小麦抗叶锈病聚合品种中间材料的分子标记辅助选择

为高效筛选多小麦抗叶锈聚合品种中间材料中含有的目的基因,加速多抗品种的培育,本研究对以生产上高产优质小麦品种、含有多个高抗叶锈病基因的抗病小麦材料为亲本杂交获得的杂交优势F2代材料进行目的基因的选择,利用Lr10、Lr24、Lr34、Lr37和Lr38的分子标记对目的基因进行分子辅助鉴定。结果从根据目标性状选择的54份材料中快速选择出含有聚合目的基因Lr10、Lr34或Lr24、Lr37、Lr38或含Lr24、Lr38的聚合中间材料47份,鉴定出表现抗叶锈病,但不含有目的基因的聚合材料7份。研究减少了假阳性的选择,缩短育种中的大量筛选与鉴定的过程,提高了目的基因的选择的准确度,并为加速育种进程奠定了基础。     

75份国内外小麦品种抗叶锈性鉴定及基因分析

为了挖掘小麦抗叶锈病基因,为我国小麦抗叶锈病遗传育种基因库提供更多的选择。选取了75份国内外小麦材料以及36份已知抗叶锈基因载体品种,将这些材料在苗期分别接种14个不同毒力的叶锈菌生理小种,分别鉴定这些不同毒力的生理小种在小麦材料上的发病严重程度;同时提取所有材料的新鲜叶片DNA,选取已经确定的抗叶锈病基因相关联的分子标记特异性引物对75份供试材料进行分子标记检测。结合以上2种方法,推测75份小麦材料中含有的抗叶锈病基因。结果显示,75份小麦材料中含有Lr1、Lr2a、Lr2c、Lr10、Lr11、Lr14a、Lr16、Lr18、Lr20、Lr26、Lr34、Lr37、Lr46这13种已知抗叶锈病基因,这些基因以单基因或多基因聚合的方式存在于小麦品种中。其中,Lr1基因和Lr46基因占比较大,分别高达43%,56%。这些基因单独存在于小麦材料中,并不能表现出良好的抗叶锈性;当几个或者多个基因共同存在于小麦材料中,可以表现出远远高于单一基因的抗叶锈性,与前人研究相符。通过分子标记检测到小麦材料大白春小麦S3中含有Lr1、Lr10和Lr46基因,温室苗期鉴定发现该材料对本研究的11种叶锈生...     

贵州小麦品种(系)中慢锈基因Lr34/Yr18的分子检测

小麦慢锈病是危害小麦生产的重要病害。为了鉴定258份贵州小麦品种(系)中慢锈基因Lr34/Yr18的组成,筛选含慢锈抗性基因Lr34/Yr18的种质资源,本研究利用STS标记csLV 34结合毛细管电泳技术对258份小麦品种(系)中慢锈抗性基因Lr34/Yr18的等位变异进行了分子检测。结果表明:毛细管电泳谱带清晰易读,可根据扩增片段分子量直接判断目标片段有无。STS标记csLV 34可在含有Lr34/Yr18基因的材料中扩增出150 bp片段,部分不含Lr34/Yrl8的材料则扩增出229 bp片段,余下大部分不含Lr34/Yrl8的材料没有扩增出150 bp和229 bp的片段;258份小麦品种(系)中有5份材料扩增出150 bp片段,可能含有Lr34/Yr18基因,占供试材料的1.9%。筛选的这些慢锈抗性种质资源可为今后贵州小麦的慢锈抗病品种选育提供参考。     

122份小麦品种(系)抗叶锈病基因Lr26、Lr34、Lr38分子标记检测

运用抗叶锈病基因Lr 26、Lr 34、Lr 38的特异性分子标记,对122份小麦品种(系)进行了检测,以明确各品种(系)的抗叶锈性.结果表明:122份供试材料中,含有Lr 26的有33个,含有Lr 34的有3个,含有Lr 38的有37个;同时含有Lr 26和Lr 38的有30个,同时含有Lr 26、Lr 34和Lr 38的有3个.本研究结果可为小麦的抗叶锈育种提供理论参考.     

四川小麦品种(系)中慢锈基因Lr 34/Yr 18/Pm 38的分子检测

慢锈性基因Lr 34/Yr 18/Pm 38对小麦叶锈病和条锈病都具有广谱持久抗性,为明确该基因在四川小麦品种(系)中的分布,对90份四川小麦品种(系)进行慢锈病基因Lr 34/Yr 18/Pm 38的分子检测。利用与Lr 34/Yr 18/Pm 38基因位点紧密连锁的STS标记cs LV 34和基于该基因第11外显子(exon 11)等位变异开发的4对功能标记cssfr 1、cssfr 2、cssfr 3、cssfr 4检测90份四川小麦品种(系)。结果表明,所检测的90份四川小麦材料中都不含Lr 34/Yr 18/Pm 38位点,功能标记cssfr 1~cssfr 4检测结果与STS标记cs LV 34检测结果一致。结论:四川小麦品种(系)含有Lr 34/Yr 18/Pm 38基因位点的材料较少,在过去育种中,Lr 34/Yr 18/Pm 38基因不被人们重视,没有得到很好的利用。     

40份CIMMYT小麦品系苗期及成株抗叶锈病基因鉴定

为了对40份来自国际玉米小麦改良中心(CIMMYT)的小麦材料进行抗叶锈病基因鉴定,试验结合系谱分析、基因推导和分子标记检测等方法在苗期对36个已知抗病基因载体品种和供试的40份小麦材料接种17个具有毒性差异的叶锈菌生理小种,通过对比供试小麦材料与已知单基因载体品种侵染型,推导出供试小麦材料中可能携带的已知抗叶锈病基因,同时利用12个与已知抗病基因紧密连锁的标记对供试材料进行标记检测,检测结果与基因推导相互验证。进一步将40份供试品系分别种植于河北保定和河南周口试验田,接种叶锈菌混合小种进行田间成株期抗叶锈性鉴定。结果表明,7个供试小麦材料含有Lr1,携带Lr10的有9个品系,携带Lr11和Lr34的分别有10个品系,含有Lr14a、Lr15和Lr26的分别有2,4,3个小麦品系;另外,经标记检测成株抗叶锈病基因Lr37和Lr46分别存在于22,39个小麦品系中,经田间鉴定有22个小麦品系表现成株抗性。     

70份国外小麦品种(系)的苗期和成株期抗叶锈病鉴定

小麦叶锈病是小麦生产中的重要病害之一,培育持久抗病品种是最经济、有效和环保的方法。本研究用19个不同毒力的叶锈菌小种苗期接种70份国外引进小麦品种(系)及36个已知抗叶锈病基因的载体品种进行抗性鉴定,同时在2016—2017年度分别于河北保定和河南周口对70份国外引进品种进行田间抗叶锈性鉴定。为进一步检测材料中所携带的苗期和成株抗叶锈病基因,利用12个与已知基因紧密连锁的分子标记进行检测,综合基因推导、系谱分析和分子标记检测的结果,在33份材料中鉴定出15个抗叶锈病基因,包括Lr1、Lr2a、Lr26、Lr3ka、Lr11、Lr17、Lr30、Lr10、Lr14a、Lr2b、Lr13、Lr15、Lr21、Lr44和Lr45,田间鉴定筛选出39份品种表现慢锈性。苗期和田间表现表明,国外品种中含有丰富的对我国叶锈菌小种有效的苗期和成株期抗叶锈病基因,可作为小麦抗叶锈病抗源在抗病育种中加以利用。     

小麦抗叶锈病基因Lr24的一个新STS标记

来源于长穗偃麦草的基因Lr24对小麦叶锈病具有很高的抗性,本研究旨在开发用于Lr24基因分子标记辅助育种的新的分子标记。从定位于小麦3D染色体的22对SSR、EST-SSR引物中筛选出4对揭示TcLr24多态性的引物,用468株F2抗感群体对这4对引物进一步检测,得到1个与Lr24共分离的EST-SSR标记Xcwem17。对该标记进行测序,并设计了STS引物。用该STS引物及已知的Lr24SCAR引物对试验群体进行验证,两对引物在该F2群体中均表现共分离,且Xcwem17可在TcLr24单基因系和已知含Lr24的农家品种泰山1号中可扩增出180bp单一条带,感病对照及其余7个近等基因系无扩增。该EST-SSR标记可直接用于分子标记辅助选择。     

28个小麦微核心种质抗叶锈性分析

选取在成株期表现高、中、低抗叶锈的28个小麦微核心种质,利用39个以Thatcher为背景的近等基因系(或单基因系)作为已知基因的鉴别寄主,接种8个小麦叶锈菌致病型进行苗期抗叶锈基因推导,结合成株期抗病鉴定,初步明确了这些品种(系)的抗性和可能携带的抗病基因。利用19个与Lr基因紧密连锁或共分离的分子标记,对28个微核心种质进行抗叶锈病基因的进一步鉴定,推测新克旱9号可能含有Lr17、Lr2b、Lr14a和Lr33;兴义4号可能含有Lr26、Lr36和Lr37;紫皮可能含有Lr2b和Lr34;大白皮含有Lr1;毕红穗含有Lr1、Lr10和Lr34;中优9507含有Lr10;小白麦、红粒当年老、老麦、蝉不吱、苏麦3号和车锏子含有Lr1和Lr34;红花早可能含有Lr1、Lr34、Lr14a和Lr2b;江西早、泡子麦、三月黄、有芒扫谷旦、阜阳红、成都光头和酱麦可能含有Lr34;敦化春麦和甘肃96可能含有Lr28;欧柔可能含有Lr34、Lr16、Lr11、Lr3bg和Lr33;此外,新克旱9号、兴义4号、红花早、红粒当年老、欧柔、有芒扫谷旦、成都光头、甘肃96、小红皮、定兴寨、中优9507和红冬麦中可能含有未知抗病基因;在这28份种质中,不含Lr9、Lr19、Lr20、Lr21、Lr24、Lr29、Lr35、Lr38和Lr47基因。研究结果表明,测试的微核心种质中含有比较丰富的抗叶锈病基因,可为育种提供丰富的抗源。     

50个小麦品种抗叶锈基因鉴定

选育抗性品种是小麦叶锈病防治举措中最经济、可行的方法。为进一步挖掘抗病基因,选取河南、河北、山东等8个省小麦产区50个小麦品种。首先在苗期将16个叶锈菌生理小种(THFS、TGTS、THJS、FHKT、FGJN、KHKS、FCJQ、RFKS、THFM、MHGT、KHGS、KBGT、FHGT、PHHT、FHJT、FCJT)接种在36份小麦抗叶锈病近等基因系材料以及50个供试小麦品种上。因各菌种带有不同毒力,可根据表现型的差异,再将已知抗病基因紧密连锁的特异性分子标记与之结合分析,进而推测50份小麦材料中可能含有的抗叶锈病基因。通过基因推导、分子标记以及系谱分析综合鉴定抗锈性基因,结果表明,在50个品种中共检测出9个(Lr1、Lr2c、Lr10、Lr16、Lr26、Lr34、Lr37、Lr45和Lr46)已知抗叶锈性基因和少量未知基因。含有Lr1基因的有淄麦12等22个品种;含有Lr2c基因的有鲁麦14等10个品种;含有Lr10基因的只有莱州9361一个品种;含有Lr16基因的有科农199等25个品种;含有Lr26基因的有徐州24等15个品种;含有Lr34基因的有宝麦3号和京冬8号;含有L...     

小麦抗叶锈基因Lr24、Lr35的STS、SCAR标记在近等基因系(NILs)上的特异性验证

用一套分别含有不同抗叶锈基因的53个以Thatcher为遗传背景的近等基因系（near-isogeniclines,NILs）对已报道的分别与抗叶锈基因Lr24和Lr35连锁的STS、SCAR进行特异性验证。结果对于与Lr24连锁的STS标记,在53个NILs中只在TcLr24亲本中扩增出片段大小与报道相同的310bp的条带,在TcLr35中也扩增出了一条片段,但片段大小不同于310bp约为270bp。对于与Lr35连锁的SCAR标记,只在TcLr35亲本中扩增出片段大小为900bp的条带,与报道片段大小一致。验证结果表明与抗病基因Lr24和Lr35连锁的STS、SCAR分子标记在NILs中特异性都较好,进一步证明了这两个分子标记可方便地用于小麦抗叶锈基因Lr24、Lr35的分子标记辅助选择育种。     

土耳其8个小麦品种农艺性状及抗叶锈性分析

为确定8个来自土耳其的普通小麦品种在我国的应用前景,对其进行全生育期农艺性状观察,并利用44个以Thatcher为背景的近等基因系(单基因系)作为已知基因的鉴别寄主,接种8个小麦叶锈菌致病型进行苗期抗叶锈基因推导,结合成株期抗病鉴定,初步明确了这些品种(系)的抗性和可能携带的抗病基因。利用20个与Lr基因紧密连锁或共分离的分子标记,对8个土耳其小麦品种进行抗叶锈病基因的进一步鉴定。推测YJ000900中可能含有Lr1、Lr3、Lr17、Lr20;YJ000906中可能含有Lr1、Lr17、Lr20;YJ000901、YJ000902、YJ000904、YJ000905、YJ000907中可能含有Lr1;8个材料中均不含Lr9、Lr19、Lr20、Lr21、Lr24、Lr26、Lr28、Lr29、Lr34、Lr35、Lr37、Lr38和Lr47基因。结果表明,来自土耳其的8个小麦材料具有较差的抗叶锈性、抗寒和抗倒伏能力,而且产量低,不适宜于大规模推广种植,也不能作为我国小麦抗叶锈的抗源使用。     

云南小麦品种（系）锈病和赤霉病抗性功能基因的KASP标记检测

利用5个锈病成株期抗性基因的KASP标记Sr2_ger9 3p、Lr34jagger、CSTM4_67G、Lr68-2、VPM_SNP和抗赤霉病基因Fhb1的KASP标记TaHRC-KASP,对云南省育成的42个小麦品种（系）进行检测,旨在筛选出含有目标基因的优异小麦种质,为云南省持久抗病小麦新品种（系）的选育提供材料。结果表明,4个材料含兼抗型成株抗锈病基因Lr34/Yr18/Sr57,频率为9.52%;6个品种（系）含兼抗型成株抗锈病基因Lr67/Yr46/Sr55,频率为14.29%;7个材料含抗慢叶锈病基因Lr68,频率为16.67%;含兼抗型成株抗锈病基因Sr2/Yr30和成株抗叶锈基因Lr37的材料各有1个,频率均为2.38%;未检测出含抗赤霉病基因Fhb1的品种（系）。云麦69、云麦75、云麦56、宜麦1号和宜麦3号等兼有2个成株期抗锈病基因,可作为今后云南持久抗锈病育种的抗源材料。     

Unveilingsources of stripe rust resistance in diverse wheat (<Emphasis Type="Italic">Triticum Aestivum</Emphasis> L.) germplasm using narrow down methodology: a proof of concept

Stripe rust of wheat caused by the fungal pathogen is a destructive foliar disease of wheat. Thus, it is crucial step to characterize the resistant germplasm for stripe rust in a diverse germplasm pool for their ultimate utilization in efficient crop rust resistance breeding. In the present study, we followed two pronged strategies involving integrated phenotypic and molecular characterization of 440 diverse wheat germplasm lines for rust resistance. The germplasm panel was extensively evaluated in field epiphytotic conditions during two consecutive years. After rigorous screening, 72 accessions were successfully revealed as resistant to moderately resistant to stripe rust. Subsequently, entries were then evaluated for their field agronomicperformances, considering prerequisites for serving as a donor germplasm,particularly for yield and 33 potential rust-resistant accessions were identified. Furthermore, to detect the sources of resistance, accessions were molecular characterized for potential race-specific resistance genes Yr5, Yr10,Yr15, and effective adult plant resistance (APR) gene Lr34/Yr18/pm38. We identified the 22 accessions possessing one or more single resistance genes and two accessions were observed with at least three of them. Moreover, Lr34/Yr18/pm38 was determined to confer resistance when observed along with any of the race-specific genes. Thus, the study not only provides proof of concept methodology to identify candidate resistant sources from large germplasm collections but simultaneouslyconfirmed the contribution of combining race-specific andnon-specific APR genes. The finding could further assist in the potential deployment of resistant genes directly into the stripe rust breeding program by involving marker-assisted approaches.     

Evaluation of seedling and adult plant resistance to leaf rust in European wheat cultivars

Summary A set of 105 European wheat cultivars, comprising 68 cultivars with known seedling resistance genes and 37 cultivars that had not been tested previously, was tested for resistance to selected Australian pathotypes of P. triticina in seedling greenhouse tests and adult plant field tests. Only 4% of the cultivars were susceptible at all growth stages. Twelve cultivars lacked detectable seedling resistance to leaf rust, and among the remaining cultivars, 10 designated genes were present either singly or in combination. Lr13 was the most frequently detected gene, present in 67 cultivars, followed by the rye-derived gene Lr26, present in 19 cultivars. Other genes present were Lr1, Lr3a, Lr3ka, Lr10, Lr14a, Lr17b, Lr20 and Lr37. There was evidence for unidentified seedling resistance in addition to known resistance genes in 11 cultivars. Field tests with known pathotypes of P. triticina demonstrated that 57% of the cultivars carried adult plant resistance (APR) to P. triticina. The genetic identity of the APR is largely unknown. Genetic studies on selected cultivars with unidentified seedling resistances as well as all of those identified to carry APR are required to determine the number and inheritance of the genes involved, to determine their relationships with previously designated rust resistance genes, and to assess their potential value in breeding for resistance to leaf rust.     

Amplified fragment length polymorphism-derived microsatellite sequence linked to the Pch1 and Ep-D1 loci in common wheat

Amplified fragment length polymorphism (AFLP) markers linked to the Aegilops ventricosa‐derived chromosome segment in ‘VPM1’ on which the eyespot resistance gene, Pch1, and the endopeptidase gene, Ep‐D1b, occur were identified. One marker was isolated from the gel, cloned and sequenced. Sequence analysis revealed a microsatellite repeat motif. Sequence‐specific primers were designed to amplify a product containing the repeat motif, and the microsatellite marker was tested for cosegregation with the Ep‐D1b allele. Distinct alleles were produced by the Pch1 sources, normal wheat and wheat containing the Lr19 translocation. A recombination frequency of 0.02 was calculated between the microsatellite marker and Ep‐D1.     

兼抗型成株抗性小麦品系的培育、鉴定与分子检测

小麦条锈病、叶锈病和白粉病是我国小麦的重要真菌病害,培育兼抗型成株抗性品种是控制病害最为经济有效和持久安全的方法。本研究选用由成株抗性育种方法培育的21份冬小麦高代品系和96份春小麦高代品系,在多个环境下进行这3种病害的成株期抗性鉴定,并利用紧密连锁的分子标记检测了兼抗型基因Lr34/Yr18/Pm38、Lr46/Yr29/Pm39和Sr2/Yr30的分布。田间鉴定表明,21份冬小麦品系中有17份兼抗3种病害,占80.9%;96份春小麦品系中有85份兼抗3种病害,占88.5%。分子标记检测发现,21份冬小麦品系均含QPm.caas-4DL,其中7份还含QPm.caas-2BS,9份还含QPm.caas-2BL;96份春小麦品系中,18份含Lr34/Yr18/Pm38,37份含Lr46/Yr29/Pm39,29份含Sr2/Yr30。以上结果表明,分子标记与常规育种相结合,可有效培育兼抗型成株抗性品种,为我国小麦抗病育种提供了新思路。     

Cytogenetic studies in wheat XIX. Chromosome location and linkage studies of a gene for leaf rust resistance in the Australian cultivar 'Harrier'

Monosomic analysis indicated that a seedling leaf rust resistance gene present in the Australian wheat cultivar ‘Harrier’(tentatively designated LrH) is located on chromosome 2A. LrH segregated independently of the stripe rust resistance gene Yr1 located in the long arm of that chromosome, but failed to recombine with Lr17 located in the short arm. LrH was therefore designated Lr17b and the allele formerly known as Lr17 was redesignated as Lr17a. The genes Lr17b and Lr37 showed close repulsion linkage. Tests of allelism indicated that Lr1 7b is also present in the English wheats ‘Dwarf A’(‘Hobbit Sib’), ‘Maris Fundin’ and ‘Norman’. Virulence for Lr17b occurs in Australia, and pathogenicity studies have also demonstrated virulence in many western European isolates of the leaf rust pathogen. Despite this, it is possible that the gene may be of value in some regions if used in combination with other leaf rust resistance genes.     

Molecular markers for the identification of resistance genes and marker-assisted selection in breeding wheat for leaf rust resistance

The resistance genes Lr9, Lr24, Lr25, Lr29, Lr35 and Lr37, which were not previously utilised in Hungary, have been incorporated into four Martonvásár winter wheat cultivars using marker-assisted selection with PCR-based markers. In the course of a backcross programme, the genes were transferred into Martonvásár wheat varieties and various BC generations were produced. Work aimed at pyramiding resistance genes is currently underway in Martonvásár, and plants containing the gene combinations Lr9 + Lr24, Lr9 + Lr25 and Lr9 + Lr29 are now available. From the BC₂F₄ generation of the ‘Mv Emma’*3/’R.L.6010’ combination (‘R.L.6010’ is the donor of the Lr9 gene) 287 lines were tested for leaf rust resistance in an artificially inoculated nursery. A co-dominant primer combination was designed to identify both resistant and susceptible offsprings. The results of resistance tests and molecular marker detection agreed in most cases. Designated leaf rust resistance genes were identified with molecular markers in wheat varieties and breeding lines. The Lr26 and Lr34 resistance genes occur frequently in the Martonvásár gene pool, and the presence of the Lr37 gene has also been detected in a number of Hungarian genotypes.