A Novel Fungal Hyperparasite of Puccinia striiformis f. sp. tritici,the Causal Agent of Wheat Stripe Rust

Puccinia striiformis f. sp. tritici (Pst), the causal fungus of wheat stripe rust, was previously reported to be infected by Lecanicillium lecanii, Microdochium nivale and Typhula idahoensis. Here, we report a novel hyperparasite on Pst. This hyperparasitic fungus was identified as Cladosporium cladosporioides (Fresen.) GA de Vries based on morphological characteristics observed by light and scanning electron microscopy together with molecular data. The hyperparasite reduced the production and viability of urediniospores and, therefore, could potentially be used for biological control of wheat stripe rust.     

Some Effects of Yellow Rust (Puccinia striiformis) on 14Carbon Assimilation and Translocation in Wheat

A well-developed infection of Yellow Rust on a leaf of springwheat (Jufy I) caused the assimilation of ¹⁴CO₂ by that leafto decrease to 43.5 per cent of that of an uninfected leaf.Over a period of three hours translocation of ¹⁴C from an infectedleaf was only 0.87 per cent of that from a control leaf. Whencontrol plants were kept in the light for periods up to 16 hoursafter assimilating ¹⁴CO₂ translocation continued at a steadyrate, whereas there was only negligible translocation from infectedleaves after the first few hours. The retention of labelledassimilates in the infected leaf could be partly, but not completely,accounted for by a conversion of assimilates to an alcohol-insolubleform. Rust infection had no effect on the distribution patternof ¹⁴C to other leaves from one which had assimilated ¹⁴CO₂.In contrast to the marked retention of assimilate by an infectedleaf, such a leaf was unable to distort the normal distributionby attracting assimilates from the other leaves.     

Some Effects of Yellow Rust (Puccinia striiformis) on the Growth and Yield of a Spring Wheat

Yellow Rust had a considerable effect on the yield of the springwheat Jufy I, that of severely infected plants being reducedby as much as 64.5 per cent. This was due to a reduction inthe number of florets per ear, the number of grains per ear,and in some cases the weight of individual grains. There wasalso a marked effect on the general growth and vigour of theplant, the growth of leaves being retarded and the final lengthand breadth greatly reduced. The roots were more seriously affectedthan any other part of the plant with reductions in dry weightof as much as 78 per cent in fully infected plants. The relatively direct effect of a late rust infection greatlyreduced the yield, while the effects of an early infection,although indirect, still caused large reductions. The variedeffects of rust on the several factors contributing to the yield,and the relative importance of the effects on these factorsare discussed.     

Race analysis of Puccinia striiformis f.sp. tritici in Iran

The wheat stripe (yellow) rust is one of the most important diseases in Iran. In this study, 41 races out of 104 isolates in greenhouse were determined from 2008 to 2010. Races 6E6A+, 6E10A+ and 6E0A+ were more common. Races 0E0A+ was less aggressive than races 166E158A+ and 134E158A+ with virulence on 11 known genes. Virulence on plant/s with gene/s Yr1, Yr2, Yr4, Yr6, Yr7, Yr8, Yr9, Yr10, Yr25, Yr27, YrSU, YrSD, YrND, Yr3, Yr2+, Yr6+, Yr9+, Yr7+, YrCV and YrA was detected. The majority of isolates with high frequency (more than 70%) showed virulence on plant/s with Yr2, Yr7, Yr9 and YrA genes. No virulence was detected on plant/s with Yr3, Yr5 and YrSP. In greenhouse test, frequency of virulence to wheat genotypes with Yr1, Yr4, Yr10, YrCV (32+) and YrSD gene was less than 7%. Frequency of virulence to other wheat genotypes was between 8 and 100%.     

Population genetic diversity of Puccinia striiformis f. sp. tritici on different wheat varieties in Tianshui, Gansu Province

Population genetic diversity in Tianshui city was analyzed with SSR markers in 605 single-pustule isolates of the stripe rust pathogen, Puccinia striiformis f. sp. tritici (Pst), obtained from 19 varieties of wheat. Significant differences in genetic diversity among populations were defected. Genetic diversity was highest in population on Tian 863-13, a highly resistant variety, whereas genetic diversity was lowest in population on Huixianhong, a highly susceptible variety. Seven populations from seven varieties that carried the common Yr18 resistance gene were clustered as one sub-group at 0.88 similarity coefficient, which showed that resistance gene selection had close relation with pathogen??s component. The results of present study can provide a theoretical basis for integrated management of wheat stripe rust and effective deployment of resistance genes in Pst over-summering zones in China.     

条形柄锈菌34号生理小种的分子标记

条形柄锈菌Puccinia striiformis f. sp. tritici 34号生理小种(CYR34)是目前我国毒性谱最宽、毒性最强的生理小种,对小麦生产和抗病品种选育造成了极大的影响。本研究采用RAPD-SCAR分子标记技术,从300条RAPD随机引物中筛选到CYR34的特异引物,通过特异性片段回收、克隆和测序(GenBank登录号为OL907303),依据序列设计出了S2008F34/S2008R34特异性引物,能够从CYR34及接种CYR34的小麦发病叶片总DNA中都扩增出417 bp的目标片段。采用该特异性引物检测2021年陕西渭南、咸阳和宝鸡地区小麦条锈菌CYR34的流行频率分别为8.6%、6.0%和10.8%。该项研究为小麦条锈菌CYR34号生理小种的快速检测提供了技术支撑。     

Phenotypic and Molecular Characterization of Wheat for Slow Rusting Resistance against Puccinia striiformis Westend. f.sp. tritici

Race‐specific resistance of wheat (Triticum aestivum L.) to yellow rust caused by Puccinia striiformis Westend. f.sp. tritici is often short‐lived. Slow‐rusting resistance has been reported to be a more durable type of resistance. A set of sixteen bread wheat varieties along with a susceptible control Morocco was tested during 2004–05 to 2006–07 in field plots at Peshawar (Pakistan) to identify slow rusting genotypes through epidemiological variables including final rust severity (FRS), apparent infection rate (r), area under disease progress curve (AUDPC), average coefficients of infection (ACI) and leaf tip necrosis (LTN). Epidemiological parameters of resistance were significantly (P < 0.01) different for years/varieties in three seasons, while variety × year interactions remained non‐significant. Sequence tagged site (STS) marker, csLV34 analyses revealed that cultivars Faisalabad‐83, Bahawalpur‐95, Suleman‐96, Punjab‐96, Bakhtawar‐93, Faisalabad‐85, Shahkar‐95 and Kohsar‐95 possessed Yr18 linked allele. Faisalabad‐83, Bahawalpur‐95, Suleman‐96, Punjab‐96, Bakhtawar‐93 and Faisalabad‐85 were relatively more stable over 3‐years where FRS, AUDPC and r values reduced by 80, 84 and 70% respectively compared to control Morocco. These six varieties therefore could be exploited for the deployment of Yr18 in breeding for slow rusting in wheat. Both FRS and ACI are suitable parameters for phenotypic selection.     

Cloning and characterization of the actin gene from Puccinia striiformis f. sp. tritici

The fungus Puccinia striiformis f. sp. tritici, the causal agent of wheat stripe rust, is an obligate biotrophic basidiomycete. Urediniospores are the most common spore type involved in the epidemiology of this disease. Tip growth of germ tubes of germinated urediniospores is a key step during infection of wheat, but few studies have investigated it so far. Recent research has found that actin is closely associated with hyphal tip growth. In this study, we have cloned and obtained the full-length actin cDNA from P. striiformis f. sp. tritici and characterized its expression. Furthermore, actin filament (F-actin) patterns were visualized microscopically during germ tube formation. The most conspicuous actin-containing structures were actin patches. They were mainly concentrated near the hyphal tip and scattered throughout the cortex. By using cytochalasin B, we observed that depolymerization of F-actin greatly reduced the germination rate of urediniospores and disrupted the transport of vesicles to the germ tube tip, indicating that F-actin played a key role in the tip growth of P. striiformis f. sp. tritici. This work helps us to understand the tip growth mechanism of P. striiformis f. sp. tritici, and may provide a theoretical framework for designing novel pesticides.     

Virulence variation of Puccinia striiformis Westend. f. sp. tritici in Pakistan

Yellow rust populations of Pakistan were characterised for their virulence pathotypes/races and pathogenetic variation using seedling evaluation of differential genotypes under glasshouse conditions in Murree (6000 feet above sea level). Differential genotypes comprised a world set, an European set, near isogenic lines and the universally susceptible bread wheat cultivar “Morocco”. Over the two-year study a total of 18 race groups were identified. Out of these 18 race groups, several (68E0, 64E0, 66E0, 70E0, 6E0, 71E0, 6E0, 2E0, 67E0, and 68E16) were found previously. The new race group 70E32 found probably evolved because of mutation from the previously existing 70E16. Virulence frequencies of yellow rust (Yr) resistance genes were also determined on near isogenic lines. The highest virulence frequencies (%) were found for Yr7 (88%), Yr9 (57%), Yr18 (70%), and Yr24 (69%). Virulence frequencies were low for Yr 1 (4%), Yr5 (7%), Yr10 (10%) and Yr15 (4%). Our studies indicated that virulence existed for almost all yr genes, necessitating regular monitoring of the yellow rust populations and intensifying efforts to identify new sources of resistance to this pathogen.     

Effect of Low Temperature and Wheat Winter-Hardiness on Survival of Puccinia striiformis f. sp. tritici under Controlled Conditions

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most important diseases of wheat worldwide. Understanding the survival of Pst during the overwintering period is critical for predicting Pst epidemics in the spring. Real-time quantitative PCR (qPCR) methods quantifying Pst DNA and RNA (cDNA) were developed and compared for the ability to quantify viable Pst in leaf tissues. Both qPCR of DNA and RNA can provide reliable measurement of viable Pst in plant tissues prior to the late sporulation stage for which qPCR of DNA gave a much higher estimate of fungal biomass than qPCR of RNA. The percentage of Pst biomass that was viable in detached and attached leaves under low temperatures decreased over time. Pst survived longer on attached leaves than on detached leaves. The survival of Pst in cultivars with strong winter-hardiness at 0°C and -5°C was greater than those with weak winter-hardiness. However, such differences in Pst survival among cultivars were negligible at -10, -15 and -20°C. Results indicated that Pst mycelia inside green leaves can also be killed by low temperatures rather than through death of green leaves under low temperatures. The relationship of Pst survival in attached leaves with temperature and winter-hardiness was well described by logistic models. Further field evaluation is necessary to assess whether inclusion of other factors such as moisture and snow cover could improve the model performance in predicting Pst overwintering potential, and hence the epidemic in spring.     

小麦条锈菌分子生态学研究进展

小麦条锈病是危害最严重的小麦流行性病害之一,小麦条锈菌的生态学研究对制定合理的防治策略和抗锈育种具有重要意义.近十几年来,DNA分子标记技术被应用于小麦条锈菌的群体遗传学研究,推动了小麦条锈菌分子生态学研究的快速发展,为揭示小麦条锈菌的群体生态特性开辟了一个新的途径.本文系统介绍小麦条锈菌分子生态学研究的主要进展,并就我国当前研究的局限性和发展趋势进行了分析.     

条形柄锈菌33号生理小种的分子标记

采用RAPD-SCAR分子标记技术,从300条RAPD随机引物中筛选到了对条形柄锈菌Puccinia striiformis f. sp. tritici 33号生理小种特异的2条引物,将特异性片段回收、克隆和测序后（GenBank注册号为AB914691和AB914692）,依据其序列设计出了2对引物S261F33/S261R33和S300F33/S300R33,能够特异性地从33号生理小种基因组DNA及发病小麦叶片总DNA中分别扩增出247bp和763bp的片段,其结果与采用常规的鉴别寄主法鉴定的结果一致。因此,这2对引物都可用于条形柄锈菌33号生理小种的快速鉴定与监测。     

小麦条锈菌CY32夏孢子萌发研究

研究夏孢子萌发过程的分子机制对于从分子水平上理解条锈菌的侵染过程及其与寄主互作的关系具有重要的理论意义。本研究以小麦条锈菌Pucciniastriiformisf.sptritici32号生理小种(CY32)为材料,研究了用水培方法萌发夏孢子的适宜条件。结果显示,CY32夏孢子萌发的最适温度为9℃,最适宜的孢子量是6mg/200mL水,适宜的溶液是无菌蒸馏水。水化能促进夏孢子的萌发,新鲜夏孢子和干燥容器中放置2d的夏孢子经水化15h后,萌发率显著提高。此方法获得的萌发夏孢子提取的RNA可以满足cDNA文库构建和基因表达分析等分子生物学研究的要求,并为小麦条锈菌的分子生物学研究奠定了物质基础。     

小麦条锈菌PsNCS1基因的克隆及转录表达特征

摘要：【目的】克隆小麦条锈菌神经钙离子感应蛋白基因PsNCS1,分析其在病菌不同发育时期的表达水平。【方法】利用文库筛选和RT-PCR技术克隆PsNCS1的cDNA序列,采用生物信息学技术预测分析该基因编码蛋白的保守结构域及基本特性,构建系统发育树;运用实时荧光定量RT-PCR技术分析PsNCS1在病菌不同生长发育时期的表达水平。【结果】PsNCS1全长cDNA为1007 bp（GenBank登录号GU134621）,开放阅读框为573 bp,编码190个氨基酸,分子量为22.17 kDa, 等电点为4.     

A Light and Electron Microscope Study of the Interaction of Yellow Rust (Puccinia striiformis) with a Susceptible Wheat Cultivar

The microscopy and ultrastructure of the interaction of Pucciniastriiformis with a susceptible wheat cultivar was examined atintervals from the time of first haustorium formation to theonset of sporulation. At any particular point in the radiallyexpanding area of infection a sequence of morphological changesoccurred in the infected host cells and the fungus which werecorrelated with successive phases of active fungal growth, accumulationof reserves and finally export of reserves to the developingreproductive structures. The observations are compared withprevious work on other host-rust interactions. yellow rust, Puccinia striiformis, wheat, host-pathogen interaction     

Early molecular diagnosis and detection of Puccinia striiformis f. sp. tritici in China

Aims: Wheat stripe (yellow) rust, caused by Puccinia striiformis f. sp. tritici (Pst), is the most important foliar disease on wheat in China. Early molecular diagnosis and detection of stripe rust will provide a useful aid to the accurate forecast and seasonal control of this destructive disease. Our objective was to develop PCR assays for the rapid identification and detection of P. striiformis. Methods and Results: The genomic DNA of P. striiformis and P. triticina were amplified by a pair of primers derived from conserved β‐tubulin gene sequence. A 235‐bp specific DNA fragment of P. striiformis was isolated and purified. Based on its sequence, another two primer sets were designed successfully to obtain new sequence‐characterized amplified region (SCAR) markers of P. striiformis, which could be amplified in all test isolates of P. striiformis, whereas no DNA fragment was obtained in other nontarget wheat pathogens. The detection limit of the primer set YR (f)/YR (r1) was 2·20 pg μl^?1. The new SCAR markers of P. striiformis can also be detected in Pst‐infected wheat leaves postinoculated for 2 days. Conclusions: Our assays are significantly faster than the conventional methods used in the identification of P. striiformis. Significance and Impact of the Study: Development of a simple, high‐throughput assay kit for the rapid diagnosis and detection of wheat stripe rust would be anticipated in a further study.     

小麦条锈菌冬孢子发生的组织学和超微结构研究

采用冷冻切片技术、光学显微镜和电子显微镜技术,系统研究了小麦条锈菌冬孢子的个体发生过程和超微结构特征。结果表明,小麦条锈菌冬孢子由排列在冬孢子堆基部的双核产孢细胞产生。在发育初期,产孢细胞一端产生突起形成冬孢子芽,随后冬孢子芽经延伸并形成隔膜,依次分化形成冬孢子原基、柄细胞和冬孢子原体。冬孢子原体经有丝分裂后产生隔膜发育形成双核双细胞冬孢子。成熟的冬孢子表面光滑,具有明显加厚的细胞壁,双核融合,原生质密度增加,富含脂肪粒和糖原类物质。在部分冬孢子堆周围还可观察到莲花状包被结构。     

Genetic analysis of resistance to Puccinia striiformis f.sp. tritici in cv. Aflak at adult plant stage

In order to investigate heritability and gene action for yellow rust resistance in wheat, a resistance yellow rust cultivar Aflak was crossed to susceptible cultivar Avocet‘s’. Parents, F₁, F₂ and F₃ generations were cultured according to randomised complete block design with two replications in the research station of Gharakhil, Iran. Parents and other generations were inoculated with 70E0A+ race. Traits including severity and infection type were recorded and then coefficient of infection was calculated. For this trait, generations mean and variance analysis were performed and results showed that there were significant differences among generations for coefficient of infection. Results showed that in addition to additive and dominance effects, at least one kind of epistasis interaction (additive × additive) control this trait. Although additive and dominance effects control this trait, but with attention to generations variance analysis, the results showed that additive variance had important role to control this trait.