A FIASCO-Based Approach for Detection and Diagnosis of Puccinia graminis f. sp. tritici in China

Stem or black rust of wheat, caused by the fungus Puccinia graminis Pers. f. sp. tritici Eriks.＆E. Henn. （Pgt）, has historically caused severe losses to wheat （Triticum aestivum L.） production worldwide. In the Fujian and Guangdong provinces of China, six moderate-to-severe epidemics of wheat stem rust have occurred, which caused destructive losses of wheat between 1949 and 1966, although these were brought under control by integrated management. A rapid and reliable detection of the pathogen will contribute to the accurate forecast and seasonal control of this disease. The objective of this study was to develop a diagnostic molecular marker generated from simple sequence repeats （SSR） for the early rapid identiifcation of P. graminis. The genomic DNA of P. graminis, Puccinia striiformis, Puccinia triticina and seven other species was ampliifed by a pair of SSR primers generated by the FIASCO （fast isolation by AFLP sequences containing repeats） enrichment protocol. The primer set Pgtw （f）/Pgtw （r） generated a polymorphic pattern displaying a 330-bp DNA fragment speciifc for P. graminis whereas no DNA fragment was obtained from other non-target wheat fungal pathogens. The detection limit of the primer was 1 ng DNA in a 25-mL PCR reaction. The SSR markers of P. graminis can also be used to detect the presence of latent hyphae in Pgt-infected wheat leaves as early as 30 h post-inoculation. A rapid approach to distinguish P. graminis from similar pathogenic fungi would be anticipated in further study.     

Evaluation of a set of introgressive spring bread wheat lines developed for resistance to stem rust race Ug99 + Sr24 (TTKST) at the Southeast Agricultural Research Institute

Under conditions of a natural stem rust epiphytotic caused by Puccinia graminis f. sp. tritici race Ug99 + Sr24 (TTKST), a set of introgressive spring bread wheat lines developed at the Southeast Agricultural Research Institute was evaluated at KARI’s international stem rust screening nursery (Kenya). New sources of resistance to the pathogen and effective resistance genes have been revealed.     

小麦秆锈病防治指标研究

为考察小麦秆锈病的防治指标,在小麦自然生长条件下,以绵阳11号、浙麦1号和龙麦30号3个具有不同抗感性的小麦品种为研究对象,通过对不同病情与产量损失关系的研究,分别构建了3个品种开花期病情(X2)与产量损失率(L)之间的回归方程,其中高感品种绵阳11号的方程为:L=-1.689+1.185X2 (R=0.981,SLx...     

44份优质春小麦品种(系)抗病性状况的分析

2001~2003年连续3年对来自国内外44份优质春小麦品种(系)进行了苗期和成株期抗秆锈、叶锈和白粉病的鉴定。结果表明:44份材料对秆锈病抗性较好,部分材料对叶锈病表现抗性,但全都感染白粉病。据此提出了选育优质、高产、抗病小麦新品种的方法和育种策略。     

加拿大小麦生产和锈病防治

小麦是加拿大种植面积最大的作物,大部分种植于加拿大西部曼尼托巴省、萨斯喀彻温省、阿尔伯塔省的草原省份。在加拿大小麦种植面积大约有1 000万hm2,包括700万hm2的六倍体春小麦,200万hm2的硬粒小麦和100万hm2的冬小麦。六倍体小麦又根据不同的质量标准和多样化的食品分类、市场需要等划分成很多类。其中最主要的一类叫加西硬红类(CWRS),是加拿大最主要用于做面包的春小麦品种系列。历史上危害小麦的病害主要是由秆锈菌(Puccinia graminisf.sp.tritici)引起的小麦秆锈病。在加拿大抗秆锈病的第一个重要品种是Thatcher,从20世纪30年代到70年代早期广泛种植。然而Thatcher对由叶锈菌(P.triticina)引起的叶锈病十分感病。多年来,随着含其他抗锈基因(主要是Sr2,Sr6,Sr7a,Sr9b,Lr13,Lr14a,Lr16,Lr34)品种的育成和推广,秆锈病得到了很好的控制,但叶锈病却由于P.triticina种群变化,导致例如Lr13和Lr16抗性基因的抗性丧失,仍然造成很大的损失。小麦条锈病主要由柄锈菌(P.striiformisf.sp.tritici)引起的,长期以来,条锈病是阿尔伯塔南部灌溉区小麦生产上的主要病害,但是自2000年以来,小麦条锈病已经在加拿大中部草原区和安大略南部发现,并造成严重危害。加拿大小麦品种中,只有少数含有中抗水平的Yr18抗条锈基因,而大多数小麦品种对条锈病的抗性基础及抗病遗传尚不清楚。展望未来,锈病仍然是加拿大小麦的主要病害,如条锈病或者是具有高致病力的秆锈病小种,Ug-99可能会是加拿大小麦及谷物生产的新威胁,目前解决这些问题的最好策略是致力于长期的抗锈病遗传育种研究,包括聚合有效耐用的基因,对基因进行有效的部署和聚合抗性基因使遗传资源最大化等手段。将科研重点放在自然遗传抗病方面旨在使加拿大农民能够按对自然环境有利的方式来生产化学残余量最少的小麦,从而在国内和国际市场竞争中占据优势。     

美国西北部59个小麦品种（系）抗条锈病基因分子检测及对中国条锈菌系抗性鉴定#br#

 【目的】了解59份美国西北部小麦品种（系）对中国当前条锈菌流行小种的抗性水平,分析其抗病基因的存在情况,为培育中国抗条锈病新品种提供资源和依据。【方法】选用中国小麦条锈菌优势小种条中31号、条中32号和条中33号对大部分具有高温成株抗性的美国59份小麦品种（系）苗期及成株期进行抗性接种鉴定,利用与当前有效的全生育期抗性基因Yr10、Yr15、成株抗性基因Yr18、高温成株抗性基因Yr39紧密连锁的分子标记对供试材料进行分子检测。【结果】抗性鉴定表明,Expresso、02W50076、ACS52610、WA008012、WA008018等5份材料苗期与成株期对条中31号、条中32号和条中33号混合菌均表现为抗病,属于全生育期抗病品种。 33份材料苗期表现感病而成株期表现抗病,属于成株期抗病品种。分子检测发现59份材料均不含有Yr10;含有Yr15、Yr18和Yr39的材料分别为12、33和29份,占20%、56%和49%。【结论】Yr18、Yr39在美国西北部小麦品（系）中的分布较普遍,并且大部分可以抵抗中国当前流行的条锈菌生理小种,可应用这些抗性材料配制中国小麦抗条锈病新品种。     

澳大利亚小麦品种Sunco抗条锈病性状的遗传分析

澳大利亚小麦品种Sunco成株期对中国条锈病小种条中32(CY32)表现高抗-免疫,而小麦品种川育12则表现为高感。利用Sunco/川育12的双单倍体(DH)群体对Sunco进行抗条锈病遗传研究,结果表明,品种Sunco可能具有持久抗性,抗性是由一对主效基因和两对微效基因联合作用的结果;主效基因的抗性反应为抗-中抗(R-MR),微效基因单独存在作用不明显,但与主效基因结合起来会加强抗条锈病能力。同时对微效多基因的持久抗性利用作了初步探讨。     

Evaluating ten spectral vegetation indices for identifying rust infection in individual wheat leaves

Ten, widely-used vegetation indices (VIs), based on mathematical combinations of narrow-band optical reflectance measurements in the visible/near infrared wavelength range were evaluated for their ability to discriminate leaves of 1 month old wheat plants infected with yellow (stripe), leaf and stem rust. Narrow band indices representing changes in non-chlorophyll pigment concentration and the ratio of non-chlorophyll to chlorophyll pigments proved more reliable in discriminating rust infected leaves from healthy plant tissue. Yellow rust produced the strongest response in all the calculated indices when compared to healthy leaves. No single index was capable of discriminating all three rust species from each other. However the sequential application of the Anthocyanin Reflectance Index to separate healthy, yellow and mixed stem rust/leaf rust classes followed by the Transformed Chlorophyll Absorption and Reflectance Index to separate leaf and stem rust classes would provide for the required species discrimination under laboratory conditions and thus could form the basis of rust species discrimination in wheat under field conditions.     

中国小麦秆锈病研究进展

小麦秆锈病是在小麦茎叶部发生的一种病害,发病时严重影响小麦产量。在中国的小麦种植区内均有秆锈病发生。通过分析对小麦秆锈病的病原菌及发病症状、环境因素对秆锈病的影响、秆锈菌生理小种变化和鉴别寄主演变以及抗病育种的方法进行了综述。     

离体叶段鉴定小麦白粉病、条锈病和叶锈病抗性方法的研究

用苯骈咪唑保绿的离体叶段对小麦白粉病、条锈病和叶锈病抗性的鉴定结果表明：小麦对白粉病的抗性,用５０ ｍｇ／ｋｇ 苯骈咪唑保绿的小麦离体叶段鉴定结果与田间成株期鉴定结果基本一致;而对小麦抗条锈病、叶锈病性的鉴定结果和田间成株期鉴定的结果有差异。说明此法可用于小麦抗白粉病性的鉴定,而对小麦抗条锈病、叶锈病的鉴定,此法尚待改进。     

243份云南普通小麦地方品种抗条锈病鉴定及分子标记检测

【目的】小麦条锈病由条形柄锈菌小麦专化型（Puccinia striiformis. f. sp. tritici,Pst）引起的气传性真菌病害,是世界范围内最具破坏性的小麦病害之一。发掘抗病资源和培育抗病品种是防治条锈病最经济有效的措施。通过鉴定和评价云南小麦地方品种对中国当前流行的条锈菌生理小种的抗性水平,综合分析可能携带的抗条锈病基因,为小麦抗病育种提供理论依据。【方法】利用当前中国小麦生产上毒性强、流行频率高的条锈菌生理小种CYR32和CYR34对243份云南小麦地方品种进行苗期抗病性鉴定,成株期使用CYR32、CYR33、CYR34和贵农致病类群等混合生理小种进行抗性鉴定,并利用小麦生产上重要抗条锈病基因Yr5、Yr10、Yr15、Yr18、Yr26、Yr28、Yr29、Yr30、Yr36、Yr39、Yr41、Yr48、Yr65、Yr67、Yr80和Yr81的紧密连锁标记对供试材料进行分子检测。【结果】在243份小麦材料中,18份种质对CYR32表现苗期抗性,32份种质对CYR34表现苗期抗性,对CYR32和CYR34均表现苗期抗病的有8份,占3.29%。结合苗期和成株期抗性鉴定,174份地方种质表现为稳定的成株期抗性,占71.6%,其中105份种质高抗条锈病。分子检测结果显示,携带Yr10、Yr18、Yr29、Yr30、Yr65和Yr81抗性基因的材料分别有48、44、4、6、4和101份。同时携带2、3和4个抗性基因的材料各有34、4和1份。所有供试材料均未检测到Yr5、Yr15、Yr26、Yr28、Yr36、Yr39、Yr41、Yr48、Yr67和Yr80。此外,74份地方种质未检测到以上所有抗性基因,其中58份具有成株期抗性,可能携带其他已知或新的条锈病抗性基因。【结论】云南小麦地方品种作为中国特有的小麦种质资源,具有优良的条锈病抗性。243份供试小麦品种对当前条锈菌流行小种抗性水平整体偏高,筛选得到174份具有稳定抗性的地方种质,同时有74份可能携带其他已知或未知的抗性基因,可作为进一步挖掘抗条锈病新基因或QTL的亲本来源。     

将秦岭黑麦遗传物质导入普通小麦的研究

用 39个小麦与秦岭黑麦杂交 ,从杂交成功的 16个组合后代中发现“中国春×秦岭黑麦”与“新中长×秦岭黑麦”的F1能够自然结实产生有生活力的杂种。其中植株 (中国春×黑麦 )F2 - 2和 (新中长×黑麦 )F2 - 2进一步自交、回交结实性较好。并从后者获得了遗传上稳定的染色体数为 42的纯合小麦新材料 99L2。秦岭黑麦的抗条锈特性已被转移到 99L2遗传背景中并得到表达。     

甘肃省小麦品种(系)抗条锈性鉴定及评价

对甘肃省40个小麦品种(系)进行抗条锈性鉴定。结果表明:全生育期抗病的参试材料较少,在苗期感病的许多材料,在成株期抗病,并且反应型非常低,说明小麦不同生育阶段抗性存在差异;而且成株期绝大部分品种(系)的严重度、普遍率及病情指数均低于苗期。并针对本研究结果对今后品种抗条锈鉴定、抗锈育种及现有品种的生产应用所涉及的有关问题进行了探讨。     

Genome-wide association analysis for stripe rust resistance in spring wheat(Triticum aestivum L.) germplasm

Stripe rust is a continuous threat to wheat crop all over the world. It causes considerable yield losses in wheat crop every year. Continuous deployment of adult plant resistance(APR) genes in newly developing wheat cultivars is the most judicious strategy to combat this disease. Herein, we dissected the genetics underpinning stripe rust resistance in Pakistani wheat germplasm. An association panel of 94 spring wheat genotypes was phenotyped for two years to score the infestation of stripe rust on each accession and was scanned with 203 polymorphic SSRs. Based on D' measure, linkage disequilibrium(LD) exhibited between loci distant up to 45 c M. Marker-trait associations(MTAs) were determined using mixed linear model(MLM). Total 31 quantitative trait loci(QTLs) were observed on all 21 wheat chromosomes. Twelve QTLs were newly discovered as well as 19 QTLs and 35 previously reported Yr genes were validated in Pakistani wheat germplasm. The major QTLs were QYr.uaf.2 AL and QYr.uaf.3 BS(PVE, 11.9%). Dissection of genes from the newly observed QTLs can provide new APR genes to improve genetic resources for APR resistance in wheat crop.     

2份春小麦种质资源成株期抗条锈病基因遗传分析

研究分析2份春小麦种质资源成株期的抗条锈病基因遗传规律,为春小麦抗条锈病基因利用提供理论依据。采用春小麦Taichung29与MY004730杂交、ZM018243与MY004730杂交的方式分别创建F_2:3代分离群体进行2 a的田间测试,对抗条锈病基因遗传进行分析。结果表明,2个F_2:3群体的病害严重度和反应型在2个试验点均未呈现连续性分布,且均不符合正态分布。初步推测,MY004730与ZM018243对春小麦条锈病的成株期抗性具有质量性状特征。Taichung29与MY004730杂交构建的F_2:3群体单株/家系抗感分离符合由1对基因作用的3R:1S的分离比,表明MY004730的成株期抗条锈性由1对显性基因控制; ZM018243与MY004730杂交构建的F_2:3群体的单株/家系抗感分离比均符合9R:7S,即符合由2对显性基因共同作用的分离比, ZM018243中可能也含有1对显性成株期抗条锈性基因。下一步可以进行抗病基因的分子标记定位,找到与抗病基因紧密连锁的标记,为分子标记辅助选择育种服务,同时还能在抗-抗杂交群体中直接筛选抗病基因聚合材料,为育种提供优良抗病材料。     

Identification of QTL for adult plant resistance to stripe rust in bread wheat line C33

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is a serious disease in bread wheat(Triticum aestivum L.). Identification and use of adult plant resistance(APR) resources are important for stripe rust resistance breeding. Bread wheat line C33 is an exotic germplasm that has shown stable APR to stripe rust for more than 10 years in Sichuan Province of China. Here, 183 recombinant inbred lines(RILs) derived from the cross between C33 and a susceptible line X440 were genotyped with diversity arrays technology(DArT) markers to identify resistance quantitative trait locus(QTL). Field trials were conducted in five years at Chengdu and Xindu of Sichuan Province, using maximum disease severity(MDS) as stripe rust reaction phenotypes. A total of four quantitative trait loci(QTLs) were detected, respectively designed as QYr.saas-3 AS, QYr.saas-5 AL, QYr.saas-5 BL, and QYr.saas-7 DS, explaining 4.14–15.21% of the phenotypic variances. QYr.saas-5 BL and QYr.saas-7 DS were contributed by C33. However, the level for stripe rust resistance contributed by them was not strong as C33, suggesting the presence of other unidentified QTLs in C33. QYr.saas-7 DS corresponded to Yr18 and QYr.saas-5 BL remains to be formally named. The RIL lines carrying combinations QYr.saas-5 AL, QYr.saas-5 BL, and QYr.saas-7 DS showed comparability resistance with C33. The present study provides resources to pyramid diverse genes into locally adapted elite germplasm to improve the stripe rust resistance of bread wheat.     

Molecular Screening and Resistance Evaluation of American Wheat Cultivars to Chinese Stripe Rust Races

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the major diseases of wheat in China. In order to asses the resistance levels and existing Yr genes among 59 wheat cultivars （lines） from the Pacific Northwest （PNW） of the United States, to provide resistance resources for genetic improvement of wheat stripe rust resistance in China, 59 wheat cultivars （lines） from PNW of the United States were infected by 3 mixed races of predominant Chinese stripe rust races CRY31, CRY32, and CRY33 to evaluate their resistance at seedling and adult plant stages, and screened with molecular markers tightly linked to currently effective all-stage resistance genes YrlO, Yrl5 and adult plant resistance genes Yrl8, Yr39. Of 59 American cultivars （lines）, five cultivars （lines）, Expresso, 02W50076, ACS52610, WA008012, and WA00801833, had all-stage resistance, showing resistance to mixed races of CRY31, CRY32, and CRY33 at both seedling and adult plant stages. 33 cultivars （lines） had adult plant resistance, only showing resistance to stripe rust at adult stage. Based on the molecular screening, none of the 59 PNW cultivars （lines） had the polymorphic bands of linked markers to YrlO. There were 12, 33 and 29 cultivars （lines） which bad polymorphic bands of linked markers to Yr15, Yr18 and Yr39, accounting for 20, 55 and 49% of the 59 PNW cultivars （lines）, respectively. All these results suggested that Yr15, Yr18 and Yr39 were widespread among PNW cultivars （cultivars） and could be utilized in Chinese wheat stripe rust resistance breeding.     

小麦-小黑麦抗条锈病杂交后代种质农艺性状及细胞分子遗传学分析

黑麦具有优良的遗传多样性,是改良小麦品质、产量和抗性的重要亲本之一,是丰富小麦种质资源的一条重要途径。在远缘杂交的基础上对16份普通小麦与小黑麦的杂交品系进行条锈病抗性鉴定、农艺性状分析、细胞分子遗传学鉴定及相关品质分析,并对可能导入小麦中的外源染色体(片段)进行了多方面定位。鉴定出9个小麦×小黑麦1BL/1RS易位系、1个代换系和2个1BL/1RS易位系/代换系;抗病鉴定结果表明9个杂交品系在苗期和成株期对条锈病均表现为高抗,可为小麦抗条锈病育种提供抗源;高分子量麦谷蛋白亚基组成分析显示16个品系中,HMW-GS组成类型以最常见的Null、7+8、2+12出现的频率最高,其中P174在16份材料中的综合得分最高,符合小麦种一级麦的标准;田间农艺性状和品质测定结果表明12个小麦品系的容重达到一级麦标准,赖氨酸含量的变化范围为2.8~4.4 g/kg。通过该结果以期鉴定出一批田间抗性优良和有价值的中间材料,为抗条锈病育种工作提供新的种质资源。     

小麦秆锈病新抗源及抗病基因所在染色体特异分子标记

【目的】小麦秆锈病是具潜在毁灭性的小麦病害之一,秆锈菌小种Ug99严重威胁全球小麦生产。本研究通过对165份小麦-近缘植物染色体系进行抗秆锈病鉴定,筛选小麦秆锈病新抗源并建立抗病基因所在染色体特异分子标记,以发掘小麦秆锈病新抗源,培育抗病品种,有效防御Ug99导致的秆锈病。【方法】将供试的165份小麦-近缘植物染色体系、3份六倍体小麦及感病对照小密穗分别播种于直径10 cm的瓦盆中,生长到一叶一心时,用中国小麦秆锈菌流行小种34MKGQM和21C3CTHSM进行接种,当感病品种小密穗充分发病时,按照0-4级标准调查记载侵染型,对供试材料的抗秆锈病级别进行统计。同时,提取免疫、近免疫、高抗秆锈病附加系/代换系及其对应的整套染色体系、中国春等材料的基因组DNA,利用101对PLUG引物进行PCR扩增,扩增产物经限制性内切酶酶切后进行电泳检测,筛选并建立抗秆锈基因所在染色体特异分子标记。【结果】在165份小麦-近缘植物染色体系中,中国春-卵穗山羊草7M^g#1附加系、中国春-卵穗山羊草7M^g#1(7A)和7M^g#1(7B)代换系、中国春-帝国黑麦1R附加系、中国春-中间偃麦草?Ai附加系（?表示外源染色体同源群未鉴定）、中国春-单芒山羊草6N附加系、中国春-易变山羊草6S^vS端体附加系和中国春-智利大麦6H^ch附加系9份材料对秆锈病表现为免疫或近免疫;ALCD-尾状山羊草7C#1附加系、中国春-卵穗山羊草7M^g#1(7D)代换系、中国春-帝国黑麦6R附加系、中国春-高大山羊草6S^l#3附加系、中国春-高大山羊草6S^l#2(6B)代换系、中国春-希尔斯山羊草3S#1附加系和中国春-拟斯卑尔脱山羊草2S^g#3附加系7份材料对秆锈病表现为高抗;其余材料均表现为中感或高感。抗秆锈性基因定位信息比较分析发现,高大山羊草6S^l#2和6S^l#3、帝国黑麦6R、智利大麦6H^ch、卵穗山羊草7M^g#1、尾状山羊草7C、中间偃麦草?Ai染色体上可能含有抗秆锈新基因。分子标记筛选、定位及特异性验证研究共获得8个新的多态性标记,其中5个（TNAC1715、TNAC1718、TNAC1737、TNAC1739和TNAC1753）和3个（TNAC1740、TNAC1751和TNAC1756）分别被定位在6R和6S^l染色体上。【结论】筛选得到8份可能含有抗秆锈新基因的材料,建立了抗秆锈基因所在染色体特异新标记8个。