Powdery mildew (Sphaerotheca fuliginea) resistance in melon is selectable at the haploid level

The major cause of powdery mildew in melons (Cucumis melo L.) is the fungus Sphaerotheca fuliginea. There are several cultivar- and season-specific races of this fungus. In order to control powdery mildew, it is important to introduce resistance to fungal infection into new cultivars during melon breeding. Haploid breeding is a powerful tool for the production of pure lines. In this study, it was investigated whether powdery mildew resistance could be manifested at the haploid level from two disease-resistant melon lines, PMR 45 and WMR 29. the effects of various races of S. fuliginea on diploid and haploid plants of PMR 45 and WMR 29 and of a disease-susceptible line, Fuyu 3 were measured. The responses of haploid and diploid plants to powdery mildew were identical. In addition, haploids that were generated from hybrids between Fuyu 3 and disease-resistant lines were examined. Seven out of 13 haploids from a Fuyu 3xPMR 45 cross and 10 out of 12 haploids from a Fuyu 3xWMR 29 cross were classified as resistant plants because they showed the same responses as their disease-resistant diploid parents to the various fungal races. These results indicate that resistance in PMR 45 and WMR 29 is selectable at the haploid level. All of the plant responses were observed by microscopy. A possible mechanism for generating powdery mildew resistance in two different melon lines is discussed.     

First report of powdery mildew caused by Podosphaera xanthii on Luffa acutangula in Odisha State,India

Abstract

Ridge gourd (Luffa acutangula) is an herbaceous perennial twining vine cultivated globally as vegetable and medicinal plant. During October to January in 2014 and 2015, 40% powdery mildew disease incidence was observed in different areas of Odisha state, India. The pathogenicity experiments confirmed the powdery mildew disease symptoms on artificially inoculated L. acutangula seedlings. Causal organism was identified as Podosphaera xanthii on the basis of morphological and molecular studies. This is the first report of powdery mildew disease on L. acutangula caused by P. xanthii.     

Identification and validation of powdery mildew (Podosphaera xanthii)-resistant loci in recombinant inbred lines of cucumber (Cucumis sativus L.)

Powdery mildew limits cucumber production worldwide. Most resistant cucumber cultivars become susceptible to powdery mildew at low temperatures. Resistance within a wide temperature range is therefore desirable for cucumber production. We constructed a cucumber genetic linkage map based on a population of 111 recombinant inbred lines derived from a cross between CS-PMR1, with strong and temperature-independent resistance, and Santou, with moderate and temperature-dependent resistance. The map spans 693.0 cM and consists of 296 markers segregating into seven linkage groups; the markers include 289 simple sequence repeats (SSRs), six sequence characterized amplified regions, and one inter simple sequence repeat. Due to the presence of 150 common SSR markers, we were able to compare our map with previously published maps obtained by using populations derived from inter- or intra-variety crosses. We also evaluated powdery mildew resistance of the recombinant inbred lines and identified seven quantitative trait loci (QTL) contributed by CS-PMR1 and two QTL contributed by Santou. Four QTL (pm3.1, pm5.1, pm5.2 and pm5.3) were successfully validated by using populations derived from residual heterozygous lines. Some of the QTL identified in our study are in good agreement with previously published results obtained with materials of different origin. The markers reported here would be useful for introducing high and temperature-independent resistance by accumulation of QTL from CS-PMR1 and Santou.     

Molecular phylogenetic status of korean strain of Podosphaera xanthii, a causal pathogen of powdery mildew on Japanese thistle (Cirsium japonicum) in Korea

Powdery mildew diseases are sensitive to climate change and spread can be favored by increased temperature and low moisture. During 2011 to 2012, a powdery mildew disease by a Podosphaera species was observed on the leaves of Japanese thistle (Cirsium japonicum) in Korea. The initial sign of this disease included scattered superficial white mycelia on leaves. As the disease progressed, abundant necrotic black spots exhibiting chasmothecia were formed on the leaves. rDNA ITS and 28S homologies of the fungus (EML-CSPW1) showed 100% identity values with those regions from many strains of P. xanthii (syn. P. fusca) via NCBI BLASTN search.     

中国长春瓜类白粉菌Podosphaera xanthii形态学和分子系统学研究

2009年长春农博园瓜类白粉病大发生,而且大量闭囊产生在葫芦科的四个属（葫芦属、黄瓜属、栝楼属、南瓜属）植物上。对病害发生的严重度和病情指数进行了调查,结果表明园中栽培的所有品种均为感病品种。形态学观察显示分子孢子串生,具纤维体,闭囊壳只含有一个子囊,内含8个子囊孢子,子囊顶端孔较宽（15.6－28.8μm）。有性和无性形态学特征表明该菌应属于Podosphaera xanthii。基于rDNA ITS序列的分子系统学分析表明该菌与GenBank中已报道的该种在其他地区瓜类上的序列以及在其他寄主上的序列聚     

Seasonal variations in the occurrence of Golovinomyces orontii and Podosphaera xanthii,causal agents of cucurbit powdery mildew in Northern Italy

Powdery mildew, caused by Golovinomyces orontii and Podosphaera xanthii, is a widespread disease that causes important losses in cucurbit production. To determine the aetiology and the epidemiology of cucurbit powdery mildew disease in the North of Italy, observations on the occurrence of the main disease‐causing fungal species were conducted during the 2010, 2011 and 2012 growing seasons. Samples of infected leaves of zucchini, melon and pumpkin plants, either from field or greenhouse crops, were collected every 15–18 days from May to September/October. To identify the fungal species, both morphological observations based on the asexual stage and molecular identifications by a Multiplex‐PCR reaction with species‐specific primers were performed. Climatic parameters of temperature and relative humidity were also monitored. Pearson's correlation coefficient and Principal Component Analysis showed a negative significant correlation between the two species, and a peculiar epidemiological behaviour was also observed: the earlier infections were caused by G. orontii, which was the predominant species till the end of June–middle of July. At this time, this species progressively decreased in frequency and was replaced by P. xanthii that became the main species infecting cucurbits till the end of the growing season. As the two species have different ecological requirements, these seasonal variations in the cucurbit powdery mildew species composition could possibly be explained by the influence of temperature and relative humidity on the pathogen epidemiology during the growing season but also by the different overwintering strategies adopted by the two species.     

No Evidence of QoI Resistance in Apple Powdery Mildew (Podosphaera leucotricha)

Sensitivity monitoring studies using detached leaf tests with isolates of Podosphaera leucotricha showed no adaptation to pyraclostrobin in the last years. Sequence analysis of the target gene of QoIs, cytochrome b, of different isolates of P. leucotricha showed the presence of an intron directly after codon 143. This makes the occurrence of the G143A mutation unlikely. On the other hand, intron sequences have not been detected in immediate vicinity to the codons 129 and 137; therefore, the occurrence of those two mutations cannot be excluded. As the effects of these mutations on field performance on QoI fungicides are rather low, the overall resistance risk of P. leucotricha to this fungicide class is estimated to be low. The amplified cytochrome b gene fragments (exons and introns) for samples from different European countries and Australia were highly conserved.     

Characterization of Reactions to Powdery Mildew (Podosphaera pannosa) in Resistant and Susceptible Rose Genotypes

Fungal development of powdery mildew Podosphaera pannosa (Wallr.: Fr.) de Bary on rose leaves depends on constitutive or induced resistance mechanisms present in attacked rose genotypes. The relationship between fungal development and plant resistance was investigated microscopically on young greenhouse leaves of four rose genotypes with different levels of resistance: Rosa wichuraiana, R. laevigata anemoides and R. hybrida cultivars ‘Excelsa’ and ‘Gomery’. Induced plant reactions, hydrogen peroxide production and cross sections through infected leaves were examined. The variation in development of the fungus on these rose genotypes depended on the relative presence of normal haustoria, abnormal haustoria, induced cell reactions, papilla formation or physical barriers. Formation of papillae could arrest up to one third of the successful penetrations. Papillae formation was often succeeded by total cell reaction. Abnormal haustoria were detected as rudimentary haustoria, haustoria with abnormal shape or haustoria without extra haustorial matrix. Post‐haustorial cell reactions, with and without cell collapse, were detected. In non‐collapsed cells, appositions were directed to both cell wall and haustorium. This was followed by accumulation of non‐identified, probably antifungal compounds. Both single and multicell reactions occurred. Hydrogen peroxide was detected during papilla formation and induced cell reactions.     

Identification of QTLs for resistance to powdery mildew and SSR markers diagnostic for powdery mildew resistance genes in melon (Cucumis melo L.)

Powdery mildew caused by Podosphaera xanthii is an important foliar disease in melon. To find molecular markers for marker-assisted selection, we constructed a genetic linkage map of melon based on a population of 93 recombinant inbred lines derived from crosses between highly resistant AR 5 and susceptible ‘Earl’s Favourite (Harukei 3)’. The map spans 877 cM and consists of 167 markers, comprising 157 simple sequence repeats (SSRs), 7 sequence characterized amplified region/cleavage amplified polymorphic sequence markers and 3 phenotypic markers segregating into 20 linkage groups. Among them, 37 SSRs and 6 other markers were common to previous maps. Quantitative trait locus (QTL) analysis identified two loci for resistance to powdery mildew. The effects of these QTLs varied depending on strain and plant stage. The percentage of phenotypic variance explained for resistance to the pxA strain was similar between QTLs (R ² = 22–28%). For resistance to pxB strain, the QTL on linkage group (LG) XII was responsible for much more of the variance (41–46%) than that on LG IIA (12–13%). The QTL on LG IIA was located between two SSR markers. Using an independent population, we demonstrated the effectiveness of these markers. This is the first report of universal and effective markers linked to a gene for powdery mildew resistance in melon.     

QTLs for tomato powdery mildew resistance (Oidium lycopersici) in Lycopersicon parviflorum G1.1601 co-localize with two qualitative powdery mildew resistance genes

Tomato (Lycopersicon esculentum) is susceptible to the powdery mildew Oidium lycopersici, but several wild relatives such as Lycopersicon parviflorum G1.1601 are completely resistant. An F2 population from a cross of Lycopersicon esculentum cv. Moneymaker x Lycopersicon parviflorum G1.1601 was used to map the O. lycopersici resistance by using amplified fragment length polymorphism markers. The resistance was controlled by three quantitative trait loci (QTLs). Ol-qtl1 is on chromosome 6 in the same region as the Ol-1 locus, which is involved in a hypersensitive resistance response to O. lycopersici. Ol-qtl2 and Ol-qtl3 are located on chromosome 12, separated by 25 cM, in the vicinity of the Lv locus conferring resistance to another powdery mildew species, Leveillula taurica. The three QTLs, jointly explaining 68% of the phenotypic variation, were confirmed by testing F3 progenies. A set of polymerase chain reaction-based cleaved amplified polymorphic sequence and sequence characterized amplified region markers was generated for efficient monitoring of the target QTL genomic regions in marker assisted selection. The possible relationship between genes underlying major and partial resistance for tomato powdery mildew is discussed.     

三裂叶豚草锈菌侵染寄主叶片的显微和超微观察

【背景】三裂叶豚草是我国重要的外来入侵植物之一,其传播速度快,已给我国造成巨大的经济损失。近年来发现的三裂叶豚草锈菌是一种对其具有生物防治潜力的病原菌。【方法】本文利用显微技术研究了三裂叶豚草锈菌的侵染过程及其对寄主结构的影响。【结果】三裂叶豚草锈菌菌丝可从多处侵入同一个叶肉细胞,胞间菌丝与叶肉细胞相接触可使部分细胞壁增厚。锈菌侵染使三裂叶豚草叶脉末梢导管分枝增多,造成三裂叶豚草水分代谢失调;叶片细胞内膜系统破碎化,细胞器结构受到不同程度的破坏,导致细胞内膜系统紊乱,细胞器结构稳定性降低。【结论与意义】豚草锈菌侵染破坏了三裂叶豚草叶片的细胞结构。本研究为深入研究豚草锈菌的致病机理奠定了基础。     

The knock‐down of the expression of MdMLO19 reduces susceptibility to powdery mildew (Podosphaera leucotricha) in apple (Malus domestica)

Varieties resistant to powdery mildew (PM; caused by Podosphaera leucotricha) are a major component of sustainable apple production. Resistance can be achieved by knocking‐out susceptibility S‐genes to be singled out among members of the MLO (Mildew Locus O) gene family. Candidates are MLO S‐genes of phylogenetic clade V up‐regulated upon PM inoculation, such as MdMLO11 and 19 (clade V) and MdMLO18 (clade VII). We report the knock‐down through RNA interference of MdMLO11 and 19, as well as the complementation of resistance with MdMLO18 in the Arabidopsis thaliana triple mlo mutant Atmlo2/6/12. The knock‐down of MdMLO19 reduced PM disease severity by 75%, whereas the knock‐down of MdMLO11, alone or in combination with MdMLO19, did not result in any reduction or additional reduction of susceptibility compared with MdMLO19 alone. The test in A. thaliana excluded a role for MdMLO18 in PM susceptibility. Cell wall appositions (papillae) were present in both PM‐resistant and PM‐susceptible plants, but were larger in resistant lines. No obvious negative phenotype was observed in plants with mlo genes knocked down. Apparently, MdMLO19 plays the pivotal role in apple PM susceptibility and its knock‐down induces a very significant level of resistance.     

Isolation and identification of eight races of powdery mildew of roses (Podosphaera pannosa) (Wallr.: Fr.) de Bary and the genetic analysis of the resistance gene Rpp1

Powdery mildew, caused by Podosphaera pannosa, is one of the most-severe diseases of roses grown under glass. The differentiation into physiological races and the genetic analysis of resistance in a segregating host population was investigated using single conidial isolates of the pathogen. Using ten rose genotypes, all eight isolates of the pathogen could be ascribed to different races. Five races were isolated from one location, which indicates that populations of P. pannosa exhibit a high racial diversity. Infection experiments in a backcross-population of 114 rose plants resulted in a 1:1 segregation, suggesting control by a single dominant gene. Rpp1 is the first resistance gene against rose powdery mildew to be described.     

Characterisation of two novel types of hexactinomyxon spores (Myxozoa) with subsidiary protrusions on their caudal processes

Two types of hexactinomyxon spores, Hexactinomyxon type 1 nov. and Hexactinomyxon type 2 nov., are reported from freshwater tubificid oligochaetes, Limnodrilus hoffmeisteri and L. udekemianus. Spores are triradially symmetrical and comprise a spore body, style and 6 caudal processes. The caudal processes arise from the division of each of the 3 valve cells into an equal pair of projections at the base of the style. One of each pair is fused conspicuously to its nearest neighbour for the initial 1/5 to 1/4 of their total length. Distally, each process possesses subsidiary protrusions which are irregularly distributed and irregularly shaped extensions of the valve cell. Scanning electron microscopy of Hexactinomyxon type 2 nov. revealed that these protrusions are a seamless extension of the valve cell wall which branch distally, occasionally laterally, and terminate in a distinct bulbous structure; they also form the terminus of each process. The small subunit ribosomal DNA gene (18S) of both hexactinomyxon types was amplified through a nested PCR, then digested with the restriction enzymes Dde I and Hha I. The resultant cleavage patterns suggested the presence of 2 forms. Subsequent partial sequencing of 18S rDNA confirmed the identification of 2 novel types.     

The complex resistance to cucumber mosaic cucumovirus (CMV) in the melon accession PI161375 is governed by one gene and at least two quantitative trait loci

The complex resistance to cucumber mosaic virus (CMV) present in the exotic melon accession Sonwang Charmi PI161375 (SC) has been studied using two populations, a near-isogenic line (NIL) collection and a doubled haploid line (DHL) collection, both generated from a cross between SC and the cultivar Piel de Sapo as resistant and susceptible parents, respectively. The NIL collection had previously allowed us to describe a single recessive gene, cmv1, which conferred full resistance to CMV strains P9 and P104.82. Screening of the whole DHL population followed by quantitative trait locus (QTL) analysis revealed that resistance to the strains M6 and TL, both non-responsive to cmv1, was quantitative and governed by at least three QTLs. One of them, cmvqw12.1, co-located with cmv1 in linkage group (LG) XII. The QTL analysis mapped another two QTLs in LGIII (cmvqw3.1) and LGX (cmvqw10.1) and showed interaction between cmvqw12.1 and cmvqw3.1. Progeny of crosses between resistant DHLs carrying the three main QTLs showed complete resistance to the strain M6, validating the accuracy of the QTL analysis. However, in our screening, there were resistant DHLs carrying only two QTLs, suggesting that there are other regions involved in resistance to M6 and required when one of the main QTLs is missing. Therefore, resistance to CMV in melon SC is qualitative for some strains and quantitative for the rest. For this late resistance, cmv1 is necessary and explains most of the phenotypic variance, but it is not sufficient, and needs the interaction with other loci.     

Antisense expression of peach mildew resistance locus O (PpMlo1) gene confers cross-species resistance to powdery mildew in Fragaria x ananassa

Powdery mildew (PM) is one of the major plant pathogens. The conventional method of PM control includes frequent use of sulfur-based fungicides adding to production costs and potential harm to the environment. PM remains a major scourge for Rosaceae crops where breeding approaches mainly resort to gene-for-gene resistance. We have tested an alternate source of PM resistance in Rosaceae. Mildew resistance locus O (MLO) has been well studied in barley due to its role in imparting broad spectrum resistance to PM. We identified PpMlo1 (Prunus persica Mlo) in peach and characterized it further to test if a similar mechanism of resistance is conserved in Rosaceae. Due to its recalcitrance in tissue culture, reverse genetic studies involving PpMloI were not feasible in peach. Therefore, Fragaria x ananassa LF9 line, a taxonomic surrogate, was used for functional analysis of PpMlo1. Agrobacterium-mediated transformation yielded transgenic strawberry plants expressing PpMlo1 in sense and antisense orientation. Antisense expression of PpMlo1 in transgenic strawberry plants conferred resistance to Fragaria-specific powdery mildew, Podosphaera macularis. Phylogenetic analysis of 208 putative Mlo gene copies from 35 plant species suggests a large number of duplications of this gene family prior to the divergence of monocots and eudicots, early in eudicot diversification. Our results indicate that the Mlo-based resistance mechanism is functional in Rosaceae, and that Fragaria can be used as a host to test mechanistic function of genes derived from related tree species. To the best of our knowledge, this work is one of the first attempts at testing the potential of using a Mlo-based resistance strategy to combat powdery mildew in Rosaceae.     

Genetic mapping of two powdery mildew resistance genes in einkorn (Triticum monococcum L.) accessions

Powdery mildew is a severe foliar disease for wheat and could cause great yield loss in epidemic years. To explore new powdery mildew resistance genes, two einkorn accessions including TA2033 and M80, both resistant to this disease, were studied for the inheritance of resistance. Each accession possessed a single but different dominant resistance gene that was designated as Mlm2033 and Mlm80, respectively. Marker mapping indicated that they are both linked to Xgwm344 on the long arm of chromosome 7A. To establish their genetic relationship with Pm1 on 7AL, five RFLP markers previously reported to co-segregate with Pm1a were converted to STS markers. Three of them detected polymorphism between the mapping parents and were mapped close to Mlm2033 or Mlm80 or both. Xmag2185, the locus determined by the STS marker derived from PSR680, one of the RFLP markers, was placed less than 2 cM away from them. The allelism test indicated that Mlm2033 and Mlm80 are likely allelic to each other. In addition, through comparative and EST mapping, more markers linked to these two genes were identified. The high density mapping of Mlm2033 and Mlm80 will contribute to map-based cloning of the Pm1 locus. The markers for both genes will also facilitate their transfer to wheat.     

亚蔬中心绿豆品种(系)抗白粉病评价

采用苗期人工接种鉴定法,在大棚种植条件下对12个亚蔬中心(AVRDC)绿豆品种白粉病抗性进行了鉴定评价。结果显示,VC1560C、V4785和VC2768A三个品种高抗(HR)白粉病,VC6173-14、V1132为中抗(MR)白粉病品种。其它品种对白粉病表现高度感病。在田间种植条件下对亚蔬中心16个抗豆象回交9代品系(BC9)进行了成株期白粉病抗性鉴定。与对照感病品种 VC1973、VC1178A 相比,VC6459-3-6-37和 VC6458-6-3-16对白粉病具有一定抗性,但白粉病感染程度仍很严重,其它14个 BC9品系均对白粉病表现高度感病。