小麦叶锈菌在感病寄主上发育的组织病理学和超微结构研究

 应用荧光显微技术、微分干涉技术和生物电镜技术,系统地研究了小麦叶锈菌在感病寄主上的发育过程及其超微结构特征。小麦叶锈菌在感病品种上的发育过程可分为几个明显的阶段,即孢子的萌发、附着胞的形成、气孔下囊的分化、初生菌丝和次生菌丝的形成和生长、吸器母细胞和吸器的形成、夏孢子床和夏孢子堆的产生以及夏孢子的形成。小麦叶锈菌的胞间菌丝呈丝状,生长和分枝通常沿寄主细胞壁进行。胞间菌丝与寄主细胞的接触诱导了吸器母细胞的分化,吸器母细胞在与寄主细胞壁的接触部位发育形成入侵栓,穿透寄主细胞壁后于细胞内形成吸器。胞间菌丝和吸器母细胞均含有双核,而成熟吸器则含有单核。经常规染色后,胞间菌丝和吸器母细胞的壁与隔膜均可分辨出由多层构成。     

Effect of interrupted leaf wetness periods on pustule development of Puccinia recondita f. sp. tritici on wheat

In growth chamber experiments, seedlings of a susceptible wheat cultivar (Rubis) were inoculated with urediospores of wheat leaf rust. Inoculated seedlings were incubated in a moist chamber. The period of exposure to leaf wetness varied in duration and progressed with or without one or more interruptions of variable duration. During the interruptions the leaves with germlings were dry. At near-optimal temperature, leaf wetness periods of six h resulted in 60 to 65% of the pustules produced with 12 or 24 h wetness periods. Interruption of a 6 h leaf wetness period by a 1 h dry period was most damaging to the rust germlings at about three to four h after inoculation. The damage became visible as a reduction in the number of resulting pustules and as a prolongation of the median latency period. Epidemiological implications of leaf wetness interruptions are discussed.     

Effect of interrupted leaf wetness periods on pustule development of Puccinia recondita f. sp. tritici on wheat

In growth chamber experiments, seedlings of a susceptible wheat cultivar (Rubis) were inoculated with urediospores of wheat leaf rust. Inoculated seedlings were incubated in a moist chamber. The period of exposure to leaf wetness varied in duration and progressed with or without one or more interruptions of variable duration. During the interruptions the leaves with germlings were dry. At near-optimal temperature, leaf wetness periods of six h resulted in 60 to 65% of the pustules produced with 12 or 24 h wetness periods. Interruption of a 6 h leaf wetness period by a 1 h dry period was most damaging to the rust germlings at about three to four h after inoculation. The damage became visible as a reduction in the number of resulting pustules and as a prolongation of the median latency period. Epidemiological implications of leaf wetness interruptions are discussed.Kiemplanten van het vatbare tarweras Rubis werden geïnoculeerd met uredosporen van de bruine roest van tarwe. De proeven vonden plaats in klimaatkamers. Geïnoculeerde planten werden geïncubeerd in incubatiehokjes met een zeer hoge luchtvochtigheid, teneinde condensatiedruppels op de bladeren en aldus bladnatperioden te krijgen. De toegediende bladnatperioden varieerden in tijdsduur en zij verliepen met of zonder één of meer onderbrekingen, ook weer van variërende duur. Tijdens de onderbrekingen stonden de planten met roestkiemlingen in het licht; het blad was dan droog. Bij ongeveer optimale temperatuur en bladnatperioden van 6 uur was het aantal verkregen puistjes 60 à 65% van het aantal verkregen puistjes na een bladnatperiode van 12 of 24 uur. Onderbreking van de 6-urige bladnatperiode door een 1-urige droge periode was voor de roestkiemlingen het schadelijkst ongeveer 3 tot 4 uur na inoculatie. De schade kwam tot uitdrukking in het kleinere aantal geproduceerde puistjes en in een verlenging van de mediane latentieperiode. De epidemiologische betekenis van onderbrekingen van de bladnatperiode wordt besproken.     

Comparative Analysis of Indices in the Study of Virulence Diversity Between and Within Populations of Puccinia recondita f. sp. tritici in Israel

ABSTRACT Isolates of Puccinia recondita f. sp. tritici (n = 260) obtained from bread, durum, and wild emmer wheat leaf collections throughout Israel during 1993 to 1997 were analyzed for virulence on a set of wheat differentials. The overall frequency of virulence increased on differentials possessing resistance genes Lr1, Lr2a, Lr3, and Lr26 and decreased on Lr17, Lr21, and Lr30. Genes Lr9 and Lr24 were resistant, while Lr18 was susceptible (98% in 1996) to all tested leaf rust isolates and Lr10 and Lr23 were susceptible to more than 78% of the isolates. Diversity between populations (years) was assessed using Kosman's H(KB) (based on degrees of similarity among distinct phenotypes) and H(KDis) (based on frequencies of individual virulences) and Nei's and Rogers' distances. The greatest difference occurred between the 1993 and 1994 populations. Phenotypic diversity within each population (year) was analyzed using the Shannon's, Simpson's, and Kosman's indices. The highest diversity within years was recorded in 1994 and significantly increased from 1993 to 1994 for all indices. The variance in the diversity between populations can be only partially explained by differences between corresponding diversities within population. The comparative analysis of diversity between and within populations over the 5 years enabled a detailed study of changes in the pathogen population. The results show that the different measures do not yield the same rank order of diversity.     

Effects of growth stage, leaf position and temperature on adult-plant resistance of wheat infected by Puccinia recondita f.sp. tritici

Growth stage, leaf position and temperature effects on components of resistance to Puccinia recondita f.sp. tritici in wheat ( Triticum aestivum ) were studied. Inoculation of Era, Glenlea, Sinton and RL6044 plants at five growth stages showed that latent period increased and uredinium numbers decreased as each adult-plant-resistant wheat matured. Growth stage had little effect on latent period in the susceptible cultivar Morocco. No significant differences in latent period determined on the flag, flag -1 and flag -2 leaves of the same plant were observed for Era, Glenlea and RL6044. However, leaf age should be considered in assessing adult-plant resistance because the latent period of P. recondita f.sp. tritici in flag leaves of Sinton was shorter than that determined for the flag-1 and flag -2 leaves. Fewer uredinia developed on the flag -2 leaf of Glenlea than on the flag leaf. The latent period of leaf rust in Era, Glenlea, RL6044 and Morocco was significantly longer at 15°C than at 21°C, but temperature did not affect Sinton to the same degree. Uredinium size was a sensitive criterion of adult-plant resistance. Uredinium development in line RL6044 was significantly restricted at 15°C, but uredinia on Era, Glenlea and Sinton at 15°C were as large as those at 21°C. On the susceptible Morocco uredinia were larger at 15°C than at 21°C.     

小麦条锈菌冬孢子生物学特性研究进展

由小麦条锈菌引起的小麦条锈病是我国广大麦区的严重病害,研究其冬孢子产生原因与作用对于解析该病菌的生活史及遗传变异机制至关重要.通过对国内外相关研究成果的整理,综述了小麦条锈菌冬孢子的形态特征、生物学特性、寄主范围及其作用与功能等方面的研究进展.     

Attempted somatic hybridization of Puccinia striiformis f. sp. tritici and P. striiformis f. sp. hordei

Attempts were made to produce somatic hybrids between isolates of Puccinia striiformis f. sp. tritici and f. sp. hordei. A mixed infection was produced on a common susceptible barley host, Fong Tien, using white-spored isolates of P. striiformis f. sp. tritici and yellow-spored isolates off. sp. hordei. Selection was made for non-parental spore colour on selective wheat and barley hosts, and variants thus isolated were analysed for virulence markers, and for isozyme and double-stranded RNA (dsRNA) markers, all of which clearly differentiated the parental isolates. Two white-spored (non-parental) isolates were found on the selective barley host which otherwise resembled the parental f. sp. hordei isolate in virulence, isozyme and dsRNA markers. The most likely explanation of the origin of these isolates is mutation to white spore colour in the f. sp. hordei isolate.     

我国小麦秆锈菌种群动态分析

１９９５～１９９６年间,在我国７省区６２个县点共收集标样２８３份,涉及品种１１７个,分离到菌株４５５份,鉴定出１５个致病类型,其出现类型的发生频率分别是２１Ｃ３ＣＫＲ为１７．５％,２１Ｃ３ＣＦＨ９．６％,２１Ｃ３ＣＫＨ８．８％,２１Ｃ３ＣＦＲ６．０％,２１Ｃ３ＣＴＲ２２．０％,２１Ｃ３ＣＰＲ９．４％,２１Ｃ３ＣＴＨ８．８％,２１Ｃ３ＣＰＨ１１．９％,３４ＭＫＧ１．３％,３４Ｃ２ＭＫＨ０．２％,３４Ｃ２ＭＫＫ０．１％,３４Ｃ２ＭＦＨ３．６％,３４Ｃ２ＭＦＫ０．４％,３４Ｃ１ＭＫＲ和３４Ｃ１ＭＦＲ为０．２％。１９９６年２１Ｃ３ＣＫＲ的出现频率上升为第１位,对Ｓｒ１１有毒力的致病类型出现频率有所下降。     

不同培养载体对小麦条锈菌夏孢子人工萌发的影响

为探索适合小麦条锈菌寄主体外萌发的载体,并建立基于不同研究目的的小麦条锈菌夏孢子人工萌发技术,采用超声波水雾培养法,以小麦条锈菌生理小种CYR32为供试材料,探讨了不同培养载体对小麦条锈菌夏孢子萌发特性的影响.在最适温度和湿度条件下,以4种不同材料为培养载体,小麦条锈菌夏孢子的萌发率、芽管生长率和外观显示情况有显著差异.在尼龙纱网上的夏孢子萌发速度和芽管伸长速度最快,12 h内萌发率最高可达93.40％,芽管最长可达446.71 μm;亲水滤膜上夏孢子的萌发率和芽管长仅次于纱网;亲水塑料膜和PVDF膜上夏孢子的萌发率均高于普通亲水载玻片,而芽管长度低于普通亲水载玻片.采用侧光照明时,在亲水滤膜和PVDF膜上极易清晰观测到夏孢子及其芽管内的黄色颗粒物.在亲水塑料膜上,还可观察到夏孢子萌发12 h后,芽管尾部膨大形成类似附着胞的结构.结果显示,超声波水雾可使条锈菌夏孢子处于水分饱和的环境,而纱网纤维组成的小格具有很好的保水结露功能,能使夏孢子萌发快,萌发效率高,芽管生长快;另一方面,使用其他培养载体还可满足观测条锈菌夏孢子内黄色颗粒物向芽管内转移和芽管尾部形成类似附着胞结构的目的.     

Comparison of Puccinia graminis f.sp. tritici from South America and Europe

Twenty isolates of Puccinia graminis f.sp. tritici from South America were compared with 19 from Europe using virulence, isozymes and random amplified polymorphic DNA (RAPD) markers. The isozyme and virulence patterns for these isolates were also compared with those of 11 isolates representative of the common race clusters in North America. All three types of marker showed a level of similarity between the South American and European isolates comparable with that between isolates from the same continent. The average similarity coefficients between the South American and European isolates were 0.65 for virulence, 0.67 for isozymes, and 0.70 for RAPD markers. Among South American isolates the values were 0.63 for virulence, 0.64 for isozymes and 0.72 for RAPDs. For the South American and European isolates, correlation between the similarity matrices based on RAPDs and on isozyme markers, respectively ( r  = 0.52), was higher than that between the RAPD and virulence matrices ( r  = 0.32) or between isozyme and virulence matrices ( r  = 0.16). The North American isolates had a comparable level of similarity for virulence and isozymes to both the South American and European populations. There was no clear distinction between the South American, North American and European isolates, which is consistent with the hypothesis that these populations may have had a common origin.     

Local dispersal of Puccinia striiformis f. sp. tritici from isolated source lesions

Understanding how disease foci arise from single source lesions has not been well studied. Here, single wheat leaves were inoculated with Puccinia striiformis f. sp. tritici urediniospores, and all wheat leaves within two intersecting 0.3 × 3.0 m transects were sampled in eight replicates over 3 years. The lesions observed on each of the top three leaves on plants within 1.5 m from the source lesion were three‐dimensionally mapped. The total number of lesions within a 1.5 m radius was estimated by dividing the number of lesions observed within each 0.025 m‐wide annulus by the fraction of the annulus sampled. The estimated total number of lesions produced within 1.5 m of a single source lesion ranged from 27 to 776, with a mean of 288 lesions. Eighty percent of the lesions were recorded within 0.69 m of the source infection. The proportion of total lesions observed at a given distance from the source was fitted well by the Lomax and Weibull distributions, reflecting the large proportion of lesions arising close to the source, and when fitted by an inverse power distribution had a slope (b) of 2.5. There were more lesions produced on leaves higher in the canopy than on lower leaves, with more lesions being detected above than below the point of inoculation. Simultaneous measurement of lesion gradients and spore dispersal in the final year of the study suggests that this pattern is due to greater susceptibility of upper leaves, rather than increased dispersal to upper leaves.     

小麦秆锈菌新型小种Ug99的研究进展

Ug99(TTKS)为1999年在乌干达首次发现的秆锈菌新小种,对小麦抗秆锈病基因Sr31具有强毒力.此小种致病性极强,传播迅速,给全球小麦生产带来威胁.在肯尼亚用Ug99对我国118个小麦生产品种和材料进行抗病性鉴定结果表明,高感品种占98.3%.该小种一旦传入我国,将对小麦生产造成严重损失.笔者对国内外关于小麦抗Ug99遗传研究、抗病基因分子标记研究现状及我国应对措施进行了概述.     

Emergence of a novel population of Puccinia striiformis f. sp. tritici in Eastern United States

The geographic range of stripe rust of wheat, caused by Puccinia striiformis f. sp. tritici, has increased dramatically since 2000 in the United States. Yield losses to the disease have been most severe in the eastern United States, where measurable yield loss had been rare prior to 2000. The objective of this study was to examine the phenotypic and genotypic variation among isolates of P. striiformis f. sp. tritici collected from populations in the eastern United States before and since 2000. Virulence phenotype and amplified fragment length polymorphism (AFLP) markers were used to examine 42 isolates collected between 1960 and 2004. In addition, the genetic structure of 59 isolates collected in 2005 using a hierarchical sampling strategy was examined. The data indicated that the contemporary isolates (collected since 2000) were very distinct from older isolates (collected before 2000) based on virulence and AFLP markers, and that the old population prevalent before 2000 may have been replaced by the contemporary population. The old and new populations appear to be genetically distinct and may represent an exotic introduction rather than a mutation in isolates of the old population.     

Uredospore production and sporulation period of Puccinia recondita f. sp. triticina on primary leaves of wheat

Uredospore production per day and per sporulation period was measured under near-optimal conditions. Pustule density influenced time and rate of pustule opening, size of pustules, time of maximum sporulation, length of the sporulation period and the time and rate of tissue necrotisation. Within limits total dry weight of spores per leaf per sporulation period was independent of pustule density; it roughly equalled the dry weight of the spore producing leaf. The longest sporulation period observed was 65 days; at low pustule densities secondary pustules replaced exhausted primary pustules. Infectivity of the spores was normal up to 46 days after inoculation. The long sporulation period was epidemiologicaly interpreted as a survival mechanism.     

云南省德宏州小麦条锈菌越夏调查初报

小麦条锈菌越夏菌源量直接影响着小麦条锈病的发生与流行。为了明确云南省德宏州是否是小麦条锈病菌的越夏菌源地,2010年和2011年在德宏州芒市江东乡海拔1 600、1 700、1 800、1 900m和2 000m处设置了小麦条锈病菌源观察圃,对观测圃中小麦品种和周围落粒自生麦苗上条锈病的发病情况进行调查,实时记录各观察点的温湿度。结果表明:(1)海拔2 000m的菌源观察圃能观测到小麦条锈菌越夏和越冬的情况,越冬菌源量和春季条锈病的发病情况存在相互影响的关系;(2)海拔1 900m及以上的地区最高日平均温度和旬平均温度均低于22℃,为小麦条锈菌的越夏区域;1 600~1 900m的区域内其最高日平均温度在22℃至25℃之间,最高旬平均温度均大于20℃小于23℃,符合小麦条锈菌越夏的理论温度要求;并且海拔1 900m及以上地区的菌源观察圃中可观察到小麦条锈菌越夏。以上结论证实了云南省德宏州芒市是小麦条锈菌的越夏菌源地之一。     

Temperature adaptation in Australasian populations of Puccinia striiformis f. sp. tritici

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the major fungal pathogens of wheat. A new pathotype was introduced to Australia in 2002 and several derivative pathotypes were detected in subsequent seasons. It has been suggested that the severity of stripe rust outbreaks in Australia since 2002 could be as a result of traits other than virulence in the pathogen population. This study was conducted to investigate the hypothesis that the stripe rust pathogen population dominant in Australia since 2002 was better adapted to warm temperature conditions compared to previous pathogen populations. Sixteen pathotypes were selected to examine the influence of two contrasting temperature regimes during the 24 h incubation (10°C and 15°C) and the subsequent post‐inoculation (17°C and 23°C) periods on latent period and infection efficiency on four susceptible wheat cultivars. In addition, the effect of two contrasting incubation temperatures on urediniospore germination was examined. The results indicated that pathotypes of P. striiformis f. sp. tritici detected after 2002 did not show evidence of adaptation to high temperatures, which suggests that other factors contributed to the observed increased aggressiveness.     

Genetic Diversity in Australian Populations of Puccinia graminis f. sp. avenae

ABSTRACT Sequence-tagged microsatellite profiling was used to develop 110 microsatellites for Puccinia graminis f. sp. tritici (causal agent of wheat stem rust). Low microsatellite polymorphism was exhibited among 10 pathogenically diverse P. graminis f. sp. tritici isolates collected from Australian cereal growing regions over a period of at least 70 years, with two polymorphic loci detected, each revealing two alleles. Limited cross-species amplification was observed for the wheat rust pathogens, P. triticina (leaf rust) and P. striiformis f. sp. tritici (stripe rust). However, very high transferability was revealed with P. graminis f. sp. avenae (causal agent of oat stem rust) isolates. A genetic diversity study of 47 P. graminis f. sp. avenae isolates collected from an Australia-wide survey in 1999, and a historical group of 16 isolates collected from Australian cereal growing regions from 1971 to 1996, revealed six polymorphic microsatellite loci with a total of 15 alleles. Genetic analysis revealed the presence of several clonal lineages and subpopulations in the pathogen population, and wide dispersal of identical races and genotypes throughout Australian cereal-growing regions. These findings demonstrated the dynamic population structure of this pathogen in Australia and concur with the patterns of diversity observed in pathogenicity studies.     

Clonality and long-distance migration of Puccinia striiformis f.sp. tritici in north-west Europe

The biotrophic fungus Puccinia striiformis f.sp. tritici , a basidiomycete that causes yellow rust on wheat, is spread by wind-dispersed spores. Analysis of amplified fragment length polymorphism (AFLP) variation showed that the fungus frequently migrates between the UK, Germany, France and Denmark. There is no biological evidence for sexual or parasexual reproduction under natural conditions, and this was supported by the lack of recombination, as revealed by AFLP, over the time and area represented by the samples in this study. A phylogeographic analysis revealed that there was effectively a single, clonal population in the four countries, up to 1700 km apart, consistent with a 'continent-island' model in which Denmark is the recipient of migrants from other countries. In five cases, specific pathogen clones were dispersed between the UK and Denmark, and on at least two recent occasions clones were also spread from the UK to Germany and France, causing outbreaks of yellow rust on wheat cultivars that were previously resistant to the disease in these countries. The agronomic consequences of migration were enhanced because of the limited genetic diversity for yellow rust resistance in wheat cultivars in the area. These results demonstrate that long-distance migration of pathogen clones, coupled with low diversity in the host species, may cause previously useful resistance genes to become ineffective for disease control on a continental scale.     

Disease severity and pathotype dynamics of Puccinia striiformis f.sp. tritici in Denmark

Disease severity of wheat yellow rust, Puccinia striiformis f.sp. tritici , was analysed in Denmark from 1985 to 1999 in relation to the effects of weather on winter survival, distribution of host cultivars and pathotype dynamics. Below-average temperatures in January and February (midwinter) reduced yellow rust on the susceptible cv. Anja, and in three of four growth seasons following cold winters no yellow rust was observed on any cultivar under natural conditions. The agronomic consequences of dispersal of yellow rust urediniospores from external sources to Denmark, in a period during which large areas were planted with relatively few wheat cultivars, were demonstrated in several cases, most evidently when the Yr9 and Yr17 resistance genes became ineffective. Yr9 was overcome by the pathogen in a period with severe yellow rust epidemics on commercial cultivars, while virulence for Yr17 was first observed in a year with almost no yellow rust. In contrast, the resistance in cv. Kraka ( Yr1, CV ) was increasingly effective in controlling yellow rust, because pathotypes with the matching combination of virulence declined in the pathogen population. Pathotype frequency dynamics were thus influenced by selection forces within the country, and by selection forces in areas where spores were spread to Denmark from outside. The importance of a sufficient level of partial resistance in the wheat germplasm to prevent too much damage by yellow rust epidemics, in the event that the resistance genes are overcome by the pathogen population, is emphasized.     

Effect of maleic hydrazide on parasitic fitness of Puccinia striiformis f. sp. tritici in wheat

As a growth inhibitor of the apical meristem in the plant stem, Maleic hydrazide plays an important role in modulation of plant growth. In this research, Puccinia striiformis f. sp. tritici and its host for reproduction was employed to characterize the effect of maleic hydrazide on parasitic fitness of the strain. Growth inhibition of the secondary leaves of wheat by maleic hydrazide was demonstrated. Results showed that root irrigation by maleic hydrazide at the seedling stage significantly increased the parasitic fitness of Puccinia striiformis f. sp. tritici, leading to the increase in sporulation amount, sporulation period, and germination rate of urediniospores. In addition, the ultrastructure of Puccinia striiformis f. sp. tritici urediniospores was not affected by maleic hydrazide treatment. Out data indicate that the optimal concentration and dose for the use of maleic hydrazide is 0.35 g?l^?1 and 1.5 ml/cm², respectively, facilitating the widespread application in wheat stripe rust studies.