Gene expression profile and physiological and biochemical characterization of hexaploid wheat inoculated with Blumeria graminis f. sp. tritici

Wheat (Triticum aestivum L.) powdery mildew has direct effects on photosynthesis- and energy-related pathways. Because it is prevalent in Sichuan Province, China, knowledge concerning Pm40-mediated wheat powdery mildew resistance is important for understanding host–pathogen interactions. The aim of this study was to understand physiological changes during the host–pathogen interaction, including gene expression, photosynthetic and chlorophyll fluorescence parameters, chlorophyll content, and antioxidant activity, specific to the resistance response to powdery mildew. The Pm40-expressing L693 and Pm40-deficient L1095 wheat lines were employed for comparison analyses. Fifty-eight and 26 expressed sequence tags (ESTs) were obtained from forward and reverse suppression subtractive hybridization cDNA libraries constructed from Bgt-infected L693 and L1095 leaves, respectively, and 69% of the ESTs in the forward library were related to photosynthesis- and energy metabolism-related pathways. Our data indicate that photosynthesis and carbon metabolism as well as nitrogen metabolism related to photosynthesis are the main pathways involved in Pm40-mediated wheat powdery mildew resistance and that an acetyl CoA-like gene might be a key regulatory factor in these pathways. This study provides new insight into the physiological and genetic mechanisms of resistance to wheat powdery mildew.     

What types of powdery mildew can infect wheat-barley introgression lines?

This work is a detailed study of the infection of fungal biotrophic pathogens causing powdery mildew diseases on introgression lines originating from the intergeneric hybridisation between wheat and barley (Triticum aestivum L. × Hordeum vulgare L.). Powdery mildew fungi are among the most widespread biotrophic pathogens of plants also and infect dicot and monocot species. Most powdery mildew species are strictly host specific. They colonize only a narrow range of species or one particular host species. The intergeneric hybridisation between wheat and barley could result in expansions of host ranges of the barley powdery mildew. Our experiments covered natural infections in the field and artificial infections under greenhouse conditions. Formae speciales of powdery mildew were identified on the basis of the sequencing results of ribosomal internal transcribed spacer sequences (rDNA-ITS). We identified Blumeria graminis f.sp. tritici isolate 14 (HM484334) on the wheat parent and all wheat-barley introgression lines and B. g. f. sp. hordei isolate MUMH1723 (AB 273556) on the barley parent, respectively. The wheat-barley introgression lines were inoculated with barley powdery mildew under greenhouse conditions. According to our results the added barley chromosomes (or segments) do not cause host range expansion of barley powdery mildew.     

小麦种质 Edinburgh-b 白粉菌侵染下的基因表达谱分析

 Edinburgh-b是从英国爱丁堡引进的小麦抗白粉病新种质,为了深入了解该种质的抗白粉病分子机制,本研究采用Agilent公司的4 SymboltB@ 44 k小麦表达谱芯片,以Edinburgh-b接种白粉菌0 h为对照,分析其在白粉菌胁迫下的基因表达谱。结果表明,在43 603个探针中筛选出差异表达探针5 835个,其中上调表达2 801个,下调表达3 034个。差异表达基因主要包括与代谢相关的酶类、转录因子、病程相关蛋白、防卫反应基因和信号传导因子等。Pathway分析显示苯丙烷类生物合成、水杨酸信号通路、茉莉酸生物合成、活性氧代谢等被激活,乙烯生物合成和生长素信号通路被抑制,推测水杨酸和茉莉酸信号通路共同参与了Edinburgh-b对白粉菌的防御反应。选取上调和下调表达共17个基因进行实时定量RT-PCR验证,表明基因芯片数据具有良好的重复性。     

普通小麦“兰考90(6)”品系对白粉病抗性的遗传研究

 普通小麦(Triticum aestivum L.)"兰考90(6)"系列品系是以六倍体小黑麦(X Triticosecale Wittmack;AABBRR)为白粉病抗源培育的新的小麦-黑麦1BL/1RS异易位系。这些品系高抗白粉病。小麦白粉病抗性基因推导试验证明,"豫麦66"携带的抗病基因与大多数已经报道的小麦抗白粉病基因不同。用白粉菌[Blumeria graminis (DC.) E. O. Speer f. sp. tritici]单孢堆分离物进行的遗传分析表明,"兰考90(6)"品系携带一个小种专化的隐性抗白粉病基因。对"中国春"和"兰考90(6)21-12"杂交F₂分离群体进行1RS染色体检测,结果证明该抗白粉病基因不在1RS染色体臂上。本研究为有效利用"兰考90(6)"系列品系中的抗白粉病基因提供了科学依据。     

Real-time PCR quantification of latent infection of wheat powdery mildew in the field

Wheat powdery mildew, caused by the fungal pathogen Blumeria graminis f. sp. tritici (Bgt), is a destructive wheat disease worldwide. The key issue for the disease forecast is to timely and accurately estimate and quantify the latent infection levels in volunteer seedlings where the pathogen over-winters or over-summers to serve as sources of initial inoculum of epidemics. To improve the conventional method, a real-time PCR assay had been established in this study to quantify latent infection level of wheat leaves. Artificially and naturally infected leaves in wheat fields at different geographical locations in China were collected and processed to determine the latent infection levels. Linear relationships between the molecular disease index (MDX) and the observed disease index (DX) were obtained from artificial inoculation experiments. Field experiments showed that the spatial distribution patterns of MDX matched well with those of DX in the most cases. This study demonstrated that the real-time PCR assay was a useful tool to rapidly and accurately quantify the latent infection levels of wheat powdery mildew and to efficiently estimate the initial inoculum potentials of epidemics in the fields.     

The role of jasmonic acid signalling in wheat (Triticum aestivum L.) powdery mildew resistance reaction

Jasmonic acid (JA) signalling plays an important role in plant resistance to pathogens. Previously, JA has been found to play a role in induced disease resistance to necrotrophic pathogens in various plant species, but current researches showed that JA also enhanced resistance to biotrophic pathogens. However, its role in wheat (Triticum aestivum L.) powdery mildew (Blumeria graminis f. sp. tritici, Bgt) resistance reaction is largely unknown. To settle this issue, several typical powdery mildew resistant and susceptible wheat varieties were employed. The sensitivity to exogenous methyl jasmonate (MeJA) to wheat powdery mildew resistance, the concentration fluctuation of endogenous JAs after Bgt inoculation, and the expression profiles of nine pathogenesis-related protein genes (PR genes) after MeJA and Bgt treatments were studied systematically. Exogenous MeJA significantly enhanced the powdery mildew resistance of the susceptible varieties. After inoculation with Bgt, endogenous JAs accumulated rapidly, reached the maxima at 2 to 5 h post-inoculation (hpi), then decreased rapidly, and the concentration was almost the same as that of un-inoculated control at 96 hpi. The expression levels of the nine PRs were measured by real time quantitative RT-PCR (qRT-PCR) at different time points after MeJA application or Bgt inoculation respectively. The MeJA and Bgt strongly activated PR1, PR2, PR3, PR4, PR5, PR9, PR10 and Ta-JA2, but almost didn’t affect Ta-GLP2a. The induced powdery mildew resistance was positively correlated with the activated PR genes. JA plays a positive role in defence against Bgt. JA is a signalling molecule in wheat powdery mildew resistance and future manipulation of this pathway may improve powdery mildew resistance in wheat breeding.     

非自然越夏区小麦白粉病模拟鉴定病圃

作者于1990～1995年根据陕西省关中麦区小麦白粉病侵染循环特点,人工模拟自然发病条件,在非自然越夏区杨陵(省农科院试验地)成功地建立起白粉病菌可周年存活,且秋、春季均能充分诱发感染的混合菌系病圃,宜对小麦种质材料进行全生育期抗白粉病鉴定。利用该病圃已对陕西省1000余份小麦品种(系)及部分国内外抗源亲本材料做了抗病性鉴定和利用评价。     

83份西农系小麦品种（系）抗性鉴定及抗病基因分子检测

为西北农林科技大学小麦新育成品种（系）在黄淮麦区的大面积推广,该研究对83份西农新育成的小麦品种（系）进行苗期抗条锈病和白粉病鉴定,成株期抗条锈病、白粉病、叶锈病和赤霉病鉴定,并在田间自然环境下对其抗性进行鉴定及对相关抗病基因进行分子检测。结果显示,在苗期人工接种鉴定中,有63、29和16份小麦品种（系）分别对条锈菌Puccinia striiformis f.sp.tritici生理小种CYR32、CYR33和CYR34表现出抗性,9份小麦品种（系）对3个条锈菌生理小种均表现出抗性;有10、3和0份小麦品种（系）分别对白粉菌Blumeria graminis f.sp.tritici生理小种E15、E09和A13表现出抗性。在成株期人工接种鉴定中,有23、15、28和62份小麦品种（系）分别对条锈病、白粉病、叶锈病和赤霉病表现出抗性。在83份小麦品种（系）中有6份在苗期和成株期均对小麦条锈病表现出抗性。在田间抗性鉴定中,有57、6、65和40份小麦品种（系）分别对条锈病、白粉病、赤霉病及叶锈病表现出抗性。在83份小麦品种（系）中,3份含有Yr5基因,22份含有Yr9基因,3份含有Yr17基因,2份含有Pm24基因,14份含有Lr1基因,所占比例分别为3.6%、26.5%、3.6%、2.4%和16.8%。     

40个小麦优良品种资源的抗白粉病基因推导

 采用21个具不同无毒性/毒性基因的小麦白粉菌菌株对40个优良品种资源进行了苗期接种,通过对其抗谱的分析发现,其中3个材料含Pm2,4个材料含Pm4b,CP91-13-4-1-4可能含Pm8,红卷芒与小白冬麦的抗谱相同,含XBD,2个具有Pm13,7个材料含2个以上的抗病基因,有11个材料含有未知的抗病基因,另外还有11个材料对所有的参试菌株都表现感病。     

Synthesis of six heterocyclic multifunctional alkylating agents and evaluation of their activity againstPyrenophora avenae andBlumeria graminis f.sp.hordei

Six heterocyclic alkylating agents were synthesized and examined for activity against the oat stripe pathogenPyrenophora avenae on agar plates and against the barley powdery mildew fungusBlumeria graminis f.sp.hordei on barley seedlings. Radial growth ofP. avenae was not significantly affected by any of the compounds, but four of them,α,α-bis[4,7-bis(2-chloroethyl)-1,4,7-triazacyclononane]-para-xylene[3], 1,4,8,11-tetra(2-chloroethyl)-1,4,8,11-tetraazacyclotetradecane[4], 8,11-bis(2-chloroethyl)-1,4,8,11-tetraaza-5,7-oxocyclotetradecane[5] and 7,16-bis(2-chloroethyl-1,4,10,13-tetraoxa-7,16-diazaoctadecane[6], gave significant reductions in biomass ofP. avenae grown in liquid culture and in powdery mildew infection on barley when used at 25μM. Polyamine biosynthesis was examined by following the incorporation of labeled ornithine into polyamines inP. avenae. The four compounds3–6 which reduced mildew infection all reduced the flux of label through to the polyamines inP. avenae. Whether the reductions in mildew infection caused by these compounds is related to reduced formation of polyamines is not known and awaits investigation. http://www.phytoparasitica.org posting May 22, 2005.     

Quantitative trait Loci mapping for adult-plant resistance to powdery mildew in bread wheat

ABSTRACT Powdery mildew, caused by Blumeria graminis f. sp. tritici, is a major disease to wheat (Triticum aestivum) worldwide. Use of adult-plant resistance (APR) is an effective method to develop wheat cultivars with durable resistance to powdery mildew. In the present study, 432 molecular markers were used to map quantitative trait loci (QTL) for APR to powdery mildew in a doubled haploid (DH) population with 107 lines derived from the cross Fukuho-komugi x Oligoculm. Field trials were conducted in Beijing and Anyang, China during 2003-2004 and 2004-2005 cropping seasons, respectively. The DH lines were planted in a randomized complete block design with three replicates. Artificial inoculation was carried out in Beijing with highly virulent isolate E20 of B. graminis f. sp. tritici and the powdery mildew severity on penultimate leaf was evaluated four times, and the maximum disease severity (MDS) on penultimate leaf was investigated in Anyang under natural inoculation in May 2004 and 2005. The heritability of resistance to powdery mildew for MDS in 2 years and two locations ranged from 0.82 to 0.93, while the heritability for area under the disease progress curve was between 0.84 and 0.91. With the method of composite interval mapping, four QTL for APR to powdery mildew were detected on chromosomes 1AS, 2BL, 4BL, and 7DS, explaining 5.7 to 26.6% of the phenotypic variance. Three QTL on chromosomes 1AS, 2BL, and 7DS were derived from the female, Fukuho-komugi, while the one on chromosome 4BL was from the male, Oligoculm. The QTL on chromosome 1AS showed high genetic effect on powdery mildew resistance, accounting for 19.5 to 26.6% of phenotypic variance across two environments. The QTL on 7DS associated with the locus Lr34/Yr18, flanked by microsatellite Xgwm295.1 and Ltn (leaf tip necrosis). These results will benefit for improving powdery mildew resistance in wheat breeding programs.     

河南省西部山区小麦白粉菌群体遗传多样性分析

为揭示河南省小麦白粉菌群体遗传结构、起源及进化关系,采用简单重复序列区间(inter-simple sequence repeats,ISSR)和扩增片段长度多态性(amplified fragment length polymorphism,AFLP)分子标记技术对河南省西部山区4个小麦产区的35个小麦白粉菌单孢分离菌株进行了群体遗传多样性分析。结果显示:ISSR和AFLP分析均将35个菌株分为3个组,组Ⅰ包括来自卢氏和灵宝的大部分菌株;组Ⅱ包括来自栾川、卢氏和巩义的菌株;组Ⅲ由4个地区的个别菌株组成,同时包含1个闭囊壳释放子囊孢子获得的菌株。ISSR分析出菌株遗传距离分布在0.0139～0.6592之间,扩增多态性比率为64.83%,各菌株间的Shannon指数为0.2749;而AFLP分析所得的各菌株遗传距离变化幅度在0.1257～0.9322之间,扩增多态性比率为82.68%,各菌株间的Shannon指数为0.5100。可见,河南省小麦白粉菌具有丰富的遗传多样性,研究所用的2种方法均可用于遗传多样性分析,其中AFLP分析小麦白粉菌群体表现出更为丰富的遗传多样性。     

苦瓜白粉病病原菌和生理小种的鉴定及苦瓜对白粉病的抗性遗传分析

为明确海南省苦瓜白粉病的病原菌、生理小种及苦瓜对白粉病的抗性遗传规律,结合形态学鉴定和分子鉴定解析白粉病菌及生理小种种类,通过显微镜观察白粉病菌侵染过程,并应用主基因+多基因混合遗传模型分析法探讨苦瓜对白粉病的主要抗性遗传规律。结果表明:采集自海南省6个市(县)的苦瓜白粉病病原菌均为单囊壳白粉菌Sphaerotheca fuliginea,属生理小种2F,该菌在侵染苦瓜叶片时有4个关键时期:接种后4 h为分生孢子萌发高峰期,8 h为附着孢形成高峰期,16～24 h为次生菌丝形成高峰期,5 d为分生孢子梗形成高峰期。将其接种于苦瓜抗、感品系,对白粉病的抗性符合2对加性-显性-上位性主基因+加性-显性多基因模型,主基因和多基因共同控制苦瓜对白粉病的抗性,其中以主基因遗传为主,且会受到环境变异的影响。根据苦瓜抗性遗传规律,F2代主基因遗传率最高,受环境影响最小,在苦瓜的白粉病抗性育种中,以早期世代F2代作为有效选择世代。研究表明白粉病菌侵染叶片的前2 d是白粉病防治的最佳时期,所以在白粉病易发的物候期,可将防治时间提前1～2 d。     

小麦品种多样性对白粉病及产量和蛋白质的影响

 对品种多样性控制小麦白粉病的效果以及对产量和蛋白质的影响进行了研究,为利用品种多样性控制白粉病提供理论依据和技术支持。采用SSR技术分析5个小麦品种的亲缘关系;2008年和2009年按组合所含品种数为2、3、4和5个进行组合,在田间种植混种组合和单播小区,人工接种白粉菌,比较各小区中小麦白粉病的AUDPC值、产量和蛋白质变化。结果表明,5个品种亲缘关系相对较近;2个品种混合时以抗感搭配较好,随着抗病品种比例增加控病效果增强;2008年混合种植中有防治效果的组合占54.55%,相对防效为10.02%~47.58%,2009年有防治效果的组合占23.08%,相对防效在1.85% 到18.96 % 之间;在适当的组合中,如京冬8号/轮选987、京双16/京411/京冬8号对小麦白粉病有控制效果,混种不会影响产量和蛋白质含量;当有大量能克服本地品种抗性的外来菌源时混合种植的控病效果降低。品种多样性可以用做防治小麦白粉病的一项措施。     

小麦新品系抗白粉病基因分析

 本文对6个小麦新品系所含的抗白粉病基因进行了遗传分析。将感病品种Liaochun10分别与SM 20121、SM 203390、SM 20125、SM 200332、SM 20126、SM 20005杂交和自交,并将这6个品系互配成半双列杂交组合。用小麦白粉菌15号小种的单孢堆菌系对各杂交组合的亲本、F₁、F₂代群体及F₃代家系进行了苗期抗病性鉴定。遗传分析表明,供试的6个品系对小麦白粉菌15号小种的抗性均由1对显性基因控制。等位性分析推断:SM 20121、SM 203390、SM 20125和SM 200332含有抗白粉病基因Pm12;SM 20126含有抗白粉病基因Pm21;SM 20005含有抗白粉病基因Pm16。建议将这6个品系作为优良抗病亲本利用。     

甘肃省主要小麦生产品种及高代品系的抗白粉基因推导

［目的］ 采用基因推导法对目前甘肃省小麦主要生产品种及高代品系进行抗白粉基因分析,为甘肃省白粉病的抗病育种和品种使用及布局提供依据。［方法］ 利用21个小麦白粉菌株,结合品种系谱对甘肃14个小麦生产品种及28个高代品系进行抗白粉基因推导。［结果］ 14个生产品种中,1个含Pm8,1个含Pm4b,4个含未知抗病基因,其余8个对所有供试菌株全部表现感病;28个高代品系中,5个含Pm8〖STBZ〗,2个含Pm〖STBX〗3〖STBZ〗a,1个含Pm〖STBX〗4〖STBZ〗b,1个含Pm〖STBX〗30〖STBZ〗,5个含未知抗病基因,其余14个对所有供试菌株全部表现感病。［结论］ 目前甘肃小麦生产品种及高代品系中缺乏抗白粉病基因,一旦条件适宜,小麦白粉病在甘肃地区极易发生和流行,应该引起生产管理部门和育种专家的注意和重视。     

小麦品种(系)抗白粉病基因推导及分子标记鉴定

利用基因推导法和分子标记对我国主要麦区的小麦品种(系)进行了抗白粉病基因的鉴定。结果表明,南30-10等15个品种(系)含有Pm8,新麦2号等9个品种(系)含有Pm4,中植4号等9个品种(系)含有Pm21,郑麦113含有Pm4b+5b,杨09-111和新紫1号含有Pm2+mld。研究发现,基因推导和分子标记相结合,可大大提高小麦品种(系)抗白粉病基因鉴定结果的准确性,鉴定结果可为抗病育种和品种布局及白粉病的防治提供依据。     

2009—2010年河南省小麦白粉菌群体毒性及其遗传多样性分析

利用40个鉴别寄主对河南省5个小麦产区的44个单孢子堆纯化分离的小麦白粉病菌进行毒性鉴定分析,并采用多基因家系法进行了遗传多样性分析。结果显示,供试群体对Pm8、Pm1、Pm3a、Pm3b、Pm3c、Pm3f、Pm3e、Pm5、Pm6、Pm7、Pm17、Pm19、Pm34等抗病基因的毒性频率分别达到90%以上,已不能再作为抗源利用;对Pm12、Pm16、Pm21、Pm18、Pm2、Pm2＋6、Pm2＋MLD、Pm5（Mli）、Pm23、Pm30、Pm4a等基因的毒性频率低于15%,仍可以在抗病育种中加以利用;河南省的小麦白粉病菌存在地理之间的差异,同时病菌在地区之间有自西南向东北/东方向的传播。表明省内群体中存在着丰富的遗传多样性。     

Powdery mildew (Oidium sp.) on tomatoes in Bulgaria

In 1989, powdery mildew (Oidium sp.) was found on tomatoes (Lycopersicon esculen-tum L. Mill.) under glass in Bulgaria. Microscopic examinations indicated that the mildew was typical ofOidium sp. Forty species, forms, breeding lines and cultivars of tomato were evaluated for their susceptibility to powdery mildew. Seven of them were immune to the disease.