茅莓与插田泡遗传关系的RAPD和ISSR分析及疑似杂种初步鉴定
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摘要
悬钩子属Rubus L.,是蔷薇科Rosaceae中的一个大属。全世界有悬钩子属植物750-1000种,中国有200多种,其中有130多种是特有种。我国西南地区是悬钩子属植物的多样性分布中心,拥有丰富的种质资源,其野生资源在各种抗性、易成活性、果实风味都优于从国外引进的栽培品种,具有重要的栽培价值和育种价值。本研究在形态学调查的基础上利用RAPD和ISSR分子标记技术对33份材料属空心莓组(Sect. Idaeobatus)属绒毛叶亚组(Subsect. Stimulantes Yu& Lu)的茅莓(R. parvifolius)16份、属柔毛叶亚组(Subsect. Pungentes (Focke) Yu&Lu)的插田泡(R. coreanus)16份以及新发现的疑似杂种1份(该杂种疑为茅莓×插田泡自然杂交所产生)进行了遗传差异分析和初步鉴定。主要结果如下：
1.通过优化反应体系,确立了茅莓和插田泡在ISSR分析中引物的最佳退火温度。即每个引物由于其碱基数目和组合的不同,退火温度有明显差异,但又与本身Tm值无规律可循,在筛选出的效果较好的引物退火温度范围在55.2-59.1℃,为ISSR技术在茅莓和插田泡中应用打下了基础。
2.从UBC#9(哥伦比亚大学开发的一套共100条ISSR引物)引物中筛选出.12个多态性强、稳定性好、重复性高的ISSR引物,用于33份茅莓和插田泡的ISSR分析。扩增DNA片段长度在200-4000bp之间,多集中于400-2000bp之间。扩增DNA谱带总数358条,多态性条带339条,多态性达94.69%。得到了12个引物对33份茅莓和插田泡材料清晰的指纹图谱,能够充分鉴定种间和种内关系。
3.利用已筛选出的18条随机引物,对供试的33份茅莓与插田泡材料进行RAPD分析。扩增DNA片段长度在200-4000bp之间,多集中于400-2000bp之间。扩增谱带总数498条,其中多态性条带462条,多态性达92.77%。得到的清晰指纹图谱,亦能够充分鉴定种间和种内关系。
4.依据RAPD和ISSR标记数据,分别计算出33份茅莓与插田泡材料的遗传相似系数,并将供试材料划分为2大类。UPGMA聚类分析显示与形态学上分类结果基本一致。茅莓和插田泡各自种内的体现出一定的地理差异。
5.通过RAPD和ISSR分析,对疑似杂种的可能亲本进行了推定,认为该疑似杂种可能为茅莓和插田泡的自然杂种。
6.通过RAPD和ISSR对茅莓和插田泡种群遗传关系分析,RAPD和ISSR的重复性和稳定性得到了进一步验证,且ISSR比RAPD表现出更好的多态性,也为更好地利用DNA分子标记提供依据。
The genus Rubus is one of the largest genera in the Rosaceae, consisting of 750-1000 species in many parts of the world, of which about 200 species with more than 130 native have been recorded in China. The southwest part of China which is abound in resources, is now the center of distribution of Rubus. The disease resistance, easily activity and fruit flavors in its wild plants which have important cultural and breeding value, are all superior than cultivars introduced from abroad. In this study, RAPD and ISSR were used to analyze the total 33 accessions with 16 taxa from Subsect. Stimulantes Yii & Lu,16 taxa from Subsect. Pungentes (Focke) Yii & Lu and 1 taxa from the progeny originated from R. parvifolius cross R. coreanus. The results were as follows:
1. Through optimizing the ISSR reaction system, the optimal annealing temperatures for ISSR primers were found. The annealing temperatures were related to the bases of each primer, while were less related to the Tm value. The selected range of temperature,55.2-59.1℃, would be helpful for utilizing ISSR on R. parvifolius and R. coreanus.
2.12 primers screened from UBC#9 primers, which amplified 358 repeatable loci, including 339 polymorphic loci, were applied on the 33 materials. The percentage of polymorphic bands was 92.77% and the long of fragments were between 200bp and 4000bp, mostly between 400bp and 2000bp. The ISSR fingerprints from 12 Primers can identify the relationships between interspecies and intraspecies exactly.
3.18 primers which amplified 498 repeatable loci, including 462 polymorphic loci, were applied on the 33 materials. The percentage of polymorphic bands was 92.77% and the long of fragments were between 200bp and 4000bp, mostly between 400bp and 2000 bp. The RAPD fingerprints from 18 Primers can identify the relationships between interspecies and intraspecies exactly.
4. According to RAPD and ISSR markers, genetic distance was computed. All materials can be divided into 2 groups which were in consistence with that of the analysis of morphology. Division of subsets in some groups were related to the geography difference of R. parvifolius and R. coreanus.
5. The possible parents of suspected hybrid was concluded by RAPD and ISSR analysis, and it was considered that the suspected hybrid was a natural hybrid between R. parvifolius and R. coreanus.
6. In this study, reproducibility and stability of RAPD and ISSR was proved. ISSR showed the better polymorphic compared with RAPD, and this would be helpful for DNA molecular maker.
1.通过优化反应体系,确立了茅莓和插田泡在ISSR分析中引物的最佳退火温度。即每个引物由于其碱基数目和组合的不同,退火温度有明显差异,但又与本身Tm值无规律可循,在筛选出的效果较好的引物退火温度范围在55.2-59.1℃,为ISSR技术在茅莓和插田泡中应用打下了基础。
2.从UBC#9(哥伦比亚大学开发的一套共100条ISSR引物)引物中筛选出.12个多态性强、稳定性好、重复性高的ISSR引物,用于33份茅莓和插田泡的ISSR分析。扩增DNA片段长度在200-4000bp之间,多集中于400-2000bp之间。扩增DNA谱带总数358条,多态性条带339条,多态性达94.69%。得到了12个引物对33份茅莓和插田泡材料清晰的指纹图谱,能够充分鉴定种间和种内关系。
3.利用已筛选出的18条随机引物,对供试的33份茅莓与插田泡材料进行RAPD分析。扩增DNA片段长度在200-4000bp之间,多集中于400-2000bp之间。扩增谱带总数498条,其中多态性条带462条,多态性达92.77%。得到的清晰指纹图谱,亦能够充分鉴定种间和种内关系。
4.依据RAPD和ISSR标记数据,分别计算出33份茅莓与插田泡材料的遗传相似系数,并将供试材料划分为2大类。UPGMA聚类分析显示与形态学上分类结果基本一致。茅莓和插田泡各自种内的体现出一定的地理差异。
5.通过RAPD和ISSR分析,对疑似杂种的可能亲本进行了推定,认为该疑似杂种可能为茅莓和插田泡的自然杂种。
6.通过RAPD和ISSR对茅莓和插田泡种群遗传关系分析,RAPD和ISSR的重复性和稳定性得到了进一步验证,且ISSR比RAPD表现出更好的多态性,也为更好地利用DNA分子标记提供依据。
The genus Rubus is one of the largest genera in the Rosaceae, consisting of 750-1000 species in many parts of the world, of which about 200 species with more than 130 native have been recorded in China. The southwest part of China which is abound in resources, is now the center of distribution of Rubus. The disease resistance, easily activity and fruit flavors in its wild plants which have important cultural and breeding value, are all superior than cultivars introduced from abroad. In this study, RAPD and ISSR were used to analyze the total 33 accessions with 16 taxa from Subsect. Stimulantes Yii & Lu,16 taxa from Subsect. Pungentes (Focke) Yii & Lu and 1 taxa from the progeny originated from R. parvifolius cross R. coreanus. The results were as follows:
1. Through optimizing the ISSR reaction system, the optimal annealing temperatures for ISSR primers were found. The annealing temperatures were related to the bases of each primer, while were less related to the Tm value. The selected range of temperature,55.2-59.1℃, would be helpful for utilizing ISSR on R. parvifolius and R. coreanus.
2.12 primers screened from UBC#9 primers, which amplified 358 repeatable loci, including 339 polymorphic loci, were applied on the 33 materials. The percentage of polymorphic bands was 92.77% and the long of fragments were between 200bp and 4000bp, mostly between 400bp and 2000bp. The ISSR fingerprints from 12 Primers can identify the relationships between interspecies and intraspecies exactly.
3.18 primers which amplified 498 repeatable loci, including 462 polymorphic loci, were applied on the 33 materials. The percentage of polymorphic bands was 92.77% and the long of fragments were between 200bp and 4000bp, mostly between 400bp and 2000 bp. The RAPD fingerprints from 18 Primers can identify the relationships between interspecies and intraspecies exactly.
4. According to RAPD and ISSR markers, genetic distance was computed. All materials can be divided into 2 groups which were in consistence with that of the analysis of morphology. Division of subsets in some groups were related to the geography difference of R. parvifolius and R. coreanus.
5. The possible parents of suspected hybrid was concluded by RAPD and ISSR analysis, and it was considered that the suspected hybrid was a natural hybrid between R. parvifolius and R. coreanus.
6. In this study, reproducibility and stability of RAPD and ISSR was proved. ISSR showed the better polymorphic compared with RAPD, and this would be helpful for DNA molecular maker.
引文
[1]曲泽洲,孙云蔚.果树种类论[M].北京：农业出版社,1990,153-16
[2]俞德浚,陆玲娣,谷粹芝,等.中国植物志.第37卷[M].北京：科学出版社,1985
[3]陆玲娣.我国悬钩子属植物的研究[J].植物分类学报,1983,21(1)：13-25
[4]Thompson M M. Survey of chromosome numbers in Rubus[J]. Annals of the Missouri Botanical Garden,1997,84(1):128-164
[5]杨燕林,唐开学,和加卫,等.悬钩子属植物分子标记技术和基因组研究进展[J].中国农学通报,2008,24(8)：93-98
[6]陈少风,叶居新,朱祥玲,等.若干悬钩子属植物的花粉形态研究[J].植物研究,1996,16(4)：463-466
[7]周丽华,韦仲新,吴征镒.国产蔷薇科蔷薇亚科的花粉形态[J].云南植物研究,1999,21(4)：455-460
[8]李维林,贺善安,顾姻,等.中国悬钩子属花粉形态观察[J].植物分类学报,2001,39(3)：234-247
[9]王小蓉,汤浩茹,黄力,等.树莓部分野生种及栽培品种花粉亚显微形态的比较[J].园艺学报,2007,34(6)：1395-1404
[10]陈瑞阳.中国主要经济植物染色体图谱(第一册)—中国果树及其野生近缘植物染色体图谱[M].北京：万国学术出版社,1993：263-279
[11]李秀兰,宋文芹,陈瑞阳.北方几种小浆果植物的核型研究[J].武汉植物学研究,1993,11(4)：289-292
[12]林盛华,张冰冰,方成泉,等.中国树莓属8个种染色体数目与核型[J].园艺学报,1994,21(4)：313-319
[13]顾姻,王传永,赵昌民,等.悬钩子属种质的评价[J].植物资源与环境,1996,5(3)：6-13
[14]王小蓉,汤浩茹,段娟,等.中国悬钩子属空心莓组与木莓组28种和变种的核型比较研究[J].植物分类学报,2008,46(4)：505-515
[15]Li W L, Gu Y, He S A et al, Applicability of RAPD to systematics and genetic diversity of Rubus L.. Acta Horticulturae,2002,585:139-142
[16]段娟.西南地区17种(33份)不同来源的野生悬钩子属植物种间遗传多样性的RAPD 分析[D].四川雅安,四川农业大学,2007
[17]李玲.RAPD对树莓栽培品种及插田泡类型的遗传差异分析[D].四川雅安,四川农业大学,2007
[18]贾静波,李维林,吴文龙,等.悬钩子属植物的RAPD分析[J].植物资源与环境学报,2008,17(3)：18-22
[19]王小蓉.悬钩子属植物遗传多样性及其特异资源的筛选[D].四川雅安,四川农业大学,2008
[20]傅承新,沈朝栋,黄爱君.浙江悬钩子属植物的综合研究—资源调查、引种及开发利用前景[J].浙江农业大学学报,1995,21(4)：393-397
[21]栾文举,肖雯,焦健.甘肃省悬钩子属植物优良种类的营养评价及利用途径[J].甘肃科学学报,1996,8(1)：51-55
[22]郭军战,陈铁山,彭少兵.岭山区黄果悬钩子种质资源分析与评价[J].西北林学院学报,2004,19(2)：41-43
[23]Gu Y, Sun Z J, Cai J H et.al.. Introduction and utilization of small fruits in China with special reference to Rubus species[J].'Acta Horticulturae,1989,262:47-55
[24]俞德浚.中国果树分类学[M].北京：中国农业出版社,1979：209-220
[25]李春奇,叶永忠,王志强,等.河南野生悬钩子属植物资源[J].果树科学,1995,12(4)：258-261
[26]栾文举,焦健.甘肃省悬钩子属植物资源分布及其开发前景初探[J].自然资源学报,1996,11(1)：41-48
[27]张志勇,俞志雄.江西悬钩子属的分类和地理分布[J].热带亚热带植物学报,2003,11(1)：27-33
[28]顾姻.悬钩子属植物资源及其利用[J].植物资源与环境,1992,1(2)：50-60
[29]何顺志.贵州悬钩子属(蔷薇科)一新种—务川悬钩子[J].植物分类学报,2006,43：345-347
[30]陆玲娣.中国蔷薇科部分属分类群的修订[J].植物分类学报,2000,38.276-281
[31]李维林,贺善安.一些悬钩子属分类群的修订[J].植物学研究,2001,21：346-249
[32]Naruhashi N. Notes on Japanese Rubus(4). Taxon,1989,37:1-4
[33]李玲,汤浩茹,段娟,等.中国悬钩子属植物的分类鉴定与开发利用[J].安徽农业科学.2006,34(8)：1513-1516
[34]王传永,顾姻,吴文龙,等.悬钩子属基因库的建立与维护[J].植物资源与环境,1996,5(1)：14-17
[35]傅承新,沈朝栋,黄爱军.浙江悬钩子属植物的综合研究[J].浙江农业大学学报,1995,21(4)：393-397
[36]金炜,黄树芝,顾姻.福建省悬钩子属植物资源的调查、收集、评价、和利用的研究[J].武汉植物学报研究，1992,10(4)：371-376
[37]贺善安,顾姻,孙醉君,等.黑莓引种的理论导向[J].植物资源与环境,1998,7(1)：1-9
[38]李维林,贺善安,顾姻.中国悬钩子属植物的利用价值概述[J].武汉植物学研究,2000,18(3)：237-243
[39]张志勇,赖小容.井冈山悬钩子属植物资源的初步研究[J].江西农业大学学报,1999，21(3)：395-398
[40]李孝苗,何家庆.覆盆子资源及其开发利用[J].安徽农业科学,2000,28(1)：128
[41]曹钰普.悬钩子人工栽培技术[J].林业月报,1999,7：19
[42]李维林.覆盆子人工栽培技术[J].中药材,1991,14(5)：10-11
[43]锁必龙,刘炳龙.树莓丰产栽培技术[J].安徽林业,2004(4)：31
[44]张福平.粤东地区野生可食悬钩子植物资源及利用研究Ⅲ[J].中国食物与营养,2003(12)：24-25
[45]田春元,姜益泉,吴金清.河南鸡公山国家级自然保护区悬钩子属植物资源开发利用研究[J].武汉植物学研究,2003,21(6)：521-525
[46]李维林.我国野生悬钩子资源及其开发利用[J].中国野生植物,1991(3)：14-18
[47]仲山民,田荆祥,吴美春.悬钩子果实的营养成分分析[J].浙江林学院学报,1993,10(4)：485-489
[48]桑建忠,顾姻.中国东南部部分悬钩子果实的营养成分Ⅲ[J].植物资源与环境,1995,4(2)：22-26
[49]陈炳华.红腺悬钩子果实营养成分研究[J].海南帅范学院学报：自然科学版,2000,12(2)：98-101
[50]张建成,屈红征,张晓伟.树莓的特性及开发利用前景[J].山两果树,2005(2)：31-32
[51]王发林.悬钩子果汁及果汁饮料的研制[J].甘肃农业科技,1991(10)：36-37
[52]和加卫,唐开学,杨静全等.云南省悬钩于属药用植物资源研究[J].中草药,2005, 36(7)：1078-1081
[53]张珂,王德群.安徽省悬钩子属药用植物资源[J].现代中药研究与实践,2005,19(3)：44-46
[54]陈炳华,刘剑秋.福建省悬钩子属药用植物资源[J].中草药,2001,32(6)：551-554
[55]孙长清,邵小明,王黎明,等.悬钩子属植物的开发利用概述[J].广西植物,2004,24(6)：578-582
[56]姚振生,杨武亮.江西省悬钩子属药用植物及利用建议[J].中药材,1995,18(11)：551-554
[57]Hummer K E, Finn C E. Recent Rubus and Ribes acquisitions at the USDA-ARS National Clonal Germplasm Repository[J]. Acta Horticulturae,1999,505:275-281
[58]李粤渤.树莓杂交育种主要技术的研究[D].辽宁沈阳,沈阳农业大学,2006
[59]Finn C E, Moore P P, Kempler C. Raspberry cultivars:What's new? What's succeeding? Where are the breeding programs headed? [J]. Acta Horticulturae,2006,777:33-40
[60]Weber C. Raspberry variety review:old reliable and new potential[J]. Berry Notes,2007, 19(2):4-7
[61]Jennings D L, McNicol R J. Rubus breeding-recent progress and problems[J]. Plant Breeding Abstracts,1991,61:755-758
[62]Peral S A, Sabit Y A. Developments in Raspberry Breeding[J]. Research Journal of Agriculture and Biological Sciences,2006,2(4):171-177
[63]Jennings D L. Raspberries and blackberries:their breeding, diseases and growth[M]. London:Academic Press,1988
[64]Hall H K. Blackberry breeding. In Janick J. (ed.) Plant Breeding Review (Vol.8) [M]. Portland:Timber Press:1990,249-312
[65]Daubeny H A. Brambles. In:J. Janick and J. N. Moore (eds.). Fruit Breeding Vol.Ⅱ., Vine and Small Fruits[M]. New York:John Wiley& Sons, Inc Press,1996
[66]Stafne E T, Clark J R. Genetic relatedness among eastern North American blackberry cultivars based on pedigree analysis[J]. Euphytica,2004,139:95-104
[67]Clark J R. Intractable Traits in eastern U. S. blackberry[J]. HortScience,2005,40(7): 1954-1955
[68]Finn C, Ballington J R, Swartz H. Use of 58 Rubus species in five North American breeding programmes-breeders notes[J]. Acta Horticulturae,2002,585:113-119
[69]Gupton C L. A marker gene in southern dewberry(Rubus trivialis Michx.)[J]. Hort Science, 1995,30(7):1457-1458
[70]Mezzetti B, Landi L, Phan H B, et al. Pegmediated fusion of Rubus idaeus(raspberry) and R.fruticosus(blackberry)protoplasts, selection and characterisation of callus lines[J]. Plant Biosystems,2001,135(1):63-69
[71]Graham J, McNicol R J, Kumar A. Use of the GUS gene as a selectable marker for Agrobacterium-mediated transformation of Rubus Plant[J]. Cell Tissue and Organ Culture, 1990,20(1):35-39
[72]Graham J, Hein I, Russell J, et al. The use of genomics technologies in contemporary Rubus and Ribes breeding programmes [J]. Acta Horticulturae,2004,649:319-322
[73]Hrazdina G, Zheng D. Expression and function of aromatic polyketide genes in Raspberry (Rubus idaeus) [J]. ACS Symposium Series 955, Washington D C:American Chemical Society:2007,128-140
[74]Zheng D, Hrazdina G. Molecular and biochemical characterization of benzalacetone synthase and chalcone synthase genes and their proteins from raspberry(Rubus idaeus) [J]. Archives of biochemistry and biophysics,2008,470(2):139-145
[75]桂明珠,胡宝忠.小浆果栽培生物学[M].北京：中国农业出版社,2002,48-72
[76]王彦辉,张清华.树莓优良品种与栽培技术[M].北京：金盾出版社,2003,1-55
[77]张庆霞.四川雅安地区树莓和黑莓引种试验研究[D].四川雅安,四川农业大学,2004
[78]吴林,张志东,李亚东,等.树莓优良品种—红宝玉[J].园艺学报,2002,29(3)：291
[79]郑德龙,李爱民,宋诗斌,等.树莓新品种—丰满红[J].中国果树,2000,(2)：7-8
[80]吴林,刘海广,张志东,等.树莓新品种‘红宝珠’[J].园艺学报,2005,32(5)：967
[81]刘海广,张志东,李亚东,等.树莓新品种‘红宝达’[J].园艺学报,2005,32(6)：1160
[82]陈贵虎,胡平正.南方红树莓新品种‘仙女红’选育简报[J].山西果树,2002,(3)：7-8
[83]浙江省农科院信息中心.树莓研究取得重大突破[J].浙江农业科学,2002,(6)：310
[84]王兆林,张德明,张国洪.树莓新品种—螺山仙子[J].北京农业,2003,(12)：21
[85]高慧敏,张颖君,宋炳彦,等.分子标记在蔬菜种质资源和育种上的应用[J].河北农业科学,2005,6(2)：78-81
[86]Williams J G K, Kubelik A R, Livak K J, et.al. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers[J]. Nucl. Acids. Res,1990,18:6531-6535
[87]Welsh J, Mc Clelland M. Fingerprinting genomes using PCR with arbitrary priers[J]. Nucleic Acids Res,1990,18:7213-7218
[88]Graham J, Marshall B, Squire G R. Genetic differentiation over a spatial environmental gradient in wild Rubus ideaus populations[J]. New Phytologist,2003,157:667-675
[89]Korpelainen H, Antonius-Klemola K, Werlermark G. Clonal structure of Rubus hamaemorus populations:comparison of different molecular methods[J]. Plant Ecology, 1999,143(1):123-128
[90]李媛媛,郭修武,代汉萍,等.31份野生树莓资源的RAPD分析[J].果树学报,2009,26(2)：240-244
[91]Weber C A. Genetic diversity in black raspberry detected by RAPD markers[J]. HortScience,2003,38(2):269-272
[92]Stafne E T, Clark J R, Pelto M C:Discrimination of Rubus cultivars using RAPD markers and pedigree analysis[J]. Acta Horticulturae. International Society for Horticultural Science(ISHS), Leuven, Belgium.2003,626:119-124
[93]Parent J G, Page D. Identification of raspberry cultivars by sequence characterized amplified region DNA analysis [J]. Horti Sci,1998,33(1):140-142
[94]Graham J, McNicol R J. Identification of red raspberry cultivars and an assessment of their relatedness using fingerprints produced by random primers[J]. Journal of Horticultural Science,1994,69(1):123-130
[95]Debnath S C. Inter Simple Sequence Repeat(ISSR)Markers and Pedigree Information to Assess Genetic Diversity and Relatedness Within Raspberry Genotypes[J]. International Journal of Fruit Science,2007, Vol.7(4):1-17
[96]Debnath S. C. Inter-simple sequence repeat (ISSR)-PCR analysis to assess genetic diversity in a collection of wild cloudberry (Rubus chamaemorus L.) clones[J]. Journal of Horticultural.Science and Biotechnology.2007,82(5):727-732
[97]Korbin M, Kuras A, Straczynska K, et al. Biotechnological Directions in Polish Breeding of Rubus. International society for Horticultural Science[J]. ISHS Acta Horticulturae 585: 9th International Rubus and Ribes Symposium, NOV 30-DEC 07,2005, Pucon, CHILE. Chile:ISHS 2008:133-139
[98]Hong Y P, Kin M J, Hong K N. genetic diversity in natural populations of two geographic isolates of Korean black raspberry[J]. Journal of horticultural science and biotechnology, 2003,78(3):350-354
[99]Keep E, Knight V H, Parker J H. Rubus coreanus as donor of resistance to cane disease and mildew in red raspberry[J]. Euphytica,1977,26:505-510
[100]Weber H E. Rubus.In:Hanelt P and Institute of Plant Genetics and Crop Plant Research (eds)Mansfeld's encyclopedia of agricultural and horticultural crops[J]. Springer, Berlin-Heidelberg-New York,2001,417-439
[101]Wang X R, Liu Y, Zhong B F, et al. Cytological and RAPD data revealed genetic relationships among nine selected populations of the wild bramble species, Rubus parvifolius and R. coreanus(Rosaceae) [J]. Genet Resour Crop Evol,2010,57:431-441
[102]Iwatsubo Y, Naruhashi N. Cytogenetical studies of natural hybrid Rubus×hiraseanus,and artificial hybrid between R. parvifolius×R. coreanus(Rosaceae) [J]. Cytologia,1998,63: 235-238
[103]周延清.DNA分子标记技术在植物研究中的应用[M].北京：化学工业出版社,2005：9-34
[104]Graham J, McNicol R J. An examination of the ability of RAPD markers to determine the relationships within and between Rubus species[J]. Springer-Verlag,1995,90 (8/7): 1128-1132
[105]董晓莉,汤浩茹,陈清,等.树莓基因组DNA的提取及ISSR反应体系的正交优化[J].中国农学通报,2009,25(14)：27-31
[106]Jaccard P. Nouvelles recherches sur la distribution florale[J]. Bull. Soc. Vaudoise Sci. Nat.1908,44:223-270
[107]Rohlf F J. NTSYS, pc. Version 2.10p[J]. Applied Biostatistics. Setauket, New York, 1998
[108]付燕,罗楠，杨芩,等.枇杷属植物ISSR反应体系的建立和优化[J].果树学报,2009,26(2)：180-185
[109]代红艳,张志宏,周传生,等.山楂ISSR分析体系的建立和优化[J].果树学报,2007,24(3)：313-318
[110]余艳,陈海山,葛学军.简单重复序列区间(ISSR)引物反应条件优化与筛选[J].热带 亚热带植物学报,2003,11(1)：15-19
[111]Tinker N A, Fondn M G, Mather D E. Random anapliflad polymorphic DNA and pedigree relationships in spring barley[J]. Theroretical& A ppliedG enetics,1993,85:976-994
[112]詹才新,朱兰宝,杨新美.RAPD技术在金针菇苗株鉴别中的应用[J].华中农业大学学报,1999,14(3)：253-258
[113]Khasa P D, Daneik B P. Rapd identification of white-Engelmann spruce species by RAPD markers[J]. Theroretical& Applied Genetics,1995,92:46-52
[114]张如莲,傅小霞,漆智平,等.菠萝17份种质的ISSR分析[J].中国农学通报,2006,22(6)：428-431
[115]王水琦,林彦,汤浩,等.果蔗种质资源的RAPD和ISSR分析[J].江西农业大学学报,2003,25(3)：412-417
[116]刘绍钦,黄上志,梁张惠,等.ISSR分子标记技术在香蕉分类上的应用[J].西南大学学报(自然科学版),2007,29(2)：70-74
[117]于晓英,张秋明,易干军,等.香蕉种质资源的扩增长度多态性鉴定与分类[J].湖南农业大学学报(自然科学版),2002,28(3)：206-211
[118]张敏,黄苏珍,仇硕,等.鸢尾属植物遗传多样性的RAPD和ISSR分析[J].植物资源与环境学报,2007,16(2)：6-1
[119]思彬彬,杨卓.ISSR标记技术及其在果树遗传育种中的应用[J].安徽农学通报,2008,14(13)：116
[120]邓明琴,景士西,洪建源,译.D.H.斯科特,F.J.劳伦斯,D.K.奥莱基,等著.草莓、悬钩子、穗醋栗和醋栗育种进展[J].农业出版社,1989,78
[121]冯斌,张希踔,解明,等.榛子种质资源遗传多样性的RAPD分析[J].辽宁师范大学学报：自然科学版,2007,30(2)：216-219
[122]李辛雷,陈发棣,崔娜欣.菊属种间杂种的鉴定[J].南京农业大学学报,2005,28(1)：24-28
[123]梁月荣,田中淳一,武田善行.应用RAPD分子标记分析“晚绿”品种的杂交亲本[J].茶叶科学,2000,20(1)：22-26
[124]齐明.运用ISSR分子标记鉴定杉木×侧柏远交杂种[J].浙江林学院学报,2008,25(5)：666-669
[125]纪德华,谢潮添,徐燕,等.坛紫菜品系间杂交子代杂种优势的ISSR分析[J].海洋 学报,2008,30(6)：147-153
[2]俞德浚,陆玲娣,谷粹芝,等.中国植物志.第37卷[M].北京：科学出版社,1985
[3]陆玲娣.我国悬钩子属植物的研究[J].植物分类学报,1983,21(1)：13-25
[4]Thompson M M. Survey of chromosome numbers in Rubus[J]. Annals of the Missouri Botanical Garden,1997,84(1):128-164
[5]杨燕林,唐开学,和加卫,等.悬钩子属植物分子标记技术和基因组研究进展[J].中国农学通报,2008,24(8)：93-98
[6]陈少风,叶居新,朱祥玲,等.若干悬钩子属植物的花粉形态研究[J].植物研究,1996,16(4)：463-466
[7]周丽华,韦仲新,吴征镒.国产蔷薇科蔷薇亚科的花粉形态[J].云南植物研究,1999,21(4)：455-460
[8]李维林,贺善安,顾姻,等.中国悬钩子属花粉形态观察[J].植物分类学报,2001,39(3)：234-247
[9]王小蓉,汤浩茹,黄力,等.树莓部分野生种及栽培品种花粉亚显微形态的比较[J].园艺学报,2007,34(6)：1395-1404
[10]陈瑞阳.中国主要经济植物染色体图谱(第一册)—中国果树及其野生近缘植物染色体图谱[M].北京：万国学术出版社,1993：263-279
[11]李秀兰,宋文芹,陈瑞阳.北方几种小浆果植物的核型研究[J].武汉植物学研究,1993,11(4)：289-292
[12]林盛华,张冰冰,方成泉,等.中国树莓属8个种染色体数目与核型[J].园艺学报,1994,21(4)：313-319
[13]顾姻,王传永,赵昌民,等.悬钩子属种质的评价[J].植物资源与环境,1996,5(3)：6-13
[14]王小蓉,汤浩茹,段娟,等.中国悬钩子属空心莓组与木莓组28种和变种的核型比较研究[J].植物分类学报,2008,46(4)：505-515
[15]Li W L, Gu Y, He S A et al, Applicability of RAPD to systematics and genetic diversity of Rubus L.. Acta Horticulturae,2002,585:139-142
[16]段娟.西南地区17种(33份)不同来源的野生悬钩子属植物种间遗传多样性的RAPD 分析[D].四川雅安,四川农业大学,2007
[17]李玲.RAPD对树莓栽培品种及插田泡类型的遗传差异分析[D].四川雅安,四川农业大学,2007
[18]贾静波,李维林,吴文龙,等.悬钩子属植物的RAPD分析[J].植物资源与环境学报,2008,17(3)：18-22
[19]王小蓉.悬钩子属植物遗传多样性及其特异资源的筛选[D].四川雅安,四川农业大学,2008
[20]傅承新,沈朝栋,黄爱君.浙江悬钩子属植物的综合研究—资源调查、引种及开发利用前景[J].浙江农业大学学报,1995,21(4)：393-397
[21]栾文举,肖雯,焦健.甘肃省悬钩子属植物优良种类的营养评价及利用途径[J].甘肃科学学报,1996,8(1)：51-55
[22]郭军战,陈铁山,彭少兵.岭山区黄果悬钩子种质资源分析与评价[J].西北林学院学报,2004,19(2)：41-43
[23]Gu Y, Sun Z J, Cai J H et.al.. Introduction and utilization of small fruits in China with special reference to Rubus species[J].'Acta Horticulturae,1989,262:47-55
[24]俞德浚.中国果树分类学[M].北京：中国农业出版社,1979：209-220
[25]李春奇,叶永忠,王志强,等.河南野生悬钩子属植物资源[J].果树科学,1995,12(4)：258-261
[26]栾文举,焦健.甘肃省悬钩子属植物资源分布及其开发前景初探[J].自然资源学报,1996,11(1)：41-48
[27]张志勇,俞志雄.江西悬钩子属的分类和地理分布[J].热带亚热带植物学报,2003,11(1)：27-33
[28]顾姻.悬钩子属植物资源及其利用[J].植物资源与环境,1992,1(2)：50-60
[29]何顺志.贵州悬钩子属(蔷薇科)一新种—务川悬钩子[J].植物分类学报,2006,43：345-347
[30]陆玲娣.中国蔷薇科部分属分类群的修订[J].植物分类学报,2000,38.276-281
[31]李维林,贺善安.一些悬钩子属分类群的修订[J].植物学研究,2001,21：346-249
[32]Naruhashi N. Notes on Japanese Rubus(4). Taxon,1989,37:1-4
[33]李玲,汤浩茹,段娟,等.中国悬钩子属植物的分类鉴定与开发利用[J].安徽农业科学.2006,34(8)：1513-1516
[34]王传永,顾姻,吴文龙,等.悬钩子属基因库的建立与维护[J].植物资源与环境,1996,5(1)：14-17
[35]傅承新,沈朝栋,黄爱军.浙江悬钩子属植物的综合研究[J].浙江农业大学学报,1995,21(4)：393-397
[36]金炜,黄树芝,顾姻.福建省悬钩子属植物资源的调查、收集、评价、和利用的研究[J].武汉植物学报研究，1992,10(4)：371-376
[37]贺善安,顾姻,孙醉君,等.黑莓引种的理论导向[J].植物资源与环境,1998,7(1)：1-9
[38]李维林,贺善安,顾姻.中国悬钩子属植物的利用价值概述[J].武汉植物学研究,2000,18(3)：237-243
[39]张志勇,赖小容.井冈山悬钩子属植物资源的初步研究[J].江西农业大学学报,1999，21(3)：395-398
[40]李孝苗,何家庆.覆盆子资源及其开发利用[J].安徽农业科学,2000,28(1)：128
[41]曹钰普.悬钩子人工栽培技术[J].林业月报,1999,7：19
[42]李维林.覆盆子人工栽培技术[J].中药材,1991,14(5)：10-11
[43]锁必龙,刘炳龙.树莓丰产栽培技术[J].安徽林业,2004(4)：31
[44]张福平.粤东地区野生可食悬钩子植物资源及利用研究Ⅲ[J].中国食物与营养,2003(12)：24-25
[45]田春元,姜益泉,吴金清.河南鸡公山国家级自然保护区悬钩子属植物资源开发利用研究[J].武汉植物学研究,2003,21(6)：521-525
[46]李维林.我国野生悬钩子资源及其开发利用[J].中国野生植物,1991(3)：14-18
[47]仲山民,田荆祥,吴美春.悬钩子果实的营养成分分析[J].浙江林学院学报,1993,10(4)：485-489
[48]桑建忠,顾姻.中国东南部部分悬钩子果实的营养成分Ⅲ[J].植物资源与环境,1995,4(2)：22-26
[49]陈炳华.红腺悬钩子果实营养成分研究[J].海南帅范学院学报：自然科学版,2000,12(2)：98-101
[50]张建成,屈红征,张晓伟.树莓的特性及开发利用前景[J].山两果树,2005(2)：31-32
[51]王发林.悬钩子果汁及果汁饮料的研制[J].甘肃农业科技,1991(10)：36-37
[52]和加卫,唐开学,杨静全等.云南省悬钩于属药用植物资源研究[J].中草药,2005, 36(7)：1078-1081
[53]张珂,王德群.安徽省悬钩子属药用植物资源[J].现代中药研究与实践,2005,19(3)：44-46
[54]陈炳华,刘剑秋.福建省悬钩子属药用植物资源[J].中草药,2001,32(6)：551-554
[55]孙长清,邵小明,王黎明,等.悬钩子属植物的开发利用概述[J].广西植物,2004,24(6)：578-582
[56]姚振生,杨武亮.江西省悬钩子属药用植物及利用建议[J].中药材,1995,18(11)：551-554
[57]Hummer K E, Finn C E. Recent Rubus and Ribes acquisitions at the USDA-ARS National Clonal Germplasm Repository[J]. Acta Horticulturae,1999,505:275-281
[58]李粤渤.树莓杂交育种主要技术的研究[D].辽宁沈阳,沈阳农业大学,2006
[59]Finn C E, Moore P P, Kempler C. Raspberry cultivars:What's new? What's succeeding? Where are the breeding programs headed? [J]. Acta Horticulturae,2006,777:33-40
[60]Weber C. Raspberry variety review:old reliable and new potential[J]. Berry Notes,2007, 19(2):4-7
[61]Jennings D L, McNicol R J. Rubus breeding-recent progress and problems[J]. Plant Breeding Abstracts,1991,61:755-758
[62]Peral S A, Sabit Y A. Developments in Raspberry Breeding[J]. Research Journal of Agriculture and Biological Sciences,2006,2(4):171-177
[63]Jennings D L. Raspberries and blackberries:their breeding, diseases and growth[M]. London:Academic Press,1988
[64]Hall H K. Blackberry breeding. In Janick J. (ed.) Plant Breeding Review (Vol.8) [M]. Portland:Timber Press:1990,249-312
[65]Daubeny H A. Brambles. In:J. Janick and J. N. Moore (eds.). Fruit Breeding Vol.Ⅱ., Vine and Small Fruits[M]. New York:John Wiley& Sons, Inc Press,1996
[66]Stafne E T, Clark J R. Genetic relatedness among eastern North American blackberry cultivars based on pedigree analysis[J]. Euphytica,2004,139:95-104
[67]Clark J R. Intractable Traits in eastern U. S. blackberry[J]. HortScience,2005,40(7): 1954-1955
[68]Finn C, Ballington J R, Swartz H. Use of 58 Rubus species in five North American breeding programmes-breeders notes[J]. Acta Horticulturae,2002,585:113-119
[69]Gupton C L. A marker gene in southern dewberry(Rubus trivialis Michx.)[J]. Hort Science, 1995,30(7):1457-1458
[70]Mezzetti B, Landi L, Phan H B, et al. Pegmediated fusion of Rubus idaeus(raspberry) and R.fruticosus(blackberry)protoplasts, selection and characterisation of callus lines[J]. Plant Biosystems,2001,135(1):63-69
[71]Graham J, McNicol R J, Kumar A. Use of the GUS gene as a selectable marker for Agrobacterium-mediated transformation of Rubus Plant[J]. Cell Tissue and Organ Culture, 1990,20(1):35-39
[72]Graham J, Hein I, Russell J, et al. The use of genomics technologies in contemporary Rubus and Ribes breeding programmes [J]. Acta Horticulturae,2004,649:319-322
[73]Hrazdina G, Zheng D. Expression and function of aromatic polyketide genes in Raspberry (Rubus idaeus) [J]. ACS Symposium Series 955, Washington D C:American Chemical Society:2007,128-140
[74]Zheng D, Hrazdina G. Molecular and biochemical characterization of benzalacetone synthase and chalcone synthase genes and their proteins from raspberry(Rubus idaeus) [J]. Archives of biochemistry and biophysics,2008,470(2):139-145
[75]桂明珠,胡宝忠.小浆果栽培生物学[M].北京：中国农业出版社,2002,48-72
[76]王彦辉,张清华.树莓优良品种与栽培技术[M].北京：金盾出版社,2003,1-55
[77]张庆霞.四川雅安地区树莓和黑莓引种试验研究[D].四川雅安,四川农业大学,2004
[78]吴林,张志东,李亚东,等.树莓优良品种—红宝玉[J].园艺学报,2002,29(3)：291
[79]郑德龙,李爱民,宋诗斌,等.树莓新品种—丰满红[J].中国果树,2000,(2)：7-8
[80]吴林,刘海广,张志东,等.树莓新品种‘红宝珠’[J].园艺学报,2005,32(5)：967
[81]刘海广,张志东,李亚东,等.树莓新品种‘红宝达’[J].园艺学报,2005,32(6)：1160
[82]陈贵虎,胡平正.南方红树莓新品种‘仙女红’选育简报[J].山西果树,2002,(3)：7-8
[83]浙江省农科院信息中心.树莓研究取得重大突破[J].浙江农业科学,2002,(6)：310
[84]王兆林,张德明,张国洪.树莓新品种—螺山仙子[J].北京农业,2003,(12)：21
[85]高慧敏,张颖君,宋炳彦,等.分子标记在蔬菜种质资源和育种上的应用[J].河北农业科学,2005,6(2)：78-81
[86]Williams J G K, Kubelik A R, Livak K J, et.al. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers[J]. Nucl. Acids. Res,1990,18:6531-6535
[87]Welsh J, Mc Clelland M. Fingerprinting genomes using PCR with arbitrary priers[J]. Nucleic Acids Res,1990,18:7213-7218
[88]Graham J, Marshall B, Squire G R. Genetic differentiation over a spatial environmental gradient in wild Rubus ideaus populations[J]. New Phytologist,2003,157:667-675
[89]Korpelainen H, Antonius-Klemola K, Werlermark G. Clonal structure of Rubus hamaemorus populations:comparison of different molecular methods[J]. Plant Ecology, 1999,143(1):123-128
[90]李媛媛,郭修武,代汉萍,等.31份野生树莓资源的RAPD分析[J].果树学报,2009,26(2)：240-244
[91]Weber C A. Genetic diversity in black raspberry detected by RAPD markers[J]. HortScience,2003,38(2):269-272
[92]Stafne E T, Clark J R, Pelto M C:Discrimination of Rubus cultivars using RAPD markers and pedigree analysis[J]. Acta Horticulturae. International Society for Horticultural Science(ISHS), Leuven, Belgium.2003,626:119-124
[93]Parent J G, Page D. Identification of raspberry cultivars by sequence characterized amplified region DNA analysis [J]. Horti Sci,1998,33(1):140-142
[94]Graham J, McNicol R J. Identification of red raspberry cultivars and an assessment of their relatedness using fingerprints produced by random primers[J]. Journal of Horticultural Science,1994,69(1):123-130
[95]Debnath S C. Inter Simple Sequence Repeat(ISSR)Markers and Pedigree Information to Assess Genetic Diversity and Relatedness Within Raspberry Genotypes[J]. International Journal of Fruit Science,2007, Vol.7(4):1-17
[96]Debnath S. C. Inter-simple sequence repeat (ISSR)-PCR analysis to assess genetic diversity in a collection of wild cloudberry (Rubus chamaemorus L.) clones[J]. Journal of Horticultural.Science and Biotechnology.2007,82(5):727-732
[97]Korbin M, Kuras A, Straczynska K, et al. Biotechnological Directions in Polish Breeding of Rubus. International society for Horticultural Science[J]. ISHS Acta Horticulturae 585: 9th International Rubus and Ribes Symposium, NOV 30-DEC 07,2005, Pucon, CHILE. Chile:ISHS 2008:133-139
[98]Hong Y P, Kin M J, Hong K N. genetic diversity in natural populations of two geographic isolates of Korean black raspberry[J]. Journal of horticultural science and biotechnology, 2003,78(3):350-354
[99]Keep E, Knight V H, Parker J H. Rubus coreanus as donor of resistance to cane disease and mildew in red raspberry[J]. Euphytica,1977,26:505-510
[100]Weber H E. Rubus.In:Hanelt P and Institute of Plant Genetics and Crop Plant Research (eds)Mansfeld's encyclopedia of agricultural and horticultural crops[J]. Springer, Berlin-Heidelberg-New York,2001,417-439
[101]Wang X R, Liu Y, Zhong B F, et al. Cytological and RAPD data revealed genetic relationships among nine selected populations of the wild bramble species, Rubus parvifolius and R. coreanus(Rosaceae) [J]. Genet Resour Crop Evol,2010,57:431-441
[102]Iwatsubo Y, Naruhashi N. Cytogenetical studies of natural hybrid Rubus×hiraseanus,and artificial hybrid between R. parvifolius×R. coreanus(Rosaceae) [J]. Cytologia,1998,63: 235-238
[103]周延清.DNA分子标记技术在植物研究中的应用[M].北京：化学工业出版社,2005：9-34
[104]Graham J, McNicol R J. An examination of the ability of RAPD markers to determine the relationships within and between Rubus species[J]. Springer-Verlag,1995,90 (8/7): 1128-1132
[105]董晓莉,汤浩茹,陈清,等.树莓基因组DNA的提取及ISSR反应体系的正交优化[J].中国农学通报,2009,25(14)：27-31
[106]Jaccard P. Nouvelles recherches sur la distribution florale[J]. Bull. Soc. Vaudoise Sci. Nat.1908,44:223-270
[107]Rohlf F J. NTSYS, pc. Version 2.10p[J]. Applied Biostatistics. Setauket, New York, 1998
[108]付燕,罗楠，杨芩,等.枇杷属植物ISSR反应体系的建立和优化[J].果树学报,2009,26(2)：180-185
[109]代红艳,张志宏,周传生,等.山楂ISSR分析体系的建立和优化[J].果树学报,2007,24(3)：313-318
[110]余艳,陈海山,葛学军.简单重复序列区间(ISSR)引物反应条件优化与筛选[J].热带 亚热带植物学报,2003,11(1)：15-19
[111]Tinker N A, Fondn M G, Mather D E. Random anapliflad polymorphic DNA and pedigree relationships in spring barley[J]. Theroretical& A ppliedG enetics,1993,85:976-994
[112]詹才新,朱兰宝,杨新美.RAPD技术在金针菇苗株鉴别中的应用[J].华中农业大学学报,1999,14(3)：253-258
[113]Khasa P D, Daneik B P. Rapd identification of white-Engelmann spruce species by RAPD markers[J]. Theroretical& Applied Genetics,1995,92:46-52
[114]张如莲,傅小霞,漆智平,等.菠萝17份种质的ISSR分析[J].中国农学通报,2006,22(6)：428-431
[115]王水琦,林彦,汤浩,等.果蔗种质资源的RAPD和ISSR分析[J].江西农业大学学报,2003,25(3)：412-417
[116]刘绍钦,黄上志,梁张惠,等.ISSR分子标记技术在香蕉分类上的应用[J].西南大学学报(自然科学版),2007,29(2)：70-74
[117]于晓英,张秋明,易干军,等.香蕉种质资源的扩增长度多态性鉴定与分类[J].湖南农业大学学报(自然科学版),2002,28(3)：206-211
[118]张敏,黄苏珍,仇硕,等.鸢尾属植物遗传多样性的RAPD和ISSR分析[J].植物资源与环境学报,2007,16(2)：6-1
[119]思彬彬,杨卓.ISSR标记技术及其在果树遗传育种中的应用[J].安徽农学通报,2008,14(13)：116
[120]邓明琴,景士西,洪建源,译.D.H.斯科特,F.J.劳伦斯,D.K.奥莱基,等著.草莓、悬钩子、穗醋栗和醋栗育种进展[J].农业出版社,1989,78
[121]冯斌,张希踔,解明,等.榛子种质资源遗传多样性的RAPD分析[J].辽宁师范大学学报：自然科学版,2007,30(2)：216-219
[122]李辛雷,陈发棣,崔娜欣.菊属种间杂种的鉴定[J].南京农业大学学报,2005,28(1)：24-28
[123]梁月荣,田中淳一,武田善行.应用RAPD分子标记分析“晚绿”品种的杂交亲本[J].茶叶科学,2000,20(1)：22-26
[124]齐明.运用ISSR分子标记鉴定杉木×侧柏远交杂种[J].浙江林学院学报,2008,25(5)：666-669
[125]纪德华,谢潮添,徐燕,等.坛紫菜品系间杂交子代杂种优势的ISSR分析[J].海洋 学报,2008,30(6)：147-153