Sporisorium reilianum possesses a pool of effector proteins that modulate virulence on maize

The biotrophic maize head smut fungus Sporisorium reilianum is a close relative of the tumour-inducing maize smut fungus Ustilago maydis with a distinct disease aetiology. Maize infection with S. reilianum occurs at the seedling stage, but spores first form in inflorescences after a long endophytic growth phase. To identify S. reilianum-specific virulence effectors, we defined two gene sets by genome comparison with U. maydis and with the barley smut fungus Ustilago hordei. We tested virulence function by individual and cluster deletion analysis of 66 genes and by using a sensitive assay for virulence evaluation that considers both disease incidence (number of plants with a particular symptom) and disease severity (number and strength of symptoms displayed on any individual plant). Multiple deletion strains of S. reilianum lacking genes of either of the two sets (sr10057, sr10059, sr10079, sr10703, sr11815, sr14797 and clusters uni5-1, uni6-1, A1A2, A1, A2) were affected in virulence on the maize cultivar ‘Gaspe Flint’, but each of the individual gene deletions had only a modest impact on virulence. This indicates that the virulence of S. reilianum is determined by a complex repertoire of different effectors which each contribute incrementally to the aggressiveness of the pathogen.     

First Report of the Occurrence of a White Smut Infecting Rice in Arkansas

False smut has recently emerged as an important disease of rice in Arkansas. In 2011, 2012 and 2013, spore balls of a white smut similar to the spore balls of false smut were observed in rice fields in eastern Arkansas. As a white false smut was previously reported in China and Japan, we examined the morphology of chlamydospores and spore balls from some of the infected heads and used selected regions of the rDNA to determine the identity of the causal agent of the disease. We also tested the virulence of an isolate of the white smut to two rice cultivars commonly grown in Arkansas. Our results indicate that the morphology of the spore balls, chlamydospores and conidia is similar to those reported for Ustilaginoidea albicans. However, sequences of ribosomal DNA amplicons indicate a high degree of similarity with both U. virens and U. albicans. The isolate of the white smut was virulent to two rice cultivars, producing spore balls similar to those observed in the field and to those previously described for U. albicans.     

Relationship of Pseudomonas syringae pv. tabaci races to the rhizosphere of Wisconsin-grown tobacco

Isolates of Pseudomonas syringae pv. tabaci, including 21 strains of the wildfire pathogen and 2 strains of the angular leafspot pathogen, were isolated from 143 rhizosphere and soil samples collected from 11 tobacco fields in Wisconsin. These pathogens were isolated by inoculating rhizosphere and soil washings into tobacco leaves and isolating the bacteria from wildfire or angular leafspot lesions that developed on the leaves. The wildfire isolates were from the rhizospheres of tobacco and Panicum capillare and from soil. While the majority of these were from wildfire-diseased fields, one isolate was from a field without disease symptoms; both angular leafspot isolates were from fields without angular leafspot symptoms. The majority of wildfire isolates were race 1, but three were race 0, and one was a new race. In three fields multiple races of wildfire were found. Both angular leafspot isolates were race 1. Two wildfire and one angular leafspot isolates were from fields where the cultivars were resistant to the races isolated.     

高粱丝黑穗病菌种内分化的RAPD分析*

应用随机引物对来自不同地理来源、不同寄主和经寄主致病力测定的高粱丝黑穗病菌2、3号生理小种的10个菌株的DNA进行分析,所产生的RAPD结果表明,丝黑穗病菌S.reilianum具有丰富的种内遗传多样性,存在明显的分化现象。经聚类分析可将供试菌株大致分成两组,辽宁清原H2和黑龙江绥化H9菌株为一组。辽宁沈阳(H3)、阜新(H1)、营口(H10),山西榆次(H4),吉林四平(H5),黑龙江哈尔滨(H6),河北张家口(H8)等高粱丝黑穗病菌株以及辽宁沈阳的玉米丝黑穗病菌株(H7)为另一组,并且同一组内的DNA多态性亦有差异。高粱丝黑穗病菌2号生理小种和3号小种间在分子水平上存在差异明显。     

高粱丝黑穗病菌种内分化的RAPD分析

应用随机引物对来自不同地理来源、不同寄主和经寄主致病力测定的高粱丝黑穗病菌2、3号生理小种的10个菌株的DNA进行分析,所产生的RAPD结果表明,丝黑穗病菌S.reilianum具有丰富的种内遗传多样性,存在明显的分化现象。经聚类分析可将供试菌株大致分成两组,辽宁清原H2和黑龙江绥化H9菌株为一组。辽宁沈阳(H3)、阜新(H1)、营口(H10),山西榆次(H4),吉林四平(H5),黑龙江哈尔滨(H6),河北张家口(H8)等高粱丝黑穗病菌株以及辽宁沈阳的玉米丝黑穗病菌株(H7)为另一组,并且同一组内的DNA多态性亦有差异。高粱丝黑穗病菌2号生理小种和3号小种间在分子水平上存在差异明显。     

Assessment of sorghum genetic resources of Ethiopia for anthracnose (Colletotrichum sublineolum Henn.) resistance and agronomic traits

Sorghum anthracnose caused by Colletotrichum sublineolum Henn. is one of the key diseases limiting sorghum production and productivity. Development of anthracnose‐resistant sorghum genotypes possessing yield‐promoting agronomic traits is an important breeding goal in sorghum improvement programs. The objective of this study was to determine the responses of diverse sorghum genetic resources for anthracnose resistance and agronomic traits to identify desirable lines for breeding. A total of 366 sorghum collections and three standard checks were field evaluated during the 2016 and 2017 cropping seasons. Lines were artificially inoculated with a virulent pure isolate of the pathogen. Anthracnose disease severity was assessed to calculate the area under disease progress curve (AUDPC). Agronomic traits such as panicle length (PL), panicle width (PW), head weight (HW) and thousand grain weight (TGW) were measured. Lines showed highly significant differences (p < .001) for anthracnose severity, AUDPC and agronomic traits. Among the collections 32 lines developed levels of disease severity between 15% and 30% in both seasons. The following sorghum landraces were selected: 71708, 210903, 74222, 73955, 74685, 74670, 74656, 74183, 234112, 69412, 226057, 214852, 71420, 71484, 200126, 71557, 75120, 71547, 220014, 228179, 16212, 16173, 16133, 69088, 238388, 16168 and 71570. These landraces had a relatively low anthracnose severity possessing farmer‐preferred agronomic traits. The selected genotypes are useful genetic resources to develop anthracnose‐resistant sorghum cultivars.     

Detection of Cochliobolus heterostrophus races in South China

Cochliobolus heterostrophus is the causal pathogen of the southern corn leaf blight (SCLB). There are three known races: race O, race C and race T. To determine which C. heterostrophus races comprise the field population in southern China and to assess diversity of these strains in terms of virulence, 200 isolates from diseased plants were collected in nine provinces/municipalities. All were race O, that is, no race T or race C isolates were found. Sixty race O isolates that sporulated well were chosen for further analysis. Virulence was measured using the integral optical density (IOD) of leaf lesions on four maize inbred lines. UPGMA cluster analysis of AFLP markers was applied to the 60 race O isolates plus control race O, T and C strains. Phylogenetic distribution, geographic location and virulence were not correlated. These results can provide valuable information for guidance in early warning and disease control.     

Resistance of Common Bean (Phaseolus vulgaris) to Bacterial Wilt Caused by Curtobacterium flaccumfaciens pv. flaccumfaciens

Bacterial wilt caused by Curtobacterium flaccumfaciens pv. flaccumfaciens is an important new disease of common bean (Phaseolus vulgaris) in western Canada. Both yellow and orange variants of the pathogen were found in the region. A controlled environment study was conducted to assess 124 common bean cultivars and lines from eight market classes for resistance to the yellow and orange variants of the pathogen, using the hilum injury/seed inoculation method. Results of the screening tests showed significant (P < 0.05) differences in resistance to bacterial wilt among the cultivars or lines. The great northern line L02E317, the great northern cultivar Resolute and pinto lines L02B662 and 999S‐2A, were highly resistant to both variants of the pathogen, with disease severity indices of 0 on a rating scale of 0 (no wilt symptoms) to 5 (dead seedling). Resistant cultivars or lines were found among black, great northern, pink, pinto, small red and Flor de Mayo bean market classes. The study concludes that new bacterial wilt‐resistant germplasm exists among Canadian bean cultivars and lines, and constitutes a valuable resource for breeding common beans for resistance to both yellow and orange variants of C. flaccumfaciens pv. flaccumfaciens.     

高粱对丝黑穗病的抗性及遗传研究

1981—1985年在人工接种的条件下,对1239份高粱品种资源,进行了抗丝黑穗病性鉴定。与此同时,用17个抗性不同的品种系,按照不完全双列杂交设计,进行了高粱对丝黑穗病的抗性遗传研究。结果表明,高粱对丝黑穗病的抗性遗传方式因品种而异。有的品种系具数量性状遗传特点,有的则具有质量性状遗传特点。抗病性属数量性状遗传的品种系,其抗性主要是受加性基因控制。     

Effects of a fraction from maize root exudates on haploid strains of Sporisorium reilianum f. sp. zeae

Sporisorium reilianum f. sp. zeae is the causal agent of head smut of maize. Although the main symptom of this disease is the formation of a black fungal sorus on the reproductive parts of the maize, the infection always occurs via the roots. Early infection stages are characterised by a hyphal proliferation of the fungus around the roots. In this paper, we describe effects of a fraction extracted from maize root exudates on growth of S. reilianum f. sp. zeae. The fungus grew as a yeast form on artificial medium, but in presence of these fractions, some yeasts switched to a hyphal form. In addition, an increased proliferation of the yeast form was also observed with exudates from a variety of maize susceptible to head smut. In the presence of exudates obtained from a tolerant variety of maize, proliferation of the yeast form was inhibited, whereas the induction of yeast-hypha transition was always observed. These results indicated that some molecules in root exudates could play a role in the pre-infectious stage between maize and S. reilianum f. sp. zeae.     

Genetic Diversity and Pathogenicity of Fusarium oxysporum f.sp. melonis Races from Different Areas of Italy

Fusarium wilt caused by Fusarium oxysporum f.sp. melonis (FOM) is a devastating disease of melon worldwide. Pathogenicity tests performed with F. oxysporum isolates obtained from Italian melon‐growing areas allowed to identify thirty‐four FOM isolates and the presence of all four races. The aims of this work were to examine genetic relatedness among FOM isolates by race determination and to perform phylogenetic analyses of identified FOM races including also other formae speciales of F. oxysporum of cucurbits. Results showed that FOM race 1,2 was the most numerous with a total of eighteen isolates, while six and nine isolates were identified as race 0 and 1, respectively, and just one isolate was assigned to race 2. Phylogenetic analysis was performed by random amplified polymorphic DNA (RAPD) profiling and by translation elongation factor‐1α (TEF‐1α) sequencing. The analysis of RAPD profiles separated FOM races into two distinct clades. Clade 1, which included races 0, 1 and 1,2, was further divided into ‘subclade a’ which grouped almost all race 1,2 isolates, and into ‘subclade b’ which included race 0 and 1 isolates. Clade 2 comprised only race 2 isolates. The phylogenetic analysis based on TEF‐1α separated FOM from the other formae speciales of F. oxysporum. Also with TEF‐1α analysis, FOM races 0, 1 and 1,2 isolates grouped in one single clade clearly separated from FOM race 2 isolates which grouped closer to F. oxysporum f.sp. cucumerinum. RAPD technique was more effective than TEF‐1α in differentiating FOM race 1,2 isolates from those belonging to the closely related races 0 and 1. Both phylogenetic analyses supported the close relationship between the three different FOM races which might imply the derivation from one another and the different origin of FOM race 2.     

Identification and molecular mapping of a resistance gene to powdery mildew from the synthetic wheat line M53

Powdery mildew disease caused by Blumeria graminis f. sp. tritici (Bgt) is an economically important disease in wheat worldwide. The identification of germplasms resistant to the disease can not only facilitate the breeding of resistant cultivars, but can also broaden the diversity of resistance genes. The Mexican M53 is a synthetic hexaploid wheat line developed at the International Maize and Wheat Improvement Center (CIMMYT) from the cross between Triticum durum and Aegilops tauschii249. Infection of M53 with 15 different pathogen races revealed that the resistance in M53 was race-dependent and effective against the majority of the tested Bgt races, including the race 15 predominant in the Beijing wheat growing area. Inoculation of the parents of M53 with the race 15 demonstrated that M53 and Ae. tauschii249 were resistant, whereas T. durum was susceptible. The inoculation of three segregating F₂ populations developed from the crosses between M53 and three susceptible Chinese wheat cultivars with the race 15 showed that the resistant gene in M53 segregated in a single dominant manner. Amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers were used to map the gene in a segregating F₂ population consisting of 213 lines developed from the cross Wan7107 × M53. Two closely linked AFLP markers, Apm109 and Apm161, were identified to flank the gene with genetic distances of 1.0 cM and 3.0 cM, respectively. The recognized gene was assigned to the long arm of chromosome 5D as determined by three linked SSR markers, Xwmc289b, Xgwm583, and Xgwm292, and by the physical mapping of Apm109 using Chinese Spring nullisomic–tetrasomic and ditelosomic stocks. The resistance gene identified in M53, temporarily designated as Pm-M53, could be used in local wheat-breeding programs to improve powdery mildew resistance.     

A region of barley chromosome 6H harbors multiple major genes associated with net type net blotch resistance

Net type net blotch (NTNB), caused by Pyrenophora teres f. teres Drechs., is prevalent in barley growing regions worldwide. A population of 118 doubled haploid (DH) lines developed from a cross between barley cultivars ‘Rika’ and ‘Kombar’ were used to evaluate resistance to NTNB due to their differential reaction to various isolates of P. teres f. teres. Rika was resistant to P. teres f. teres isolate 15A and susceptible to isolate 6A. Conversely, Kombar was resistant to 6A, but susceptible to 15A. A progeny isolate of a 15A × 6A cross identified as 15A × 6A#4 was virulent on both parental lines. The Rika/Kombar (RK) DH population was evaluated for disease reactions to the three isolates. Isolate 15A induced a resistant:susceptible ratio of 78:40 (R:S) whereas isolate 6A induced a resistant:susceptible ratio of 40:78. All but two lines had opposite disease reactions indicating two major resistance genes linked in repulsion. Progeny isolate 15A × 6A#4 showed a resistant:susceptible ratio of 1:117 with the one resistant line also being the single line that was resistant to both 15A and 6A. An RK F₂ population segregated in a 1:3 (R:S) ratio for both 15A and 6A indicating that resistance is recessive. Molecular markers were used to identify a region on chromosome 6H that harbors the two NTNB resistance genes. This work shows that multiple NTNB resistance genes exist at the locus on chromosome 6H, and the recombinant DH line harboring the resistance alleles from both parents will be useful for the development of NTNB-resistant barley germplasm.     

New Races of Sunflower Downy Mildew (Plasmopara Halstedii) in Germany

Plasmopara halstedii, the causal agent of downy mildew of cultivated sunflower (Helianthus annuus), was documented in Germany for the first time in commercial fields. The pathogen was first observed in the Württemberg area, where races 1 and 4 were identified using a set of differential lines. Later, commercial fields near Baden were found to be infected by race 5, which is the first occurrence of that race outside of North America. With the discovery of race 5, there are now eight races of the sunflower downy mildew fungus that have been found in Europe. The sunflower cultivars most frequently grown in Germany were investigated for resistance to race 1, 4 and 5; while all were resistant to race 1, none were resistant to either race 4 or 5.     

New Races of Sunflower Downy Mildew (Plasmopara Halstedii) in Germany

Plasmopara halstedii, the causal agent of downy mildew of cultivated sunflower (Helianthus annuus), was documented in Germany for the first time in commercial fields. The pathogen was first observed in the Württemberg area, where races 1 and 4 were identified using a set of differential lines. Later, commericial fields near Baden were found to be infected by race 5, which is the first occurrence of that race outside of North America. Withthe discovery of race 5, there are now eight races of the sunflower downy mildew fungus that have been found in Europe. The sunflower cultivars most frequently grown in Germany were investigated for resistance to race 1, 4 and 5; while all were resistant to race 1, none were resistant to either race, 4 or 5.     

Molecular Detection of Stripe Rust Resistance Gene(s) in 115 Wheat Cultivars (lines) from the Yellow and Huai River Valley Wheat Region

Stripe rust (yellow rust), caused by Puccinia striiformis f.sp. tritici (Pst), is a serious disease of wheat worldwide, including China. Growing resistant cultivars is the most cost‐effective and environmentally friendly approach to control the disease. To assess the stripe rust resistance in commercial wheat cultivars and advanced lines in the Yellow and Huai River Valley Wheat Region, 115 wheat cultivars (lines) collected from 13 provinces in this region were evaluated with the most prevalent Chinese Pst races CYR32, CYR33 and the new race V26 at seedling stage. In addition, these wheat entries were inoculated with the mixed races of CYR32 and CYR33 at the adult‐plant stage in the field. The results indicated that 53 (46.1%) cultivars (lines) had all‐stage resistance to all the three races, and 16 (13.9%) cultivars (lines) showed adult‐plant resistance. The possible stripe rust resistance genes in these entries were postulated by the closely linked markers of all‐stage resistance genes Yr5, Yr9, Yr10, Yr15 and Yr26 and adult‐plant resistance gene Yr18. Molecular analysis indicated that resistance genes Yr5, Yr9, Yr10, Yr18 and Yr26 were found in 5 (4.3%), 38 (33.0%), 1 (0.9%), 2 (1.7%) and 8 (7.0%) entries, respectively. No entry was found to carry the Yr15 gene. In future breeding programs, Yr5, Yr15 and Yr18 should be used to pyramid with other effective genes to develop wheat cultivars with high‐level and durable resistance to stripe rust, whereas Yr9, Yr10 and Yr26 should not be used or used in a limited way due to the virulent races present in China.     

Characterization of emerging populations of Fusarium oxysporum f. sp. conglutinans causing cabbage wilt in China

Fusarium oxysporum f. sp. conglutinans (FOC) causes Fusarium wilt, a disease of cabbage that has brought about significant economic loss throughout northern China since it was first detected in 2001. To characterize the Chinese FOC isolates, we compared the cultural characteristics, pathogenicity and races between the Chinese isolates and the type strains (race 1: 52,557 and race 2: 58,385). The Chinese FGL‐03‐6 isolate had cultural characteristics similar to those of strain 52,557, including colony growth rate, colony and spore characteristics and responses to temperature changes, while the strain 58,385 grew faster, produced more pigment and spores and was more adaptable to temperature fluctuations. The lethal temperature for all strains was 60°C, and the optimal temperatures for pathogen growth on potato dextrose agar and pathogenicity on plants were 25°C and 25 to 30°C, respectively. Tests for race and pathogenicity indicated that different cabbage cultivars had similar resistance reactions to FGL‐03‐6 and 52,557. However, the pathogenicity of FGL‐03‐6 was similar to that of 58,385, which infected quickly and caused more severe disease symptoms. This study further provides information regarding characterizing different strains of F. oxysporum f. sp. conglutinans.