2003-2013年小麦品种(系)抗条锈性鉴定及评价

2003-2013年在甘肃省农业科学院植物保护研究所兰州温室和甘谷试验站,分别对来自国内35个相关育种单位的冬春小麦品种(系)5001份,其中冬小麦4291份、春小麦710份,进行苗期混合菌、成株期分小种和混合菌抗条锈性接种鉴定,结果表明:全生育期表现免疫近免疫的有兰天31号等479份,高抗的有兰天23号等76份,中抗的有天选49等291份,分别占9.58％、1.52％和5.82％;成株期表现免疫近免疫的有天选50号等840份,高抗的有兰天27号等47份,中抗的有天选52等311份,分别占16.80％、0.94％和6.22％;苗期表现免疫近免疫的有兰天30号等964份,高抗的有天98102等122份,中抗的有00-30等273份,分别占19.28％、2.44％和5.46％.冬小麦有天选49号等914份材料表现全生育期抗病,占18.28％;有97-473等906份成株期表现抗病,占18.12％;有兰天20号等1225份苗期表现抗病,占24.50％.春小麦有定西41号等113份材料全生育期表现抗病,占2.26％;有陇春28号等125份成株期表现抗病,占2.50％;有0109-1等114份苗期对混合菌表现抗病,占2.28％.先后在甘肃天水汪川良种场对相关材料进行成株期抗条锈性评价,结果发现:1154份从小种圃筛选出的抗病材料中,表现抗病的有兰天31号等745份,占64.56％;105份甘肃陇南生产品种中,到2013年表现抗病的仅有兰天28号、中梁31号等30份材料,占28.57％;后备品系中,00-30-2-1、CP04-20、00127-2-3等抗性表现优异;抗源材料中,仅有贵农775、中四、T.Spelta albun、贵协1、贵协3等少数材料表现抗病,重要抗源材料贵农21、贵农22、南农92R、川麦42、Moro从2011年开始在田间表现感病,逐步失去利用价值.其衍生系品种材料如陇鉴9343、天选43号、中梁29号、兰天17号、兰天24号等也在田间逐步感病,条锈病发生流行压力持续增大.     

Characterization of non-host resistance in broad bean to the wheat stripe rust pathogen

ABSTRACT: BACKGROUND: Non-host resistance (NHR) confers plant species immunity against the majority of microbialpathogens and represents the most robust and durable form of plant resistance in nature. Asone of the main genera of rust fungi with economic and biological importance, Puccinia infects almost all cereals but is unable to cause diseases on legumes. Little is known about themechanism of this kind of effective defense in legumes to these non-host pathogens. RESULTS: In this study, the basis of NHR in broad bean (Vicia faba L.) against the wheat stripe rustpathogen, Puccinia striiformis f. sp. tritici (Pst), was characterized. No visible symptomswere observed on broad bean leaves inoculated with Pst. Microscopic observations showedthat successful location of stomata and haustoria formation were significantly reduced in Pstinfection of broad bean. Attempted infection induced the formation of papillae, cell wallthickening, production of reactive oxygen species, callose deposition and accumulation ofphenolic compounds in plant cell walls. The few Pst haustoria that did form in broad beancells were encased in reactive oxygen and callose materials and those cells elicited cell death.Furthermore, a total of seven defense-related genes were identified and found to be upregulatedduring the Pst infection. CONCLUSIONS: The results indicate that NHR in broad bean against Pst results from a continuum of layereddefenses, including basic incompatibility, structural and chemical strengthening of cell wall,posthaustorial hypersensitive response and induction of several defense-related genes,demonstrating the multi-layered feature of NHR. This work also provides useful informationfor further determination of resistance mechanisms in broad bean to rust fungi, especially theadapted important broad bean rust pathogen, Uromyces viciae-fabae, because of strongsimilarities and association between NHR of plants to unadapted pathogens and basalresistance of plants to adapted pathogens.     

两系杂交小麦恢复系MR168抗条锈病基因遗传分析及分子标记定位

小麦条锈病是影响杂交小麦普及推广的重要因素。文章利用基因推导法和SSR分子标记技术,研究了温光型两系杂交小麦恢复系MR168的抗条锈性遗传规律及其控制基因染色体位置。结果表明,MR168对CY29、CY31、CY32、CY33等条锈菌生理小种表现高抗至免疫;对SY95-71/MR168杂交组合的正反交F1、BC1、F2和F3群体分单株接种鉴定显示,MR168对CY32号小种的抗性受1对显性核基因控制,该抗病基因来源于春小麦品种辽春10号。利用集群分离分析法(Bulked segregant analysis,BSA)和简单重复序列(Simple sequence repeat,SSR)分子标记分析抗病亲本MR168、感病亲本SY95-71及183个F2代单株,发现了与MR168抗条锈病基因连锁的5个微卫星标记Xgwm273、Xgwm18、Xbarc187、Xwmc269、Xwmc406,并将该基因初步定位在1BS着丝粒附近,暂命名为YrMR168;构建了包含YrMR168的SSR标记遗传图谱,距离YrMR168最近的两个微卫星位点是Xgwm18和Xbarc187,遗传距离分别为1.9 cM和2.4 cM,这两个微卫星标记可用于杂交小麦抗条锈病分子标记辅助育种。     

Introgression of chromosome 3Ns from Psathyrostachys huashanica into wheat specifying resistance to stripe rust

Wheat stripe rust is a destructive disease in the cool and humid wheat-growing areas of the world. Finding diverse sources of stripe rust resistance is critical for increasing genetic diversity of resistance for wheat breeding programs. Stripe rust resistance was identified in the alien species Psathyrostachys huashanica, and a wheat-P. huashanica amphiploid line (PHW-SA) with stripe rust resistance was reported previously. In this study, a P. huashanica 3Ns monosomic addition line (PW11) with superior resistance to stripe rust was developed, which was derived from the cross between PHW-SA and wheat J-11. We evaluated the alien introgressions PW11-2, PW11-5 and PW11-8 which were derived from line PW11 for reaction to new Pst race CYR32, and used molecular and cytogenetic tools to characterize these lines. The introgressions were remarkably resistant to CYR32, suggesting that the resistance to stripe rust of the introgressions thus was controlled by gene(s) located on P. huashanica chromosome 3Ns. All derived lines were cytologically stable in term of meiotic chromosome behavior. Two 3Ns chromosomes of P. huashanica were detected in the disomic addition line PW11-2. Chromosomes 1B of substitution line PW11-5 had been replaced by a pair of P. huashanica 3Ns chromosomes. In PW11-8, a small terminal segment from P. huashanica chromosome arm 3NsS was translocated to the terminal region of wheat chromosomes 3BL. Thus, this translocated chromosome is designated T3BL-3NsS. These conclusions were further confirmed by SSR analyses. Two 3Ns-specific markers Xgwm181 and Xgwm161 will be useful to rapidly identify and trace the translocated fragments. These introgressions, which had significant characteristics of resistance to stripe rust, could be utilized as novel germplasms for wheat breeding.     

小麦抗条锈基因Yr69的连锁标记开发

抗条锈病基因Yr69对我国小麦条锈菌(Puccinia striiformis f.sp.tritici)小种具有广谱抗性,在小麦抗条锈病育种中具有重要价值.为提高分子标记辅助选择育种的效率,加快Yr69在小麦抗病育种中的应用,本研究利用条锈菌小种CYR34对包含340个小麦家系的'Taichung29/CH7086'...     

Molecular mapping of a non-host resistance gene YrpstY1 in barley (Hordeum vulgare L.) for resistance to wheat stripe rust

Cultivated barley (Hordeum vulgare L.) is considered as a non-host or inappropriate host species for wheat stripe rust caused by Puccinia striiformis f. sp. tritici. Most barley cultivars show a broad-spectrum resistance to wheat stripe rust. To determine the genes for resistance to wheat stripe rust in barley, a cross was made between a resistant barley line Y12 and a susceptible line Y16. The two parents, F(1) and 147 BC(1) plants were tested at seedling stage with Chinese prevalent race CYR32 of Puccinia striiformis f. sp. tritici by artificial inoculation in greenhouse. The results indicated that Y12 possessed one dominant resistance gene to wheat stripe rust, designated YrpstY1 provisionally. A total of 388 simple sequence repeat (SSR) markers were used to map the resistance gene in Y12 using bulked segregant analysis. A linkage map, including nine SSR loci on chromosome 7H and YrpstY1, was constructed using the BC(1) population, indicating that the resistance gene YrpstY1 is located on chromosome 7H. It is potential to transfer the resistance gene into common wheat for stripe rust resistance.     

Role of an endogenous nitric oxide burst in the resistance of wheat to stripe rust

The stripe rust (or yellow rust) disease caused by Puccinia striiformis Westend is a serious disease of wheat in many areas of the world. The role of NO, which is an important redox‐active signalling molecule in plants, was investigated in the wheat‐stripe rust system. The phenotypes from interactions of the same wheat variety, Lovrin10, with two different clones of stripe rust strains (P. striiformis Westend), namely China yellow rust (CY)22‐2 and CY29‐1, which are immune and susceptible reaction types, respectively. The time course of host endogenous NO detected by electron spin resonance indicated that recognition of an avirulent strain was associated with two peaks of NO production. The first peak of NO accumulated in the early infection stage whereas the second peak accumulated in the latent period; however, only a single peak of NO was observed in the latent period for the virulent strain. Furthermore, the activity of pathogen‐related protein‐phenylalanine ammonia‐lyase was higher in the resistant system than in the susceptible system, which suggested that the first NO production was associated with resistance. Exogenous NO improved the activity of phenylalanine ammonia‐lyase and induced a resistant response of Lovrin10 to the virulent strain CY29‐1, thereby providing further evidence that the first peak of NO production was associated with resistance. These results indicate that the first NO burst in the immune system plays an important role in the resistant reaction of wheat to strip rust.     

Molecular mapping of stripe rust resistance gene YrSN104 in Chinese wheat line Shaannong 104

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is a serious yield-limiting factor for wheat production worldwide. The objective of this study was to identify and map a stripe rust resistance gene in wheat line Shaannong 104 using SSR markers. F(1) , F(2) and F(3) populations from Shaannong 104/Mingxian 169 were inoculated with Chinese Pst race CYR32 in a greenhouse. Shaannong 104 carried a single dominant gene, YrSN104. Six potential polymorphic SSR markers identified in bulk segregant analysis were used to genotype F(2) and F(3) families. YrSN104 was closely linked with all six SSR markers on chromosome 1BS with genetic distances of 2.0 cM (Xgwm18, Xgwm273, Xbarc187), 2.6 cM (Xgwm11, Xbarc137) and 5.9 cM (Xbarc240). Pedigree analysis, pathogenicity tests using 26 Pst races, haplotyping of associated markers on isogenic lines carrying known stripe rust resistance genes, and associations with markers suggested that YrSN104 was a new resistance gene or an allele at the Yr24/Yr26 locus on chromosome 1BS. Deployment of YrSN104 singly or in combination to elite genotypes could play an effective role to lessen yield losses caused by stripe rust.     

Isolation and characterization of a wheat IF2 homolog required for innate immunity to stripe rust

Key message

The wheat eIF2 homolog, TaIF2, is induced by the stripe rust pathogen CYR 32 at an early stage of inoculation and is related to the innate immunity resistance level in wheat.

Abstract

The initiation of translation represents a critical control point in the regulation of gene expression in all organisms. We previously identified an upregulated EST S186 (EL773056) from an SSH-cDNA library of the Shaanmai 139 strain of wheat (Triticum aestivum) infected with Puccinia striiformis (Pst). In the present work, we isolated a cDNA clone and identified it as a wheat IF2 homolog. This cDNA consisted of 1,314 nucleotides and contained an open reading frame of 795 nucleotides encoding a polypeptide of 254 amino acids. The amino acids represent a conserved domain in EF-Tu, mtIF2-II, and mtIF2-Ivc. The alignment result showed that it maybe a partial cDNA of the initiation factor 2/eukaryotic initiation factor 5B (IF2/eIF5B) superfamily gene. Paradoxically, results of a Swiss-model analysis suggesting a low QMEAN Z-score implied that it was a membrane protein. Quantitative RT-PCR studies confirmed that the wheat eIF2 (TaIF2) homolog was differentially expressed in three near-isogenic lines. Critical time points for the induction of resistance by inoculation with Pst CYR32 in YrSM139-1B + YrSM139-2D immune resistance genotype occurred at 1 and 3 dpi (days post-infection). RNAi test showed that the inoculated BSMV-IF2 leaves of Shaanmai 139 showed obvious cell death after 15 days of inoculation with CYR 32. qRT-PCR analysis of the target gene in cDNA samples isolated from BSMV-IF2-Pst, BSMV-0-Pst and Pst infected leaves confirmed that the expression of TaIF2 is suppressed by BSMV-IF2 at 3 dpi. This suggested that TaIF2/eIF5B plays an important role in the mechanism of innate immunity to stripe rust pathogen.     

小麦条锈菌国际鉴别寄主Vilmorin23的抗条锈病基因YrV23微卫星标记

Vilmorin23是小麦条锈菌国际鉴别寄主和国际上重要抗源材料。采用SSR技术,利用由Vilmorin23为基因供体转育而成的小麦抗条锈近等基因系Taichung29*6/YrV23,选用YrV23所在2B染色体上的55对SSR引物,对Taichung29*6/ YrV23及其轮回亲本Taichung29和抗性基因供体Vilmorin23的基因组DNA进行PCR扩增和聚丙烯酰胺凝胶电泳分析。结果显示,引物Xwmc356在近等基因系与轮回亲本间扩增出特异性DNA片段,经F₂代群体150个抗、感单株检测证实,该片段位点与抗条锈病基因YrV23有连锁关系,遗传距离为9.4 cM。Xwmc356可作为抗条锈基因YrV23的SSR标记。      

转录因子基因TuGTγ-3参与乌拉尔图小麦对条锈病的抗性

小麦条锈病是由条形柄锈菌小麦专化型(Puccinia striiformis West. f. sp. tritici Eriks.＆Henn., Pst)引起的一种严重的真菌病害,发掘新的抗条锈病相关基因对于小麦抗病育种和抗病机理研究都具有重要意义。Trihelix是植物特有的转录因子家族,参与调控生长发育、形态建成、胁迫应答等过程。迄今,小麦属Trihelix家族与抗条锈病相关的研究尚未见报道。本研究从乌拉尔图小麦(Triticum urartu Tum., 2n=2x=14, AA)中克隆了Trihelix家族GTγ亚家族中的一个基因,命名为TuGTγ-3。序列分析表明,TuGTγ-3基因具有完整的开放阅读框(ORF),编码序列(CDS)全长1329 bp,编码442个氨基酸,推测其编码蛋白的分子量为50.31 kDa,理论等电点为6.12。生物信息学预测TuGTγ-3蛋白有一个单分型核定位信号(GLPMQKKMRYT),没有信号肽和跨膜结构域。TuGTγ-3蛋白的保守trihelix结构域的氨基酸序列位置为Q115~R187,第四α-螺旋位置为F234~Y241,CC结构域的位置为K362~K436。二级结构分析显示,TuGTγ-3蛋白由43.89%的α-螺旋、9.51%的伸展链、9.95%的β-转角和36.65%的不规则卷曲构成。利用普通小麦的基因组数据库BLAST分析表明,TuGTγ-3被定位于5A染色体长臂上。瞬时表达实验显示,TuGTγ-3蛋白主要定位在细胞核中,但细胞质中也有少量分布。表达谱分析表明,TuGTγ-3基因在叶片中的表达量显著高于根和叶鞘,且受小麦条锈菌小种CYR32的诱导而强烈上调表达。进一步通过大麦条纹花叶病毒诱导的基因沉默(BSMV-VIGS)实验证明,转录因子TuGTγ-3正向调控了乌拉尔图小麦对条锈病的抗性。     

Histological and cytological characterization of adult plant resistance to wheat stripe rust

Wheat cultivar Xingzi 9104 (XZ) possesses adult plant resistance (APR) to stripe rust caused by Puccinia striiformis f. sp. tritici (Pst). In this study, histological and cytological experiments were conducted to elucidate the mechanisms of APR in XZ. The results of leaf inoculation experiments indicated that APR was initiated at the tillering stage, gradually increased as the plant aged and highly expressed after boot stage. The histology and oxidative burst in infected leaves of plants at seedling, tillering and boot stages were examined using light microscopic and histochemical methods. Subcellular changes in the host–pathogen interactions during the seedling and boot stages were analyzed by transmission electron microscopy. The results showed that haustorium formation was retarded in the adult plants and that the differentiation of secondary intercellular hyphae was significantly inhibited, which decreased the development of microcolonies in the adult plants, especially in plants of boot stage. The expression of APR to stipe rust during wheat development was clearly associated with extensive hypersensitive cell death of host cells and localized production of reactive oxygen species, which coincided with the restriction of fungal growth in infection sites in adult plants. At the same time, cell wall-related resistance in adult plants prevented ingression of haustorial mother cells into plant cells. Haustorium encasement was coincident with malformation or death of haustoria. The results provide useful information for further determination of mechanisms of wheat APR to stripe rust. Key message The expression of APR to stipe rust in wheat cultivar Xingzi 9104 (XZ) was clearly associated with extensive hypersensitive cell death of host cells and the localized production of reactive oxygen species.     

Over-expression of the cell death regulator BAX inhibitor-1 in barley confers reduced or enhanced susceptibility to distinct fungal pathogens

BAX inhibitor-1 (BI-1) is a conserved cell death regulator protein that inhibits mammalian BAX-induced cell death in yeast, animals and plants. Additionally, HvBI-1 suppresses defense responses and resistance to the powdery mildew fungus Blumeria graminis f.sp. hordei (Bgh) when over-expressed in single epidermal cells of barley. To test the potential of ectopic expression of BI-1 to influence fungal interactions with crop plants, we produced stable transgenic barley plants expressing a green fluorescing protein (GFP) fusion of HvBI-1 under control of the cauliflower mosaic virus 35S promoter. GFP-HvBI-1 plants were fertile and did not display obvious developmental alterations when compared to wild type parents. GFP-HvBI-1 plants were more resistant to single cell death induced by ballistic delivery of a mammalian proapototic BAX expression construct and more susceptible to biotrophic Bgh. Microscopic observation of the interaction phenotype revealed that enhanced susceptibility, i.e. a higher degree of successful establishment of haustoria in epidermal cells, was associated with a reduced frequency of hypersensitive cell death reactions. In contrast, young seedlings of GFP-HvBI-1 barley were more resistant to Fusarium graminearum than wild type or azygous controls. Hence the effect of GFP-HvBI-1 on the outcome of a particular plant–fungus interaction appeared dependent on the lifestyle of the pathogen. V. Babaeizad and J. Imani contributed equally to this study.     

Development and characterization of wheat-Psathyrostachys huashanica partial amphiploids for resistance to stripe rust

Two partial amphiploid lines, B113 (32 plants) and B21 (13 plants), derived from a wheat-Psathyrostachys huashanica intergeneric cross were characterized by Giemsa C-banding and SDS-PAGE and evaluated for stripe rust resistance. All 15 partial amphiploid plants were aneuploids with either 50 (8 plants), 51 (6 plants) or 54 (1 plant) chromosomes. Some showed regular meiosis and all the P. huashanica chromosomes were included, although not in a single plant. Of 45 plants 34 showed specific bands on SDS-PAGE representing high molecular weight glutenin subunit (HMW-GS) and 41 had bands representing P. huashanica low molecular weight glutenin subunit (LMW-GS), including two new subunits. All 45 plants were highly resistant (10) or immune (35) to stripe rust mixed races CYR-30, CYR-31, Shuiyuan 7 and Shuiyuan 14. These amphiploid plants could be useful germplasm for enhancing stripe rust resistance and might improve wheat grain quality.     

小麦抗条锈病基因YrCN19的诊断检测和遗传分析

用小麦条锈病抗性基因YrCN19的诊断标记Xgwm410对19个小麦品种或品系进行PCR筛选,其中川农19、新抗5号、爱民5号和爱民6号扩增出与条锈病抗性基因YrCN19共分离的特征片断,其大小为391个碱基,而在其他的小麦品种或品系中未能检测到该片断。系谱分析和抗性鉴定结果表明川农19、新抗5号、爱民5号和爱民6号含有小麦条锈病基因YrCN19。抗性遗传分析发现小麦条锈病抗性在川农19,新抗5号和爱民5号中的遗传符合单个显性基因的遗传规律(3抗:1感);杂交组合烟辐188/爱民6号的抗性遗传也符合单个显性基因的遗传规律,而另外一些杂交组合(如R25/爱民6号,鲁955159/爱民6号和苏3110/爱民6号)中的抗性分离则符合两对基因互补的遗传规律(9抗:7感)。本研究揭示了小麦条锈病抗性基因YrCN19在不同遗传背景和杂交组合的抗性表达和分离有差异,从而加速YrCN19在小麦抗条锈育种中的开发与利用。     

The 3Ns chromosome of Psathyrostachys huashanica carries the gene(s) underlying wheat stripe rust resistance

Stripe rust (Puccinia striiformis tritici (Pst)) is one of the most destructive diseases of wheat in the world. Exploiting and utilizing stripe rust resistance genes of wild species has become an essential strategy for resistance breeding. Psathyrostachyshuashanica Keng ex Kuo is a wild species in Triticeae that has been used for wheat improvement because of its high resistance or immunity to stripe rust. In this study, 9 wheat-P. huashanica addition lines were characterized by Giemsa C-banding, genomic in situ hybridization (GISH), and disease resistance evaluation. Giemsa C-banding and GISH demonstrated that lines 163-5, 165-1, 183-5, 240-3, and 240-4 are P. huashanica 3Ns chromosome monosomic addition lines; lines 183-1 and 183-20 are P. huashanica 3Ns chromosome disomic addition lines; line 165-20 is a P. huashanica 3Ns and 4Ns chromosomes double disomic addition line, and line 219-1 is a P. huashanica 1Ns and 3Ns/5A chromosomes double disomic addition-substitution line. All these addition lines with P. huashanica 3Ns chromosome(s) expressed high resistance or immunity to stripe rust. By comparing the series of wheat-P. huashanica chromosome addition lines, we concluded that the P. huashanica 3Ns chromosome carries the gene(s) for resistance or immunity to stripe rust. These addition lines can be used as a donor source of novel stripe rust resistance to wheat breeding programs.     

The Bax lnhibitor-1 needs a functional electron transport chain for cell death suppression

Bax inhibitor-1 (BI-1) is an evolutionarily conserved cell death suppresser in animals, yeast, and plants. In this study, yeast strains carrying single-gene deletions were screened for factors related to cell death suppression by Arabidopsis BI-1 (AtBI-1). Our screen identified mutants that failed to survive Bax-induced lethality even with AtBI-1 coexpression (Bax suppressor). The Deltacox16 strain was isolated as a BI-1-inactive mutant; it was disrupted in a component of the mitochondrial cytochrome c oxidase. Other mutants defective in mitochondrial electron transport showed a similar phenotype. ATP levels were markedly decreased in all these mutants, suggesting that BI-1 requires normal electron transport activity to suppress cell death in yeast.     

1994 - 2002年小麦品种(系)抗条锈性鉴定与监测

1994—2002年经对3822份小麦品种(系)材料抗条锈性鉴定结果表明,冬小麦抗条锈性优于春小麦,甘肃品种抗条锈性优于国内其它省区品种。田间抗条锈性监测结果表明,我国主要生产品种均表现感病,甘肃主要生产品种仅陇鉴127等少数几个品种抗病,抗源材料中也仅有中四等少数品种表现抗病,结合抗病性鉴定、监测结果及田间综合农艺性状观察,筛选出20余份可供育种利用的抗源材料。同时在针对今后抗条中31、32号等主要小种类型的抗病育种、抗病性监测等方面进行了讨论。     

小麦与条锈菌互作中过氧化物酶的细胞化学研究

采用细胞化学方法对小麦与条锈菌互作过程中过氧化物酶的分布及其活性大小进行了研究,结果表明：过氧化物酶主要分布于细胞壁和细胞间隙中;在未行接种的小麦叶片中,抗病品种和感病品种的过氧化物酶活性均比较低;条锈菌侵染后,诱导抗、感病品种叶片中的过氧化物酶活性升高,且抗病品种升高的幅度明显大于感病品种;感病品种中过氧化物酶活性在侵染位点附近细胞壁上表现升高,而抗病品种中该酶的活性在侵染点细胞以及远离侵染点的叶肉细胞的细胞壁和细胞间隙中均显著升高。高活性的过氧化物酶是小麦抗条锈性的生化标记和重要机制之一。