小麦抗条锈病一致性数量性状位点(MQTL)图谱构建

 小麦条锈病是造成小麦减产和品质劣化的最重要病害,定位小麦染色体上一致性条锈病抗性基因/位点/区段是小麦条锈病抗性分子育种的重要基础。本研究对至今分子标记和遗传定位的342个条锈病抗性基因/位点/区段进行数据搜集整理,借助Maccaferr和Andrzej的参考图谱,基于元分析技术进行Meta-QTL(MQTL)检测,共获得194个小麦抗条锈病MQTL,包括74个与严重度(Disease severity, DS)相关,46个与反应型(Infection type, IT)相关、19个与病程曲线下面积相关(Area under disease progress curve, AUDPC)、28个与DS和IT共相关、6个与DS和AUDPC共相关、15个与IT和AUDPC共相关、6个与其他条锈病抗性性状相关。这些抗条锈病一致性QTL定位于小麦21条染色体上,呈非均匀分布,且部分MQTL集中成簇。通过与已发表的正式命名抗条锈病基因比较分析,发现大多数正式命名基因定位于MQTL簇区段,说明这些MQTL簇区段很可能是控制小麦条锈病抗性热点区域。控制小麦抗条锈病一致性QTL遗传图谱的构建为小麦条锈病抗性基因精细定位及抗病育种提供了遗传信息参考依据。     

来自簇毛麦抗条锈病新基因的SSR标记

 用小麦条锈菌条中30号生理小种,对小麦抗病种质小麦-簇毛麦易位系V9128-1和铭贤169的杂交后代进行抗条锈性遗传分析,小麦-簇毛麦易位系V9128-1的抗病性符合1对显性抗条锈病基因控制。并根据F₂抗、感病单株分离比例组建抗感池,用SSR技术寻找与抗病基因连锁的分子标记。从121个SSR引物组合中筛选到2个与抗病基因YrV1(暂命名)紧密连锁的微卫星标记Xgwm566和Xgwm376,遗传距离分别为3.6和5.5cM;因此,该抗条锈病基因位于小麦3B染色体短臂上。这2个标记不仅能在小麦-簇毛麦易位系V9128-1中检测到,而且在抗病基因供体亲本簇毛麦中也能检测到。综合抗病基因来源和分子生物学试验结果,可以推断,YrV1很可能是1个来自簇毛麦并与已知抗条锈病基因不同的新基因。     

小麦条锈病新抗源的抗谱鉴定初析

 利用南京农业大学细胞遗传研究所育成的一套涉及不同簇毛麦染色体的异附加系和代换系以及5个6VS/6AL易位系,经1997、1998、1999连续3年在陕西、北京、四川进行小麦条锈病抗性接种鉴定,结果表明普通小麦-簇毛麦6V异附加系,6V(6A)异代换系和6VS/6AL易位系高抗条锈病菌条中29、条中31、水源11-2、水源11-5、水源11-13和杂46等强毒小种。考虑到含整组V染色体的硬粒小麦-簇毛麦双倍体不抗水源11-13小种,上述普通小麦-簇毛麦6V异附加系、异代换系和6VS/6AL易位系的条锈病抗性可能还与其所涉及的小麦亲本基因的作用有关。     

小麦条锈病抗病遗传及菌源基地基因布局研究进展

小麦条锈病是世界范围内严重影响小麦生产安全的重要病害。我国是世界上最大的小麦条锈病流行区,自成独立的流行体系。培育和种植抗病品种是防治病害最有效的措施。然而,小麦品种对条锈病的抗性常常由于病菌新小种的产生而丧失,这既是一个重大科学问题,也是一个亟待研究解决的生产实际问题。如何有效、合理地利用小麦的抗病性,植病学家和育种学家进行了一个多世纪的研究与探索,提出了各种理论与策略,开展了各种实践与探索。该文就小麦抗条锈病遗传及其基因布局研究进展进行综述,主要包括抗性鉴定评价、抗病基因发掘与利用、数量抗性位点定位、抗病基因克隆与功能解析、近等基因系创建与应用,以及抗源创制、抗病生态育种和大区基因布局等,并对深入开展抗条锈病基因发掘与利用和大区基因布局进行展望,以期为抗病育种和病害持续治理提供参考。     

22份中国小麦生产主栽品种抗条锈病基因解析

为挖掘新的小麦抗条锈病基因,掌握小麦生产主栽品种的抗条锈病基因携带情况,有效防控小麦条锈病,采用抗性鉴定、基因推导分析和分子标记技术对22份小麦生产上主栽品种进行了研究,通过抗性鉴定比较22份小麦主栽品种与已知基因载体品种的抗谱。结果显示,共推测出14份供试品种携带已知抗条锈基因,8份供试品种携带未知抗条锈基因,是新的抗锈基因资源;聚类分析结果显示,供试22份小麦品种可分为2个大类6个亚类;利用SSR分子标记检测抗条锈病基因Yr1、Yr10和Yr24的携带情况发现,11份品种携带Yr1基因,2份品种携带Yr10基因,22份品种均不携带Yr24基因。部分生产主栽品种携带新的抗条锈病基因,表明小麦品种选育中避免了抗性基因单一化,并加强了未知基因的利用。     

小麦抗条锈病近等基因系Taichung29*6/Strubes Dickkopf抗病主效基因的单体分析

小麦品种Strubes Dickkopf是小麦条锈菌国际鉴别寄主,通过对以其为基因供体与完全感病品种Taichung29杂交转育而成的近等基因品系Taichung29*6/Strubes Dickkopf的单体分析,检测和定位Taichung29*6/Strubes Dickkopf中所含的抗条锈病主效基因,并明确其抗性特点。结果表明,Taichung29*6/Strubes Dickkopf对CY26小种的抗性由1对显性主效抗条锈基因所控制,定位在4B染色体上,暂定名为YrSD。同时说明Strubes Dickkopf中含有与Yr25不同的新的抗条锈病基因YrSD。     

小麦品系ICA56抗条锈病鉴定及其抗性基因SSR标记

 小麦品系ICA56对条锈菌优势生理小种CYR30、CYR31和CYR32均表现免疫反应;遗传分析表明,ICA56携带一个显性抗条锈病基因。基因等位性测定显示,ICA56所含抗条锈病基因不同于已知抗锈基因Yr5、Yr10、Yr15和Yr26,暂将该基因定名为YrICA56。利用川麦28/ICA56的F₂群体及抗感亲本筛选到5对SSR引物WMC503、Xgwm261、Xgwm296、WMC112和Xgwm210与YrICA56连锁,遗传距离分别为16.6、10.4、7.0、4.5和14.1cM。根据Mapmaker3.0确定标记、YrICA56和着丝点在染色体上的顺序为:-WMC503-Xgwm261-Xgwm296-YrICA56-WMC112-Xgwm210-着丝点-。根据作图结果,将YrICA56定位在2DS。目前定位在2DS上的抗条锈病基因有Yr16和YrKat。Yr16为成株期抗性,YrKat属温敏抗性,而YrICA56在苗期和成株期对条锈病均表现免疫,由此推测YrICA56是一个新的抗条锈病基因。     

小麦条锈菌中国鉴别寄主阿夫的抗条锈病基因单体分析

应用单体分析技术,用2E16单孢菌系对小麦条锈菌中国鉴别寄主阿夫进行抗条锈病主效基因分析及染色体定位。结果表明,阿夫对2E16菌系的抗性是由1对显性抗条锈基因控制,未发现其中含有与Sonalike相同的抗条锈病基因,确认阿夫中除含YrA外至少还含有1对未知的显性抗条锈病基因,并将其定位在3B染色体上,暂定名为YrFun。     

普通小麦-柔软滨麦草易位系M852-1抗条锈基因的遗传分析和分子作图

 M852-1是经杂交和回交培育的普通小麦-柔软滨麦草易位系,苗期对我国小麦条锈菌流行小种均表现良好抗性。为明确其抗条锈性遗传规律,本研究选用条锈菌流行小种（类型）CYR29、CYR32、CYR33和Su11-7的单孢菌系对其与铭贤169杂交F₁、F₂、F₃及BC₁代群体进行遗传分析, 同时应用420对SSR引物对接种CYR32的M852-1/铭贤169 F₂代144个单株作图群体进行抗病基因定位。结果表明,M852-1对供试小种均表现免疫或近免疫,对CYR29的抗锈性由1对显性基因控制,对CYR32、CYR33和Su11-7的抗锈性均由1对隐性基因控制。筛选到3个与抗CYR32基因连锁的SSR标记Xbarc124、Xbarc200和Xgwm429,遗传距离分别为6.3、5.6 和 9.7 cM。根据SSR标记锚定性将该基因定位于小麦2BS染色体,暂命名为YrM852。基因来源、分子标记检测及染色体位点分析表明,YrM852很可能是1个不同于目前已知抗条锈病基因的新基因。     

源于柔软滨麦草抗小麦条锈病基因YrElm的遗传分析与SSR标记

 M852-1是由柔软滨麦草和普通小麦7182经杂交和回交培育的易位系。苗期抗病性鉴定结果表明,M852-1对CYR29、CYR31、CYR32、CYR33、Su11-4、Su11-7和V26等7个中国小麦条锈菌主要生理小种或新的致病类型均表现免疫至高抗,是一个较好的抗条锈资源材料。用条锈菌流行小种CYR33对M852-1与铭贤169杂交F₁、F₂、F₃和BC₁代进行抗性鉴定与遗传分析,发现M852-1对CYR33的抗条锈性由1对隐性基因控制,暂定名为YrElm。以F₂代分离群体构建作图群体,利用集群分离分析法,筛选到与YrElm连锁的5个SSR标记:Xcfd35、Xgwm161、Xwmc630、Xgwm533和Xcfd34,并将YrElm定位于小麦染色体3DS上。YrElm两侧最近2个SSR标记Xcfd35与Xgwm161的遗传距离分别为6.5 cM和4.2 cM。抗锈性鉴定、系谱分析以及分子标记检测结果表明,该抗病基因来源于柔软滨麦草。综合基因来源、分子检测及染色体位点等方面的分析,认为YrElm可能是一个新的抗条锈病基因。用该基因两侧最近两个标记Xcfd35和Xgwm161 检测68个甘肃和黄淮麦区小麦品种(系),10个(14.7%)品种能扩增出与M852-1相同的条带。进一步进行抗病性及系谱分析表明,这10个品种均不含YrElm。本研究结果为利用YrElm进行分子标记辅助育种和进一步的精细定位奠定了基础。     

The dissection and SSR mapping of a high-temperature adult-plant stripe rust resistance gene in American spring wheat cultivar Alturas

Stripe rust is one of major diseases in wheat production worldwide. The best economic and efficient method is to utilize resistant varieties. Alturas has high-temperature adult-plant resistance. In order to determine stripe rust resistance characteristics, resistance gene combination and molecular map of the resistance gene(s), Alturas was crossed with Chinese susceptible cultivar Taichung29. The parents, F₁, F₂ progenies were tested with Chinese predominant mixed races CYR31, CYR32 and CYR33 in field experiments in 2010 and F₃ progenies were evaluated at one site in Beijing, the other site in Langfang, Hebei Province. Infection type (IT) and disease severity (DS) were recorded three times for each plant for F₁ and F₂, and each progeny for F₃ during each growing season. The DS data were used to calculate relative area under the disease progress curve (AUDPC) values. Both IT and AUDPC data showed continuous distributions, indicating that the Alturas HTAP resistance was controlled by quantitative trait loci (QTLs). A major HTAP QTL, designated as QYrAlt.syau-3BS, was consistently detected across environments and was located on chromosome 3BS. The gene contributed to 34.28?% of the phenotypic variation for average AUDPC and 50.20?% for average IT. Markers Xgwm389 and Xbarc238 flanking the major QTL, should be useful in breeding for obtaining durable and high-level resistance by combinations with other non-race-specific resistance genes.     

Microsatellite markers for genes lr34/yr18 and other quantitative trait Loci for leaf rust and stripe rust resistance in bread wheat

ABSTRACT Leaf rust and stripe rust, caused by Puccinia triticina and P. striiformis, respectively, are important diseases of wheat in many countries. In this study we sought to identify molecular markers for adult plant resistance genes that could aid in incorporating such durable resistance into wheat. We used a doubled haploid population from a Japanese cv. Fukuho-komugi x Israeli wheat Oligoculm cross that had segregated for resistance to leaf rust and stripe rust in field trials. Joint and/or single-year analyses by composite interval mapping identified two quantitative trait loci (QTL) that reduced leaf rust severity and up to 11 and 7 QTLs that might have influenced stripe rust severity and infection type, respectively. Four common QTLs reduced stripe rust severity and infection type. Except for a QTL on chromosome 7DS, no common QTL for leaf rust and stripe rust was detected. QTL-7DS derived from 'Fukuho-komugi' had the largest effect on both leaf rust and stripe rust severities, possibly due to linked resistance genes Lr34/Yr18. The microsatellite locus Xgwm295.1, located almost at the peak of the likelihood ratio contours for both leaf and stripe rust severity, was closest to Lr34/Yr18. QTLs located on 1BL for leaf rust severity and 3BS for stripe rust infection type were derived from 'Oligoculm' and considered to be due to genes Lr46 and Yr30, respectively. Most of the remaining QTLs for stripe rust severity or infection type had smaller effects. Our results indicate there is significant diversity for genes that have minor effects on stripe rust resistance, and that successful detection of these QTLs by molecular markers should be helpful both for characterizing wheat genotypes effectively and combining such resistance genes.     

Identification of a quantitative trait locus for high-temperature adult-plant resistance against Puccinia striiformis f. sp. hordei in 'Bancroft' barley

Sustainable control of plant diseases can be achieved by developing cultivars with durable resistance. 'Bancroft' barley has durable high-temperature, adult-plant (HTAP) resistance to stripe rust caused by Puccinia striiformis f. sp. hordei. The objectives of this study were to determine the inheritance of the HTAP resistance in Bancroft, develop molecular markers for the HTAP resistance using the resistance gene analog polymorphism (RGAP) technique, map the HTAP resistance quantitative trait locus or loci (QTL) on barley chromosomes, and determine the usefulness of the RGAP markers in other barley cultivars for marker-assisted selection. The parents and F(4) recombinant inbred lines (RIL) and the parents and F(5) RIL were evaluated in 2004 and 2005 in one and three field sites, respectively, in Washington State. Infection type (IT) and disease severity (DS) were recorded three times at each location during each growing season. Area under the disease progress curve (AUDPC) was calculated for each parent and RIL based on the DS data. Genetic analyses of IT data of the parents, F(1), and F(2) tested in the adult-plant stage under controlled high-temperature cycle in the greenhouse and the parents, F(4), and F(5) RIL in the field indicated that one dominant gene controlled the HTAP resistance in Bancroft. Using 119 F(5:6) RIL and IT data, a linkage map on chromosome arm 3HL was constructed with eight RGAP markers and three simple sequence repeat (SSR) markers. Using the QTL analysis, a QTL for HTAP resistance was mapped with the DS and AUDPC data on the same chromosome location as with the IT data. The QTL explained >70% of the total phenotypic variation for the DS and AUDPC. The heritability of the HTAP resistance based on the AUDPC data was 76%. The two markers most close to the QTL peak detected polymorphisms in 84 and 88% of 25 barley genotypes that do not have the Bancroft HTAP resistance when used individually, and detected polymorphism in 100% of the genotypes when used in combination, indicating that the markers could be used in incorporating the HTAP resistance into these barley genotypes to improve the level and durability of resistance to stripe rust.     

Genetic architecture of adult plant resistance to leaf rust in a wheat association mapping panel

Leaf rust caused by Puccina triticina is one of the most destructive fungal diseases of wheat (Triticum aestivum). Adult plant resistance (APR) is an effective strategy to achieve long‐term protection from the disease. In this study, findings are reported from a genome‐wide association study (GWAS) using a panel of 96 wheat cultivars genotyped with 874 Diversity Arrays Technology (DArT) markers and tested for adult leaf rust response in six field trials. A total of 13 quantitative trait loci (QTL) conferring APR to leaf rust were identified on chromosome arms 1BL, 1DS, 2AS, 2BL, 2DS, 3BS, 3BL, 4AL, 6BS (two), 7DS, 5BL/7BS and 6AL/6BS. Of these, seven QTLs mapped close to known resistance genes and QTLs, while the remaining six are novel and can be used as additional sources of resistance. Accessions with a greater number of combined QTLs for APR showed lower levels of disease severity, demonstrating additive and significant pyramiding effects. All QTLs had stable main effects and they did not exhibit a significant interaction with the experiments. These findings could help to achieve adequate levels of durable resistance through marker‐assisted selection and pyramiding resistance QTLs in local germplasm.     

Tagging and validation of a major quantitative trait locus for leaf rust resistance and leaf tip necrosis in winter wheat cultivar forno

ABSTRACT A major leaf rust (Puccinia triticina) resistance quantitative trait locus (QTL) (QLrP.sfr-7DS) previously has been described on chromosome 7DS in the winter wheat (Triticum aestivum) cv. Forno. It was detected in a population of single-seed descent (SSD) lines derived from the cross Arina x Forno. QLrP.sfr-7DS conferred a durable and slow-rusting resistance phenotype, co-segregated with a QTL for leaf tip necrosis (LTN) and was mapped close to Xgwm295 at a very similar location as the adult plant leaf rust resistance gene Lr34 found in some spring wheat lines. Here, we describe the validation of this QTL by mapping it to the same chromosomal region close to Xgwm295 on chromosome 7DS in a population of SSD lines from the winter wheat x spelt (T. spelta) cross Forno x Oberkulmer. In both populations, the log of the likelihood ratio curves for leaf rust resistance and LTN peaked at identical or very similar locations, indicating that both traits are due to the same gene. We have improved the genetic map in the target region of QLrP.sfr-7DS using microsatellite and expressed sequence tag (EST) markers. Two EST loci (Xsfr.BF473324 and Xsfr.BE493812) define a genetic interval of 7.6 centimorgans containing QLrP.sfr-7DS, a considerably more precise genetic location for this QTL than previously described both in spring and winter wheat. The identified genetic interval is physically located in the distal 39% of chromosome 7DS. Single-marker analysis identified Xsfr.BF473324 and Xgwm1220 as the most informative loci for QLrP.sfr-7DS and QLtn.sfr-7DS. In the rice genome, the two ESTs flanking the QLrP.sfr-7DS/QLtn.sfr-7DS chromosomal segment in wheat are conserved on chromosome 6S in a region colinear with wheat chromosome 7DS. There, they define a physical region of three rice bacterial artificial chromosomes spanning approximately 300 kb.     

Quantitative trait loci for high-temperature adult-plant and slow-rusting resistance to Puccinia striiformis f. sp. tritici in wheat cultivars

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most damaging diseases of wheat (Triticum aestivum) globally. High-temperature adult-plant resistance (HTAPR) and slow-rusting have great potential for sustainable management of the disease. The wheat cultivars Luke and Aquileja have been previously reported to possess HTAPR and slow-rusting to stripe rust, respectively. Aquileja displayed less number of stripes per unit leaf area than Luke, while Luke showed lower infection type than Aquileja at adult-plant stages of growth under high-temperature conditions. The objectives of this study were to confirm the resistances and to map the resistance genes in Luke and Aquileja. Luke was crossed with Aquileja, and 326 of the F(2) plants were genotyped using 282 microsatellite primer pairs. These F(2) plants and their derived F(3) families were evaluated for resistance to stripe rust by inoculation in the fields and greenhouses of high- and low-temperatures. Infection type was recorded for both seedlings and adult plants, and stripe number was recorded for adult plants only. Two quantitative trait loci (QTL) were identified, on the short arm of chromosome 2B, to be significantly associated with infection type at adult-plant stages in the fields and in the high-temperature greenhouse. The locus distal to centromere, referred to as QYrlu.cau-2BS1, and the locus proximal to centromere, referred to as QYrlu.cau-2BS2, were separated by a genetic distance of about 23 cM. QYrlu.cau-2BS1 was flanked by the microsatellite markers Xwmc154 and Xgwm148, and QYrlu.cau-2BS2 was flanked by Xgwm148 and Xabrc167. QYrlu.cau-2BS1 and QYrlu.cau-2BS2 explained up to 36.6 and 41.5% of the phenotypic variation of infection type, respectively, and up to 78.1% collectively. No significant interaction between the two loci was detected. Another QTL, referred to as QYraq.cau-2BL, was detected on the long arm of chromosome 2B to be significantly associated with stripe number. QYraq.cau-2BL was flanked by the microsatellite markers Xwmc175 and Xwmc332, and it explained up to 61.5% of the phenotypic variation of stripe number. It is possible that these three QTL are previously unmapped loci for resistance to stripe rust.     

Mapping of QTL lengthening the latent period of Puccinia striiformis in winter wheat at the tillering growth stage

The winter wheat lines Luke and AQ24788-83 are respectively susceptible and slow-rusting at tillering stage to yellow (stripe) rust, caused by Puccinia striiformis f. sp. tritici (Pst). A mapping population consisting of 206 recombinant inbred lines was developed from the cross Luke?×?AQ24788-83. These lines were evaluated at the tillering stage in the field trials for infection type (IT) and disease incidence (DI) and in greenhouse trials for IT and latent period (LP). A significant negative correlation was found between LP and DI. A genetic map with 473 marker loci was constructed and used for identifying QTL associated with LP and IT. Two QTL, QYr.cau-1BS and QYr.cau-5AS, were mapped on 1BS and 5AS respectively, explaining collectively up to 46.4 % of LP phenotypic variance. QYr.cau-5AS was clearly distinct, in terms of mapping position, from all six yellow rust resistance genes/QTL previously reported on 5A. QYr.cau-1BS could not be spatially differentiated from three (i.e. YrAlp, Yr15, and YrH52) of the six genes/QTL known on 1BS and centromere-vicinity regions, but was determined to be different from these three genes based on phenotype. The two QTL identified here, therefore, are likely to be novel to the currently known Pst resistance genes/QTL. A minor QTL on 3AL was detected to be associated with both IT and LP. Expression of quantitative resistance at early wheat growth stages and usefulness of the QTL are discussed for the wheat-Pst system.     

甘肃陇南感病小檗在小麦条锈病发生中起提供(初始)菌源作用的直接证据

 陇南是中国小麦条锈菌易变区、小麦条锈病的常发流行区和防治的关键地区。明确陇南小麦条锈菌转主寄主小檗在小麦条锈病发生中的作用,对阐释该地区小麦条锈菌新小种产生的来源和指导小麦条锈病的综合防控具有重要意义。本研究从陇南春季自然受锈菌侵染的堆花小檗及其邻近的小麦上分离获得小麦条锈菌菌系,19个来自发病小檗的单夏孢子堆菌系在中国小麦条锈菌鉴别寄主上产生5个不同毒力类型VP1~VP5,均为新小种;在近等基因系和载体品种组成的鉴别寄主上产生17个不同毒力类型。29个来自邻近发病小檗的小麦上条锈菌菌系在中国小麦条锈菌鉴别寄主上产生了10个不同的毒力类型VP1~VP10,均为新小种;在近等基因系和载体品种组成的鉴别寄主上产生了24个不同的毒力类型。两个小麦条锈菌群体享有共同的毒力类型VP1~VP5,同时来自发病小檗上菌系的一些毒力类型不同于小麦上的毒力类型。主坐标分析(PCoA)和毒性表型聚类分析表明来自于自然受锈菌侵染小檗及其邻近小麦上的小麦条锈菌存在菌源交流。综合研究表明在自然条件下,陇南小麦条锈菌在野生感病小檗上进行有性生殖是常年发生的,感病小檗在新小种产生和在陇南小麦条锈病的发生中起提供菌源的作用。因此,在陇南小麦条锈病的综合治理策略上加强对小檗的处理,降低新小种产生速率和品种抗性丧失速率,从而延长品种使用年限,同时减少新小种向东部广大麦区传播,保障小麦生产安全。