基于单核苷酸多态性的甜瓜枯萎病抗性基因Fom-2功能性分子标记开发与利用
|
摘要
甜瓜链孢菌性枯萎病(Fusarium oxysporum f.sp.Melonis,Fom)是由尖孢镰刀菌(Fusarium oxysporum f.sp.melonis Snyder and Hansen)导致的一种最难控制的土传病害,成为影响甜瓜品质和产量的主要因素。本研究以两个厚皮甜瓜(Cucumis melo)品种为实验材料,分别为美国甜瓜(Cantaloupe)和仙果027-5(xianguo 027-5)。通过基因克隆和序列比对获得3个单核苷酸多态性位点。在它们的F_1与F_2代群体中,采用限制性内切酶酶切(CAPS)和等位基因特异性PCR(AS-PCR)两种DNA分子标记验证这些单核苷酸多态性位点的有效性和可靠性,进而将验证的标记转化为功能性分子标记,应用于常规品种的筛选。
我们克隆得到了Fom-2 LRR区DNA序列,长为1311 bp。采用生物信息学软件对其进行比对分析,发现高抗和高感枯萎病材料的第281、1076和1216碱基处存在3个单核苷酸多态性位点,并且这些碱基的突变导致了相应氨基酸的改变。
以高抗和高感枯萎病材料杂交得到的F_1(30株)与F_2(100株)代群体为研究材料,采用限制性内切酶酶切方法对其单核苷酸多态性位点进行分析。筛选得到特异性的扩增引物对CAPS-2F与CAPS-2R,CAPS-3F与CAPS-3R,特异性PCR后经限制性内切酶酶切,结果发现SNP2与SNP3两个位点在F_2群体中纯合抗病类型、杂合抗病类型与纯合感病类型呈现1∶2∶1的分离比例,符合孟德尔单基因遗传学规律,成功将CAPS标记转化为功能性分子标记。依据CAPS方法获得的结果,我们设计筛选得到AS-PCR特异性引物2-3F1与2-3R1引物对,在退火温度为62℃,35个循环条件下进行特异性PCR,F_2群体中抗性材料和感病材料呈现3∶1的孟德尔单基因分离比例。因此CAPS和AS-PCR两种方法都证明了SNP2与SNP3的有效性与可靠性。
采用CAPS与AS-PCR方法对34份常规栽培品种进行分析,筛选得到2个纯合枯萎病抗病材料和9个杂合抗病材料,为甜瓜枯萎病抗性育种提供了良好的种质资源。
Fusarium wilt of melon (Fusarium oxysporum f. sp. Melonis, Fom) caused by the fungus Fusarium oxysporum f. sp. melonis Snyder and Hansen, has become one of the most destructive diseases of melon. It severely limited the melon yeild production and quality. This research focused on the development and utilization of the SNP (single nucleotide polymorphism)-based FM associated with resistance of fusarium wilt in melon. The DNA moleculer marker, CAPS (Cleaved amplified polymorphic sequences, CAPS) and AS-PCR (Allele-specific PCR, AS-PCR), were employed to validate the effectiveness and reliability of the markers in F_1 and F_2 population, which derived from a cross between Cantaloupe and xianguo 027-5. Then the candidate markers were converted into the functional marker and used in the selection of cultivars.
A 1311bp leucine-rich repeat (LRR) region of the Fom-2 gene was cloned. Three single nucleotide polymorphism (SNP) sites were found and located at 281st, 1076th and 1216th of the cloned LRR region, respectively. These three single nucleotide polymorphism sites were analysed by CAPS method in F_1 (30 plants) and F_2 populations (100 plants) derived from cross between wilt resistant and susceptible germplasms. According to CAPS method, we used CAPS-2F and CAPS-2R, CAPS-3F and CAPS-3R as two pairs of specific primers to amplify fragments of interest, then the PCR products were digested by EcoR I and Xba I. The result showed the 1:2:1 Mendelian single-dominant segregation pattern for site 2 and 3 in the F_2 population. The CAPS makers were sucessfully converted into the fuctional markers. As for the AS-PCR method, the specific primers 2-3F1 and 2-3R1 were used to amplify fragments of interest. When the annealing tempreture was 62℃and cycle number was 35, it could be easily distinguished the resistant and susceptible plants. The genetic segregation ratio in F_2 population was 3:1 among the resistant and susceptible plants. The results of CAPS and AS-PCR methods showed that site 2 and 3 were reliable and effective.
The further screening of 34 cultivars were conducted by CAPS and AS-PCR markers. Among the 34 tested varieties, 11 were identified as resistant genotypes including 2 homozygous and 9 heterozygous genotypes. These resistant plants can be used as good germplasm resources in resistant breeding.
我们克隆得到了Fom-2 LRR区DNA序列,长为1311 bp。采用生物信息学软件对其进行比对分析,发现高抗和高感枯萎病材料的第281、1076和1216碱基处存在3个单核苷酸多态性位点,并且这些碱基的突变导致了相应氨基酸的改变。
以高抗和高感枯萎病材料杂交得到的F_1(30株)与F_2(100株)代群体为研究材料,采用限制性内切酶酶切方法对其单核苷酸多态性位点进行分析。筛选得到特异性的扩增引物对CAPS-2F与CAPS-2R,CAPS-3F与CAPS-3R,特异性PCR后经限制性内切酶酶切,结果发现SNP2与SNP3两个位点在F_2群体中纯合抗病类型、杂合抗病类型与纯合感病类型呈现1∶2∶1的分离比例,符合孟德尔单基因遗传学规律,成功将CAPS标记转化为功能性分子标记。依据CAPS方法获得的结果,我们设计筛选得到AS-PCR特异性引物2-3F1与2-3R1引物对,在退火温度为62℃,35个循环条件下进行特异性PCR,F_2群体中抗性材料和感病材料呈现3∶1的孟德尔单基因分离比例。因此CAPS和AS-PCR两种方法都证明了SNP2与SNP3的有效性与可靠性。
采用CAPS与AS-PCR方法对34份常规栽培品种进行分析,筛选得到2个纯合枯萎病抗病材料和9个杂合抗病材料,为甜瓜枯萎病抗性育种提供了良好的种质资源。
Fusarium wilt of melon (Fusarium oxysporum f. sp. Melonis, Fom) caused by the fungus Fusarium oxysporum f. sp. melonis Snyder and Hansen, has become one of the most destructive diseases of melon. It severely limited the melon yeild production and quality. This research focused on the development and utilization of the SNP (single nucleotide polymorphism)-based FM associated with resistance of fusarium wilt in melon. The DNA moleculer marker, CAPS (Cleaved amplified polymorphic sequences, CAPS) and AS-PCR (Allele-specific PCR, AS-PCR), were employed to validate the effectiveness and reliability of the markers in F_1 and F_2 population, which derived from a cross between Cantaloupe and xianguo 027-5. Then the candidate markers were converted into the functional marker and used in the selection of cultivars.
A 1311bp leucine-rich repeat (LRR) region of the Fom-2 gene was cloned. Three single nucleotide polymorphism (SNP) sites were found and located at 281st, 1076th and 1216th of the cloned LRR region, respectively. These three single nucleotide polymorphism sites were analysed by CAPS method in F_1 (30 plants) and F_2 populations (100 plants) derived from cross between wilt resistant and susceptible germplasms. According to CAPS method, we used CAPS-2F and CAPS-2R, CAPS-3F and CAPS-3R as two pairs of specific primers to amplify fragments of interest, then the PCR products were digested by EcoR I and Xba I. The result showed the 1:2:1 Mendelian single-dominant segregation pattern for site 2 and 3 in the F_2 population. The CAPS makers were sucessfully converted into the fuctional markers. As for the AS-PCR method, the specific primers 2-3F1 and 2-3R1 were used to amplify fragments of interest. When the annealing tempreture was 62℃and cycle number was 35, it could be easily distinguished the resistant and susceptible plants. The genetic segregation ratio in F_2 population was 3:1 among the resistant and susceptible plants. The results of CAPS and AS-PCR methods showed that site 2 and 3 were reliable and effective.
The further screening of 34 cultivars were conducted by CAPS and AS-PCR markers. Among the 34 tested varieties, 11 were identified as resistant genotypes including 2 homozygous and 9 heterozygous genotypes. These resistant plants can be used as good germplasm resources in resistant breeding.
引文
岸国平.野菜病虫害诊断与防治.全国农业教育协会.1977:654
陈幼源,陈绯翔.不同南瓜砧木品种对网纹甜瓜生长发育的影响.上海农业学报,2002,18(4):47-51
川出武夫.甜瓜对枯萎病的抗性,杨鼎新译.中国西瓜甜瓜,2002,(4):43-45
杜玮南,孙红霞,方福德.单核苷酸多态性的研究进展.中国医学科学院学报,2000,(8):392-394
侯栋,闫秀玲,李浩,高艳霞.几种不同砧木在哈密瓜嫁接无土栽培中的表现.中国西瓜甜瓜,2002
黄振文.利用土壤添加物防治作物之土壤传播性病害.植物保护学会会刊,1991,33:113-123
吉加兵,徐润芳.西瓜种质资源抗枯萎病性的苗期鉴定.中国西瓜甜瓜,1990,(2):19-20
蒋平,谢青,唐华.施宝灵防治西瓜枯萎病药效试验.1992,16(5):18-19
杰弗里.植物分类学入门.胡征宇译.1990
李红霞,周明国.用等位基因特异性寡核苷酸(ASO)-PCR快速检测抗多菌灵的油菜菌核病菌.中国农业科学.2004,37(9):1396-1399
李瑞琴,甜瓜枯萎病病原学及防治技术研究.西北农林科技大学硕士论文,2004
林德佩.国外甜瓜西瓜的研究动态.中国西瓜甜瓜,1989(2):1-8;1990(1):7-8
刘君璞,马跃.我国西瓜甜瓜种业的现状与发展对策.中国西瓜甜瓜,2000,(3):2-6
刘连杰,董凤珍.吉林省早熟甜瓜枯萎病的发生与防治.中国西瓜甜瓜,1995,(1):28
刘秀芳,刘芳政,王叶霜.西瓜甜瓜主要病虫害的识别及其防治.北京:农业出版社,1993
卢文经.薄皮甜瓜嫁接抗病增产技术研究.辽宁农业科学,2002,(1):16-20
陆致平,项继忠,吴雄飞.恶霉灵防治西瓜枯萎病试验研究.现代农药,2002,(3):44
马国斌,林德佩,王叶筠,郭庆元.西瓜枯萎病菌镰刀菌酸对西瓜苗作用机制的初步探讨.植物病理学报,1999,30:373-374
马国斌,王鸣,郑学勤.西瓜和甜瓜镰刀菌酸抗性变异体的诱导.中国园艺学 会第六届青年学术讨论会论文集,2004
戚配坤.瓜类枯萎病菌专化型研究简介.华南农业大学学报,1994,9(4):81-86
齐三魁,吴大康,林德佩.中国甜瓜.北京:科学普及出版社,1991
乔凤,陈泽良,王玉飞,赵瑾,杜昕颖,于雅琴,黄留玉.pUC19K质粒的构建及其在布鲁氏菌突变株构建中的应用.中国生物工程杂志,2007,27(12):1-5
王坚.中国西瓜甜瓜.中国农业出版社,2000,08
王贤磊,李群,方勇,高小明,杨艳,李冠.单核苷酸多态性与甜瓜抗枯萎病分子育种研究.生物技术,2007,17(4):31-34
王叶筠,林德佩,郭庆元.新疆甜瓜抗病育种研究.中国西瓜甜瓜,1993,(4):17-20
温玲.枯萎灵等药剂对西瓜枯萎病防治效果.黑龙江农业科学,2000,6:28
谢大森,何晓明,彭庆务,骆焕彬.瓜类枯萎病发生机理研究进展.仲恺农业技术学院学报,2006,19(3):65-70
徐胜利,李新民.厚皮甜瓜嫁接育苗栽培的防病增产效果.中国蔬菜,2000(4):16-18
许勇,朱其杰,陈晓莉,梁根庆.镰刀菌酸对黄瓜不同品种的致萎力与枯萎病菌致病力的相关性.中国蔬菜,1993,1:19-20
颜志强,杨胜利,龚毅.PCR及其衍生技术在基因突变检测中的应用.遗传,2003,25(2):198-200
张蓓,沈立松.实时荧光定量PCR的研究进展及其应用.国外医学临床生物化学与检验学分册,2003,24(6):327-329
周真,孙芙蓉,曹长余,于建美,韩冰.西瓜枯萎病的田间施药方式及药效对比试验.山东蔬菜,2000,(3):36-37
朱天圣,戚配坤.苦瓜枯萎病病原菌研究.华南农业大学学报,1998,19(4):14-18
Abea F,Saitob K,Miurac K,Toriyamaa K.A single nucleotide polymorphism in the alternative oxidase gene among rice varieties differing in low temperature tolerance.FEBS Letters,2002,527:181-185
Andersen JR,L(u|¨)bberstedt T.Functional markers in plants.Trends in Plant Science,2003,8(11):554-560
Amholdt-Schmitt B,Costa JH,Melo DF.AOX-a functional marker for efficient cell reprogram ruing under stress? Trends in Plant Science,2006,11(6):282-287
Arnholdt-Schmitt B.Functional markers and a 'systemic strategy':convergency between plant breeding, plant nutrition and molecular biology. Plant Physiology and Biochemistry, 2005, 43: 817-820
Bang H, Kim S, Leskovar D, King S. Development of a codominant CAPS marker for allelic selection between canary yellow and red watermelon based on SNP in lycopene beta-cyclase (LCYB) gene. Moleculer Breeding, 2007, 20: 63-72
Bang HJ, Kim SG, Leskovar D, King S. Development of a codominant CAPS marker for allelic selection between canary yellow and red watermelon based on SNP in lycopene beta-cyclase (LCYB) gene. Molecular breeding, 2007, 20(1): 63-72
Baumler S, Felsenstein FG, Schwarz G. CAPS and DHPLC analysis of a single nucleotide polymorphism in the cytochrome b gene conferring resistance to strobilurins in field isolates of Blumeria gra minis f. sp hordei. Phytopathology phytopathologische zeitschrift. 2003,151(3): 149-152
Booth C. 陈其瑛译.镰刀菌属.北京: 农业出版社, 1988
Caranta C, Thabuis A, Palloix A. Development of a CAPS marker for the Pvr4 locus: A tool for pyramiding potyvirus resistance genes in pepper. Genome, 1999, 42(6): 1111-1116
Chasman D; Adams RM. Predicting the functional consequences of non-synonymous single nucleotide polymorphisms: Structure-based assessment of amino acid variation. Molecular Biology, 2001, 307(2): 683-706
Chiapparino E, Lee D, Donini P. Genotyping single nucleotide polymorphisms in barley by tetra-primer ARMS-PCR. Genome, 2004,47: 414-420
Danin-Poleg, Y, Burger Y, Schreiber S, Katzir N, Cohen R. Identification of the gene for resistance to Fusarium wilt races 0 and 2 in Cucumis melo. Cucurbit Genetic Cooperate Report, 1999, 22: 19-20
David D. Fusaric acid in selective pathogenicity of Fusarium oxysporum. Phytopathology, 1969, 59: 1391-1395
Filatov DA, Charlesworth D. DNA polymorphism, haplotype structure and balancing selection in the Leavenworthia PgiC locus. Genetics, 1999, 153: 1423-1434
Fiorani F, Umbach AL, Siedow JN. The alternative oxidase of plant mitochondria is involved in the acclimation of shoot growth at low temperature. A study of Arabidopsis AOXla transgenic plants. Plant Physiology, 2005, 139: 1795-1805.
Fraaije BA, Butters JA, Coelho JM, Jones DR, Hollomon DW. Following the dynamics of strobilurin resistance in Blumeria gra minis f.sp tritici using quantitative allele-specific real-time PCR measurements with the fluorescent dye SYBR Green I. Plant pathology, 2002, 51(1): 45-54
Fraaije BA, Cools HJ, Fountaine J, Lovell DJ, Motteram J, West JS, Lucas JA. Role of ascospores in further spread of QoI-resistant cytochrome b alleles (G143A) in field populations of Mycosphaerella gra minicola. Phytopathology, 2005, 95 (8): 933-941
Gao ZS, Van de Weg WE, Schaart JG, Van der Meer IM, Kodde L, Laimer M, Breiteneder H, Hoffmann-Sommergruber K, Gilissen L J. Linkage map positions and allelic diversity of two Mal d 3 (non-specific lipid transfer protein) genes in the cultivated apple (Malus domestica). Theoretical and Applied Genetics, 2005, 110:479-491
Gordon TR, Martyn RD. The evolutionary biology of Fusarium oxysporum. Annual review of Phytopathology, 1997,35:111-128
Grodzicker T, Anderson C, Sharp PA, Sambrook J. Conditional lethal mutants of adenovirus 2-simian virus 40 hybrids. I. Host range mutants of Ad2+ND1. The Jounal of virology, 1974, 13(6): 1237-1244
Gubler WD, Grogan RG. Fusarium wilt of muskmelon in San Joaquin Valley of California. Plant disease report, 1976,60(9): 742-744
Gupta PK, Varshney RK, Prasad M. Molecular markers: principles and methodology. In: Jain S M. Molecular Techniques in Crop Improvement. Kluwer Academic Publishers, 2002: 9-54
Hackauf B, Wehling P. Identification of microsatellite polymorphisms in an expressed portion of the rye genome. Plant Breeding, 2002,121: 17-25
Hagenblad J, Nordborg M. Sequence variation and haplotype structure surrounding the flowering time locus FR1 in Arabidopsis thaliana. Genetics, 2002, 161:289-298
Hughes AL, Nei M. Pattern of nucleotide substitution at major histocompatibility complex class I loci reveals overdominant selection. Nature, 1988, 335: 167-170
Ilic K, Berleth T, Provart NJ. BlastDigester-a web-based program for efficient CAPS marker design. Trends in Genetics, 2004, 20(7): 280-283
Ishii H, Fraaije BA, Sugiyama T, Noguchi K, Nishimura K, Takeda T, Amano T, Hollomon DW. Occurrence and molecular characterization of strobilurin resistance in cucumber powdery mildew and downy mildew. Phytopathology, 2001,91(12): 1166-1171
Iyer-Pascuzzi AS, McCouch SR. Functional markers for xa5-mediated resistance in rice (Oryza sativa L.). Molecular Breeding, 2007, 19: 291-296
Joobeur T, King JJ, Nolin SJ, Thomas CE, Dean RA. The fusarium wilt resistance locus Fom-2 of melon contains a single resistance gene with complex features. Plant Journal, 2004, 39(3): 283-297
Kim DS, Kim DH, Yoo JH, Kim BD. Cleaved amplified polymorphic sequence and amplified fragment length polymorphism markers linked to the fertility restorer gene in chili pepper (Capsicum annuum L.). Molecular Cells, 2006,21: 135-40
Kobe B, Deisenhofer J. The leucine rich repeata versatile binding motif. Trends in Biochemical Sciences, 1994,19(10): 415-421
Konieczny A, Ausubel FM. A procedure for mapping Arabidopsis mutations using co-dominant ecotype-specific PCR-based markers. Plant Journal, 1993, 4: 403-410
Lander, ES. The new genomics: Global views of biology. Science, 1996, 274(5287): 536-539
Ma Z, Michailides TJ. A real-time PCR assay for detecting azoxystrobin-resistant Alternaria isolates from pistachio orchards in Chalifornia. Crop protection, 2004B, 23: 1259-1263
Ma Z, Michailides TJ. An allele-specific PCR assay for detecting azoxystrobin-resistant Alternaria isolates from pistachio in Chalifornia. Phytopathology, 2004A, 152: 118-121
Martin GB, Bogdanove AJ, Sessa G. Understanding the functions of plant disease resistance proteins. Annual Review of Plant Biology, 2003, 54: 23-61
Martyn RD, Banes LW, Amador J. Fusarium wilt (F. oxysporum f. sp. melonis race 0) of muskmelon in Texas. Plant Disease, 1987, 70: 233-236
Martyn RD, Miller ME. Monosporascus root rot and vine decline:An emerging disease of melons worldwide. Plant Disease, 1996, 80: 716-725
Martyn RD. An initial survey of the United States for races of Fusarium oxysporum f. sp melonis. Science, 1989, 24: 696-698
Meyers, BC, Dickerman AW, Michelmore RW, Sivaramakrishnan S, Sobral BW, Young ND. Plant disease resistance genes encode members of an ancient and diverse protein family within the nucleotide-binding superfamily. Plant Journal. 1999,20:317 332
Michaels SD, Amasino RM. A robust method for detecting single-nucleotide changes as polymorphic markers by PCR. Plant Journal, 1998, 14: 381-385
Moore AL, Albury MS, Crichton PG, Affourtit C. Function of the alternative oxidase:is it still a scavenger? Trends in Plant Science, 2003, 7: 478-481
Nam HG, Giraudat J, Den BB, Moonan F, Loos W, Hauge BM, Goodman HM. Restriction Fragment Length Polymorphism Linkage Map of Arabidopsis thaliana. Plant Cell, 1989, 1: 699-705
Neff MM, Turk E, Kalishman M. Web-based primer design for single nucleotide polymorphism analysis. Trends in genetics, 2002,18 (12): 613-615
Orita M, Wahana H, Kanazawa H. Detection of polymorphisms of human DNA by gel electrophorosis as single-strand conformation polymorphisms. Proceedings of the National Academy of Science of the United State of America, 1989, 86(8): 2766-2770
Osterberg MK, Shavorskaya O, Lascoux M. Naturally occurring indel variation in the Brassica nigra COL1 gene is associated with variation in flowering time. Genetics, 2002, 161:299-306
Pollak E. On the theory of partially inbreeding finite populations I. Partial selfing. Genetics, 1987, 117:353-360
Porter DR. Some environmental relations of watermelon wilt. Phytopathology, 1932, 22:813-825
Powell W, Morgante M, Andre C, Hanafey M, Vogel J, Tingey S, Rafalski A. The comparison of RFLP, RAPDs, AFLP and SSR (microsatellite) markers for germplasm analysis. Molecular Breeding, 1996, 2: 225-238
Pritchard JK, Stephens M, Rosenberg NA, Donnelly P. Association mapping in structured populations. The American Journal of Human Genetics, 2000, 67: 170-181
Risser G, Banihashemi Z, Davis DW. A proposed nomenclature of Fusarium oxysporum f. sp melonis races and resistance genes. Phytopathology, 1976, 66: 1105-1106
Risser G. Study of resistance of melon (Cucumis melo) to races 1 and 2 of fusarium-oxysporum f. sp melonis. African Journal of Biotechnology Annales del amelioration des plant, 1973,23: 259-263
Semagn K, Bj(?)rnstad A, Ndjiondjop MN. An overview of molecular marker methods for plants. African Journal of Biotechnology, 2006, 5(25): 2540-2568
Shepard KA, Purugganan MD. Molecular population genetics of the Arabidopsis CLAVATA2 region. Genetics, 2003, 163: 1083-1095
Sierotzki H, Baumler S, Gisi U, Volker M, Friedrich GF, Gerhard S. Evaluation of Erysiphe gra minis f. sp tritici field isolates for resistance to strobilurin fungicides with different SNP detection systems. Pest management science, 2003, 59(3): 310-314
Small RL, Ryburn JA, Wendel JF. Low levels of nucleotide diversity at homoeologous Adh loci in allotetraploid cotton (Gossypium L.). Molecular Biology Evolution, 1999, 16: 491-501
Sumner DR, Johnson AW. Effect of Root-knot nematodes on fusarium wilt of watermelon. Phytopathology, 1973, 63: 857-861
Tarek J, Joseph JK, Shelly JN, Claude ET, Ralph AD. The fusarium wilt resistance locus Fom-2 of melon contains a single resistance gene with complex features. The Plant Journal, 2004, 39: 283-297
Tenaillon M, Sawkins MC, Long AD, Anderson LK, Gaut RL, Doebley JF, Gaut BS. Patterns of DNA sequence polymorphism along chromosome 1 of maize (Zea mays ssp. mays L.). Proceedings of the National Academic Science of the USA, 2001,98:9161-9166
Thiel T, Kota R, Grosse I, Stein N, Graner A. SNP2CAPS: a SNP and INDEL analysis tool for CAPS marker development. Nucleic acids research, 2004, 32(1): 1-5
Thiel T, Michalek W, Varshney RK, Graner A. Exploiting EST databases for the development and characterization of gene-derived SSR-markers in barley (Hordeum vulgare L.). Theoretical and Applied Genetics, 2003,106: 411-422.
Thornsberry JM, Goodman MM, Doebley J, Kresovich S, Nielsen D, Buckler ES. Dwarf8 polymorphisms associate with variation in flowering time. Nature Genetics, 2001, 28: 286-289
Van Tienderen PH, De Haan AA, Van der Linden CG, Vosman B. Biodiversity assessment using markers for ecologically important traits. Trends in Ecology and Evolution, 2002, 17: 577-582
Wang YH, Thomas CE, Dean RA. Genetic mapping of a fusarium wilt resistance gene (Fom-2) in melon (Cucumis melo L.). Molecular Breeding, 2000, 6: 379-389
Warren RF, Henk A, Mowery P. A mutation within the leucine-rich repeat domain of the Arabidopsis disease resistance gene RPS5 partially suppresses multiple bacterial and downy mildew resistance genes.The Plant Cell, 1998, 10: 1439-1452
Wechter P, Michael W, Whitehead P, Claude ET. Identification of a Randomly Amplfied Polymorphic DNA Marker Linked to the Fom-2 Fusarium wilt Resistance Gene in Muskmelon MR-1. Phytopathology, 1995, 8: 1245-1249
Yeam I, Kang BC, Lindeman W, Frantz JD, Faber N, Jahn MM. Allele-specific CAPS markers based on point mutations in resistance alleles at the pvrl locus encoding eIF4E in Capsicum. Theoretical and Applied Genetics, 2005, 112: 178-86
Yu J, Hu S, Wang J, Wong G K, Li S, Liu B, Deng Y, Dai L, Zhou Y, Zhang X, Cao M, Liu J, Sun J, Tang J, Chen Y, Huang X, Lin W, Ye C, Tong W, Cong L, Geng J, Han Y, Li L, Li W, Hu G, Huang X, Li W, Li J, Liu Z, Li L, Liu J, Qi Q, Liu J, Li L, Li T, Wang X, Lu H, Wu T, Zhu M, Ni P, Han H, Dong W, Ren X, Feng X, Cui P, Li X, Wang H, Xu X, Zhai W, Xu Z, Zhang J, He S, Zhang J, Xu J, Zhang K, Zheng X, Dong J, Zeng W, Tao L, Ye J, Tan J, Ren X, Chen X, He J, Liu D, Tian W, Tian C, Xia H, Bao Q, Li G, Gao H, Cao T, Wang J, Zhao W, Li P, Chen W, Wang X, Zhang Y, Hu J, Wang J, Liu S, Yang J, Zhang G, Xiong Y, Li Z, Mao L, Zhou C, Zhu Z, Chen R, Hao B, Zheng W, Chen S, Guo W, Li G, Liu S, Tao M, Wang J, Zhu L, Yuan L, Yang H. A draft sequence of the rice genome (Oryza sativa L. ssp. indica). Science, 2002,296: 79-92.
Zheng XY, Wolff DW, Baudracco-Arnas S, Pitrat M. Development and utility of cleaved amplified polymorphic sequences (CAPS) and restriction fragment length polymorphisms (RFLPs) linked to the Fom-2 fusarium wilt resistance gene in melon (Cucumis melo L.). Theoretical and Applied Genetics, 1999, 99: 453-463
Zhu YL, Song QJ, Hyten DL, Van Tossell CP, Matukumalli LK, Grimm DR, Hyatt SM, Fickus EW, Yong ND, Cregan PB. Single-nucleotide polymorphisms in soybean. Genetics, 2003, 163: 1123-1134
Zink FW, Gubler WD. Inheritance of resistance in muskmelon to Fusarium wilt. Journal of the American Society for Horticultural Science, 1985,110: 600-604
陈幼源,陈绯翔.不同南瓜砧木品种对网纹甜瓜生长发育的影响.上海农业学报,2002,18(4):47-51
川出武夫.甜瓜对枯萎病的抗性,杨鼎新译.中国西瓜甜瓜,2002,(4):43-45
杜玮南,孙红霞,方福德.单核苷酸多态性的研究进展.中国医学科学院学报,2000,(8):392-394
侯栋,闫秀玲,李浩,高艳霞.几种不同砧木在哈密瓜嫁接无土栽培中的表现.中国西瓜甜瓜,2002
黄振文.利用土壤添加物防治作物之土壤传播性病害.植物保护学会会刊,1991,33:113-123
吉加兵,徐润芳.西瓜种质资源抗枯萎病性的苗期鉴定.中国西瓜甜瓜,1990,(2):19-20
蒋平,谢青,唐华.施宝灵防治西瓜枯萎病药效试验.1992,16(5):18-19
杰弗里.植物分类学入门.胡征宇译.1990
李红霞,周明国.用等位基因特异性寡核苷酸(ASO)-PCR快速检测抗多菌灵的油菜菌核病菌.中国农业科学.2004,37(9):1396-1399
李瑞琴,甜瓜枯萎病病原学及防治技术研究.西北农林科技大学硕士论文,2004
林德佩.国外甜瓜西瓜的研究动态.中国西瓜甜瓜,1989(2):1-8;1990(1):7-8
刘君璞,马跃.我国西瓜甜瓜种业的现状与发展对策.中国西瓜甜瓜,2000,(3):2-6
刘连杰,董凤珍.吉林省早熟甜瓜枯萎病的发生与防治.中国西瓜甜瓜,1995,(1):28
刘秀芳,刘芳政,王叶霜.西瓜甜瓜主要病虫害的识别及其防治.北京:农业出版社,1993
卢文经.薄皮甜瓜嫁接抗病增产技术研究.辽宁农业科学,2002,(1):16-20
陆致平,项继忠,吴雄飞.恶霉灵防治西瓜枯萎病试验研究.现代农药,2002,(3):44
马国斌,林德佩,王叶筠,郭庆元.西瓜枯萎病菌镰刀菌酸对西瓜苗作用机制的初步探讨.植物病理学报,1999,30:373-374
马国斌,王鸣,郑学勤.西瓜和甜瓜镰刀菌酸抗性变异体的诱导.中国园艺学 会第六届青年学术讨论会论文集,2004
戚配坤.瓜类枯萎病菌专化型研究简介.华南农业大学学报,1994,9(4):81-86
齐三魁,吴大康,林德佩.中国甜瓜.北京:科学普及出版社,1991
乔凤,陈泽良,王玉飞,赵瑾,杜昕颖,于雅琴,黄留玉.pUC19K质粒的构建及其在布鲁氏菌突变株构建中的应用.中国生物工程杂志,2007,27(12):1-5
王坚.中国西瓜甜瓜.中国农业出版社,2000,08
王贤磊,李群,方勇,高小明,杨艳,李冠.单核苷酸多态性与甜瓜抗枯萎病分子育种研究.生物技术,2007,17(4):31-34
王叶筠,林德佩,郭庆元.新疆甜瓜抗病育种研究.中国西瓜甜瓜,1993,(4):17-20
温玲.枯萎灵等药剂对西瓜枯萎病防治效果.黑龙江农业科学,2000,6:28
谢大森,何晓明,彭庆务,骆焕彬.瓜类枯萎病发生机理研究进展.仲恺农业技术学院学报,2006,19(3):65-70
徐胜利,李新民.厚皮甜瓜嫁接育苗栽培的防病增产效果.中国蔬菜,2000(4):16-18
许勇,朱其杰,陈晓莉,梁根庆.镰刀菌酸对黄瓜不同品种的致萎力与枯萎病菌致病力的相关性.中国蔬菜,1993,1:19-20
颜志强,杨胜利,龚毅.PCR及其衍生技术在基因突变检测中的应用.遗传,2003,25(2):198-200
张蓓,沈立松.实时荧光定量PCR的研究进展及其应用.国外医学临床生物化学与检验学分册,2003,24(6):327-329
周真,孙芙蓉,曹长余,于建美,韩冰.西瓜枯萎病的田间施药方式及药效对比试验.山东蔬菜,2000,(3):36-37
朱天圣,戚配坤.苦瓜枯萎病病原菌研究.华南农业大学学报,1998,19(4):14-18
Abea F,Saitob K,Miurac K,Toriyamaa K.A single nucleotide polymorphism in the alternative oxidase gene among rice varieties differing in low temperature tolerance.FEBS Letters,2002,527:181-185
Andersen JR,L(u|¨)bberstedt T.Functional markers in plants.Trends in Plant Science,2003,8(11):554-560
Amholdt-Schmitt B,Costa JH,Melo DF.AOX-a functional marker for efficient cell reprogram ruing under stress? Trends in Plant Science,2006,11(6):282-287
Arnholdt-Schmitt B.Functional markers and a 'systemic strategy':convergency between plant breeding, plant nutrition and molecular biology. Plant Physiology and Biochemistry, 2005, 43: 817-820
Bang H, Kim S, Leskovar D, King S. Development of a codominant CAPS marker for allelic selection between canary yellow and red watermelon based on SNP in lycopene beta-cyclase (LCYB) gene. Moleculer Breeding, 2007, 20: 63-72
Bang HJ, Kim SG, Leskovar D, King S. Development of a codominant CAPS marker for allelic selection between canary yellow and red watermelon based on SNP in lycopene beta-cyclase (LCYB) gene. Molecular breeding, 2007, 20(1): 63-72
Baumler S, Felsenstein FG, Schwarz G. CAPS and DHPLC analysis of a single nucleotide polymorphism in the cytochrome b gene conferring resistance to strobilurins in field isolates of Blumeria gra minis f. sp hordei. Phytopathology phytopathologische zeitschrift. 2003,151(3): 149-152
Booth C. 陈其瑛译.镰刀菌属.北京: 农业出版社, 1988
Caranta C, Thabuis A, Palloix A. Development of a CAPS marker for the Pvr4 locus: A tool for pyramiding potyvirus resistance genes in pepper. Genome, 1999, 42(6): 1111-1116
Chasman D; Adams RM. Predicting the functional consequences of non-synonymous single nucleotide polymorphisms: Structure-based assessment of amino acid variation. Molecular Biology, 2001, 307(2): 683-706
Chiapparino E, Lee D, Donini P. Genotyping single nucleotide polymorphisms in barley by tetra-primer ARMS-PCR. Genome, 2004,47: 414-420
Danin-Poleg, Y, Burger Y, Schreiber S, Katzir N, Cohen R. Identification of the gene for resistance to Fusarium wilt races 0 and 2 in Cucumis melo. Cucurbit Genetic Cooperate Report, 1999, 22: 19-20
David D. Fusaric acid in selective pathogenicity of Fusarium oxysporum. Phytopathology, 1969, 59: 1391-1395
Filatov DA, Charlesworth D. DNA polymorphism, haplotype structure and balancing selection in the Leavenworthia PgiC locus. Genetics, 1999, 153: 1423-1434
Fiorani F, Umbach AL, Siedow JN. The alternative oxidase of plant mitochondria is involved in the acclimation of shoot growth at low temperature. A study of Arabidopsis AOXla transgenic plants. Plant Physiology, 2005, 139: 1795-1805.
Fraaije BA, Butters JA, Coelho JM, Jones DR, Hollomon DW. Following the dynamics of strobilurin resistance in Blumeria gra minis f.sp tritici using quantitative allele-specific real-time PCR measurements with the fluorescent dye SYBR Green I. Plant pathology, 2002, 51(1): 45-54
Fraaije BA, Cools HJ, Fountaine J, Lovell DJ, Motteram J, West JS, Lucas JA. Role of ascospores in further spread of QoI-resistant cytochrome b alleles (G143A) in field populations of Mycosphaerella gra minicola. Phytopathology, 2005, 95 (8): 933-941
Gao ZS, Van de Weg WE, Schaart JG, Van der Meer IM, Kodde L, Laimer M, Breiteneder H, Hoffmann-Sommergruber K, Gilissen L J. Linkage map positions and allelic diversity of two Mal d 3 (non-specific lipid transfer protein) genes in the cultivated apple (Malus domestica). Theoretical and Applied Genetics, 2005, 110:479-491
Gordon TR, Martyn RD. The evolutionary biology of Fusarium oxysporum. Annual review of Phytopathology, 1997,35:111-128
Grodzicker T, Anderson C, Sharp PA, Sambrook J. Conditional lethal mutants of adenovirus 2-simian virus 40 hybrids. I. Host range mutants of Ad2+ND1. The Jounal of virology, 1974, 13(6): 1237-1244
Gubler WD, Grogan RG. Fusarium wilt of muskmelon in San Joaquin Valley of California. Plant disease report, 1976,60(9): 742-744
Gupta PK, Varshney RK, Prasad M. Molecular markers: principles and methodology. In: Jain S M. Molecular Techniques in Crop Improvement. Kluwer Academic Publishers, 2002: 9-54
Hackauf B, Wehling P. Identification of microsatellite polymorphisms in an expressed portion of the rye genome. Plant Breeding, 2002,121: 17-25
Hagenblad J, Nordborg M. Sequence variation and haplotype structure surrounding the flowering time locus FR1 in Arabidopsis thaliana. Genetics, 2002, 161:289-298
Hughes AL, Nei M. Pattern of nucleotide substitution at major histocompatibility complex class I loci reveals overdominant selection. Nature, 1988, 335: 167-170
Ilic K, Berleth T, Provart NJ. BlastDigester-a web-based program for efficient CAPS marker design. Trends in Genetics, 2004, 20(7): 280-283
Ishii H, Fraaije BA, Sugiyama T, Noguchi K, Nishimura K, Takeda T, Amano T, Hollomon DW. Occurrence and molecular characterization of strobilurin resistance in cucumber powdery mildew and downy mildew. Phytopathology, 2001,91(12): 1166-1171
Iyer-Pascuzzi AS, McCouch SR. Functional markers for xa5-mediated resistance in rice (Oryza sativa L.). Molecular Breeding, 2007, 19: 291-296
Joobeur T, King JJ, Nolin SJ, Thomas CE, Dean RA. The fusarium wilt resistance locus Fom-2 of melon contains a single resistance gene with complex features. Plant Journal, 2004, 39(3): 283-297
Kim DS, Kim DH, Yoo JH, Kim BD. Cleaved amplified polymorphic sequence and amplified fragment length polymorphism markers linked to the fertility restorer gene in chili pepper (Capsicum annuum L.). Molecular Cells, 2006,21: 135-40
Kobe B, Deisenhofer J. The leucine rich repeata versatile binding motif. Trends in Biochemical Sciences, 1994,19(10): 415-421
Konieczny A, Ausubel FM. A procedure for mapping Arabidopsis mutations using co-dominant ecotype-specific PCR-based markers. Plant Journal, 1993, 4: 403-410
Lander, ES. The new genomics: Global views of biology. Science, 1996, 274(5287): 536-539
Ma Z, Michailides TJ. A real-time PCR assay for detecting azoxystrobin-resistant Alternaria isolates from pistachio orchards in Chalifornia. Crop protection, 2004B, 23: 1259-1263
Ma Z, Michailides TJ. An allele-specific PCR assay for detecting azoxystrobin-resistant Alternaria isolates from pistachio in Chalifornia. Phytopathology, 2004A, 152: 118-121
Martin GB, Bogdanove AJ, Sessa G. Understanding the functions of plant disease resistance proteins. Annual Review of Plant Biology, 2003, 54: 23-61
Martyn RD, Banes LW, Amador J. Fusarium wilt (F. oxysporum f. sp. melonis race 0) of muskmelon in Texas. Plant Disease, 1987, 70: 233-236
Martyn RD, Miller ME. Monosporascus root rot and vine decline:An emerging disease of melons worldwide. Plant Disease, 1996, 80: 716-725
Martyn RD. An initial survey of the United States for races of Fusarium oxysporum f. sp melonis. Science, 1989, 24: 696-698
Meyers, BC, Dickerman AW, Michelmore RW, Sivaramakrishnan S, Sobral BW, Young ND. Plant disease resistance genes encode members of an ancient and diverse protein family within the nucleotide-binding superfamily. Plant Journal. 1999,20:317 332
Michaels SD, Amasino RM. A robust method for detecting single-nucleotide changes as polymorphic markers by PCR. Plant Journal, 1998, 14: 381-385
Moore AL, Albury MS, Crichton PG, Affourtit C. Function of the alternative oxidase:is it still a scavenger? Trends in Plant Science, 2003, 7: 478-481
Nam HG, Giraudat J, Den BB, Moonan F, Loos W, Hauge BM, Goodman HM. Restriction Fragment Length Polymorphism Linkage Map of Arabidopsis thaliana. Plant Cell, 1989, 1: 699-705
Neff MM, Turk E, Kalishman M. Web-based primer design for single nucleotide polymorphism analysis. Trends in genetics, 2002,18 (12): 613-615
Orita M, Wahana H, Kanazawa H. Detection of polymorphisms of human DNA by gel electrophorosis as single-strand conformation polymorphisms. Proceedings of the National Academy of Science of the United State of America, 1989, 86(8): 2766-2770
Osterberg MK, Shavorskaya O, Lascoux M. Naturally occurring indel variation in the Brassica nigra COL1 gene is associated with variation in flowering time. Genetics, 2002, 161:299-306
Pollak E. On the theory of partially inbreeding finite populations I. Partial selfing. Genetics, 1987, 117:353-360
Porter DR. Some environmental relations of watermelon wilt. Phytopathology, 1932, 22:813-825
Powell W, Morgante M, Andre C, Hanafey M, Vogel J, Tingey S, Rafalski A. The comparison of RFLP, RAPDs, AFLP and SSR (microsatellite) markers for germplasm analysis. Molecular Breeding, 1996, 2: 225-238
Pritchard JK, Stephens M, Rosenberg NA, Donnelly P. Association mapping in structured populations. The American Journal of Human Genetics, 2000, 67: 170-181
Risser G, Banihashemi Z, Davis DW. A proposed nomenclature of Fusarium oxysporum f. sp melonis races and resistance genes. Phytopathology, 1976, 66: 1105-1106
Risser G. Study of resistance of melon (Cucumis melo) to races 1 and 2 of fusarium-oxysporum f. sp melonis. African Journal of Biotechnology Annales del amelioration des plant, 1973,23: 259-263
Semagn K, Bj(?)rnstad A, Ndjiondjop MN. An overview of molecular marker methods for plants. African Journal of Biotechnology, 2006, 5(25): 2540-2568
Shepard KA, Purugganan MD. Molecular population genetics of the Arabidopsis CLAVATA2 region. Genetics, 2003, 163: 1083-1095
Sierotzki H, Baumler S, Gisi U, Volker M, Friedrich GF, Gerhard S. Evaluation of Erysiphe gra minis f. sp tritici field isolates for resistance to strobilurin fungicides with different SNP detection systems. Pest management science, 2003, 59(3): 310-314
Small RL, Ryburn JA, Wendel JF. Low levels of nucleotide diversity at homoeologous Adh loci in allotetraploid cotton (Gossypium L.). Molecular Biology Evolution, 1999, 16: 491-501
Sumner DR, Johnson AW. Effect of Root-knot nematodes on fusarium wilt of watermelon. Phytopathology, 1973, 63: 857-861
Tarek J, Joseph JK, Shelly JN, Claude ET, Ralph AD. The fusarium wilt resistance locus Fom-2 of melon contains a single resistance gene with complex features. The Plant Journal, 2004, 39: 283-297
Tenaillon M, Sawkins MC, Long AD, Anderson LK, Gaut RL, Doebley JF, Gaut BS. Patterns of DNA sequence polymorphism along chromosome 1 of maize (Zea mays ssp. mays L.). Proceedings of the National Academic Science of the USA, 2001,98:9161-9166
Thiel T, Kota R, Grosse I, Stein N, Graner A. SNP2CAPS: a SNP and INDEL analysis tool for CAPS marker development. Nucleic acids research, 2004, 32(1): 1-5
Thiel T, Michalek W, Varshney RK, Graner A. Exploiting EST databases for the development and characterization of gene-derived SSR-markers in barley (Hordeum vulgare L.). Theoretical and Applied Genetics, 2003,106: 411-422.
Thornsberry JM, Goodman MM, Doebley J, Kresovich S, Nielsen D, Buckler ES. Dwarf8 polymorphisms associate with variation in flowering time. Nature Genetics, 2001, 28: 286-289
Van Tienderen PH, De Haan AA, Van der Linden CG, Vosman B. Biodiversity assessment using markers for ecologically important traits. Trends in Ecology and Evolution, 2002, 17: 577-582
Wang YH, Thomas CE, Dean RA. Genetic mapping of a fusarium wilt resistance gene (Fom-2) in melon (Cucumis melo L.). Molecular Breeding, 2000, 6: 379-389
Warren RF, Henk A, Mowery P. A mutation within the leucine-rich repeat domain of the Arabidopsis disease resistance gene RPS5 partially suppresses multiple bacterial and downy mildew resistance genes.The Plant Cell, 1998, 10: 1439-1452
Wechter P, Michael W, Whitehead P, Claude ET. Identification of a Randomly Amplfied Polymorphic DNA Marker Linked to the Fom-2 Fusarium wilt Resistance Gene in Muskmelon MR-1. Phytopathology, 1995, 8: 1245-1249
Yeam I, Kang BC, Lindeman W, Frantz JD, Faber N, Jahn MM. Allele-specific CAPS markers based on point mutations in resistance alleles at the pvrl locus encoding eIF4E in Capsicum. Theoretical and Applied Genetics, 2005, 112: 178-86
Yu J, Hu S, Wang J, Wong G K, Li S, Liu B, Deng Y, Dai L, Zhou Y, Zhang X, Cao M, Liu J, Sun J, Tang J, Chen Y, Huang X, Lin W, Ye C, Tong W, Cong L, Geng J, Han Y, Li L, Li W, Hu G, Huang X, Li W, Li J, Liu Z, Li L, Liu J, Qi Q, Liu J, Li L, Li T, Wang X, Lu H, Wu T, Zhu M, Ni P, Han H, Dong W, Ren X, Feng X, Cui P, Li X, Wang H, Xu X, Zhai W, Xu Z, Zhang J, He S, Zhang J, Xu J, Zhang K, Zheng X, Dong J, Zeng W, Tao L, Ye J, Tan J, Ren X, Chen X, He J, Liu D, Tian W, Tian C, Xia H, Bao Q, Li G, Gao H, Cao T, Wang J, Zhao W, Li P, Chen W, Wang X, Zhang Y, Hu J, Wang J, Liu S, Yang J, Zhang G, Xiong Y, Li Z, Mao L, Zhou C, Zhu Z, Chen R, Hao B, Zheng W, Chen S, Guo W, Li G, Liu S, Tao M, Wang J, Zhu L, Yuan L, Yang H. A draft sequence of the rice genome (Oryza sativa L. ssp. indica). Science, 2002,296: 79-92.
Zheng XY, Wolff DW, Baudracco-Arnas S, Pitrat M. Development and utility of cleaved amplified polymorphic sequences (CAPS) and restriction fragment length polymorphisms (RFLPs) linked to the Fom-2 fusarium wilt resistance gene in melon (Cucumis melo L.). Theoretical and Applied Genetics, 1999, 99: 453-463
Zhu YL, Song QJ, Hyten DL, Van Tossell CP, Matukumalli LK, Grimm DR, Hyatt SM, Fickus EW, Yong ND, Cregan PB. Single-nucleotide polymorphisms in soybean. Genetics, 2003, 163: 1123-1134
Zink FW, Gubler WD. Inheritance of resistance in muskmelon to Fusarium wilt. Journal of the American Society for Horticultural Science, 1985,110: 600-604