRAP primer combinations were used for SRAP amplification and about 1228 fragments were obtained from male and female DNA pool. A marker of about 230bp linked to female was generated by Me5Em14 and was named Me5Em14-230. This marker could be used to identify different sex of Ficus awkeotsang Makino at the seedling stage. SRAP marker was imployed to identify different strains of Ficus awkeotsang Makino in this study, the result showed that a primer combination MelEm8 could identify the 16 male strains, and two primer combinations Me6Em6 and Me3Em2 could identify the 24 female strains. On the basis of the result, specific fingerprint of each strains was produced. More over, the result of Nei's genetic distance and UPGMA cluster analysis indicated that the relationship between strains was very complicated, and the diversity of cultivated Ficus awkeotsang Makino was high.
通过对CTAB法进行改良,建立了适合爱玉子DNA提取的方法,获得了浓度和纯度都较高的基因组DNA。利用128个SRAP引物组合对爱玉子雌、雄DNA池进行扩增,从1228个扩增条带中找到一个雌性特异的标记Me5Em14-230,可用于爱玉子的早期性别鉴别。对福建省主栽的爱玉子品系(24个雌性品系,16个雄性品系)进行品系鉴别研究,结果表明:Me1Em8引物组合能够区分全部雄性品系,Me6Em6和Me3Em2双引物组合可以区分全部雌性品系;并在此基础上建立了雌、雄爱玉子各品系独特的数字化核酸指纹。进行Nei's遗传距离的计算,并以UPGMA法聚类,结果表明,雌、雄爱玉子各品系之间存在着较为复杂的亲缘关系,具有较丰富的遗传多样性。
Ficus awkeotsang Makino is a species of Moraceae in Ficus, and it is a dioecism and a unique woody vine growing in China. Its achenes can be utilized to produce jelly curd, which is an excellent food in the market. So, its artificial cultivation has broad market prospects. But it is hard to discriminate Ficus awkeotsang Makino's sex and strain from its morphologic characteristics when they are not in blossom. A simple, rapid and accurate method of identification should be developed and applied in practical production. Sex and strains identification and the genetic diversity of Ficus awkeotsang Makino were studied in this work using SRAP(Sequence-related amplified polymorphism) molecular marker. The result could provide a theoretical basis for the cultivation and the improvement of germplasm resources in Ficus awkeotsang Makino.
The genomic DNA of Ficus awkeotsang Makino with high concentration and purity could be obtained with an improved CTAB method, which was suitable for molecular analysis. 128 SRAP primer combinations were used for SRAP amplification and about 1228 fragments were obtained from male and female DNA pool. A marker of about 230bp linked to female was generated by Me5Em14 and was named Me5Em14-230. This marker could be used to identify different sex of Ficus awkeotsang Makino at the seedling stage. SRAP marker was imployed to identify different strains of Ficus awkeotsang Makino in this study, the result showed that a primer combination MelEm8 could identify the 16 male strains, and two primer combinations Me6Em6 and Me3Em2 could identify the 24 female strains. On the basis of the result, specific fingerprint of each strains was produced. More over, the result of Nei's genetic distance and UPGMA cluster analysis indicated that the relationship between strains was very complicated, and the diversity of cultivated Ficus awkeotsang Makino was high.
引文
[1]张秀实,吴征缢,曹子余.中国植物志(第21卷,第1分册)[M].北京:科学出版社.1998.
[2]林赞标.爱玉子专论[M].台湾:台湾省林业试验所,1991.
[3]陈俊森,许信玲,罗美娟.台湾爱玉子研究与开发及福建引种栽培现况[J].防护林科技,2004,(6):52-54.
[4]陈勇.爱玉子及其传粉昆虫的研究[J].宁德师范高等专科学校学报(自然科学版),2001,13(3):204-207.
[5]陈勇,李宏庆,马炜梁.爱玉隐头花序的生物学特性[J].吉首大学学报(自然科学版),2002,23(4):4.
[6]陈勇,李宏庆,马炜梁.爱玉及其传粉昆虫的共生关系[J].武汉植物学研究,2002,20(4):315-319.
[7]黄永传,陈文彬.爱玉冻原料植物--爱玉之回顾与前瞻[J].中国园艺,1979,25(4):103-111.
[8]Makino.Observation on the flora of Japan[J].Bot Mag,1904,(18):151-152.
[9]高尾与一郎.爱玉子ノ凝胶质二就イテ.台湾总督府研究报告No.49,pp1-6.1917.
[10]三宅捷,大野成雄.爱玉子粘性物质の研究第一报[J].热带农学会志,1930,2(1):1-13.
[11]三宅捷,大野成雄.爱玉子粘性物质の研究第二报[J].热带农学会志,1931,2(3):317-327.
[12]三宅捷,大野成雄.爱玉子粘性物质の研究第三报[J].热带农学会志,1932,4(3):295-299.
[13]三宅捷,大野成雄.爱玉子粘性物质の研究第四报[J].热带农学会志,1933,5(3):270-272.
[14]#12
[15]#12
[16]黄永传,蔡纹州,王芬芳.台湾果胶资源之调查Ⅱ,蔬菜及其它特殊植物中各部位之果胶含量[J].中国园艺,1967,13:1-11.
[17]黄永传.陈文彬,邵云屏.爱玉冻凝胶机构之研究[J].中国园艺,1980.26(4):117-126.
[18]Lin T.P,Liu C.C,Chen S.W,et al.Purification and Characterization of Pectinmethylesterase from Ficus awkeotsang Makino Achenes[J].Plant physiology,1989,91(4):1445-1453.
[19]Miyazaki Y,Yakou S,Takayama K.Study on jelly fig extract as a potential hydrophilic matrix for controlled drug delivery[J].International journal of pharmaceutics,2004,287(1-2):39-46.
[20]Peng C.C,Hsiao E.S,Ding J.L,et al.Functional expression in pichia pastoris of an acidic pectin methylesterase from jelly fig(Ficus awkeotsang)[J].Journal of agricultural and food chemistry,2005,53(14):5612-5616.
[21]Li Y.C,Yang Y.C,Hsu J.S,et al.Cloning and immunolocalization of an antifungal chitinase in jelly fig (Ficus awkeotsang) achenes[J].Phytochemistry,2005,66(8):879-886.
[22]Chang J.H,Wang Y.T,Chang H.M.Pectinesterase inhibitor from jelly fig(Ficus awkeotsang Makino)achene induces apoptosis of human leukemic U937 cells[J].Annals of the New York Academy of Sciences,2005,1042:506-515.
[23]胡大维,刘哲政.爱玉的选种、繁殖、栽培[J].丰年(台湾),1985,35(5):32-38.
[24]李志生,林原,陈躬国,等.台湾爱玉子引种与种植技术[J].中国林副特产,2006,(1):15-17.
[25]徐俊森,许信玲,罗美娟.台湾爱玉子研究与开发及福建引种栽培现况[J].防护林科技,2004,(6):52-54.
[26]Bemardo R R-SJ,Ziegle J,Hauser J,et al.Parental contribution and coefficient among maize inbreds:Predigree,RFLP and SSR data[J],Theoretical and Applied Genetics,2000,100(3-4):552-556.
[27]Paull J G CKJ,Karakousis A,Kretschrner J M,et al.Genetic diversity in Australian wheat varieties and breeding material based on RFLP data[J].Thero Appl Genet,1998,96(3-4):435-446.
[28]Yaneshita M KS,Sasakuma T.Allotetraploidy of zoysia species with 2n=40 base on a RFLP genetic map[J].Theoretical and Applied Genetics,1999,98(5):751-756.
[29]Galili SAY,Millet E,Feldman.RFLP-based analysis of three RbcS subfamilies in diploid and polyploid species of wheat[J].Molecular and General Genetics,2000,263(4):674-680.
[30]汪小全,邹喻苹,张大明,等.RAPD应用于遗传多样性和系统学研究中的问题[J].植物学报,1996,38(12):954-962.
[31]许明辉,郑民慧,刘广田.烟草品种RAPD分子标记遗传差异研究[J].农业生物技术学报,1998,6(3):281-285.
[32]何川生,何兴金,葛颂,等.烤烟品种资源的RAPD分析[J].植物学报,2001,43(6):610-614.
[33]王志德,牟建民,戴培刚,等.部分烟草核心种质RAPD分析[J].中国烟草学报,2003,9(4):20-26.
[34]Mailer R.J,et al.Discrimination among cuitivars of rape seed(Bassica napus L.) using DNA polymorphisms amplified from arbitrary primers[J].TAG,1994,87(697-704).
[35]吴敏生,戴景瑞,王守才,等.RAPD在玉米品种鉴定和纯度分析中的应用[J].作物学报,1999,25(4):489-493.
[36]李宽钰,黄敏仁,王明庥,等.用RAPD探讨毛白杨起源[J].植物分类学报,1997,35(1):24-31.
[37]吴燕民,李嘉瑞.用RAPD分析麻核桃起源与分类地位[J].林业科学,1999,35(4):25-30.
[38]张继益,董玉琛.早麦草属种质资源的随机扩增多态性DNA(RAPDs)分析[J].遗传学报,1999,26(1):54-60.
[39]Carlson J,et al.Segregation of random amplified DNA markers in F1 progeny of conifers[J].TAG,1991,83:194-200.
[40]朱立煌.用分子标记定位一个未知的抗瘟病基因[J].中国科学(B辑),1994,24(10):1048-1052.
[41]Monna L,et al.Determination of RAPD markers in rice and their conversion into sequence-tagged sites (STSs) and STS-specific primers[J].DNA Res,1994,1:139-148.
[42]Becker J,et al.Combined mapping of AFLP and RFLP markers in Barley[J].MGG,1995,249:65-73.
[43]Cho Y,et al.Integrated map of AFLP、SSLP and RFLP of markers using a recombinant inbred population rice(Oryza sativa L.)[J].TAG,1998,97:370-380.
[44]Maheswaran M,et al.Polymorphism distribution and segregation of AFLP markers in a doubled haploid rice population[J].TAG,1997,94:39-45.
[45]Panaud O,et al.Frequency of microsatellite sequences in rice(Oryza Sativa L.)[J].Genome,1995,38:1170-1176.
[46]Hearne C,Ghosh S,Todd J.Microsattelites for linkage analysis of genetic traits[J].Trends in Genetics,1993,8:288-294.
[47]Dayanadan S,Bawa K.S,Kesseli R.Conservation of microsattelites among tropical tree(leguminosae)[J].American Journal of Botany,1997,84(12):1658-1663.
[48]Primmer C.R,Ellegren H.Patterns of molecular evolution in avian microsattelites[J].Mol.Biol.Evol,1998,15(8):997-1008.
[49]Jain A,Apparanda C,Bhalla P.L.Evaluation of genetic diversity and genome fingerprinting of Pandorea(Bignoniaceae) by PAPD and inter-SSR PCR[J].Genome,1999,42(4):714-719.
[50]吴晓雷,贺超英,陈受宜,等.用SSR分子标记研究大豆属种间亲缘关系[J].遗传学报,2001,28(4):359-366.
[51]Wu K,et al.Abundance polymorphism and genetic mapping of microsaellites in rice[J].MGG,1993,241:225-245.
[52]Beckmann J,Soller M.Toward a unified approach to genetic mapping of eukaryotes based on sequence tagged microsateilite sites[J].Bio/tech- nologys,1990,930-932.
[53]江树业,陈启锋,方宣钧.水稻农垦58与农垦58s的RAPD和ISSR分析[J].农业生物技术学报,2000,8(1):63-66.
[54]祁建民,周东新,吴为人,等.用ISSR标记检测黄麻野生种与栽培种遗传多样性[J].应用生态学报,2003,14(9):1473-1477.
[55]邱英雄,傅承新,吴斐捷.明党参与川明参群体遗传结构及分子鉴定的ISSR分析[J].中国中药杂 志,2003,28(7):598-603.
[56]Kojima T,Nagaoka T,Noda K,et al.Genetic linkage map of ISSR and RAPD markers in Einkorn wheat in relation to that of RFLP Markers[J].TAG,1998,96:37-45.
[57]钱韦,葛颂,洪德元.采用RAPD和ISSR标记探讨中国疣粒野生稻的遗传多样性[J].植物学报,2000,42(7):741-750.
[58]Li G,Quiros C.F.Sequence-related amplified polymorphism(SRAP),a new marker system based on a simple PCR reaction:Its application to mapping and gene tagging in Brassica[J].Theor Appl Genet,2001,103:455-461
[59]Budak H,Shearman R.C,Parmaksiz I,et al.Molecular characterization of Bufalograss germplasm using sequence-related amplified polymorphism markers[J].Theor Appl Genet,2004,108(2):328-334
[60]Ferriol M,Pico B,Nuez F.Genetic diversity of germplasm collection of Cucurbita pepo using SRAP and AFLP markers[J].Theoretical and Applied Genetics,2003,107:271-282.
[61]柳李旺,龚义勤,黄浩,等.新型分子标记--SRAP与TRAP及其应用[J].遗传,2004,26(5):777-781.
[62]Rennet S.S,Ricklefs R.E.Dioecy and its correlates in the flowering plants[J].Amer J Bot,1995,82:596-606.
[63]陈中海,陈晓静.雌雄异株果树的性别决定及性别鉴定的研究进展[J].福建农业大学学报,2000,29(1):429-434.
[64]马铁华.植物的性别决定[J].农业与技术,2001,21(2):52-54.
[65]李瑞丽.芦笋性别差异的比较研究与雌性SCAR标记的建立.[河南师范大学硕士学位论文].2006:2-3
[66]Yin L,Zhan Y,Li C,et al.Appraisal of researching ways in sex identification between female and male plants[J].Bull Bot Res,2003,23(1):123-128.
[67]Bugala W.Sex determination of Poplars from the colour of leaves[J].Forestry Abstracts,1951,52:013-02835.
[68]龚文楼.毛白杨雌雄株的直观鉴别[J].天津农林科技,1995,(2):40.
[69]廉永善,卢顺光,薛顺康,等.沙棘属植物生物学和化学[M].兰州:甘肃科学技术出版社,2000.
[70]Jaiswal VS,Prpatap N,Madan L.Activities of acid and alkaline phosphatases in relation to sex diferentiation in Carica papaya[J].Biochem Physioi Pflanzen,1984,179:799-801.
[71]赵云云,刘捷平.雌雄异株植物的生理生化特性及性别鉴定[J].北京师范学院学报,1991,12(4):27-33.
[72]菜汝,陶俊,陈鹏.银杏雌雄株叶片光合特征、蒸腾速率及产量的比较研究[J].落叶果树,2000,(1):14-1 5.
[73]周国章.香榧雌雄株叶片酚类物质的比较研究[J].林业科技,1985,(4):16-19.
[74]李教社,巩宝星,杨云,等.不同时期中国沙棘雌雄株叶中总黄酮含量的测定.沙棘[J].2000,13(1):39-40.
[75]王丙武,王雅清,莫华,等.杜仲雌雄株细胞学、顶芽及叶含胶量的比较.植物学报[J].1999,41(1):11-15.
[76]张谷曼.千年桐不同性别植株组织在生化特性上的差异.植物生理学通讯[J].1964,19(58):1.
[77]范怡梅,谭仁祥.银杏叶中萜类内酯含量与生长季节及植株性别的相关性研究.中国中药杂志[J].1999,24(2):82-83.
[78]王白坡,杨飞峰.银杏雌雄株内源激素和核酸的变化.浙江林学院学报[J].1999,16(2):114-118.
[79]李正理.最近十年(1949-1959)关于银杏的形态解剖学及细胞学上的研究.植物学报[J].1959,8(4):262-270.
[80]Pollock E.G.The sex chromosomes of the maidenhair tree[J].Hereditary,1957,48(6):290-294
[81]陈瑞阳,宋文芹,李秀兰.植物染色体标本制备的去壁、低渗法及其在细胞遗传学中的意义[J].遗传学报,1982,9(2):151-159.
[82]陈学森,张艳敏.银杏雌雄株核型及性别早期鉴定[J].果树科学,1997,14(2):87-90.
[83]何梅,李懋学.银杏雌株染色体Ag-NOR的研究[J].遗传,1996,18(3):3-5.
[84]Penel C.L,Greppin H.Evolution of the auxin-oxidase and peroxidase activity during the spinach's photoperiodic induction and sexualistion[J].Plant & cell physiology,1972,13(1):151-156.
[85]范双喜,宋学锋.石刁柏性别表现与同工酶的关系[J].华北农学报,1995,10(2):67-71.
[86]赵云云,闫毓秀.雌雄构树过氧化物同工酶的比较研究[J].首都师范大学学报:自然科学版,1996,17(2):84-87.
[87]袁仕禄,贾卫国.华中五味子雌雄株过氧化物酶同工酶性别鉴定研究[J].东北农业大学学报,1999,30(2):195-198.
[88]钟诲文,杨中汉,朱广廉.根据过氧化物酶同工酶图谱鉴定银杏植株的性别[J].林业科学,1982,18(1):1-5.
[89]李国梁,林伯年.杨梅雌雄株同工酶和酚类物质的鉴别[J].浙江农业大学学报,1995,21(1):22-26.
[90]张立平,林柏年.雌雄异株葡萄的性别鉴定研究[J].植物学通报,1998,15(4):63-67.
[91]艾辛,何光存.黄瓜植株性别表现与3种氧化酶同工酶的关系[J].武汉植物学研究,2000,18(3):184-188.
[92]Bracale M,Galli M.G,Falavigna A,et al.Sexual differentiation in Asparagus officinalis L[J].Sexual Plant Reproduction,1990,3(1):23-30.
[93]Caporali E,Carboni A,Galli MG,et al.Development of male and female flower in Asparagus officinalis.Search for point of transition from hermaphroditic to unisexual developmental pathway[J].Sexual Plant Reproduction,1994,7(4):239-249.
[94]汪俏梅,曾广文.苦瓜性别分化的特异蛋白质研究[J].Acta Botanica Sinica(植物学报:英文版),1998,40(3):241-246.
[95]于凤池,孙耘子,袁高峰,等.栝楼性别分化特异大分子标记物研究[J].浙江农业学报,2003,15(6):332-335.
[96]Harvey C.F,Gill G.P,Fraser L.G,et al.Sex determination in Actinidia.1.Sex-linked markers and progeny sex ratio in diploid Achinensis[J].Sexual Plant Reproduction,1997,10(3):149-154.
[97]王晓梅,宋文芹.与银杏性别相关的RAPD标记[J].南开大学学报:自然科学版,2001,34(3):116-117.
[98]Cai L,Tian X.C,Li M,et al.Application of RAPD in discrimination of Podocarpus macrophyllus's sex[J].Journal of Fudan University(Nature Science),2002,41(6):635-640.
[99]谭冬梅,韩海涛.阿月浑子性别鉴定的RAPD分析[J].果树学报,2003,20(2):124-126.
[100]Jiang L,You R,Li M,et al.Identification of a sex-Associated RAPD Marker in Ginkgo biloba[J].Acta botanica Sinica,2003,45(6):742-747.
[101]王晓梅,宋文芹,刘松等.与银杏性别相关的RAPD标记[J].南开大学学报:自然科学版,2001,34(3):116-117.
[102]Gill G.P,Harvey C.F,Gardner R.C,et al.Development of sex-linked PCR markers for gender identification in Actinidia[J].TAG Theoretical and Applied Genetics,1998,97(3):439-445.
[103]Mandolino G,Carboni A,Forapani S,et al.Identification of DNA markers linked to the male sex in dioecious hemp(Cannabis sativa L.)[J].TAG Theoretical and Applied Genetics,1999,98(1):86-92.
[104]陈其军.韩玉珍.大麻性别的RAPD和SCAR分子标记[J].植物生理学报,2001.27(2):173-178.
[105]Urasaki N,Tokumoto M,Tarora K,et al.A male and hermaphrodite specific RAPD marker for papaya (Carica papaya L.)[J].TAG Theoretical and Applied Genetics,2002,104(2):281-285.
[106]Xu W.J,Wang B.W,Cui K.M.RAPD and SCAR markers linked to sex determination in Eucommia ulmoides Oiiv[J].Euphytica,2004,136(3):233-238.
[107]任朝兴,黄建昌,肖艳,等.番木瓜雄性性别的RAPD和SCAR标记[J].果树学报,2007,24(1):72-75.
[108]Terauchi R,Kahl G.Mapping of the Dioscorea tokoro genome:AFLP markers linked to sex[J].Genome, 1999,42(4):752-762.
[109]Reamon-Büttner SM,Jung C.AFLP-derived STS markers for the identification of sex in Asparagus officinalis L[J].TAG Theoretical and Applied Genetics,2000,100(3):432-438.
[110]陶莉,王跃进,尤敏,等.AFLP用于构建罗汉果DNA指纹图谱及其幼苗雌雄鉴别[J].武汉植物学研究,2005,23(001):77-80.
[111]王晓梅,宋文芹,等.利用AFLP技术筛选与银杏性别相关的分析标记[J].南开大学学报:自然科学版,2001,34(1):5-9.
[112]Peil A,Flachowsky H,Schumann E,et ai.Sex-linked AFLP markers indicate a pseudoautosomal region in hemp(Cannabis sativa L.)[J].TAG Theoretical and Applied Genetics,2003,107(1):102-109.
[113]Parasnis A.S,Ramakrishna W,Chowdari K.V,et al.Microsateilite(GATA) n reveals sex-specific differences in Papaya[J].TAG Theoretical and Applied Genetics,1999,99(6):1047-1052.
[114]周劲松,汤泳萍,罗绍春,等.DNA分子标记早期快速鉴别芦笋雌雄株[J].分子植物育种,2007,5(3):363-366.
[115]蒋谦才,黄悦朝,李增祥.广东榕属观赏植物资源及其开发利用[J].亚热带植物科学,2004,33(4):48-51.
[116]张今今,张莹,李文燕.榕属植物基因组DNA的提取及RAPD体系的优化[J].西北农林科技大学学报,2006,34(5):147-152.
[117]林忠旭,张献龙,聂以春等.棉花SRAP遗传连锁图构建[J].科学通报,2003,48(15):1676-1679.
[118]宋显军,张伟,曹萍等.大豆SSR标记PAGE银染方法的改进[J].安徽农业科学,2006,34(16):3922-3923.
[119]李志生,林原,陈躬国,等.台湾爱玉子引种与种植技术[J].中国林副特产,2006,(1):15-17.
[120]Gill G.P,Harvey C.F,Gardner R.C,et al.Development of sex-linked PCR markers for gender identification in Actinidia[J].Theor Appl Genet,1998,97(3):439-445.
[121]Mandolino G,Carboni A,Forapani S,et al.Identification of DNA markers linked to the male sex in dioecious hemp(Cannabis sativa L)[J].Theor Appl Genet,1999,98(1):86-92.
[122]陈其军,韩玉珍,等.大麻性别的RAPD和SCAR分子标记[J].植物生理学报,2001,27(2):173-178.
[123]Korpelainen H.A genetic method to resolve gender complements investigations on sex ratios in Rumex acetosa[J].Molecular ecology,2002,11(10):2151-2156.
[124]李瑞丽,卢龙斗,高武军,等.雌雄异株植物性别鉴定的研究进展[J].广西植物,2006,26(4):387-391,351.
[125]Marziani G,Caporali E,pada.A Search for genes involved in asparagus sex determination in plants[J]. Oxford:Bios Scientific Publishers,1999:149-162.
[126]Alstrom R.C,Lascoux M,Wand Y.C,et al.Identification of a RAPD marker linked to sex determination in the basket wiiow(Salix viminalis L)[J].Journal of Heredity,1998,89:44-49.
[127]Gunter L.E,Roberts G.T,Lee K,et al.The development of two flanking SCAR markers linked to a sex determination locus in Salix viminalis L[J].J Hered JT-The Joural of heredity,2003,94(2):185-189.
[128]Ainsworth C.Boys and girls come out to play:The molecular biology of dioecious plants[J].Annuls Bot,2000,86:211-221.
[129]林忠旭,张献龙,聂以春.新型标记SRAP在棉花F2分离群体及遗传多样性评价中的适用性分析.[J]遗传学报.2004,31(6):622-626.
[130]于拴仓,柴敏,姜立纲.主要番茄品种的分子鉴别研究[J].农业生物技术科学,2005,21(5):84-89.
[2]林赞标.爱玉子专论[M].台湾:台湾省林业试验所,1991.
[3]陈俊森,许信玲,罗美娟.台湾爱玉子研究与开发及福建引种栽培现况[J].防护林科技,2004,(6):52-54.
[4]陈勇.爱玉子及其传粉昆虫的研究[J].宁德师范高等专科学校学报(自然科学版),2001,13(3):204-207.
[5]陈勇,李宏庆,马炜梁.爱玉隐头花序的生物学特性[J].吉首大学学报(自然科学版),2002,23(4):4.
[6]陈勇,李宏庆,马炜梁.爱玉及其传粉昆虫的共生关系[J].武汉植物学研究,2002,20(4):315-319.
[7]黄永传,陈文彬.爱玉冻原料植物--爱玉之回顾与前瞻[J].中国园艺,1979,25(4):103-111.
[8]Makino.Observation on the flora of Japan[J].Bot Mag,1904,(18):151-152.
[9]高尾与一郎.爱玉子ノ凝胶质二就イテ.台湾总督府研究报告No.49,pp1-6.1917.
[10]三宅捷,大野成雄.爱玉子粘性物质の研究第一报[J].热带农学会志,1930,2(1):1-13.
[11]三宅捷,大野成雄.爱玉子粘性物质の研究第二报[J].热带农学会志,1931,2(3):317-327.
[12]三宅捷,大野成雄.爱玉子粘性物质の研究第三报[J].热带农学会志,1932,4(3):295-299.
[13]三宅捷,大野成雄.爱玉子粘性物质の研究第四报[J].热带农学会志,1933,5(3):270-272.
[14]#12
[15]#12
[16]黄永传,蔡纹州,王芬芳.台湾果胶资源之调查Ⅱ,蔬菜及其它特殊植物中各部位之果胶含量[J].中国园艺,1967,13:1-11.
[17]黄永传.陈文彬,邵云屏.爱玉冻凝胶机构之研究[J].中国园艺,1980.26(4):117-126.
[18]Lin T.P,Liu C.C,Chen S.W,et al.Purification and Characterization of Pectinmethylesterase from Ficus awkeotsang Makino Achenes[J].Plant physiology,1989,91(4):1445-1453.
[19]Miyazaki Y,Yakou S,Takayama K.Study on jelly fig extract as a potential hydrophilic matrix for controlled drug delivery[J].International journal of pharmaceutics,2004,287(1-2):39-46.
[20]Peng C.C,Hsiao E.S,Ding J.L,et al.Functional expression in pichia pastoris of an acidic pectin methylesterase from jelly fig(Ficus awkeotsang)[J].Journal of agricultural and food chemistry,2005,53(14):5612-5616.
[21]Li Y.C,Yang Y.C,Hsu J.S,et al.Cloning and immunolocalization of an antifungal chitinase in jelly fig (Ficus awkeotsang) achenes[J].Phytochemistry,2005,66(8):879-886.
[22]Chang J.H,Wang Y.T,Chang H.M.Pectinesterase inhibitor from jelly fig(Ficus awkeotsang Makino)achene induces apoptosis of human leukemic U937 cells[J].Annals of the New York Academy of Sciences,2005,1042:506-515.
[23]胡大维,刘哲政.爱玉的选种、繁殖、栽培[J].丰年(台湾),1985,35(5):32-38.
[24]李志生,林原,陈躬国,等.台湾爱玉子引种与种植技术[J].中国林副特产,2006,(1):15-17.
[25]徐俊森,许信玲,罗美娟.台湾爱玉子研究与开发及福建引种栽培现况[J].防护林科技,2004,(6):52-54.
[26]Bemardo R R-SJ,Ziegle J,Hauser J,et al.Parental contribution and coefficient among maize inbreds:Predigree,RFLP and SSR data[J],Theoretical and Applied Genetics,2000,100(3-4):552-556.
[27]Paull J G CKJ,Karakousis A,Kretschrner J M,et al.Genetic diversity in Australian wheat varieties and breeding material based on RFLP data[J].Thero Appl Genet,1998,96(3-4):435-446.
[28]Yaneshita M KS,Sasakuma T.Allotetraploidy of zoysia species with 2n=40 base on a RFLP genetic map[J].Theoretical and Applied Genetics,1999,98(5):751-756.
[29]Galili SAY,Millet E,Feldman.RFLP-based analysis of three RbcS subfamilies in diploid and polyploid species of wheat[J].Molecular and General Genetics,2000,263(4):674-680.
[30]汪小全,邹喻苹,张大明,等.RAPD应用于遗传多样性和系统学研究中的问题[J].植物学报,1996,38(12):954-962.
[31]许明辉,郑民慧,刘广田.烟草品种RAPD分子标记遗传差异研究[J].农业生物技术学报,1998,6(3):281-285.
[32]何川生,何兴金,葛颂,等.烤烟品种资源的RAPD分析[J].植物学报,2001,43(6):610-614.
[33]王志德,牟建民,戴培刚,等.部分烟草核心种质RAPD分析[J].中国烟草学报,2003,9(4):20-26.
[34]Mailer R.J,et al.Discrimination among cuitivars of rape seed(Bassica napus L.) using DNA polymorphisms amplified from arbitrary primers[J].TAG,1994,87(697-704).
[35]吴敏生,戴景瑞,王守才,等.RAPD在玉米品种鉴定和纯度分析中的应用[J].作物学报,1999,25(4):489-493.
[36]李宽钰,黄敏仁,王明庥,等.用RAPD探讨毛白杨起源[J].植物分类学报,1997,35(1):24-31.
[37]吴燕民,李嘉瑞.用RAPD分析麻核桃起源与分类地位[J].林业科学,1999,35(4):25-30.
[38]张继益,董玉琛.早麦草属种质资源的随机扩增多态性DNA(RAPDs)分析[J].遗传学报,1999,26(1):54-60.
[39]Carlson J,et al.Segregation of random amplified DNA markers in F1 progeny of conifers[J].TAG,1991,83:194-200.
[40]朱立煌.用分子标记定位一个未知的抗瘟病基因[J].中国科学(B辑),1994,24(10):1048-1052.
[41]Monna L,et al.Determination of RAPD markers in rice and their conversion into sequence-tagged sites (STSs) and STS-specific primers[J].DNA Res,1994,1:139-148.
[42]Becker J,et al.Combined mapping of AFLP and RFLP markers in Barley[J].MGG,1995,249:65-73.
[43]Cho Y,et al.Integrated map of AFLP、SSLP and RFLP of markers using a recombinant inbred population rice(Oryza sativa L.)[J].TAG,1998,97:370-380.
[44]Maheswaran M,et al.Polymorphism distribution and segregation of AFLP markers in a doubled haploid rice population[J].TAG,1997,94:39-45.
[45]Panaud O,et al.Frequency of microsatellite sequences in rice(Oryza Sativa L.)[J].Genome,1995,38:1170-1176.
[46]Hearne C,Ghosh S,Todd J.Microsattelites for linkage analysis of genetic traits[J].Trends in Genetics,1993,8:288-294.
[47]Dayanadan S,Bawa K.S,Kesseli R.Conservation of microsattelites among tropical tree(leguminosae)[J].American Journal of Botany,1997,84(12):1658-1663.
[48]Primmer C.R,Ellegren H.Patterns of molecular evolution in avian microsattelites[J].Mol.Biol.Evol,1998,15(8):997-1008.
[49]Jain A,Apparanda C,Bhalla P.L.Evaluation of genetic diversity and genome fingerprinting of Pandorea(Bignoniaceae) by PAPD and inter-SSR PCR[J].Genome,1999,42(4):714-719.
[50]吴晓雷,贺超英,陈受宜,等.用SSR分子标记研究大豆属种间亲缘关系[J].遗传学报,2001,28(4):359-366.
[51]Wu K,et al.Abundance polymorphism and genetic mapping of microsaellites in rice[J].MGG,1993,241:225-245.
[52]Beckmann J,Soller M.Toward a unified approach to genetic mapping of eukaryotes based on sequence tagged microsateilite sites[J].Bio/tech- nologys,1990,930-932.
[53]江树业,陈启锋,方宣钧.水稻农垦58与农垦58s的RAPD和ISSR分析[J].农业生物技术学报,2000,8(1):63-66.
[54]祁建民,周东新,吴为人,等.用ISSR标记检测黄麻野生种与栽培种遗传多样性[J].应用生态学报,2003,14(9):1473-1477.
[55]邱英雄,傅承新,吴斐捷.明党参与川明参群体遗传结构及分子鉴定的ISSR分析[J].中国中药杂 志,2003,28(7):598-603.
[56]Kojima T,Nagaoka T,Noda K,et al.Genetic linkage map of ISSR and RAPD markers in Einkorn wheat in relation to that of RFLP Markers[J].TAG,1998,96:37-45.
[57]钱韦,葛颂,洪德元.采用RAPD和ISSR标记探讨中国疣粒野生稻的遗传多样性[J].植物学报,2000,42(7):741-750.
[58]Li G,Quiros C.F.Sequence-related amplified polymorphism(SRAP),a new marker system based on a simple PCR reaction:Its application to mapping and gene tagging in Brassica[J].Theor Appl Genet,2001,103:455-461
[59]Budak H,Shearman R.C,Parmaksiz I,et al.Molecular characterization of Bufalograss germplasm using sequence-related amplified polymorphism markers[J].Theor Appl Genet,2004,108(2):328-334
[60]Ferriol M,Pico B,Nuez F.Genetic diversity of germplasm collection of Cucurbita pepo using SRAP and AFLP markers[J].Theoretical and Applied Genetics,2003,107:271-282.
[61]柳李旺,龚义勤,黄浩,等.新型分子标记--SRAP与TRAP及其应用[J].遗传,2004,26(5):777-781.
[62]Rennet S.S,Ricklefs R.E.Dioecy and its correlates in the flowering plants[J].Amer J Bot,1995,82:596-606.
[63]陈中海,陈晓静.雌雄异株果树的性别决定及性别鉴定的研究进展[J].福建农业大学学报,2000,29(1):429-434.
[64]马铁华.植物的性别决定[J].农业与技术,2001,21(2):52-54.
[65]李瑞丽.芦笋性别差异的比较研究与雌性SCAR标记的建立.[河南师范大学硕士学位论文].2006:2-3
[66]Yin L,Zhan Y,Li C,et al.Appraisal of researching ways in sex identification between female and male plants[J].Bull Bot Res,2003,23(1):123-128.
[67]Bugala W.Sex determination of Poplars from the colour of leaves[J].Forestry Abstracts,1951,52:013-02835.
[68]龚文楼.毛白杨雌雄株的直观鉴别[J].天津农林科技,1995,(2):40.
[69]廉永善,卢顺光,薛顺康,等.沙棘属植物生物学和化学[M].兰州:甘肃科学技术出版社,2000.
[70]Jaiswal VS,Prpatap N,Madan L.Activities of acid and alkaline phosphatases in relation to sex diferentiation in Carica papaya[J].Biochem Physioi Pflanzen,1984,179:799-801.
[71]赵云云,刘捷平.雌雄异株植物的生理生化特性及性别鉴定[J].北京师范学院学报,1991,12(4):27-33.
[72]菜汝,陶俊,陈鹏.银杏雌雄株叶片光合特征、蒸腾速率及产量的比较研究[J].落叶果树,2000,(1):14-1 5.
[73]周国章.香榧雌雄株叶片酚类物质的比较研究[J].林业科技,1985,(4):16-19.
[74]李教社,巩宝星,杨云,等.不同时期中国沙棘雌雄株叶中总黄酮含量的测定.沙棘[J].2000,13(1):39-40.
[75]王丙武,王雅清,莫华,等.杜仲雌雄株细胞学、顶芽及叶含胶量的比较.植物学报[J].1999,41(1):11-15.
[76]张谷曼.千年桐不同性别植株组织在生化特性上的差异.植物生理学通讯[J].1964,19(58):1.
[77]范怡梅,谭仁祥.银杏叶中萜类内酯含量与生长季节及植株性别的相关性研究.中国中药杂志[J].1999,24(2):82-83.
[78]王白坡,杨飞峰.银杏雌雄株内源激素和核酸的变化.浙江林学院学报[J].1999,16(2):114-118.
[79]李正理.最近十年(1949-1959)关于银杏的形态解剖学及细胞学上的研究.植物学报[J].1959,8(4):262-270.
[80]Pollock E.G.The sex chromosomes of the maidenhair tree[J].Hereditary,1957,48(6):290-294
[81]陈瑞阳,宋文芹,李秀兰.植物染色体标本制备的去壁、低渗法及其在细胞遗传学中的意义[J].遗传学报,1982,9(2):151-159.
[82]陈学森,张艳敏.银杏雌雄株核型及性别早期鉴定[J].果树科学,1997,14(2):87-90.
[83]何梅,李懋学.银杏雌株染色体Ag-NOR的研究[J].遗传,1996,18(3):3-5.
[84]Penel C.L,Greppin H.Evolution of the auxin-oxidase and peroxidase activity during the spinach's photoperiodic induction and sexualistion[J].Plant & cell physiology,1972,13(1):151-156.
[85]范双喜,宋学锋.石刁柏性别表现与同工酶的关系[J].华北农学报,1995,10(2):67-71.
[86]赵云云,闫毓秀.雌雄构树过氧化物同工酶的比较研究[J].首都师范大学学报:自然科学版,1996,17(2):84-87.
[87]袁仕禄,贾卫国.华中五味子雌雄株过氧化物酶同工酶性别鉴定研究[J].东北农业大学学报,1999,30(2):195-198.
[88]钟诲文,杨中汉,朱广廉.根据过氧化物酶同工酶图谱鉴定银杏植株的性别[J].林业科学,1982,18(1):1-5.
[89]李国梁,林伯年.杨梅雌雄株同工酶和酚类物质的鉴别[J].浙江农业大学学报,1995,21(1):22-26.
[90]张立平,林柏年.雌雄异株葡萄的性别鉴定研究[J].植物学通报,1998,15(4):63-67.
[91]艾辛,何光存.黄瓜植株性别表现与3种氧化酶同工酶的关系[J].武汉植物学研究,2000,18(3):184-188.
[92]Bracale M,Galli M.G,Falavigna A,et al.Sexual differentiation in Asparagus officinalis L[J].Sexual Plant Reproduction,1990,3(1):23-30.
[93]Caporali E,Carboni A,Galli MG,et al.Development of male and female flower in Asparagus officinalis.Search for point of transition from hermaphroditic to unisexual developmental pathway[J].Sexual Plant Reproduction,1994,7(4):239-249.
[94]汪俏梅,曾广文.苦瓜性别分化的特异蛋白质研究[J].Acta Botanica Sinica(植物学报:英文版),1998,40(3):241-246.
[95]于凤池,孙耘子,袁高峰,等.栝楼性别分化特异大分子标记物研究[J].浙江农业学报,2003,15(6):332-335.
[96]Harvey C.F,Gill G.P,Fraser L.G,et al.Sex determination in Actinidia.1.Sex-linked markers and progeny sex ratio in diploid Achinensis[J].Sexual Plant Reproduction,1997,10(3):149-154.
[97]王晓梅,宋文芹.与银杏性别相关的RAPD标记[J].南开大学学报:自然科学版,2001,34(3):116-117.
[98]Cai L,Tian X.C,Li M,et al.Application of RAPD in discrimination of Podocarpus macrophyllus's sex[J].Journal of Fudan University(Nature Science),2002,41(6):635-640.
[99]谭冬梅,韩海涛.阿月浑子性别鉴定的RAPD分析[J].果树学报,2003,20(2):124-126.
[100]Jiang L,You R,Li M,et al.Identification of a sex-Associated RAPD Marker in Ginkgo biloba[J].Acta botanica Sinica,2003,45(6):742-747.
[101]王晓梅,宋文芹,刘松等.与银杏性别相关的RAPD标记[J].南开大学学报:自然科学版,2001,34(3):116-117.
[102]Gill G.P,Harvey C.F,Gardner R.C,et al.Development of sex-linked PCR markers for gender identification in Actinidia[J].TAG Theoretical and Applied Genetics,1998,97(3):439-445.
[103]Mandolino G,Carboni A,Forapani S,et al.Identification of DNA markers linked to the male sex in dioecious hemp(Cannabis sativa L.)[J].TAG Theoretical and Applied Genetics,1999,98(1):86-92.
[104]陈其军.韩玉珍.大麻性别的RAPD和SCAR分子标记[J].植物生理学报,2001.27(2):173-178.
[105]Urasaki N,Tokumoto M,Tarora K,et al.A male and hermaphrodite specific RAPD marker for papaya (Carica papaya L.)[J].TAG Theoretical and Applied Genetics,2002,104(2):281-285.
[106]Xu W.J,Wang B.W,Cui K.M.RAPD and SCAR markers linked to sex determination in Eucommia ulmoides Oiiv[J].Euphytica,2004,136(3):233-238.
[107]任朝兴,黄建昌,肖艳,等.番木瓜雄性性别的RAPD和SCAR标记[J].果树学报,2007,24(1):72-75.
[108]Terauchi R,Kahl G.Mapping of the Dioscorea tokoro genome:AFLP markers linked to sex[J].Genome, 1999,42(4):752-762.
[109]Reamon-Büttner SM,Jung C.AFLP-derived STS markers for the identification of sex in Asparagus officinalis L[J].TAG Theoretical and Applied Genetics,2000,100(3):432-438.
[110]陶莉,王跃进,尤敏,等.AFLP用于构建罗汉果DNA指纹图谱及其幼苗雌雄鉴别[J].武汉植物学研究,2005,23(001):77-80.
[111]王晓梅,宋文芹,等.利用AFLP技术筛选与银杏性别相关的分析标记[J].南开大学学报:自然科学版,2001,34(1):5-9.
[112]Peil A,Flachowsky H,Schumann E,et ai.Sex-linked AFLP markers indicate a pseudoautosomal region in hemp(Cannabis sativa L.)[J].TAG Theoretical and Applied Genetics,2003,107(1):102-109.
[113]Parasnis A.S,Ramakrishna W,Chowdari K.V,et al.Microsateilite(GATA) n reveals sex-specific differences in Papaya[J].TAG Theoretical and Applied Genetics,1999,99(6):1047-1052.
[114]周劲松,汤泳萍,罗绍春,等.DNA分子标记早期快速鉴别芦笋雌雄株[J].分子植物育种,2007,5(3):363-366.
[115]蒋谦才,黄悦朝,李增祥.广东榕属观赏植物资源及其开发利用[J].亚热带植物科学,2004,33(4):48-51.
[116]张今今,张莹,李文燕.榕属植物基因组DNA的提取及RAPD体系的优化[J].西北农林科技大学学报,2006,34(5):147-152.
[117]林忠旭,张献龙,聂以春等.棉花SRAP遗传连锁图构建[J].科学通报,2003,48(15):1676-1679.
[118]宋显军,张伟,曹萍等.大豆SSR标记PAGE银染方法的改进[J].安徽农业科学,2006,34(16):3922-3923.
[119]李志生,林原,陈躬国,等.台湾爱玉子引种与种植技术[J].中国林副特产,2006,(1):15-17.
[120]Gill G.P,Harvey C.F,Gardner R.C,et al.Development of sex-linked PCR markers for gender identification in Actinidia[J].Theor Appl Genet,1998,97(3):439-445.
[121]Mandolino G,Carboni A,Forapani S,et al.Identification of DNA markers linked to the male sex in dioecious hemp(Cannabis sativa L)[J].Theor Appl Genet,1999,98(1):86-92.
[122]陈其军,韩玉珍,等.大麻性别的RAPD和SCAR分子标记[J].植物生理学报,2001,27(2):173-178.
[123]Korpelainen H.A genetic method to resolve gender complements investigations on sex ratios in Rumex acetosa[J].Molecular ecology,2002,11(10):2151-2156.
[124]李瑞丽,卢龙斗,高武军,等.雌雄异株植物性别鉴定的研究进展[J].广西植物,2006,26(4):387-391,351.
[125]Marziani G,Caporali E,pada.A Search for genes involved in asparagus sex determination in plants[J]. Oxford:Bios Scientific Publishers,1999:149-162.
[126]Alstrom R.C,Lascoux M,Wand Y.C,et al.Identification of a RAPD marker linked to sex determination in the basket wiiow(Salix viminalis L)[J].Journal of Heredity,1998,89:44-49.
[127]Gunter L.E,Roberts G.T,Lee K,et al.The development of two flanking SCAR markers linked to a sex determination locus in Salix viminalis L[J].J Hered JT-The Joural of heredity,2003,94(2):185-189.
[128]Ainsworth C.Boys and girls come out to play:The molecular biology of dioecious plants[J].Annuls Bot,2000,86:211-221.
[129]林忠旭,张献龙,聂以春.新型标记SRAP在棉花F2分离群体及遗传多样性评价中的适用性分析.[J]遗传学报.2004,31(6):622-626.
[130]于拴仓,柴敏,姜立纲.主要番茄品种的分子鉴别研究[J].农业生物技术科学,2005,21(5):84-89.