黑龙江省83份小麦品种抗秆锈病基因的分子检测
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摘要
为了明确黑龙江省小麦品种(系)对中国秆锈菌的抗性水平和了解抗秆锈病基因在该区域的分布情况,本研究选用中国小麦秆锈菌流行小种21C3CTHQM、34MKGQM和34C3RTGQM对从该区域征集到的83份主要小麦品种(系)进行了苗期抗秆锈病的评价,并利用与抗秆锈病基因Sr2、Sr24、Sr25、Sr26、Sr31和Sr38紧密连锁的分子标记分别进行了分子检测,结合苗期表型及系谱,推测这些品种(系)可能含有的抗病基因。结果表明,83份小麦品种(系)对供试秆锈菌小种均表现抗性,对21C3CTHQM、34MKGQM和34C3RTGQM表现免疫或近免疫的分别为57、53和60份,各占供试材料数量的68.68%、63.85%和72.29%,其他剩余材料对3个供试秆锈菌小种表现中抗或高抗。分子标记分析表明,83份主要小麦品种(系)中有12份可能含有Sr2;克旱3号可能含有Sr25; 6份小麦品种可能含有Sr31; 19份小麦品种可能含有Sr38;没有检测出含有Sr24和Sr26的品种。因此,黑龙江省小麦品种对中国小麦秆锈病抗性水平相对较高,含有抗秆锈病基因Sr2以及对我国小麦秆锈病表现良好抗性的基因Sr31和Sr38,可能含有其他未知抗秆锈病基因,这些优良抗源材料可作为未来小麦生产育种的种质资源。
For clarifying the resistance level of wheat cultivars or lines developed in Heilongjiang to Puccinia graminis f. sp. tritici( Pgt) and finding out the stem rust resistance genes utilization in them,the seedling stem rust resistance of 83 major wheat cultivars or lines collected from Heilongjiang were evaluated with predominant Chinese Pgt races 21 C3 CTHQM,34 MKGQM and 34 C3 RTGQM in this study. Besides,molecular markers closely linked with genes Sr2,Sr24,Sr25,Sr26,Sr31,and Sr38 were used for molecular identification. Resistance genes possibly carried in the cultivars were analyzed based on the results of molecular identification,seedling reaction types as well as cultivar pedigrees. The results showed that all tested wheat cultivars or lines were resistant to the three tested races. Among them,57,53 and 60 of the 83 cultivars were immune or nearly immune to21 C3 CTHQM,34 MKGQM and 34 C3 RTGQM,namely accounting for 68.68%,63.85% and 72.29% of the total cultivars,respectively. The rest others remained moderately or highly resistant to the tested races. With molecular marker analysis,12 of 83 wheat cultivars or lines possibly carry gene Sr2,Sr25 tested in Kehan 3,and Sr31 and Sr38 in 6 and 19 wheat cultivars respectively. Both genes Sr24 and Sr26 were not detected in any tested cultivar.Therefore,wheat cultivars developed in Heilongjiang Province was relatively highly resistant to stem rust. Most of them carry resistance gene Sr2,and/or Sr31 and Sr38 being resistant to all Chinese Pgt races,and/or possible other unknown resistance genes. These highly resistant wheat cultivars can be used as effective resistance resources for future wheat breeding.
For clarifying the resistance level of wheat cultivars or lines developed in Heilongjiang to Puccinia graminis f. sp. tritici( Pgt) and finding out the stem rust resistance genes utilization in them,the seedling stem rust resistance of 83 major wheat cultivars or lines collected from Heilongjiang were evaluated with predominant Chinese Pgt races 21 C3 CTHQM,34 MKGQM and 34 C3 RTGQM in this study. Besides,molecular markers closely linked with genes Sr2,Sr24,Sr25,Sr26,Sr31,and Sr38 were used for molecular identification. Resistance genes possibly carried in the cultivars were analyzed based on the results of molecular identification,seedling reaction types as well as cultivar pedigrees. The results showed that all tested wheat cultivars or lines were resistant to the three tested races. Among them,57,53 and 60 of the 83 cultivars were immune or nearly immune to21 C3 CTHQM,34 MKGQM and 34 C3 RTGQM,namely accounting for 68.68%,63.85% and 72.29% of the total cultivars,respectively. The rest others remained moderately or highly resistant to the tested races. With molecular marker analysis,12 of 83 wheat cultivars or lines possibly carry gene Sr2,Sr25 tested in Kehan 3,and Sr31 and Sr38 in 6 and 19 wheat cultivars respectively. Both genes Sr24 and Sr26 were not detected in any tested cultivar.Therefore,wheat cultivars developed in Heilongjiang Province was relatively highly resistant to stem rust. Most of them carry resistance gene Sr2,and/or Sr31 and Sr38 being resistant to all Chinese Pgt races,and/or possible other unknown resistance genes. These highly resistant wheat cultivars can be used as effective resistance resources for future wheat breeding.
引文
[1]Cao Y Y,Han J D,Zhu G Q,et al.Ug99,a new virulent race of Puccinia graminis f.sp.tritici,and its effect on China(in Chinese)[J].Plant Protection(植物保护),2007,33(6):86-89.
[2]Pretorius Z A,Singh RP,Wagoire WW,et al.Detection of virulence to w heat stem rust resistance gene Sr31 in Puccinia graminis f.sp.tritici in Uganda[J].Plant Disease,2000,84:203.
[3]BGRI.A global wheat rust monitoring system.2017.Available at:http://rusttracker.cimmyt.org/?page_id=22.
[4]Jin Y,Szabo L J,Pretorius Z A,et al.Detection of virulence to resistance gene Sr24 w ithin race TTKS of Puccinia graminis f.sp.tritici[J].Plant Disease,2008,92:923-926.
[5]Jin Y,Szabo L J,Rouse M N,et al.Detection of virulence to resistance gene Sr36 w ithin the TTKS race lineage of Puccinia graminis f.sp.tritici[J].Plant Disease,2009,93:367-370.
[6]Pretorius Z A,Szabo G,Boshoff W H P,et al.First report of a new TTKSF race of w heat stem rust(Puccinia graminis f.sp.tritici)in South Africa and Zimbabw e[J].Plant Disease,2012,96:590.
[7]Rouse M N,Nirmala J,Jin Y,et al.Characterization of Sr9h,a w heat stem rust resistance allele effective to Ug99[J].Theoretical and Applied Genetics,2014,127:1681-1688.
[8]Olivera P,Newcomb M,Szabo L J,et al.Phenotypic and genotypic characterization of race TKTTF of Puccinia graminis f.sp.tritici that caused a w heat stem rust epidemic in Southern Ethiopia in 2013-2014[J].Phytopathology,2015,105:917-928.
[9]Bhattacharya S.Deadly new wheat disease threatens Europe’s crops[J].Nature,2017,542:145-146.
[10]Umesh G,Sarvjeet K,Rakesh Y,et al.Recent trends and perspectives of molecular markers against fungal diseases in w heat[J].Frontiers in M icrobiology,2015,6:1-13.
[11]Yu L X,Liu S X,Anderson J A,et al.Haplotype diversity of stem rust resistance loci in uncharacterized w heat lines[J].M olecular Breeding,2010,26:667-680.
[12]Khan R R,Bariana H S,Dholakia B B,et al.Molecular mapping of stem and leaf rust resistance in w heat[J].Theoretical and Applied Genetics,2005,111:846-850.
[13]Haile J K,Hammer K,Badebo A,et al.Genetic diversity assessment of Ethiopian tetraploid w heat landraces and improved durum w heat varieties using microsatellites and markers linked w ith stem rust resistance[J].Genetic Resources&Crop Evolution,2013,60(2):513-527.
[14]Kokhmetova A M,Atishova M N.Identification of sources of resistance to w heat stem rust using molecular markers[J].Russian Journal of Genetics Applied Research,2012,2(6):486-493.
[15]Pretorius Z A,Jin Y,Bender C M,et al.Seedling resistance to stem rust race Ug99 and marker analysis for Sr2,Sr24 and Sr31 in South African w heat cultivars and lines[J].Euphytica,2012,186:15-23.
[16]Wu X X,Li T Y,Chen S,et al.Stem rust resistance evaluation and Ug99-resistance gene detection of 139w heat cultivars(in Chinese)[J].Scientia Agricultura Sinica(中国农业科学),2014,47(23):4618-4626.
[17]Xu X F,Li D D,Liu Y,et al.Evaluation and identification of stem rust resistance genes Sr2,Sr24,Sr25,Sr26,Sr31 and Sr38 in w heat lines from Gansu Province in China[J].Peer J,2017,5:e4146.DOI:10.7717/peerj.4146.
[18]Ma Y.Preliminary molecular detection for the stem rust resistant gene Sr22 in part of the w heat germplasms(in Chinese)[J].Heilongjiang Agricultural Science(黑龙江农业科学),2013,1:7-10.
[19]Fan Y,Yao Z Y,Ding Z Z,et al.Study on the dynamics of small w heat stalk rust pathogen in Heilongjiang Province(in Chinese)[J].Heilongjiang Agricultural Science(黑龙江农业科学),1980,5:42-45.
[20]Stakman E C,Stewart D M,Loegering W Q.Identification of physiologic races of Puccinia graminis var.tritici[M].United States:US Department of Agriculture Press,1962:53.
[21]Mago R,Brown-Guedira G,Dreisigacker S,et al.An accurate DNA marker assay for stem rust resistance gene Sr2 in w heat[J].Theoretical and Applied Genetics 2011,122:735-744.
[22]Mago R,Bariana H S,Dundas I S,et al.Development of PCR markers for the selection of w heat stem rust resistance genes Sr24 and Sr26 in diverse w heat germplasm[J].Theoretical and Applied Genetics,2005,111:496-504.
[23]Liu S,Yu L X,Singh R P,et al.Diagnostic and co-dominant PCR markers for w heat stem rust resistance genes Sr25 and Sr26[J].Theoretical and Applied Genetics,2010,120:691-697.
[24]Mago R,Spielmeyer W,Lawrence G J,et al.Identification and mapping of molecular markers linked to rust resistance genes located on chromosome 1RS of rye using w heat-rye translocation lines[J].Theoretical and Applied Genetics,2002,104:1317-1324.
[25]Helguera M,Khan I A,Kolmer J,et al.PCR assays for the Lr37-Yr17-Sr38 cluster of rust resistance genes and their use to develop isogenic hard red spring w heat lines[J].Crop Science,2003,43:1839-1847.
[26]Smith E L,Schlehuber A M,Young H C Jr,et al.Registration of agent w heat[J].Crop Science,1968,8:511-512.
[27]Cao Y Y.On epiphytotic pattern,long dispersion of Puccinia graminis f.sp.tritici and its gene control through systematic engineering in China(in Chinese)[D].Shenyang:Shenyang Agricultural University(沈阳:沈阳农业大学),1994.
[28]Singh R P,Huerta-Espino J,Bhavani S,et al.Race non-specific resistance to rust diseases in CIM M YTspring w heats[J].Euphytica,2011,179:175-186.
[29]Han J D,Cao Y Y,Sun Z G.2007-2008 Race dynamics of Puccinia graminis f.sp.tritici in China and the virulence of CIM M YT w heat germplasm resistant to Ug99(in Chinese)[J].Journal of Triticeae Crops(麦类作物学报),2010,30:163-166.
[30]FAO.Spread of damaging wheat rust continues:new races found in Europe,Africa,Central Asia.3 February,2017.Available at http://w w w.fao.org/new s/story/en/item/46946 7/icode/.
[31]Li T Y,Cao Y Y,Wu X X,et al.Seedling resistance to stem rust and molecular marker analysis of resistance genes in w heat cultivars of Yunnan,China[J].PLoSONE,2016,11:e0165640.DOI:10.1371/journal.pone.0165640.
[32]Jiang Y Y,Chen W Q,Zhao Z H,et al.Threat of new w heat stem rust race Ug99 to w heat production in China and counter measure(in Chinese)[J].Plant Protection(植物保护),2007,27:14-16.
[2]Pretorius Z A,Singh RP,Wagoire WW,et al.Detection of virulence to w heat stem rust resistance gene Sr31 in Puccinia graminis f.sp.tritici in Uganda[J].Plant Disease,2000,84:203.
[3]BGRI.A global wheat rust monitoring system.2017.Available at:http://rusttracker.cimmyt.org/?page_id=22.
[4]Jin Y,Szabo L J,Pretorius Z A,et al.Detection of virulence to resistance gene Sr24 w ithin race TTKS of Puccinia graminis f.sp.tritici[J].Plant Disease,2008,92:923-926.
[5]Jin Y,Szabo L J,Rouse M N,et al.Detection of virulence to resistance gene Sr36 w ithin the TTKS race lineage of Puccinia graminis f.sp.tritici[J].Plant Disease,2009,93:367-370.
[6]Pretorius Z A,Szabo G,Boshoff W H P,et al.First report of a new TTKSF race of w heat stem rust(Puccinia graminis f.sp.tritici)in South Africa and Zimbabw e[J].Plant Disease,2012,96:590.
[7]Rouse M N,Nirmala J,Jin Y,et al.Characterization of Sr9h,a w heat stem rust resistance allele effective to Ug99[J].Theoretical and Applied Genetics,2014,127:1681-1688.
[8]Olivera P,Newcomb M,Szabo L J,et al.Phenotypic and genotypic characterization of race TKTTF of Puccinia graminis f.sp.tritici that caused a w heat stem rust epidemic in Southern Ethiopia in 2013-2014[J].Phytopathology,2015,105:917-928.
[9]Bhattacharya S.Deadly new wheat disease threatens Europe’s crops[J].Nature,2017,542:145-146.
[10]Umesh G,Sarvjeet K,Rakesh Y,et al.Recent trends and perspectives of molecular markers against fungal diseases in w heat[J].Frontiers in M icrobiology,2015,6:1-13.
[11]Yu L X,Liu S X,Anderson J A,et al.Haplotype diversity of stem rust resistance loci in uncharacterized w heat lines[J].M olecular Breeding,2010,26:667-680.
[12]Khan R R,Bariana H S,Dholakia B B,et al.Molecular mapping of stem and leaf rust resistance in w heat[J].Theoretical and Applied Genetics,2005,111:846-850.
[13]Haile J K,Hammer K,Badebo A,et al.Genetic diversity assessment of Ethiopian tetraploid w heat landraces and improved durum w heat varieties using microsatellites and markers linked w ith stem rust resistance[J].Genetic Resources&Crop Evolution,2013,60(2):513-527.
[14]Kokhmetova A M,Atishova M N.Identification of sources of resistance to w heat stem rust using molecular markers[J].Russian Journal of Genetics Applied Research,2012,2(6):486-493.
[15]Pretorius Z A,Jin Y,Bender C M,et al.Seedling resistance to stem rust race Ug99 and marker analysis for Sr2,Sr24 and Sr31 in South African w heat cultivars and lines[J].Euphytica,2012,186:15-23.
[16]Wu X X,Li T Y,Chen S,et al.Stem rust resistance evaluation and Ug99-resistance gene detection of 139w heat cultivars(in Chinese)[J].Scientia Agricultura Sinica(中国农业科学),2014,47(23):4618-4626.
[17]Xu X F,Li D D,Liu Y,et al.Evaluation and identification of stem rust resistance genes Sr2,Sr24,Sr25,Sr26,Sr31 and Sr38 in w heat lines from Gansu Province in China[J].Peer J,2017,5:e4146.DOI:10.7717/peerj.4146.
[18]Ma Y.Preliminary molecular detection for the stem rust resistant gene Sr22 in part of the w heat germplasms(in Chinese)[J].Heilongjiang Agricultural Science(黑龙江农业科学),2013,1:7-10.
[19]Fan Y,Yao Z Y,Ding Z Z,et al.Study on the dynamics of small w heat stalk rust pathogen in Heilongjiang Province(in Chinese)[J].Heilongjiang Agricultural Science(黑龙江农业科学),1980,5:42-45.
[20]Stakman E C,Stewart D M,Loegering W Q.Identification of physiologic races of Puccinia graminis var.tritici[M].United States:US Department of Agriculture Press,1962:53.
[21]Mago R,Brown-Guedira G,Dreisigacker S,et al.An accurate DNA marker assay for stem rust resistance gene Sr2 in w heat[J].Theoretical and Applied Genetics 2011,122:735-744.
[22]Mago R,Bariana H S,Dundas I S,et al.Development of PCR markers for the selection of w heat stem rust resistance genes Sr24 and Sr26 in diverse w heat germplasm[J].Theoretical and Applied Genetics,2005,111:496-504.
[23]Liu S,Yu L X,Singh R P,et al.Diagnostic and co-dominant PCR markers for w heat stem rust resistance genes Sr25 and Sr26[J].Theoretical and Applied Genetics,2010,120:691-697.
[24]Mago R,Spielmeyer W,Lawrence G J,et al.Identification and mapping of molecular markers linked to rust resistance genes located on chromosome 1RS of rye using w heat-rye translocation lines[J].Theoretical and Applied Genetics,2002,104:1317-1324.
[25]Helguera M,Khan I A,Kolmer J,et al.PCR assays for the Lr37-Yr17-Sr38 cluster of rust resistance genes and their use to develop isogenic hard red spring w heat lines[J].Crop Science,2003,43:1839-1847.
[26]Smith E L,Schlehuber A M,Young H C Jr,et al.Registration of agent w heat[J].Crop Science,1968,8:511-512.
[27]Cao Y Y.On epiphytotic pattern,long dispersion of Puccinia graminis f.sp.tritici and its gene control through systematic engineering in China(in Chinese)[D].Shenyang:Shenyang Agricultural University(沈阳:沈阳农业大学),1994.
[28]Singh R P,Huerta-Espino J,Bhavani S,et al.Race non-specific resistance to rust diseases in CIM M YTspring w heats[J].Euphytica,2011,179:175-186.
[29]Han J D,Cao Y Y,Sun Z G.2007-2008 Race dynamics of Puccinia graminis f.sp.tritici in China and the virulence of CIM M YT w heat germplasm resistant to Ug99(in Chinese)[J].Journal of Triticeae Crops(麦类作物学报),2010,30:163-166.
[30]FAO.Spread of damaging wheat rust continues:new races found in Europe,Africa,Central Asia.3 February,2017.Available at http://w w w.fao.org/new s/story/en/item/46946 7/icode/.
[31]Li T Y,Cao Y Y,Wu X X,et al.Seedling resistance to stem rust and molecular marker analysis of resistance genes in w heat cultivars of Yunnan,China[J].PLoSONE,2016,11:e0165640.DOI:10.1371/journal.pone.0165640.
[32]Jiang Y Y,Chen W Q,Zhao Z H,et al.Threat of new w heat stem rust race Ug99 to w heat production in China and counter measure(in Chinese)[J].Plant Protection(植物保护),2007,27:14-16.