QTL Mapping Reveals Both All-Stage and Adult-Plant Resistance to Powdery Mildew in Chinese Elite Wheat Cultivars.

Powdery mildew caused by Blumeria graminis f. sp. tritici is a threat to wheat production in China. Mapping quantitative trait loci (QTL) for resistance to powdery mildew and developing breeder-friendly markers are important initial steps in breeding resistant cultivars. An all-stage resistance gene and several QTL were identified using a population of 254 recombinant inbred lines developed from a Jingdong 8/Aikang 58 cross. The population was evaluated for powdery mildew resistance across six field environments over three consecutive growing seasons utilizing two different mixtures of B. graminis f. sp. tritici isolates, named #Bgt-HB and #Bgt-BJ. Using genotypic data obtained from the Wheat TraitBreed 50K single-nucleotide polymorphism array, seven stable QTL were identified on chromosome arms 1DL, 2AL, 2DS, 4DL, 5AL, 6BL.1, and 6BL.2. The QTL on 2AL conferred all-stage resistance to B. graminis f. sp. tritici race E20 in greenhouse tests and explained up to 52% of the phenotypic variance in field trials but was resistant only against #Bgt-HB. The gene involved in this QTL was predicted to be Pm4a based on genome location and gene sequence. QPmja.caas-1DL, QPmja.caas-4DL, and QPmja.caas-6BL.1 were identified as potentially new QTL for powdery mildew resistance. QPmja.caas-2DS and QPmja.caas-6BL.1 were effective against both B. graminis f. sp. tritici mixtures, indicating their probable broad-spectrum resistance. A Kompetitive allele-specific PCR marker closely linked to QPmja.caas-2DS was developed and validated in a panel of 286 wheat cultivars. Because both Jingdong 8 and Aikang 58 have been leading cultivars and breeding parents, the QTL and marker reported represent valuable resources for wheat researchers and breeders.

VqBGH40a isolated from Chinese wild Vitis quinquangularis degrades trans-piceid and enhances trans-resveratrol.

Resveratrol (3,5,4'-trihydroxy-stilbene) is a phytoalexin that can prevent plants from pathogen attacks. Piceid is the glycosylation product of resveratrol and the main storage form of stilbenes in grapevines. Here, we reported the function of a ß-glycoside hydrolase gene, VqBGH40a, from the Chinese wild grapevine Vitis quinquangularis accession Danfeng-2 in the regulation of plant resistance to powdery mildew (Uncinula necator). VqBGH40a belonging to ß-glycoside hydrolase family 1 encoded 506 amino acids and was located on the cytomembrane. Its optimal induction condition was 28 or 30℃, for 4 h, with 0.1 mM IPTG in a prokaryotic expression system. Enzyme activity detection showed that purified VqBGH40a could hydrolyze trans-piceid to form trans-resveratrol in vitro. VqBGH40a was transiently overexpressed in Danfeng-2 leaves and then artificially inoculated with powdery mildew showed that VqBGH40a protein could hydrolyze trans-piceid in vivo. Additionally, a comparative family analysis between VqBGH40a and 38 VviBGHs was performed. Overall, these results demonstrate that VqBGH40a can hydrolyze trans-piceid, enhance trans-resveratrol content, and participate in the defense mechanism of grapevine against powdery mildew.

Investigation of long non-coding RNAs as regulatory players of grapevine response to powdery and downy mildew infection.

BACKGROUND: Long non-coding RNAs (lncRNAs) are regulatory transcripts of length > 200 nt. Owing to the rapidly progressing RNA-sequencing technologies, lncRNAs are emerging as considerable nodes in the plant antifungal defense networks. Therefore, we investigated their role in Vitis vinifera (grapevine) in response to obligate biotrophic fungal phytopathogens, Erysiphe necator (powdery mildew, PM) and Plasmopara viticola (downy mildew, DM), which impose huge agro-economic burden on grape-growers worldwide. RESULTS: Using computational approach based on RNA-seq data, 71 PM- and 83 DM-responsive V. vinifera lncRNAs were identified and comprehensively examined for their putative functional roles in plant defense response. V. vinifera protein coding sequences (CDS) were also profiled based on expression levels, and 1037 PM-responsive and 670 DM-responsive CDS were identified. Next, co-expression analysis-based functional annotation revealed their association with gene ontology (GO) terms for 'response to stress', 'response to biotic stimulus', 'immune system process', etc. Further investigation based on analysis of domains, enzyme classification, pathways enrichment, transcription factors (TFs), interactions with microRNAs (miRNAs), and real-time quantitative PCR of lncRNAs and co-expressing CDS pairs suggested their involvement in modulation of basal and specific defense responses such as: Ca2+-dependent signaling, cell wall reinforcement, reactive oxygen species metabolism, pathogenesis related proteins accumulation, phytohormonal signal transduction, and secondary metabolism. CONCLUSIONS: Overall, the identified lncRNAs provide insights into the underlying intricacy of grapevine transcriptional reprogramming/post-transcriptional regulation to delay or seize the living cell-dependent pathogen growth. Therefore, in addition to defense-responsive genes such as TFs, the identified lncRNAs can be further examined and leveraged to candidates for biotechnological improvement/breeding to enhance fungal stress resistance in this susceptible fruit crop of economic and nutritional importance.

Comparative proteomic analysis reveals molecular differences between incompatible and compatible interaction of Erysiphe pisi in garden pea.

Comparative proteome analysis of Erysiphe pisi-infected pea genotypes; JI-2480 carrying er2 resistant gene and Arkel, the susceptible genotype by liquid chromatography- mass spectrometry (LCMS/MS QTOF) at 72 h post inoculation (hpi) revealed several differentially abundant proteins (DAPs) of both the host and the pathogen. The functional annotation of proteins through gene enrichment and KEGG pathway analyses revealed strong up-regulation of pathogenesis related protein NPR1, proteins related to defense, transportation and signal transduction, hypersensitive response, cell wall modifications, phenylpropanoid and metabolic pathways in J-72. Significant abundance of membrane-related polypeptides, kinase domains and small GTPase signal transduction-related proteins suggested their major role in plant defense. The abundance of cellular antioxidant protein, catalase and its isozyme along with calreticulin-1 and 2 in J-72 confirmed their intervention in maintaining a redox balance in powdery mildew defense. High abundance levels of Glycolysis-related proteins indicated it as a major pathway for energy source during fungal growth. The majority of pathogenicity and virulence genes were downregulated in J-72 compared to A-72, while four EKA (Effectors homologues to Avk1 and Avra10) like avirulence proteins were significantly upregulated in incompatible interaction suggesting their role in eliciting hypersensitive response in pea against E. pisi.

Phylogeny and taxonomy of powdery mildew caused by Erysiphe species on Corylus hosts.

Erysiphe species (powdery mildews) on Corylus and Ostrya hosts (Betulaceae subfam. Coryloideae) in Asia and North America are widespread pathogens on these economically and ecologically valuable nut crops. An improved understanding of their phylogeny and taxonomy is of ecological and applied importance. Phylogenetic analyses and morphological reexaminations conducted in this study revealed a higher degree of diversity and cryptic speciation than reflected in earlier species concepts. North American collections on C. cornuta, which were previously assigned to E. corylacearum, proved to constitute a species of its own and are herein introduced as E. cornutae, sp. nov. Two additional North American species, E. coryli-americanae, sp. nov. and E. ostryae, sp. nov., have been detected on C. americana and O. virginiana and are described. They are morphologically similar to E. cornutae, but genetically distinct. Based on phylogenetic analyses, E. corylacearum is an Asian species confined to various Asian Corylus species. Sequence data retrieved from Japanese type material of E. corylicola revealed that this species clusters with sequences from E. elevata on Catalpa species, distant from all other Erysiphe species on Corylus. Morphologically similar, yet distinct, specimens on C. sieboldiana, which were previously assigned to E. corylicola, form a distinct, distant clade. The species involved is described herein as E. pseudocorylacearum, sp. nov. Additionally, an unusual infection of C. sieboldiana in Japan by E. syringae has been shown by means of sequence data. The phylogeny and taxonomy of Erysiphe species belonging to the Corylioideae are discussed in detail, and a key to the species concerned is provided.

One Crop Disease, How Many Pathogens? Podosphaera xanthii and Erysiphe vignae sp. nov. Identified as the Two Species that Cause Powdery Mildew of Mungbean (Vigna radiata) and Black Gram (V. mungo) in Australia.

Powdery mildew is a significant threat to mungbean (Vigna radiata) and black gram (V. mungo) production across Australia and overseas. Although they have been present in Australia for at least six decades and are easily recognized in the field, the precise identification of the pathogens causing this disease has remained unclear. Our goal was to identify the powdery mildew species infecting mungbean, black gram, and wild mungbean (V. radiata ssp. sublobata) in Australia. The internal transcribed spacer (ITS) and large subunit sequences of the ribosomal DNA and/or morphology of 57 Australian specimens were examined. Mungbean and black gram were infected by two species: Podosphaera xanthii and a newly recognized taxon, Erysiphe vignae sp. nov. Wild mungbean was infected only with P. xanthii. Mungbean and black gram powdery mildew ITS sequences from China, India, and Taiwan revealed the presence of only P. xanthii on these crops despite controversial reports of an Erysiphe species on both crops in India. Sequence analyses indicated that the closest relative of E. vignae is E. diffusa, which infects soybean (Glycine max) and other plants. E. vignae did not infect soybean in cross-inoculation tests. In turn, E. diffusa from soybean infected black gram and provoked hypersensitive response in mungbean. The recognition of a second species, E. vignae, as another causal agent of mungbean and black gram powdery mildew in Australia may complicate plant breeding efforts and control of the disease with fungicide applications.

Alternative products to control powdery mildew in soybeans culture in field

The occurrence of powdery mildew (Microsphaera diffusa) in soybean (Glycine max L.) has increased in the last harvests. In order to study the efficiency of powdery mildew control due to the application of alternative products and conventional fungicide, trials were conducted in Ponta Grossa, PR, Brazil, during the 2013/2014 and 2014/2015 growing seasons. The design used was randomized blocks with four replications. The treatments for the experiments were: 1 - control; 2 - acibenzolar-S-methyl (Bion 500 WG®); 3 - calcium (Max Fruit®); 4 - Micronutrients: copper, manganese and zinc (Wert Plus®); 5 - Micronutrients: manganese, zinc and molybdenum (V6®); 6 - NK fertilizer (Hight Roots®); 7 - Ascophyllum nodosum (Acadian®) and 8 - fungicide (azoxystrobin + cyproconazole) (Priori XTRA®) with the addition of the adjuvant. Four applications of alternative products (phenological stages V3, V6, R1 and R5.1) and two of fungicide (phenological stages R1 and R5.1) were carried out. The parameters evaluated were powdery mildew severity and productivity. The severity data made it possible to calculate the area under the disease progress curve (AUDPG). Alternative products didn't reduce powdery mildew in the two harvests. The conventional fungicide treatment was the only one that controlled powdery mildew and didn't reduce the productivity in both experiments.

VvERD6l13 is a grapevine sucrose transporter highly up-regulated in response to infection by Botrytis cinerea and Erysiphe necator.

The Early-Response to Dehydration six-like (ERD6l) is one of the largest families of sugar transporters in plants, however, is also one of the less studied with very few members characterized. In this work, we identified 18 members of the grapevine ERD6l family, analyzed their promoters and putative topology and additionally functionally characterized the member VvERD6l13. VvERD6l13 was strongly up-regulated in grape berries infected with Botrytis cinerea and Erysiphe necator in cv. Trincadeira and Carignan, respectively, suggesting an important role in grape berry-pathogen interaction, as we had hypothesized. In Cabernet Sauvignon Berry suspension cultured cells, VvERD6l13 was also up-regulated, by 4-fold, 48 h after elicitation with mycelium extract of B. cinerea. Besides being expressed in grape berries from various developmental stages, VvERD6l13 is also expressed in leaves, canes, flowers and, noticeably, in roots. Using tobacco and an hxt-null Saccharomyces cerevisiae strain as heterologous expression models, we showed that VvERD6l13 is localized at the plasma membrane and mediates the H+-dependent transport of sucrose (Km = 33 mM) thus confirming VvERD6l13 as a bona fide sugar transporter involved in sugar mobilization in grapevine and transcriptionally induced in response to biotic stress.

Dual RNA-Seq analysis of Medicago truncatula and the pea powdery mildew Erysiphe pisi uncovers distinct host transcriptional signatures during incompatible and compatible interactions and pathogen effector candidates.

Powdery mildew (PM) is a serious fungal disease of legumes. To gain novel insights into PM pathogenesis and host resistance/susceptibility, we used dual RNA-Seq to simultaneously capture host and pathogen transcriptomes at 1 d post-inoculation of resistant and susceptible Medicago truncatula genotypes with the PM Erysiphe pisi (Ep). Differential expression analysis indicates that R-gene mediated resistance against Ep involves extensive transcriptional reprogramming. Functional enrichment of differentially expressed host genes and in silico analysis of co-regulated promoters suggests that amplification of PTI, activation of the JA/ET signaling network, and regulation of growth-defense balance correlate with resistance. In contrast, processes that favor biotrophy, including suppression of defense signaling and programmed cell death, and weaker cell wall defenses are important susceptibility factors. Lastly, Ep effector candidates and genes with known/putative virulence functions were identified, representing a valuable resource that can be leveraged to improve our understanding of legume-PM interactions.

Danos causados pela infecção de oídio em diferentes estádios fenológicos da soja / Damages caused by infection of powdery mildews in different phenological stages of soybeans

Ainda não há estudos precisos que quantifiquem os prejuízos decorrentes de infecção por oídio e/ou outras doenças foliares, para a maioria das culturas de importância econômica no Brasil. O objetivo foi quantificar as perdas causadas por oídio (Microsphaera diffusa) infectando a cultura da soja em diferentes estádios fenológicos e relacioná-las ao desenvolvimento e produtividade da cultura. O experimento foi desenvolvido em ambiente protegido, e os tratamentos foram testemunha controlada, testemunha sem controle, infecção iniciada em R1 ­ R2, infecção iniciada em R ­ R, infecção iniciada em R ­ R e infecção iniciada em R ­ R. A avaliação foi feita 345.15.25.35.4 semanalmente, considerando a porcentagem da área foliar infectada. Os resultados mostraram que, no tratamento em que houve infecções iniciadas em R1-R2 e R3-R4, a porcentagem de área foliar afetada foi maior (41% e 38%, respectivamente), com consequente menor produtividade (1.186,6 e 1.309,5 kg.ha-1 respectivamente). No tratamento em que a infecção ocorreu em R ­ R, houve 5.35.4 a menor média de área foliar afetada pela doença (24%) e a produtividade teve queda de 26%. Os resultados mostraram que as perdas de produtividade pelo oídio na cultivar Embrapa 48 variaram ao redor de 26 a 50%, e que a recomendação oficial para o início de controle do oídio da soja, quando esta se apresentar entre 40 e 50% de severidade, deve ser questionada e outros trabalhos neste âmbito devem ser desenvolvidos para determinação das perdas ocasionadas por esta doença na cultura.

At present there are no precise studies quantifying the damages caused by powdery mildews and other foliar diseases for the majority of economically important crops in Brazil. The objective of the present study was to quantify the losses caused by powdery mildews (Microsphaera diffusa) in soybeans in different phenological stages, and to correlate them with the development and yield of the crop. The trials was carried out in the greenhouse and the treatments were: controlled check, noncontrolled check, infection initiated at stage R1-R2, infection initiated at stage R3-R4, infection initiated at stage R5.1-R5.2, infection initiated at stage R5.3-R5.4. The evaluation was done weekly considering the percentage of infected leaf area. The results showed that for the infection beginning at stages R1-R2 and R3-R4 the percentage of affected leaf area was higher (41% and 38%), with consequently lower yields (1,200 and 1,240 kg ha-1). When the infection occurred later at stage R5.3-R5.4, a lower affected leaf area (24%) was observed, and the yield decreased 26%. The results showed that the loss of yield by powdery mildew in cultivar Embrapa 48 ranged from around 26 to 50%, and that the official recommendation for the beginning of control of powdery mildew of soybean, where it presents between 40 and 50% of severity, should be questioned, and other work in this area should be undertaken to determine the loss caused by this disease in the crop.