Developmental characteristics of sporogenous hyphae: a new observation between Brassica juncea var. tumida and Albugo candida

Brassica juncea var. tumida (tumorous stem mustard) is widely cultivated as a feature vegetable in Ningbo, Zhejiang Province, and is used as a raw material for pickles in China. White blister rust, caused by Albugo candida, has severe negative effects on tumorous stem mustard, resulting in significant economic losses. In this study, hypha colonization and extension, haustorial formation, and the development process of sporogenous hyphae and sporangia in A. candida-infected tissues were studied using light and transmission electron microscopy. Additionally, the complex and unique sporogenous hypha developmental process was revealed. During A. candida infection, cytological and ultrastructural characteristics were similar to those of cruciferous plants. During sporogenous hyphae development, an initially cylindrical sporogenous hypha with a thin inner wall at the apex developed from a cell with a thick inner wall. At the apical end of the sporogenous hypha, an electron-dense complex gathered, causing the disintegration of the inner and outer walls and the formation of new inner and outer walls. With the formation of a clavate sporogenous hypha, a deep constriction of the cell wall occurred at its subapex, in which a septum was produced and a primary sporangium was separated from the sporogenous hypha. Both layers of the sporogenous hypha wall took part in the formation of the sporangial wall but were not related the formation of the sporangial outer wall in a septum, providing direct evidence of the holoblastic formation of sporangia. Furthermore, ultrastructural observation demonstrated that the development of the primary sporangial walls within a septum occurred later than that of their lateral walls, including the degradation of constricted outer walls around the septum, formation of the inner wall and subsequent outer walls of a sporangium in the upper septum layers, confirming the earlier observation. Therefore, this study provided the model of sporogenous hyphae development and sporangial formation in chain.

An improved assembly of the Albugo candida Ac2V genome reveals the expansion of the “CCG” class of effectors

Albugo candida is an obligate oomycete pathogen that infects many plants in the Brassicaceae family. We re-sequenced the genome of isolate Ac2V using PacBio long reads and constructed an assembly augmented by Illumina reads. The Ac2VPB genome assembly is 10% larger and more contiguous compared to a previous version. Our annotation of the new assembly, aided by RNASeq information, revealed a 175% expansion (40 to 110) in the CHxC effector class, which we redefined as “CCG” based on motif analysis. This class of effectors consist of arrays of phylogenetically related paralogs residing in gene sparse regions, and shows signatures of positive selection and presence/absence polymorphism. This work provides a resource that allows the dissection of the genomic components underlying A. candida adaptation and particularly the role of CCG effectors in virulence and avirulence on different hosts.

An improved assembly of the Albugo candida Ac2V genome reveals the expansion of the “CCG” class of effectors

Albugo candida is an obligate oomycete pathogen that infects many plants in the Brassicaceae family. We re-sequenced the genome of isolate Ac2V using PacBio long reads and constructed an assembly augmented by Illumina reads. The Ac2VPB genome assembly is 10% larger and more contiguous compared to a previous version. Our annotation of the new assembly, aided by RNASeq information, revealed a dramatic 250% expansion (40 to 110) in the CHxC effector class, which we redefined as “CCG” based on motif analysis. This class of effectors consist of arrays of phylogenetically related paralogs residing in gene sparse regions, and shows signatures of positive selection and presence/absence polymorphism. This work provides a resource that allows the dissection of the genomic components underlying A. candida adaptation and particularly the role of CCG effectors in virulence and avirulence on different hosts.

Evolutionary trade-offs at the Arabidopsis WRR4A resistance locus underpin alternate Albugo candida recognition specificities

The oomycete Albugo candida causes white rust of Brassicaceae, including vegetable and oilseed crops, and wild relatives such as Arabidopsis thaliana. Novel White Rust Resistance (WRR)-genes from Arabidopsis enable new insights into plant/parasite co-evolution. WRR4A from Arabidopsis accession Col-0 provides resistance to many but not all white rust races, and encodes a nucleotide-binding (NB), leucine-rich repeat (LRR) (NLR) immune receptor protein. Col-0 WRR4A resistance is broken by a Col-0-virulent isolate of A. candida race 4 (AcEx1). We identified an allele of WRR4A in Arabidopsis accession Oy-0 and other accessions that confers full resistance to AcEx1. WRR4AOy-0 carries a C-terminal extension required for recognition of AcEx1, but reduces recognition of several effectors recognized by the WRR4A_Col-0 allele. WRR4A_Oy-0 confers full resistance to AcEx1 when expressed as a transgene in the oilseed crop Camelina sativa.

The Arabidopsis WRR4A and WRR4B paralogous NLR proteins both confer recognition of multiple Albugo candida effectors

The oomycete Albugo candida causes white blister rust, an important disease of Brassica crops. Distinct races of A. candida are defined by their specificity for infecting different host species. The White Rust Resistance 4 (WRR4) locus in Col-0 accession of Arabidopsis thaliana contains three genes that encode TIR-NLR resistance proteins. The Col-0 alleles of WRR4A and WRR4B confer resistance to at least four A. candida races (2, 7 and 9 from B. juncea, B. rapa and B. oleracea, respectively, and Race 4 from Capsella bursa-pastoris). Resistance mediated by both paralogs can be overcome by Col-0-virulent isolates of Race 4. After comparing repertoires of candidate effectors in resisted and resistance-breaking strains, we used transient co-expression in tobacco or Arabidopsis to identify effectors recognized by WRR4A and WRR4B. A library of CCG effectors from four A. candida races was screened for WRR4A- or WRR4B- dependent elicitation of hypersensitive response (HR). These CCG genes were validated for WRR-dependent HR by bombardment assays in wild type Col-0, wrr4A or wrr4B mutants. Our analysis revealed eight WRR4A-recognized CCGs and four WRR4B-recognized CCGs. Remarkably, the N-terminal region of 100 amino acids after the secretion signal is sufficient for WRR4A recognition of these eight recognized effectors. This multiple recognition capacity potentially explains the broad-spectrum resistance to many A. candida races conferred by WRR4 paralogs.

Downy mildew of lavender caused by Peronospora belbahrii in Israel

Peronospora belbahrii is one of the most destructive downy mildew diseases that has emerged throughout the past two decades. Due to the lack of quarantine regulations and its possible seed-borne nature, it has spread globally and is now present in most areas in which basil is produced. While most obligate biotrophic, plant parasitic oomycetes are highly host-specific, there are a few that have a wider host range, e.g. Albugo candida, Bremia tulasnei, and Pseudoperonospora cubensis. Recently, it was shown that Peronospora belbahrii is able to infect Rosmarinus, Nepetia, and Micromeria in Israel in cross-infection trials, hinting an extended host range for also this pathogen. In this study, a newly occurring downy mildew pathogen on lavender was investigated with respect to its morphology and phylogeny, and it is shown that it belongs to Peronospora belbahrii as well. Thus, it seems that Peronospora belbahrii is currently extending its host range to additional members of the tribe Mentheae and Ocimeae. Therefore, it seems advisable to scrutinise all commonly used members of these tribes in order to avoid further spread of virulent genotypes.