基于簇毛麦No.1026转录组的SSR序列分析及其PCR标记开发
|
摘要
【目的】探究从前苏联引进的簇毛麦No.1026(Dv#4)的EST-SSR序列特征及其在染色体的分布,分析它们在不同簇毛麦间及与小麦间的多态性,为其进一步的研究与利用提供依据。【方法】通过Illumina HiSeq测序获得No.1026植株的转录组序列,利用MISA软件分析转录组SSR序列及特征,采用Primer 3设计SSR引物,随机合成238对引物,对小麦中国春与簇毛麦Dv#4和引自英国剑桥的簇毛麦Dv#2的基因组DNA进行扩增,在琼脂糖凝胶上分离评价扩增产物的多态性,并利用一套小麦-簇毛麦异染色体系进行扩增分析。【结果】检测了No.1026总长62.76 Mb的转录组序列,发现10 497个SSR位点,它们分布于8 735条Unigene上。在1—6个核苷酸的重复单元中,单、双、三碱基的重复占95.85%,其中三核苷酸串联重复数量最多,占SSR总数的50.33%,而CCG/CGG基序的重复占三核苷酸串联重复的41.66%;单核苷酸重复是第二大类型,出现频率为27.13%,其中A/T重复占单核苷酸重复的74.58%。二核苷酸重复类型的数量位列第三,占SSR总数的18.39%。在238对EST-SSR引物中,88对在中国春与簇毛麦(包括Dv#2和Dv#4)之间的扩增产物显示多态性;8对只在簇毛麦和单一异染色体系扩增;4对可在簇毛麦和多个异染色体系中特异扩增;但多数可在簇毛麦中清晰扩增的引物不能在任何异染色体系中扩增,推测可能与簇毛麦基因组及染色体导入小麦过程中的变异有关;47对(19.74%)在2份不同来源的簇毛麦Dv#2和Dv#4之间的扩增产物呈现多态性。利用1对EST-SSR引物和1个EST-PCR标记检测48个簇毛麦植株,证明多态性的SSR引物可有效用于检测簇毛麦的异质性。【结论】簇毛麦No.1026(Dv#4)的转录组中存在丰富的SSR序列,其中CCG/CGG、A/T和AG/CT等三核苷酸、单核苷酸和二核苷酸是其最主要的串联重复基序。部分EST-SSR的侧翼保守序列与单一或若干外源染色体特异相关联,据此开发特异的分子标记,可用于跟踪检测小麦背景中特异的簇毛麦染色体或染色体片段。Dv#4与Dv#2间的部分EST-SSR具有多态性,提示2份不同来源簇毛麦的表达序列间存在一定程度的遗传差异,因此,簇毛麦Dv#4值得进一步的发掘研究。
【Objective】The aim of this study is to explore the characteristics of the EST-SSR sequences of a Dasypyrum villosum accession No.1026(Dv#4) introduced from the former Soviet Union, and their distributions on chromosomes and polymorphism in different D. villosum accessions and between Dv#4 and common wheat. 【Method】Transcriptome sequences of Dv#4 plants were obtained by Illumina HiSeq sequencing and used to search and analyze the SSR sequences using MISA software and design primers by Primer 3. In total, 238 pairs of primers were selected randomly for synthesis and used to amplify the genomic DNAs of wheat Chinese Spring(CS) and the two different D. villosum accessions. The polymorphisms of the PCR products on agarose gel were evaluated. Further, the features of their chromosome distributions in Dv#4 were studied by using a set of wheat-D. villosum alien chromosome lines(including disomic addition lines and disomic substitution lines). 【Result】 A total sequence length of 62.76 Mb was detected and 10 497 SSR loci were found in the transcriptome data. They are involved in 8 735 unigenes. Repeated unit of mono-, di-, tri-nucleotides are the main type, holding 95.85% of all loci among 1-6 nucleotides repeats, among which tri-nucleotide is the richest component that makes up of 50.33% and contains CCG/CGG motif by 41.66%. The next component is mono-nucleotide tandem repeat, and its occurrence frequency is 18.39%, and A/T repeats occupy 74.58% in this type. Di-nucleotide ranks the 3 rd, and it holds 18.39% in the total SSR loci. Among 238 pairs of randomly synthetized EST-SSR primers, 88 pairs amplified polymorphic fragments between CS and D. villosum(including Dv#2 and Dv#4); 8 pairs only had amplifications in D. villosum and some single alien chromosome lines; 4 pairs could specifically amplify bands in D. villosum and multiple alien chromosome lines. But, many primers which had amplification in both D. villosum accessions had no amplification in any alien chromosome lines. Therefore, it can be inferred that variations on the flanking conserved sequences of SSR might occur during the transferring of the exogenous genome or chromosomes into wheat. Additionally, 47 pairs of primers(19.74%) showed polymorphisms between Dv#2 and Dv#4. By using a pair of EST-SSR primer and a PCR marker, respectively, polymorphic amplicons were detected in 48 D. villosum plants, indicating that polymorphic SSR primer can be used to detect the heterogeneity of D. villosum effectively.【Conclusion】 D. villosum Dv#4 has abundant SSR sequences in its transcriptome, of which, CCG/CGG, A/T, AG/CT and other tri-, mono-, and di-nucleotide are the main tandem repeats. The flanking conserved sequences of partial EST-SSR in Dv#4 are associated with a single or several chromosomes specifically, which provides an ideal sequence resource for the development of specific molecular markers to track and detect D. villosum chromosomes or chromosome fragments in wheat background. Some of the EST-SSR polymorphism between Dv#4 and Dv#2 indicates that there is a certain degree of genetic diversity in some of the expressed sequences between the two different origin D. villosum accessions. Therefore, Dv#4 is valuable to be further investigated.
【Objective】The aim of this study is to explore the characteristics of the EST-SSR sequences of a Dasypyrum villosum accession No.1026(Dv#4) introduced from the former Soviet Union, and their distributions on chromosomes and polymorphism in different D. villosum accessions and between Dv#4 and common wheat. 【Method】Transcriptome sequences of Dv#4 plants were obtained by Illumina HiSeq sequencing and used to search and analyze the SSR sequences using MISA software and design primers by Primer 3. In total, 238 pairs of primers were selected randomly for synthesis and used to amplify the genomic DNAs of wheat Chinese Spring(CS) and the two different D. villosum accessions. The polymorphisms of the PCR products on agarose gel were evaluated. Further, the features of their chromosome distributions in Dv#4 were studied by using a set of wheat-D. villosum alien chromosome lines(including disomic addition lines and disomic substitution lines). 【Result】 A total sequence length of 62.76 Mb was detected and 10 497 SSR loci were found in the transcriptome data. They are involved in 8 735 unigenes. Repeated unit of mono-, di-, tri-nucleotides are the main type, holding 95.85% of all loci among 1-6 nucleotides repeats, among which tri-nucleotide is the richest component that makes up of 50.33% and contains CCG/CGG motif by 41.66%. The next component is mono-nucleotide tandem repeat, and its occurrence frequency is 18.39%, and A/T repeats occupy 74.58% in this type. Di-nucleotide ranks the 3 rd, and it holds 18.39% in the total SSR loci. Among 238 pairs of randomly synthetized EST-SSR primers, 88 pairs amplified polymorphic fragments between CS and D. villosum(including Dv#2 and Dv#4); 8 pairs only had amplifications in D. villosum and some single alien chromosome lines; 4 pairs could specifically amplify bands in D. villosum and multiple alien chromosome lines. But, many primers which had amplification in both D. villosum accessions had no amplification in any alien chromosome lines. Therefore, it can be inferred that variations on the flanking conserved sequences of SSR might occur during the transferring of the exogenous genome or chromosomes into wheat. Additionally, 47 pairs of primers(19.74%) showed polymorphisms between Dv#2 and Dv#4. By using a pair of EST-SSR primer and a PCR marker, respectively, polymorphic amplicons were detected in 48 D. villosum plants, indicating that polymorphic SSR primer can be used to detect the heterogeneity of D. villosum effectively.【Conclusion】 D. villosum Dv#4 has abundant SSR sequences in its transcriptome, of which, CCG/CGG, A/T, AG/CT and other tri-, mono-, and di-nucleotide are the main tandem repeats. The flanking conserved sequences of partial EST-SSR in Dv#4 are associated with a single or several chromosomes specifically, which provides an ideal sequence resource for the development of specific molecular markers to track and detect D. villosum chromosomes or chromosome fragments in wheat background. Some of the EST-SSR polymorphism between Dv#4 and Dv#2 indicates that there is a certain degree of genetic diversity in some of the expressed sequences between the two different origin D. villosum accessions. Therefore, Dv#4 is valuable to be further investigated.
引文
[1]GR?DZIELEWSKA A.The genus Dasypyrum-part 2.Dasypyrum villosum-a wild species used in wheat improvement.Euphytica,2006,152:441-454.
[2]赵万春,董剑,陈其皎,李晓燕,高翔,石引刚,陈良国.簇毛麦-用于小麦改良的一种野生植物.草业科学,2012,29(10):1613-1621.ZHAO W C,DONG J,CHEN Q J,LI X Y,GAO X,SHI Y G,CHENL G.Advance in Dasypyrum villosum-a valuable wild species used in wheat improvement.Protacultural Science,2012,29(10):1613-1621.(in Chinese)
[3]CHEN Q,CONNER R L,LI H,LAROCHE A,GRAF R J,KUZYK AD.Expression of resistance to strip rust,powdery mildew and the wheat curl mite in Triticum aestivum×Haynaldia villosa lines.Canadian Journal of Plant Science,2002,82(2):451-456.
[4]刘大钧,陈佩度,吴沛良,王耀南,邱伯行,王苏玲.硬粒小麦-簇毛麦双二倍体.作物学报,1986,12(3):155-162.LIU D J,CHEN P D,WU P L,WANG Y N,QIU B X,WANG S L.Triticum durum-Haynadia villosa amphidiploid.Acta Agronomica Sinica,1986,12(3):155-162.(in Chinese)
[5]陈孝,徐惠君,杜丽璞,尚立民,韩彬,施爱农,肖世和.利用组织培养技术向普通小麦导入簇毛麦抗白粉病基因的研究.中国农业科学,1996,29(5):1-8.CHEN X,XU H J,DU L P,SHANG L M,HAN B,SHI A N,XIAO SH.Transfer of gene resistant to powdery mildew from H.villosa to common wheat by tissue culture.Scientia Agricultura Sinica,1996,29(5):1-8.(in Chinese)
[6]陈佩度,周波,齐莉莉,刘大钧.用分子原位杂交(GISH)鉴定小麦-簇毛麦双倍体、附加系、代换系和易位系.遗传学报,1995,22(5):380-386.CHEN P D,ZHOU B,QI L L,LIU D J.Identification of wheat-Haynaldia villosa amphiloid,addition,substitution and translocation lines by in situ hybridization using biotin-labelled genomic DNA as a probe.Acta Genetica Sinica,1995,22(5):380-386.(in Chinese)
[7]CHEN P D,QI L L,ZHOU B,ZHANG S Z,LIU D J.Development and molecular cytogenetic analysis of wheat-Haynaldia villosa6VS/6AL translocation lines specifying resistance to powdery mildew.Theoretical and Applied Genetics,1995,91:1125-1128.
[8]齐莉莉,陈佩度,刘大钧,周波,张守中,盛宝钦,向齐君,段霞渝,周益林.小麦白粉病新抗源--基因Pm21.作物学报,1995,21(3):257-262.QI L L,CHEN P D,LIU D J,ZHOU B,ZHANG S Z,SHEENG B Q,XIANG Q J,DUAN X Y,ZHOU Y L.The gene Pm21-a new source for resistance to wheat powdery mildew.Acta Agronomica Sinica,1995,21(3):257-262.(in Chinese)
[9]LI H,CHEN X,XIN Z Y,MA Y Z,XU H J,CHEN X Y,JIA X.Development and identification of wheat-Haynaldia villosa T6DL.6VS chromosome translocation lines conferring resistance to powdery mildew.Plant Breeding,2005,124:203-205.
[10]LIU C,QI L L,LIU W X,ZHAO W C,WILSON J,FRIEBE B,GILLB S.Development of a set of compensating Triticum aestivumDasypyrum villosum Robertsonian translocation lines.Genome,2011,54:836-844.
[11]XING L,HU P,LIU J.,WITEK K,ZHOU S,XU J,ZHOU W,GAO L,HUANG Z,ZHANG R,WANG X,CHEN P,WANG H,JONES J D G,KARAFIATOVA M,VRANA J,BARTOS J,DOLEZEL J,TIAN Y,WU Y,CAO A.Pm21 from Haynaldia villosa encodes a CC-NBS-LRR that confers powdery mildew resistance in wheat.Molecular Plant,2018,doi:10.1016/j.molp.2018.02.013.
[12]HE H,ZHU S,ZHAO R,JIANG Z,JI Y,JI J,QIU D,LI H,BIE T.Pm21,encoding a typical CC-NBS-LRR protein,confers broadspectrum resistance to wheat powdery mildew disease.Molecular Plant,2018,doi:10.1016/j.molp.2018.03.004.
[13]ZHANG Q P,LI Q,WANG X E,WANG H Y,LANG S P,WANG Y N,WANG S L,CHEN P D,LIU D J.Development and characterization of a Triticum aestivum-Haynaldia villosa translocation line T4VS·4DLconferring resistance to wheat spindle streak mosaic virus.Euphytica,2005,145:317-320.
[14]ZHAO R,WANG H,XIAO J,BIE T,CHENG S,JIA Q,YUAN C,ZHANG R,CAO A,CHEN P,WANG X.Induction of 4VSchromosome recombinants using the CS ph1b mutant and mapping of the wheat yellow mosaic virus resistance gene from Haynaldia villosa.Theoretical and Applied Genetics,2013,126:2921-2930.
[15]李淑梅,徐川梅,周波,陈佩度.普通小麦-簇毛麦2V染色体端体异附加系的选育与鉴定.南京农业大学学报,2009,32(1):1-5.LI S M,XU C M,ZHOU B,CHEN P D.Development and identification of Triticum-Haynadia villosa ditelesomic addition lines involving chromosome 2V of H.villosa.Journal of Nanjing Agricultural University,2009,32(1):1-5.(in Chinese)
[16]董剑,杨华,赵万春,李晓燕,陈其皎,高翔.普通小麦中国春-簇毛麦易位系T1DL·1VS和T1DS·1VL的农艺和品质特性.作物学报,2013,39(8):1386-1390.DONG J,YANG H,ZHAO W C,LI X Y,CHEN Q J,GAO X.Agronomic traits and grain quality of Chinese Spring-Dasypyrum villosum translocation lines T1DL·1VS and T1DS·1VL.Acta Agronomica Sinica,2013,39(8):1386-1390.(in Chinese)
[17]张瑞奇,冯祎高,侯富,陈树林,别同德,陈佩度.普通小麦-簇毛麦T5VS.5DL易位染色体对小麦主要农艺性状、品质和白粉病抗性的遗传效应分析.中国农业科学,2015,48(6):1041-1051.ZHANG R Q,FENG Y G,HOU F,CHEN S L,BIE T D,CHEN P D.The genetic effect of wheat-H.villosa T5VS.5DL translocated chromosome on agronomic characteristics,quality,and powdery mildew resistance of common wheat.Scientia Agricultura Sinica,2015,48(6):1041-1051.(in Chinese)
[18]ZHANG R,SUN B,CHEN J,CAO A,XING L,FENG Y,LAN C,CHEN P.Pm55,a developmental-stage and tissue-specific powdery mildew resistance gene introgressed from Dasypyrum villosum into common wheat.Theoretical and Applied Genetics,2016,129:1975-1984.
[19]QI L L,PUMPHREY M O,FRIEBE B,ZHANG P,QIAN C,BOWDEN R L,ROUSE M N,JIN Y,GILL B S.A novel Robertsonian translocation event leads to transfer of a stem rust resistance gene(Sr52)effective against race Ug99 from Dasypyrum villosum into bread wheat.Theoretical and Applied Genetics,2011,123(1):159-167.
[20]LI H J,CONNER R L,CHEN Q,JIA X,LI H,GRAF R J,LAROCHE A,KUZYK A D.Different reactions to the wheat curl mite and wheat streak mosaic virus in various wheat-Haynaldia villosa 6V and 6VS lines.Plant Disease,2002,86(4):423-428.
[21]LIU C,Ye X G,Wang M J,LI S J,LIN Z S.Genetic behavior of Triticum aestivum-Dasypyrum villosum translocation chromosomes T6V#4S·6DL and T6V#2S·6AL.Journal of Integrative Agriculture,2017,16(10):2136-2144.
[22]张云龙,王美蛟,张悦,褚翠萍,林志珊,徐琼芳,叶兴国,陈孝,张宪省.不同簇毛麦6VS染色体臂的白粉病抗性特异功能标记的开发及应用.作物学报,2012,38(10):1827-1832.ZHANG Y L,WANG M J,ZHANF Y,CHU C P,LIN Z S,XU Q F,YE X G,CHEN X,ZHANG X S.Development and application of functional markers specific to powdery mildew resistance on chromosome arm 6VS from different origins of Haynaldia villosa.Acta Agronomica Sinica,2012,38(10):1827-1832.(in Chinese)
[23]LIN Z S,ZHANG Y L,WANG K,LI J R,XU Q F,CHEN X,ZHANG X S,YE X G.Isolation and molecular analysis of genes Stpk-V2 and Stpk-V3 homologous to powdery mildew resistance gene Stpk-V in a Dasypyrum villosum accession and its derivatives.Journal of Applied Genetics,2013,54:417-426.
[24]BIE T D,ZHAO R H,ZHU S Y,CHEN S L,HE H G.Development and characterization of marker MBH1 simultaneously tagging genes Pm21 and Pm V conferring resistance to powdery mildew in wheat.Molecular Breeding,2015,35:189.
[25]刘畅,李仕金,王轲,叶兴国,林志珊.簇毛麦6VS特异转录序列P21461及P33259的获得及其分子标记在鉴定小麦-簇毛麦抗白粉病育种材料中的应用.作物学报,2017,43(7):983-992.LIU C,LI S J,WANG K,YE X G,LIN Z S.Developing of specific transcription sequences P21461 and P33259 on Dasypyrum villosum6VS and application of molecular markers in identifying wheat-D.villosum breeding materials with powdery mildew resistance.Acta Agronomica Sinica,2017,43(7):983-992.(in Chinese)
[26]LI S J,LIN Z S,LIU C,WANG K,DU L P,YE X G.Development and comparative genomic mapping of Dasypyrum villosum 6V#4S-specific PCR markers using transcriptome data.Theoretical and Applied Genetics,2017,130(10):2057-2068.
[27]PLASCHKE J,GANAL M E,RODER M S.Detection of genetic diversity in closely related bread wheat using microsatellite markers.Theoretical and Applied Genetics,1995,91:1001-1007.
[28]BRYAN G J,COLLINS A J,STEPHENSON P,ORRY A,SMITH J B,GALE M D.Isolation and characterization of microsatellites from hexaploid bread wheat.Theoretical and Applied Genetics,1997,94(5):557-563.
[29]谢皓,陈学珍,杨柳,王建立.EST-SSR标记的发展和在植物遗传研究中的应用.北京农学院学报,2005,20(4):73-76.XIE H,CHEN X J,YANG L,WANG J L.Development and application of EST-SSR marker in plant genetic.Journal of Beijing Agricultural College,2005,20(4):73-76.(in Chinese)
[30]曹亚萍,曹爱忠,王秀娥,陈佩度.基于EST-PCR的簇毛麦染色体特异分子标记筛选及应用.作物学报,2009,35(1):1-10.CAO Y P,CAO A Z,WANG X E,CHEN P D.Screening and application of EST-based PCR markers specific to individual chromosomes of Haynaldia villosa.Acta Agronomica Sinica,2009,35(1):1-10.(in Chinese)
[31]李珊珊,曾艳飞,何彩云,张建国.基于沙棘转录组序列开发EST-SSR分子标记.林业科学研究,2017,30(1):69-74.LI S S,ZENG Y F,HE C Y,ZHANG J G.Development of EST-SSRmarkers based on Seabuckthorn transcriptomic sequences.Forest Research,2017,30(1):69-74.(in Chinese)
[32]陈岳,张微微,莫海波,付乃峰,田代科.EST-SSR标记构建莲(Nelumbo Adans)遗传连锁图谱.分子植物育种,2017,15(6):2265-2273.CHEN Y,ZHANG W W,MO H B,FU N F,TIAN D K.Construction of a genetic linkage map of lotus(Nelumbo Adans)using EST-SSRmarkers.Molecular Plant Breeding,2017,15(6):2265-2273.(in Chinese)
[33]陈勋,龚自明.基于EST-SSR标记的湖北茶树种质资源遗传多样性分析.分子植物育种,2017,15(5):1831-1838.CHEN X,GONG Z M.Analysis of genetic diversity with EST-SSRmarkers for tea germplasm in Hubei province.Molecular Plant Breeding,2017,15(5):1831-1838.(in Chinese)
[34]李翔,张颖,许俊强,宋婷,李文璐,董玉梅,张应华.基于EST-SSR标记的云南黄瓜种质资源遗传多样性分析.分子植物育种,2016,14(5):1179-1188.LI X,ZHANG Y,XU J Q,SONG T,LI W L,DONG Y M,ZHANG YH.Analysis on genetic diversity of local cucumber germplasm resources in Yunnan based on EST-SSR markers.Molecular Plant Breeding,2016,14(5):1179-1188.(in Chinese)
[35]QI L L,WANG S L,CHEN P D,LIU D J,GILL B S.Identification and physical mapping of three Haynaldia villosa chromosome-6Vdeletion lines.Theoretical and Applied Genetics,1998,97(7):1042-1046.
[36]ZHANG L D,YUAN D J,YU S W,LI Z G,CAO Y F,MIAO Z Q,QIAN H M,TANG K X.Preference of simple sequence repeats in coding and non-coding regions of Arebidopsis thaliana.Bioinformations,2004,20(7):1081-1086.
[37]张利达,唐克轩.植物EST-SSR标记开发及其应用.基因组学与应用生物学,2010,29(3):534-541.ZHANG L D,TANG K X.Development of plant EST-SSR markers and its application.Genomics and Applied Biology,2010,29(3):534-541.(in Chinese)
[38]GAO L F,TANG J F,LI H W,JIA J Z.Analysis of microsatellites in major crops assessed by computational and experimental approaches.Molecular Breeding,2003,12:245-261.
[39]文亚峰,韩文军,周宏,徐刚标.杉木转录组SSR挖掘及EST-SSR标记规模化开发.林业科学,2015,51(11):40-49.WEN Y F,HAN W J,ZHOU H,XU G B.SSR mining and development of EST-SSR markers for Cunninghamia lanceolata based on transcriptome sequences.Scientia Silvae Sinicae,2015,51(11):40-49.(in Chinese)
[40]KASHKUSH K,FELDMAN M,LEVY A A.Silencing and activation in a newly synthesized wheat allotetraploid.Genetics,2002,160(4):1651-1659.
[2]赵万春,董剑,陈其皎,李晓燕,高翔,石引刚,陈良国.簇毛麦-用于小麦改良的一种野生植物.草业科学,2012,29(10):1613-1621.ZHAO W C,DONG J,CHEN Q J,LI X Y,GAO X,SHI Y G,CHENL G.Advance in Dasypyrum villosum-a valuable wild species used in wheat improvement.Protacultural Science,2012,29(10):1613-1621.(in Chinese)
[3]CHEN Q,CONNER R L,LI H,LAROCHE A,GRAF R J,KUZYK AD.Expression of resistance to strip rust,powdery mildew and the wheat curl mite in Triticum aestivum×Haynaldia villosa lines.Canadian Journal of Plant Science,2002,82(2):451-456.
[4]刘大钧,陈佩度,吴沛良,王耀南,邱伯行,王苏玲.硬粒小麦-簇毛麦双二倍体.作物学报,1986,12(3):155-162.LIU D J,CHEN P D,WU P L,WANG Y N,QIU B X,WANG S L.Triticum durum-Haynadia villosa amphidiploid.Acta Agronomica Sinica,1986,12(3):155-162.(in Chinese)
[5]陈孝,徐惠君,杜丽璞,尚立民,韩彬,施爱农,肖世和.利用组织培养技术向普通小麦导入簇毛麦抗白粉病基因的研究.中国农业科学,1996,29(5):1-8.CHEN X,XU H J,DU L P,SHANG L M,HAN B,SHI A N,XIAO SH.Transfer of gene resistant to powdery mildew from H.villosa to common wheat by tissue culture.Scientia Agricultura Sinica,1996,29(5):1-8.(in Chinese)
[6]陈佩度,周波,齐莉莉,刘大钧.用分子原位杂交(GISH)鉴定小麦-簇毛麦双倍体、附加系、代换系和易位系.遗传学报,1995,22(5):380-386.CHEN P D,ZHOU B,QI L L,LIU D J.Identification of wheat-Haynaldia villosa amphiloid,addition,substitution and translocation lines by in situ hybridization using biotin-labelled genomic DNA as a probe.Acta Genetica Sinica,1995,22(5):380-386.(in Chinese)
[7]CHEN P D,QI L L,ZHOU B,ZHANG S Z,LIU D J.Development and molecular cytogenetic analysis of wheat-Haynaldia villosa6VS/6AL translocation lines specifying resistance to powdery mildew.Theoretical and Applied Genetics,1995,91:1125-1128.
[8]齐莉莉,陈佩度,刘大钧,周波,张守中,盛宝钦,向齐君,段霞渝,周益林.小麦白粉病新抗源--基因Pm21.作物学报,1995,21(3):257-262.QI L L,CHEN P D,LIU D J,ZHOU B,ZHANG S Z,SHEENG B Q,XIANG Q J,DUAN X Y,ZHOU Y L.The gene Pm21-a new source for resistance to wheat powdery mildew.Acta Agronomica Sinica,1995,21(3):257-262.(in Chinese)
[9]LI H,CHEN X,XIN Z Y,MA Y Z,XU H J,CHEN X Y,JIA X.Development and identification of wheat-Haynaldia villosa T6DL.6VS chromosome translocation lines conferring resistance to powdery mildew.Plant Breeding,2005,124:203-205.
[10]LIU C,QI L L,LIU W X,ZHAO W C,WILSON J,FRIEBE B,GILLB S.Development of a set of compensating Triticum aestivumDasypyrum villosum Robertsonian translocation lines.Genome,2011,54:836-844.
[11]XING L,HU P,LIU J.,WITEK K,ZHOU S,XU J,ZHOU W,GAO L,HUANG Z,ZHANG R,WANG X,CHEN P,WANG H,JONES J D G,KARAFIATOVA M,VRANA J,BARTOS J,DOLEZEL J,TIAN Y,WU Y,CAO A.Pm21 from Haynaldia villosa encodes a CC-NBS-LRR that confers powdery mildew resistance in wheat.Molecular Plant,2018,doi:10.1016/j.molp.2018.02.013.
[12]HE H,ZHU S,ZHAO R,JIANG Z,JI Y,JI J,QIU D,LI H,BIE T.Pm21,encoding a typical CC-NBS-LRR protein,confers broadspectrum resistance to wheat powdery mildew disease.Molecular Plant,2018,doi:10.1016/j.molp.2018.03.004.
[13]ZHANG Q P,LI Q,WANG X E,WANG H Y,LANG S P,WANG Y N,WANG S L,CHEN P D,LIU D J.Development and characterization of a Triticum aestivum-Haynaldia villosa translocation line T4VS·4DLconferring resistance to wheat spindle streak mosaic virus.Euphytica,2005,145:317-320.
[14]ZHAO R,WANG H,XIAO J,BIE T,CHENG S,JIA Q,YUAN C,ZHANG R,CAO A,CHEN P,WANG X.Induction of 4VSchromosome recombinants using the CS ph1b mutant and mapping of the wheat yellow mosaic virus resistance gene from Haynaldia villosa.Theoretical and Applied Genetics,2013,126:2921-2930.
[15]李淑梅,徐川梅,周波,陈佩度.普通小麦-簇毛麦2V染色体端体异附加系的选育与鉴定.南京农业大学学报,2009,32(1):1-5.LI S M,XU C M,ZHOU B,CHEN P D.Development and identification of Triticum-Haynadia villosa ditelesomic addition lines involving chromosome 2V of H.villosa.Journal of Nanjing Agricultural University,2009,32(1):1-5.(in Chinese)
[16]董剑,杨华,赵万春,李晓燕,陈其皎,高翔.普通小麦中国春-簇毛麦易位系T1DL·1VS和T1DS·1VL的农艺和品质特性.作物学报,2013,39(8):1386-1390.DONG J,YANG H,ZHAO W C,LI X Y,CHEN Q J,GAO X.Agronomic traits and grain quality of Chinese Spring-Dasypyrum villosum translocation lines T1DL·1VS and T1DS·1VL.Acta Agronomica Sinica,2013,39(8):1386-1390.(in Chinese)
[17]张瑞奇,冯祎高,侯富,陈树林,别同德,陈佩度.普通小麦-簇毛麦T5VS.5DL易位染色体对小麦主要农艺性状、品质和白粉病抗性的遗传效应分析.中国农业科学,2015,48(6):1041-1051.ZHANG R Q,FENG Y G,HOU F,CHEN S L,BIE T D,CHEN P D.The genetic effect of wheat-H.villosa T5VS.5DL translocated chromosome on agronomic characteristics,quality,and powdery mildew resistance of common wheat.Scientia Agricultura Sinica,2015,48(6):1041-1051.(in Chinese)
[18]ZHANG R,SUN B,CHEN J,CAO A,XING L,FENG Y,LAN C,CHEN P.Pm55,a developmental-stage and tissue-specific powdery mildew resistance gene introgressed from Dasypyrum villosum into common wheat.Theoretical and Applied Genetics,2016,129:1975-1984.
[19]QI L L,PUMPHREY M O,FRIEBE B,ZHANG P,QIAN C,BOWDEN R L,ROUSE M N,JIN Y,GILL B S.A novel Robertsonian translocation event leads to transfer of a stem rust resistance gene(Sr52)effective against race Ug99 from Dasypyrum villosum into bread wheat.Theoretical and Applied Genetics,2011,123(1):159-167.
[20]LI H J,CONNER R L,CHEN Q,JIA X,LI H,GRAF R J,LAROCHE A,KUZYK A D.Different reactions to the wheat curl mite and wheat streak mosaic virus in various wheat-Haynaldia villosa 6V and 6VS lines.Plant Disease,2002,86(4):423-428.
[21]LIU C,Ye X G,Wang M J,LI S J,LIN Z S.Genetic behavior of Triticum aestivum-Dasypyrum villosum translocation chromosomes T6V#4S·6DL and T6V#2S·6AL.Journal of Integrative Agriculture,2017,16(10):2136-2144.
[22]张云龙,王美蛟,张悦,褚翠萍,林志珊,徐琼芳,叶兴国,陈孝,张宪省.不同簇毛麦6VS染色体臂的白粉病抗性特异功能标记的开发及应用.作物学报,2012,38(10):1827-1832.ZHANG Y L,WANG M J,ZHANF Y,CHU C P,LIN Z S,XU Q F,YE X G,CHEN X,ZHANG X S.Development and application of functional markers specific to powdery mildew resistance on chromosome arm 6VS from different origins of Haynaldia villosa.Acta Agronomica Sinica,2012,38(10):1827-1832.(in Chinese)
[23]LIN Z S,ZHANG Y L,WANG K,LI J R,XU Q F,CHEN X,ZHANG X S,YE X G.Isolation and molecular analysis of genes Stpk-V2 and Stpk-V3 homologous to powdery mildew resistance gene Stpk-V in a Dasypyrum villosum accession and its derivatives.Journal of Applied Genetics,2013,54:417-426.
[24]BIE T D,ZHAO R H,ZHU S Y,CHEN S L,HE H G.Development and characterization of marker MBH1 simultaneously tagging genes Pm21 and Pm V conferring resistance to powdery mildew in wheat.Molecular Breeding,2015,35:189.
[25]刘畅,李仕金,王轲,叶兴国,林志珊.簇毛麦6VS特异转录序列P21461及P33259的获得及其分子标记在鉴定小麦-簇毛麦抗白粉病育种材料中的应用.作物学报,2017,43(7):983-992.LIU C,LI S J,WANG K,YE X G,LIN Z S.Developing of specific transcription sequences P21461 and P33259 on Dasypyrum villosum6VS and application of molecular markers in identifying wheat-D.villosum breeding materials with powdery mildew resistance.Acta Agronomica Sinica,2017,43(7):983-992.(in Chinese)
[26]LI S J,LIN Z S,LIU C,WANG K,DU L P,YE X G.Development and comparative genomic mapping of Dasypyrum villosum 6V#4S-specific PCR markers using transcriptome data.Theoretical and Applied Genetics,2017,130(10):2057-2068.
[27]PLASCHKE J,GANAL M E,RODER M S.Detection of genetic diversity in closely related bread wheat using microsatellite markers.Theoretical and Applied Genetics,1995,91:1001-1007.
[28]BRYAN G J,COLLINS A J,STEPHENSON P,ORRY A,SMITH J B,GALE M D.Isolation and characterization of microsatellites from hexaploid bread wheat.Theoretical and Applied Genetics,1997,94(5):557-563.
[29]谢皓,陈学珍,杨柳,王建立.EST-SSR标记的发展和在植物遗传研究中的应用.北京农学院学报,2005,20(4):73-76.XIE H,CHEN X J,YANG L,WANG J L.Development and application of EST-SSR marker in plant genetic.Journal of Beijing Agricultural College,2005,20(4):73-76.(in Chinese)
[30]曹亚萍,曹爱忠,王秀娥,陈佩度.基于EST-PCR的簇毛麦染色体特异分子标记筛选及应用.作物学报,2009,35(1):1-10.CAO Y P,CAO A Z,WANG X E,CHEN P D.Screening and application of EST-based PCR markers specific to individual chromosomes of Haynaldia villosa.Acta Agronomica Sinica,2009,35(1):1-10.(in Chinese)
[31]李珊珊,曾艳飞,何彩云,张建国.基于沙棘转录组序列开发EST-SSR分子标记.林业科学研究,2017,30(1):69-74.LI S S,ZENG Y F,HE C Y,ZHANG J G.Development of EST-SSRmarkers based on Seabuckthorn transcriptomic sequences.Forest Research,2017,30(1):69-74.(in Chinese)
[32]陈岳,张微微,莫海波,付乃峰,田代科.EST-SSR标记构建莲(Nelumbo Adans)遗传连锁图谱.分子植物育种,2017,15(6):2265-2273.CHEN Y,ZHANG W W,MO H B,FU N F,TIAN D K.Construction of a genetic linkage map of lotus(Nelumbo Adans)using EST-SSRmarkers.Molecular Plant Breeding,2017,15(6):2265-2273.(in Chinese)
[33]陈勋,龚自明.基于EST-SSR标记的湖北茶树种质资源遗传多样性分析.分子植物育种,2017,15(5):1831-1838.CHEN X,GONG Z M.Analysis of genetic diversity with EST-SSRmarkers for tea germplasm in Hubei province.Molecular Plant Breeding,2017,15(5):1831-1838.(in Chinese)
[34]李翔,张颖,许俊强,宋婷,李文璐,董玉梅,张应华.基于EST-SSR标记的云南黄瓜种质资源遗传多样性分析.分子植物育种,2016,14(5):1179-1188.LI X,ZHANG Y,XU J Q,SONG T,LI W L,DONG Y M,ZHANG YH.Analysis on genetic diversity of local cucumber germplasm resources in Yunnan based on EST-SSR markers.Molecular Plant Breeding,2016,14(5):1179-1188.(in Chinese)
[35]QI L L,WANG S L,CHEN P D,LIU D J,GILL B S.Identification and physical mapping of three Haynaldia villosa chromosome-6Vdeletion lines.Theoretical and Applied Genetics,1998,97(7):1042-1046.
[36]ZHANG L D,YUAN D J,YU S W,LI Z G,CAO Y F,MIAO Z Q,QIAN H M,TANG K X.Preference of simple sequence repeats in coding and non-coding regions of Arebidopsis thaliana.Bioinformations,2004,20(7):1081-1086.
[37]张利达,唐克轩.植物EST-SSR标记开发及其应用.基因组学与应用生物学,2010,29(3):534-541.ZHANG L D,TANG K X.Development of plant EST-SSR markers and its application.Genomics and Applied Biology,2010,29(3):534-541.(in Chinese)
[38]GAO L F,TANG J F,LI H W,JIA J Z.Analysis of microsatellites in major crops assessed by computational and experimental approaches.Molecular Breeding,2003,12:245-261.
[39]文亚峰,韩文军,周宏,徐刚标.杉木转录组SSR挖掘及EST-SSR标记规模化开发.林业科学,2015,51(11):40-49.WEN Y F,HAN W J,ZHOU H,XU G B.SSR mining and development of EST-SSR markers for Cunninghamia lanceolata based on transcriptome sequences.Scientia Silvae Sinicae,2015,51(11):40-49.(in Chinese)
[40]KASHKUSH K,FELDMAN M,LEVY A A.Silencing and activation in a newly synthesized wheat allotetraploid.Genetics,2002,160(4):1651-1659.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |