Molecular Detection of Resistance Genes to Stripe Rust and Powdery Mildew in Common Wheat Cultivar Yunmai52

Yunmai52, developed by crossing with common wheat-Haynaldia villosa6AL/6VS translocation line 92R149 as a resistant parent in 1992, was a common wheat cultivar approved and released in 2007 in Yunnan Province, China, which is characterized by high resistance to powdery mildew and stripe rust. In this study,an F_2 population derived from a cross K78S/Yunmai52 was constructed to investigate the resistance genes, where K78 S is a wheat male sterile line susceptible to powdery mildew and stripe rust. Phenotypic identification of the parents, F_1 and F_2 populations and chi-square analyses showed that F_1 population was immune to stripe rust and powdery mildew; the segregation ratio of resistance and susceptibility to powdery mildew（χ~2=1.10χ~2_（1,0.05）=3.84） and stripe rust（χ~2=0.15χ~2_（1,0.05）=3.84） fit to a 3：1 ratio in F_2 population, indicating that Yunmai52 harbors a dominant stripe rust resistance gene and a dominant powdery mildew resistance gene. The individuals were further detected with a marker co-segregated with Pm21（SCAR_（1400）） and two markers closely linked with Yr26（XWe173 and Xbarc181）. The results showed that polymorphic bands could be amplified between the parents and between resistance and susceptibility gene pools at the same locus. Randomly 96 individuals of F_2 population were selected for verification. The results showed that the phenotype was significantly correlated with the genotype. The detection accuracy of markers SCAR_（1400）, XWe173 and Xbarc181 was 100%, 97.91% and 92.70%, respectively.Yunmai52 harbored powdery mildew resistance gene Pm21 and stripe rust resistance gene Yr26, which were both derived from 6AL/6VS translocation line 92R149.In addition, the results also demonstrate that Pm21 and Yr26 are two genes conferring durable resistance to powdery mildew and stripe rust in wheat.

Identification of Co-Segregating RAPD Marker Linked to Powdery Mildew Resistance Gene Pm18 in Wheat

The Pm18 gene of wheat confers resistance to the powdery mildew which is one of the mostserious diseases in many regions of the world. In this study, bulked segregant analysis(BSA) was used to develop randomly amplified polymorphic DNA (RAPD) markers linked toPm18 gene. Three hundred and twenty decamer primers were screened and one of them wasidentified as RAPD marker (S411600) linked to Pm18. Using the F2 mapping population fromthe cross Pm18Chancellor, the marker S411600 was shown to co-segregate with the genePm18. This marker can be conveniently used for marker-assisted selection in wheatbreeding programs for the identification or pyramiding of Pm18 with other resistancegenes.

IDENTIFICATION OF RAPD MARKER LINKED TO POWDERY MILDEW RESISTANCE GENE Pm12 IN WHEAT

Powdery mildew is one of the most serious diseases of wheat in China. In this paper,bulked segregant analysis (BSA) was used to search for randomly amplified polymorphic DNA (RAPD) markers linked to the Pm12 gene,which confers resistance to the powdery mildew in wheat. 200 decamer primers were screened and one RAPD marker (S107 1900 ) was identified to be linked to Pm12 in coupling phase,and their genetic distance is 11.98± 4.00cM. This marker can be used for marker-assisted selection in wheat breeding for the identification or pyramiding of Pm12 with other resistance genes.

The Relationship of Methyl Jasmonate Enhanced Powdery Mildew Resistance in Wheat and the Expressions of 9 Disease Resistance Related Genes

[Objective] This study was carried out to determine the induction effect of jasmonic acid（JA）on powdery mildew resistance in wheat,the activation effect on the expressions of plant disease resistance related genes,and to investigate the relationship between the induced resistance and the gene expression patterns.[Method] Three powdery mildew susceptible cultivars of ＂Chinese Spring＂,＂Pumai 9＂ and ＂Zhoumai 18＂ typically representing different phenotypes in the field were employed.The powdery mildew was assessed by detached leaf assay,and real time quantitative RT-PCR was used to determine the expression patterns of 9 disease resistance related genes of PR1（PR1.1）,PR2（β,1-3 glucanase）,PR3（chitinase）,PR4（wheatwin1）,PR5（thaumatin-like protein）,PR9（TaPERO,peroxidase）,PR10,TaGLP2a（germin-like）and Ta-JA2（jasmonate-induced protein）in leaf of the three cultivars.[Result] MeJA application enhanced the powdery mildew resistances of ＂Chinese Spring＂,＂Pumai 9＂ and ＂Zhoumai 18＂.The induced powdery mildew resistance could be detected from 12 h to 96 h after MeJA treatment,and the peak value was at 24 h.Though there were differences between the three cultivars,MeJA significantly effect on the expressions of the 8 disease resistance related genes except TaGLP2a,and the peak values were at 12 h,24 h or 48 h after treatments.The strongest activation of MeJA was on PR9 and PR1 that their expressions could reach more than 100 times of the untreated samples.MeJA strongly activated PR2、PR4、PR5、PR3、PR10 and Ta-JA2,their expression could reach 10 to 70 times,and there was almost no activation effect on TaGLP2a.The induced powdery mildew resistance positively correlated with the induced expressions of the 8 disease related genes.[Conclusion] The induced powdery mildew resistance positively correlated with the induced expressions of the disease related genes.Jasmonate signalling plays a role in defence against Blumeria graminis f.sp.tritici.and future manipulation of this pathway may improve powdery mildew resistance in wheat.

Genetics,resistance mechanism,and breeding of powdery mildew resistance in cucumbers(Cucumis sativus L.)

Cucumber is an important vegetable worldwide,and powdery mildew(PM)is a common and serious disease of cucumbers.Breeding disease-resistant cucumber varieties is the most advantageous strategy to control this disease.In recent years,exploration and identification of cucumber PM resistance genes have achieved great advancement,and many genes have been cloned and verified using different methods.However,the resistance mechanism of cucumber PM is still unclear,and many ambiguities need to be elucidated urgently.In this review,we summarized the research advances in PM resistance in cucumbers,including genetic analysis,quantitative trait locus mapping,map-based cloning,transcriptomics,mlo-mediated PM resistance,and mining of noncoding RNAs involved in resistance.Finally,the research directions and the problems that need to be solved in the future were discussed.

Molecular and Physical Mapping of Powdery Mildew Resistance Genes and QTLs in Wheat: A Review

Wheat powdery mildew （Pro） is a major disease of wheat worldwide. During the past years, numerous studies have been published on molecular mapping of Pm resistance gene（s） in wheat. We summarized the relevant findings of 89 major re- sistance gene mapping studies and 25 quantitative trait loci （QTL） mapping studies. Major Pm resistance genes and QTLs were found on all wheat chromosomes, but the Pm resistance genes/QTLs were not randomly distributed on each chromosome of wheat. The summarized data showed that the A or B genome has more major Pm resistance genes than the D genome and chromosomes 1A, 2A, 2B, 5B, 5D, 6B, 7A and 7B harbor more major Pm resistance genes than the other chromosomes. For adult plant resistance （APR） genes/QTLs, B genome of wheat harbors more APR genes than A and D genomes, and chromo- somes 2A, 4A, 5A, 1B, 2B, 3B, 5B, 6B, 7B, 2D, 5D and 7D harbor more Pm resistance QTLs than the other chromosomes, suggesting that A genome except 1A, 3A and 6A, B genome except 4B, D genome except 1D, 3D, 4D, and 6D play an impor- tant role in wheat combating against powdery mildew. Furthermore, Pm resistance genes are derived from wheat and its rela- tives, which suggested that the resistance sources are diverse and Pm resistance genes are diverse and useful in combating against the powdery mildew isolates. In this review, four APR genes, Pm38/Lr34/Yr18/Sr57, Pm46/Lr67/Yr46/Sr55, Pm？/Lr27/Yr30/ SY2 and Pm39/Lr46/Yr29, are not only resistant to powdery mildew but also effective for rust diseases in the field, indicating that such genes are stable and useful in wheat breeding programmes. The summarized data also provide chromosome locations or linked markers for Pm resistance genes/QTLs. Markers linked to these genes can also be utilized to pyramid diverse Pm resis- tance genes/QTLs more efficiently by marker-assisted selection.

Genetic dissection of the powdery mildew resistance in wheat breeding line LS5082 using BSR-Seq

Powdery mildew of wheat is a destructive disease seriously threatening yield and quality worldwide.Comprehensive dissection of new resistance-related loci/genes is necessary to control this disease.LS5082 is a Chinese wheat breeding line with resistance to powdery mildew.Genetic analysis,using the populations of LS5082 and three susceptible parents(Shannong 29,Shimai 22 and Huixianhong),indicated that a single dominant gene,tentatively designated PmLS5082,conferred seedling resistance to different Blumeria graminis f.sp.tritici(Bgt)isolates.Bulked segregant RNA-Seq was carried out to map PmLS5082 and to profile differentially expressed genes associated with PmLS5082.PmLS5082 was mapped to a 0.7 cM genetic interval on chromosome arm 2BL,which was aligned to a 0.7 Mb physical interval of 710.3–711.0 Mb.PmLS5082 differs from the known powdery mildew(Pm)resistance genes on chromosome arm 2BL based on their origin,chromosome positions and/or resistance spectrum,suggesting PmLS5082 is most likely a new Pm gene/allele.Through clusters of orthologous groups and kyoto encyclopedia of genes and genomes analyses,differentially expressed genes(DEGs)associated with PmLS5082 were profiled.Six DEGs in the PmLS5082 interval were confirmed to be associated with PmLS5082 via qPCR analysis,using an additional set of wheat samples and time-course analysis postinoculation with Bgt isolate E09.Ten closely linked markers,including two kompetitive allele-specific PCR markers,were confirmed to be suitable for marker-assisted selection of PmLS5082 in different genetic backgrounds,thus can be used to detect PmLS5082 and pyramid it with other genes in breeding programs.

Fine mapping of powdery mildew resistance gene PmTm4 in wheat using comparative genomics

Powdery mildew,caused by Blumeria graminis f.sp.tritici,is one of the most severe wheat diseases.Mining powdery mildew resistance genes in wheat cultivars and their appliance in breeding program is a promising way to control this disease.Genetic analysis revealed that a single dominant resistance gene named PmTm4 originated from Chinese wheat line Tangmai 4 confers resistance to prevailing isolates of B.graminis f.sp.tritici isolate E09.Detailed comparative genomics analyses helped to develop closely linked markers to PmTm4 and a fine genetic map was constructed using large F2population,in which PmTm4 was located into a 0.66-c M genetic interval.The orthologous subgenome region of PmTm4in Aegilops tauschii was identified,and two resistance gene analogs（RGA）were characterized from the corresponding sequence scaffolds of Ae.tauschii draft assembly.The closely linked markers and identified Ae.tauschii orthologs in the mapping interval provide an entry point for chromosome landing and map-based cloning of PmTm4.

The Potential Role of Powdery Mildew-Resistance Gene Pm40 in Chinese Wheat-Breeding Programs in the Post-Pro21 Era

Powdery mildew, which is caused by Blumeria graminis f. sp. tritici （Bgt）, is an important leaf disease that affects wheat yield. Powdery mildew-resistance （Pro） gene Pro21 was first transferred into wheat in the 1980s, by translocating the Heuchera villosa chromosome arm 6VS to the wheat chromosome arm 6AL （6VS.6AL）. Recently, new Bgt isolates that are virulent to Pm21 have been identified in some wheat fields, indicating that wheat breeders should be aware of the risk of deploying Pm21, although pathological details regarding these virulent isolates still remain to be discovered. Pm40 was identified and mapped on the wheat chromosome arm 7BS from several wheat lines developed from the progenies of a wild cross between wheat and Thinopyrum intermedium. Pm40 offers a broad spectrum of resistance to Bgt, which suggests that it is likely to provide potentially durable resistance. Cytological methods did not detect any large alien chromosomal segment in the wheat lines carrying Pm40. Lines with Pm40 and promising agronomical traits have been released by several wheat-breeding programs in the past several years. Therefore, we believe that Pm40 will play a role in powdery mildew-resistance wheat breeding after Pm21 resistance is overcome by Bgt isolates. In addition, both Prn21 and Pm40 were derived from alien species, suggesting that the resistance genes derived from alien species are potentially more durable or effective than those identified from wheat.

Characterization of RAPD Markers,and the RFLP Marker Linked to Powdery Mildew Resistance Gene Derived from Different Accessions of H.villosa

The analysis was carried out on performance of the resistance gene from Haynaldia villosa accession of the former Soviet Union to different isolates of Bluemerie graminis. Polymorphisms were revealed between 6D/6V substitution line Pm930640 and its pedigree parents using five RAPD markers of OPAN031700, OPAI01700, OPAL03750, OPAD07480 and OPAG15580 screened out from 120 random 10-mers primers. Three RAPD markers of OPAN03, OPAI01 and OPAL03 were linked with the resistance gene by analysis of F2 population of Chancellor×Pm930640. Analysis of 29 wheat lines including part of lines conferring the known genes from Pm1 to Pm20 respectively, lines conferring resistance gene from two H. villosa accessions and the related wheat parents, were analyzed and the results showed that these markers not only linked to the gene resistant to powdery mildew from H. villosa, but also detected different genetic backgrounds. OPAL03750 can be used as the marker to distinguish the different resistant lines from two H. villosa accessions because it was only observed in the materials from H. villosa of the former Soviet Union. RFLP analysis also showed the polymorphisms between two H. villosa accessions and their derived resistant lines.

Fine mapping of an adult-plant resistance gene to powdery mildew in soybean cultivar Zhonghuang 24

Powdery mildew(PM),caused by the fungus Microsphaera diffusa,causes severe yield losses in soybean[Glycine max(L.)Merr.]under suitable environmental conditions.Identifying resistance genes and developing resistant cultivars may prevent soybean PM damage.In this study,analysis of F_(1),F_(2),and F8:11 recombinant inbred line(RIL)populations derived from the cross between Zhonghuang 24(ZH24)and Huaxia 3(HX3)indicated that adult-plant resistance(APR)to powdery mildew in the soybean cultivar(cv.)ZH24 was controlled by a single dominant locus.A high-density genetic linkage map of the RIL population was used for fine mapping.The APR locus in ZH24 was mapped to a 281-kb genomic region on chromosome 16.Using 283 susceptible plants of another F2 population,the candidate region was finemapped to a 32.8-kb genomic interval flanked by the markers InDel14 and Gm16_428.The interval harbored five genes,including four disease resistance(R)-like genes,according to the Williams 82.a2.v1 reference genome.Quantitative real-time PCR assays of candidate genes revealed that the expression levels of Glyma.16g214300 and Glyma.16g214500 were changed by M.diffusa infection and might be involved in disease defense.Rmd_B13 showed all-stage resistance(ASR)to PM in soybean cv.B13.An allelism test in the F2 segregating population from the cross of ZH24 × B13 suggested that the APR locus Rmd_ZH24 and the ASR locus Rmd_B13 may be allelic or tightly linked.These results provide a reference marker-assisted selection in breeding programs.

Comparative genetic mapping revealed powdery mildew resistance gene MlWE4 derived from wild emmer is located in same genomic region of Pm36 and Ml3D232 on chromosome 5BL

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most devastating wheat diseases. Wild emmer wheat（Triticum turgidum ssp. dicoccoides） is a promising source of disease resistance for wheat. A powdery mildew resistance gene conferring resistance to B. graminis f. sp. tritici isolate E09, originating from wild emmer wheat, has been transferred into the hexaploid wheat line WE4 through crossing and backcrossing. Genetic analyses indicated that the powdery mildew resistance was controlled by a single dominant gene, temporarily designated Ml WE4. By mean of comparative genomics and bulked segregant analysis, a genetic linkage map of Ml WE4 was constructed, and Ml WE4 was mapped on the distal region of chromosome arm 5BL. Comparative genetic linkage maps showed that genes Ml WE4, Pm36 and Ml3D232 were co-segregated with markers XBD37670 and XBD37680, indicating they are likely the same gene or alleles in the same locus. The co-segregated markers provide a starting point for chromosome landing and map-based cloning of Ml WE4, Pm36 and Ml3D232.

Resistance Analysis of Wheat Cultivars against Powdery Mildew in Anhui Province of China

[ Objective ] The paper was to determine the resistance level of the tried and pre-examination wheat cultivars against powdery mildew in Anhui Province of China. [ Method ] By using artificial inoculation and identification method in fields, the resistance of wheat cultivars was identified in consecutive three years from 2010 to 2012. [ Result] The highly susceptible （HS） cultivar accounted for 30%, 42% and 11% of total tested cultivars in the years of 2010, 2011 and 2012, respectively; moderately susceptible （MS） cultivar accounted for 53% of total tested cuhivars in 2010, which accounted for 47% and 57% in 2011 and 2012, respectively; moderately resistant （MR） cuhivar accounted for 17% of total tested cultivars in 2010, which accounted for 11% and 32% in 2011 and 2012, respectively. [ Conclusion] The paper can guide breeding direction, and also provide scientific basis for variety approval.

Identification of Resistance of Powdery Mildew and Stripe Rust of Winter Wheat Strains in Xinjiang

Wheat powdery mildew and stripe rust are the major diseases in wheat producing area in Xinjiang.To obtain wheat germplasm resources and varieties resistant to powdery mildew and rust,36 high-generation stable strains of Xinjiang winter wheat were evaluated using the method of natural inducement from 2018 to 2020.A total of 5 strains with high resistance to powdery mildew,4 strains with slow stripe rust and 1 strain with resistance to powdery mildew and adult plant slow stripe rust were obtained.And the parental combination of disease-resistant varieties was analyzed.These studies will provide theoretical basis for the breeding of resistant wheat varieties in Xinjiang.

Genome Analysis in Wheat Breeding for Disease Resistance

A brief review on the development of wheat germplasm with introduced powdery mildew and scab resistance from Haynaldia villosa Sch. and Leymus racemosus Lam., Roegneria ciliaris (Trin.) Nevski as well as R. kamoji C. Koch respectively was made. In the course of germplasm development, genome analysis by means of chromosome banding, genomic in situ hybridization (GISH) or fluorescence in situ hybridization (FISH), molecular markers, particularly restriction fragment length polymorphism (RFLP) coupled with aneuploid analysis was employed for the purpose of improving breeding efficiency. Potential use of such germplasm in wheat breeding practice, basic studies and some related problems were also discussed.

Gene expression profiling related to powdery mildew resistance in wheat with the method of suppression subtractive hybridization

'Bainong 3217 × Mardler' BC5F4 wheat line at the initial stage of inoculation with powdery mildew pathogen (Erysiphe graminis DC) was used to construct a suppression subtractive hybridization (SSH) cDNA library. Totally 760 ESTs were obtained through sequencing. Similarity analysis of ESTs based on BLASTn and BLASTx with the sequences in GenBank, in combination with macroarray differential screening, revealed that 199 ESTs of 65 kinds were known to be functionally disease resistance related. Based on the gene expression profiling in the present study, it is postulated that salicylic acid (SA) and MAP-related signal transduction pathways were involved in powdery mildew resistance in wheat. System acquired resistance genes were predominant in terms of kinds and quantity. With the initiation of cell defense reaction, the genes conferring anti-oxidation substances were largely expressed and thus cell protection mechanism was activated. Much evidence revealed that phenylpropanes metabolic pathway was

小麦四个抗白粉病基因的KASP标记检测

为提高Pm13的检测效率,了解Pm21、PmV、Pm12和Pm13共4个小麦抗白粉病基因在山东小麦育种中的利用情况,本研究通过测序和引物设计成功转化了Pm13的KASP标记,并采用4个基因的KASP标记对2022—2023年山东省小麦高产区试组和强筋区试组的140份参试品系和2份区试对照品种进行检测,结果显示140份材料均不含上述4个基因,表明它们在山东小麦育种中应用较少。本研究可为利用KASP标记开展小麦抗白粉病基因的分子标记辅助选择提供参考,促进其育种应用。

腥掷孢酵母17wy1的分离、鉴定及对小麦白粉病防效初探

采用26S rDNA序列对分离自异常生长小麦白粉菌分生孢子上的菌株17wy1进行鉴定,明确其分类地位;通过孢子萌发抑制试验、室内盆栽和田间小麦白粉病防治试验测定17wy1发酵液对小麦白粉菌分生孢子萌发的影响及对小麦白粉病的生防效果,并通过定量PCR技术分析小麦病程相关蛋白基因PR1、PR2、PR5、TaPAL的表达模式,解析17wy1的拮抗机制,为小麦白粉病的生物防治提供依据。结果表明,菌株17wy1为腥掷孢酵母(Golubevia albescens),其发酵液(5×107个/mL)对小麦白粉菌分生孢子萌发抑制率为85.1%,对室内盆栽麦苗和田间小麦白粉病的防效分别为58.5%和69.8%;喷施酵母17wy1发酵液后,室内显症麦苗体内的病程相关蛋白基因PR1、PR5被诱导显著上调表达。可见,酵母菌17wy1可抑制小麦白粉菌分生孢子萌发,对室内盆栽麦苗和田间小麦白粉病具有较好的防治作用,是小麦白粉病的潜在生防菌。

辣椒白粉病抗性主基因+多基因混合遗传分析

辣椒白粉病是一种常见的真菌病害,该病严重影响辣椒生产。以辣椒白粉病高抗材料(NSR和NVD)为母本,高感材料(20c89、20c93)为父本,分别构建两组P_(1)、P_(2)、F_(1)、F_(2)四世代遗传群体(2021组和2022组),通过调查统计辣椒不同世代群体内单株的白粉病病情指数,利用植物数量性状主基因+多基因混合遗传分析法对辣椒白粉病抗性进行遗传分析。结果表明:两组群体辣椒白粉病抗性遗传均受两对加性-显性上位性主基因+加性-显性多基因控制,并且基因遗传效应和遗传力等参数在两组群体中表现出较高的一致性。2021组第1对主基因加性效应和显性效应值分别为12.7065和-34.7107,第2对主基因加性效应和显性效应值分别为-9.1706和-16.9432,主基因遗传率为86.11%;2022组第1对主基因加性效应和显性效应值分别为20.0431和-18.0682,第2对主基因加性效应和显性效应值分别为-1.0668和-6.0904,主基因遗传率为97.16%。

小麦新品种品育8012白粉病抗性研究

品育8012是由山西农业大学小麦研究所选育,2018年通过山西省审定,亲本组合临优20165/济麦22,2020-2022年连续三年被列为山西省农业生产主推品种,该品种近免疫白粉病。为了解品育8012白粉病抗性,先进行亲本已知抗病基因检测,继而进行其它抗病基因检测,随后在山西、山东和河北3省15点田间鉴定进行验证试验。得出品育8012携带13个白粉病抗性基因,在山西、山东和河北多点对白粉病表现中抗乃至近免疫。品育8012可作为选育抗白粉病小麦品种的抗源加以利用。