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.

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.

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.

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.

The Effect of Wheat Mixtures on the Powdery Mildew Disease and Some Yield Components

Mixtures composed of five wheat cultivars,Jingshuang 16,Jing 411,Jingdong 8,Lunxuan 987,and Baofeng 104,with different levels of resistance against powdery mildew were tested for their potential containment of the disease development in the field and for the influence on grain yield and the content of crude protein in the years 2007 and 2010.The plots were inoculated artificially with mixed isolates collected in the fields and propagated in the greenhouse and the disease was scored in 7 d interval during the two growing seasons.It was indicated that certain combinations,e.g.,Jingdong 8：Lunxuan 987,Jingdong 8：Baofeng 104,and Jing 411：Jingdong 8：Baofeng 104,showed positive efficacy on the mildew.The cultivar combinations tested in 2007 showed increase of grain yield,while most of the combinations tested in 2010 did not show the increase.The differences of the increases or decreases were not statistically significant except combinations Jing 411：Jingdong 8：Baofeng104,Jingshuang16：Jingdong8：Lunxuang 987 and Jingshuang 16：Jingdong 8：Lunxuan 987：Baofeng 104,which showed the decrease of the grain yield.The mixtures did not show influence on the content of crude protein in grain.More cultivar combinations need to be tested.

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.

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 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.

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.

栽培小麦Brock中抗白粉病相关基因TaRPP13-1B的克隆及功能分析

白粉病是小麦生产中的主要病害之一,发掘抗病基因并实现抗病基因转育是提高作物抗病性的最经济有效的方法。本研究克隆了一个位于小麦染色体1B上、具有典型CC、NBS和LRR结构域的基因TaRPP13-1B。接种白粉菌后,TaRPP13-1B基因在抗病小麦Brock和BJ-1中表达量虽然出现上下调波动,但平均表达水平一直高于感病小麦品种京411。采用病毒诱导的基因沉默和转基因过表达技术进行功能分析,发现抑制目标基因表达,抗病小麦品种Brock对白粉菌的抗性显著降低;过表达TaRPP13-1B的转基因小麦津强5号对白粉菌抗性明显提高。说明TaRPP13-1B基因参与小麦抗白粉病的防御反应过程。该研究为小麦抗病品种的选育提供了有价值的备选基因。

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。

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。

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。

127份甘肃农家小麦品种兼抗条锈病与白粉病鉴定与评价

小麦条锈病、白粉病是发生于甘肃陇南乃至全国小麦生产上的最主要病害之一,种植抗病品种是防治病害最经济有效且绿色环保的措施。兼抗种质资源材料的匮乏是制约甘肃陇南小麦条锈病和白粉病品种选育和病害有效防控的主要因素。农家品种是小麦抗病种质资源重要的基因库,开展兼抗病害材料鉴定是准确评价供试材料抗病性的基础性工作。2021年和2022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,对127份小麦农家品种进行了苗期、成株期抗条锈病和白粉病性鉴定,结果发现:供试材料中苗期对条锈菌和白粉菌混合菌表现抗病的分别有36份和24份,占28.35%和18.90%;兼抗材料15份,占11.81%。成株期在接种及自然诱发条锈菌和白粉菌条件下,表现抗病的分别有39份和72份,占30.71%和56.69%;兼抗材料有27份,占21.26%。全生育期兼抗材料有11份,占8.66%。抗白粉病基因分子检测结果发现,含有抗病基因Pm2、Pm4a、Pm8、Pm21的分别占30.71%、86.61%、25.20%、3.15%;含有2个及以上基因组合品种有60个,占47.24%。结合前期抗条锈病研究结果,将会为筛选出的优异材料利用和持续解决甘肃陇南兼抗品种匮乏难题提供材料支撑。

波斯小麦Cypa35-3成株期抗白粉病基因定位

源自苏联的波斯小麦Cypa35-3对陕西关中地区白粉菌优势小种表现为成株期高抗白粉病。为明确Cypa35-3抗白粉病基因所在染色体位置及其抗白粉病基因的遗传规律,利用Cypa35-3与高感白粉病的加拿大二粒小麦Flavescens进行杂交,在成株期自然发病条件下对亲本及其杂交F_(1)、F_(2)、F_(2)∶3群体进行白粉病抗性评价与遗传分析;选用分布于A、B染色体组14对染色体上共计601对SSR标记,利用分离群体分组分析法(BSA)对Cypa35-3的F_(2)群体进行多态性标记筛选。结果表明,Cypa35-3的成株期白粉病抗性受1对显性基因控制,暂命名为PmCypa35-3。通过群体筛选,获得4对位于1B染色体上的SSR标记,分别为Xbarc81、Xcfd48、Xbarc61、Xbarc302,其中Xbarc81、Xcfd48位于PmCypa35-3两侧,遗传距离分别为25.2 cM、8.6 cM。由此将成株期抗白粉病基因PmCypa35-3初步定位于1B染色体。通过与1B染色体上及波斯小麦中正式命名的抗白粉病基因的基因来源和标记位置进行对比,发现PmCypa35-3与它们均不相同,推测PmCypa35-3可能是一个新的小麦成株期抗白粉病基因,可作为小麦抗病育种的新抗源。

四倍体小麦成株抗白粉病基因定位

四倍体小麦是普通小麦的祖先种,也是重要的粮食作物。发掘四倍体小麦抗病种质并鉴定其携带的抗病基因,旨在为小麦新品种选育提供新抗源。TDI-1是栽培二粒小麦,在多年的田间种植过程中表现出对白粉病免疫的表型。为了确定TDI-1携带的抗病基因,为小麦白粉病抗性遗传提供理论依据,首先将其与表现为感白粉病表型的硬粒小麦TDU-1进行杂交并构建了遗传群体,然后对亲本及其杂交F_(1)、F_(2)、F_(2:3)群体进行了白粉菌小种E09接种试验和抗病性分析,最后利用混合分离群体分析法(BSA)对抗白粉病基因进行了定位。结果表明,TDI-1在苗期对E09表现为感病,在成株期对E09表现为抗病;TDI-1和TDU-1的F_(1)植株在成株期对E09表现为抗病;F_(2)群体中,表现为抗病的单株数和感病的单株数的比例符合3∶1(χ^(2)_(3∶1)=0.11,P=0.74);F_(2∶3)家系中,纯合抗病、抗病性分离、纯合感病的家系数目之比符合1∶2∶1(χ^(2)_(1∶2∶1)=0.47,P=0.79),表明TDI-1成株期白粉病抗性由1个显性基因控制,暂将其命名为PmTDI-1。对TDI-1和TDU-1及其杂交后代F_(2)群体进行分子标记筛选,发现4个位于2A染色体短臂上的分子标记Xwmc407、NRM-2AS29、NRM-2AS45和NRM-2AS84与PmTDI-1紧密连锁,其中,NRM-2AS45和NRM-2AS84位于两侧,遗传距离分别为1.8,4.6 cM。由此,将成株期抗白粉病基因PmTDI-1初步定位于2A染色体短臂上。研究结果显示,从四倍体小麦TDI-1中鉴定到1个新的显性成株抗白粉病基因PmTDI-1。

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.

普通小麦Brock中一个抗白粉病新基因AFLP标记P13/M13_(250)的筛选

用225对AFLP引物组合,对白粉病敏感小麦品种京411,抗病品种Brock和以京411为轮回亲本、Brock为抗源供体育成的近等基因系(NILs)进行分子标记筛选,其中,引物组合P13/M13能在抗、感材料间稳定产生P13/M13-250 bp的多态性片段.用Brock与京411杂交F2代抗、感分离单株,对P13/M13250进行了初步连锁性鉴定,该标记对小麦植物抗病育种分子标记辅助选择有一定用处.

小麦Brock抗白粉菌侵染早期防御应答基因分析

为了探究小麦对禾本科布氏白粉菌的防御机制,以强抗白粉病小麦品种Brock为研究对象,利用RNA-seq技术分析了白粉菌侵染早期Brock中的应答基因,共鉴定出4625个差异表达基因(DEGs).利用KEGG分析重点注释了小麦与白粉菌互作的20条主要途径,通过qRT-PCR技术检测了来自不同途径的9个差异表达基因.其中,PR1和RPM1基因的表达量变化明显.这2个基因是PTI和ETI反应的重要参与基因,PR1表达量在2 hpi时显著升高,RPM1基因表达量在12 hpi时显著升高.以上结果说明白粉菌侵染早期(2 hpi和12 hpi),小麦通过PTI和ETI的协同作用抵抗白粉菌侵染.

栽培小麦Brock中与新抗白粉病基因紧密连锁的AFLP分子标记筛选与分析(英文)

本研究用225对引物对农艺性状优良但对白粉菌敏感的栽培小麦京411、抗白粉病栽培小麦Brock以及京411与Brock配制的近等基因系进行AFLP分子标记筛选,结果发现只有2对引物组合Pst+GAC/Mse+TCT(P1)和Pst+AGC/Mse+ACC(P2)在上述抗感材料中表现出多态性,分别扩增到2个特异片段,将特异片段克隆并测序发现,P1扩增的特异片段长268bp,P2扩增的特异片段长227bp,命名为AFLP标记P1268和P2227。用106个京411×Brock的F2单株进行连锁性分析表明,P1268和P2227与抗白粉病基因的遗传距离分别为3.6和1.9cM,与Brock中的抗白粉病基因呈紧密连锁。该两个AFLP标记对小麦抗白粉病基因的积累和分子标记辅助选择育种有重要意义。