Study on the Biology and Epidemiology of Uncinula necator---The Causal Agent of Grape Powdery Mildew Disease

This study was carried out during 2007-2009 in the vineyards of Ardebil province of lran to study the biology and the epidemiology of Uncinula necator--the causal agent of grape powdery mildew disease. The study concentrated on the survival and the initiation of primary inoculum of the fungal causal agent. Results of histopathological experiments indicated that U. necator survived as mycelium in the dormant buds of the grapes during winter season. Results of study on the effect of environmental factors on fungus biology showed that the pathogenic activity of the fungus began when the temperature was between 16-19 ℃ with a relative humidity more than 50%. It was also found that optimum temperature and relative humidity for the sporulation of U.necator was 20-25 ~C and 50%-100% respectively. According to the results, fungal conidia were trapped during formation of 5-6 true leaves and first disease symptoms were observed on the clusters on late June after fruit formation. Fungal cleistothecia were observed abundantly at the end of season on the leaves, petioles and twigs but they were not able to survive during winter. Formation of ascospores on young leaves was proved but their role as the primary inoculum was not supported by the results of this study. Results of this study and the new findings on the biology and epidemiology of U.necator may be of national and international interests for the management of powdery mildew disease which is one of the most destructive diseases around the world including Iran.

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.

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.

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.

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.

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.

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.

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.

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.

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.

基于长尖叶蔷薇与大花香水月季转录组数据的MLO unigenes生物信息学及表达分析

在对长尖叶蔷薇和大花香水月季转录组测序的基础上,利用生物信息学方法,从其转录组数据中共获得23条MLO unigenes并对其进行了分析。结果显示它们之间的氨基酸数量、碱基数、分子量差异较小,多数为疏水性蛋白,平均亲水系数为-0.157~0.190,富含亮氨酸和丝氨酸。亚细胞定位主要分布于质膜中,包含5~9个跨膜结构;二级结构主要由α-螺旋组成。其中,等电点小于7的仅有1条,2条蛋白具有信号肽。这23条MLO蛋白具有15个长度在15~50个氨基酸间的保守基序。它们的系统进化关系分析结果揭示了MLO基因在长尖叶蔷薇与大花香水月季间具有较高的同源性和保守性。将这23条unigenes与来自于月季和拟南芥的MLO基因进行聚类分析,确定了6条可能参与抗白粉病的候选基因,对这6条候选基因进一步分析发现仅有2条unigenes符合MLO型白粉病基因的典型结构特征。最后,通过q RT-PCR试验验证,结果表明:这2条候选基因的相对表达量,在受到白粉病菌侵染后呈积极上调趋势。上述研究结果表明这2条候选基因很可能参与了寄主—白粉病菌的互作过程。

Genome-wide identification and evolutionary analysis of MLO gene family in Rosaceae plants

Mutants lacking wild-type MLO(Mildew resistance Locus O)proteins show broad-spectrum resistance to the powdery mildew fungus,and dysregulated cell death control,with spontaneous cell death in response to developmental or abiotic stimuli.In order to understand the evolution and divergence patterns of the MLO gene family in Rosaceae plants,we analysed systematically genome-wide data from Fragaria vesca,Prunus persica,Prunus mume,Malus domestica,Pyrus bretschneideri and Rubus occidentalis based on bioinformatics methods.Using three phylogenetic methods(the neighbour-joining,maximum likelihood,and Bayesian methods),we identified 117 MLO genes from 6 Rosaceae species.The results of all three phylogenetic analysis methods supported that these genes were divided into six clades.Conserved motif analysis found that only motif 2 was present in all MLO proteins and had 3 nearly invariant amino acid residues.The findings indicated that motif 2 might be shared by the MLO gene family.The structural features of these genes showed large variations in sequence length among different species,although the lengths and the numbers of exons exhibited high degrees of similarity.Selective pressure analysis showed extremely significant differences in all 6 clades,with 2,1,and 1 site(s)under significant positive selection detected in clades III,IV,and VI,respectively.These positive selection sites were important driving forces for the promotion of the functional differentiation of the MLO genes.Functional divergence analysis showed that the significantly divergent sites were located within the domains of the MLO genes.Functional distance analysis showed that the clade V had more conservative functions and might have retained more original functions during the evolutionary process.However,clade I may have undergone extensive altered functional constraints as a specialised functional role.Moreover,the most original function of the MLO genes in Rosaceae could be related to the evolution of their resistance to powdery mildew,which then gradually evolved into functions such as the regulation of flower development,the control of root morphology,and seed evolution due to the different evolutionary rates after gene duplication.These results provide a theoretical basis for further studies of the molecular evolutionary patterns of the plant MLO gene family.

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。

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。

俄罗斯春小麦抗白粉病基因检测及其组成分析

为了解俄罗斯春小麦抗白粉病基因的组成及分布规律,利用已开发的Pm2、Pm3b、Pm4、Pm8、Pm13和Pm21标记对外引俄罗斯251份春小麦品种进行了检测和分析。结果表明,在251份小麦品种中,分布5种抗白粉病基因,其中Pm2和Pm3分布频率较高,均为90.44%;Pm13为57.77%,抗病基因Pm4分布频率为10.36%,Pm8基因的分布频率为5.58%,抗病基因Pm21在引入的俄罗斯春小麦中没有分布,有6份材料没有检测出含有上述基因标记。俄罗斯251份春小麦品种中小麦抗白粉病基因组合共有15种类型,其中116份小麦品种以Pm2/Pm3/Pm13类型所占比例为46.22%;其他14种类型所占比例依次为,Pm2/Pm3类型比例为23.90%;Pm2/Pm3/Pm4类型比例为4.78%;Pm2/Pm3/Pm4/Pm13类型比例为4.38%;Pm2类型比例为3.58%;Pm3和Pm2/Pm13类型比例均为2.79%;Pm3/Pm13和Pm2/Pm3/Pm8类型比例均为2.39%;Pm2/Pm3/Pm4/Pm8和Pm2/Pm3/Pm8/Pm13类型比例均为1.20%;Pm13类型比例为0.79%;Pm2/Pm8、Pm3/Pm4/Pm13和Pm3/Pm8/Pm13比例为0.40%;本研究明确了俄罗斯春小麦品种抗白粉病基因的组成和分布频率,可进一步利用外引材料开展抗白粉病育种。

农家豌豆品种白粉病抗性基因分子标记筛查

为了获得农家豌豆抗白粉病品种,利用分子标记技术,对29份农家豌豆品种白粉病的4个抗性基因连锁的7个标记做了筛查,并对其抗性进行预测。结果表明,经PCR扩增,与豌豆白粉病抗性基因连锁的7个分子标记获得了650,1600,1400,637,880,1200和874 bp的目的条带;7个标记的分布频率均较高,在65%~90%之间,但分布频率不一;29份供试豌豆材料,只有清水河山豌豆不含豌豆白粉病抗性基因连锁标记,其余28个品种均检测到含1~7个标记,其中,和林灰绿野豌豆和张掖民乐箭筈豌豆检测到仅含抗病基因er1;和林粉红野豌豆只含抗病基因Er3;丰镇三棱豌豆只含抗病基因er2、Er3;凉城三棱豌豆同时含3个抗病基因er1、er2、Er3,而不含感病基因Er1;其余的23份材料,均是既含抗病基因er1、er2、Er3,也含感病基因Er1。不同产地来源的豌豆品种所含的分子标记具有一定的地域性,各品种所含分子标记有一定区别;不同产地来源的同名品种的抗性分子标记具多样性,相同地区来源的同名品种其抗性标记均相同。

西瓜抗白粉病研究进展

白粉病是西瓜生产中的重要病害之一,严重影响西瓜产量和品质。介绍了西瓜白粉病生理小种的分类以及白粉病发生规律和危害,统计了我国白粉病发生的地区;对近年来报道的西瓜抗白粉病种质资源进行了归类,同时对西瓜抗白粉病基因的遗传规律、分子标记开发和抗病调控网络等方面的研究进展进行了综述,并对西瓜抗白粉病研究提出了新的思路和见解,旨在为西瓜抗白粉病品种选育提供参考。