HISTONE DEACETYLASE 6 represses pathogen defence responses in Arabidopsis thaliana

Plant defence mechanisms are suppressed in the absence of pathogen attack to prevent wasted energy and growth inhibition. However, how defence responses are repressed is not well understood. Histone deacetylase 6 (HDA6) is a negative regulator of gene expression, and its role in pathogen defence response in plants is not known. In this study, a novel allele of hda6 (designated as shi5) with spontaneous defence response was isolated from a forward genetics screening in Arabidopsis. The shi5 mutant exhibited increased resistance to hemibiotrophic bacterial pathogen Pst DC3000, constitutively activated expression of pathogen‐responsive genes including PR1, PR2, etc. and increased histone acetylation levels at the promoters of most tested genes that were upregulated in shi5. In both wild type and shi5 plants, the expression and histone acetylation of these genes were upregulated by pathogen infection. HDA6 was found to bind to the promoters of these genes under both normal growth conditions and pathogen infection. Our research suggests that HDA6 is a general repressor of pathogen defence response and plays important roles in inhibiting and modulating the expression of pathogen‐responsive genes in Arabidopsis.     

Analysis of expressed sequence tags derived from pea leaves infected by Peronospora viciae f. sp. pisi

A cDNA library was constructed from field pea leaves infected by the downy mildew pathogen, Peronospora viciae f. sp. pisi, using a suppression subtractive hybridisation approach. The library consists of 399 expressed sequence tags, from which 207 unisequences were obtained after sequence assembly. Of the unisequences, six were shown to be of Peronospora viciae f. sp. pisi origin. The remaining unisequences were subjected to gene ontology analysis and their functions were predicted in silico. Eleven of these unisequences (representing 24 clones) shared significant sequence similarities with Arabidopsis genes known to be involved in downy mildew resistance, including the well‐characterised genes RPP5, RPP6 and RPP27. Expression analysis of five selected unisequences by real‐time PCR indicated that all five were up‐regulated during downy mildew pathogenesis, suggesting a significant role for these genes in the host response to downy mildew infection.     

Immunolocalization and Histocytopathological Effects of Xanthomonas arboricola pv. pruni on Naturally Infected Leaf and Fruit Tissues of Peach (Prunus persica L. Batsch)

Immunofluorescence and cytohistochemical studies have been performed to understand the host–parasite relationships in the pathosystem: peach–Xanthomonas arboricola pv. pruni (Xap). Using a commercial immunodetection kit, Xap cells were specifically identified in tissues from infected leaves and fruits. Sections from infected leaves showed that the pathogen penetrates the mesophyll via stomata and develops in the intercellular spaces where it degrades the cell wall components. This leads to cell collapse and consequently to the formation of necrotic lesions. The same events have been noted in sections from infected fruits. However, the contaminated zones of mesocarp parenchyma exhibited cell dedifferentiation and generated somatic embryo‐like structures. Sections from midrib samples collected at different distances from infected lamina revealed the presence of Xap cells in the sieve tubes and xylem suggesting a systemic trafficking of the pathogen. The results are discussed in terms of cytological effects and epidemiology of Xap.     

Transcriptome analysis of sugar beet root maggot (Tetanops myopaeformis) genes modulated by the Beta vulgaris host

Sugar beet root maggot (SBRM, Tetanops myopaeformis von Röder) is a major but poorly understood insect pest of sugar beet (Beta vulgaris L.). The molecular mechanisms underlying plant defense responses are well documented, however, little information is available about complementary mechanisms for insect adaptive responses to overcome host resistance. To date, no studies have been published on SBRM gene expression profiling. Suppressive subtractive hybridization (SSH) generated more than 300 SBRM ESTs differentially expressed in the interaction of the pest with a moderately resistant (F1016) and a susceptible (F1010) sugar beet line. Blast2GO v. 3.2 search indicated that over 40% of the differentially expressed genes had known functions, primarily driven by fruit fly D. melanogaster genes. Expression patterns of 18 selected EST clones were confirmed by RT‐PCR analysis. Gene Ontology (GO) analysis predicted a dominance of metabolic and catalytic genes involved in the interaction of SBRM with its host. SBRM genes functioning during development, regulation, cellular process, signaling and under stress conditions were annotated. SBRM genes that were common or unique in response to resistant or susceptible interactions with the host were identified and their possible roles in insect responses to the host are discussed.     

β‐Aminobutyric Acid Primed Expression of WRKY and Defence Genes in Brassica carinata Against Alternaria Blight

Foliar spray with BABA led to a significant reduction of lesion development in Brassica carinata caused by Alternaria brassicae. To get better insight into molecular mechanisms underlying priming of defence responses by BABA, expression pattern of BcWRKY genes and marker genes for the SA and JA pathway namely PR‐1 and PDF 1.2 was examined. Q‐RT‐PCR analysis revealed priming of BcWRKY70, BcWRKY11 and BcWRKY53 gene expression in BABA‐pretreated Brassica plants challenged with pathogen. However, the expression of BcWRKY72 and BcWRKY18 remained unchanged. Furthermore, BcWRKY7 gene was found to be upregulated in water‐treated plants in response to pathogen indicating its role in susceptibility. In addition, BABA application potentiated expression of defence genes PR‐1, PDF1.2 and PAL in response to the pathogen. In conclusion, BABA‐primed expression of BcWRKY70, BcWRKY11 and BcWRKY53 genes is strongly correlated with enhanced expression of PR‐1, PDF1.2 and PAL hence suggesting their role in BABA‐induced resistance.     

Silencing of SlPL,which encodes a pectate lyase in tomato,confers enhanced fruit firmness,prolonged shelf‐life and reduced susceptibility to grey mould

Pectate lyase genes have been documented as excellent candidates for improvement of fruit firmness. However, implementation of pectate lyase in regulating fruit postharvest deterioration has not been fully explored. In this report, 22 individual pectate lyase genes in tomato were identified, and one pectate lyase gene SlPL (Solyc03g111690) showed dominant expression during fruit maturation. RNA interference of SlPL resulted in enhanced fruit firmness and changes in pericarp cells. More importantly, the SlPL‐RNAi fruit demonstrated greater antirotting and pathogen‐resisting ability. Compared to wild‐type, SlPL‐RNAi fruit had higher levels of cellulose and hemicellulose, whereas the level of water‐soluble pectin was lower. Consistent with this, the activities of peroxidase, superoxide dismutase and catalase were higher in SlPL‐RNAi fruit, and the malondialdehyde concentration was lower. RNA‐Seq results showed large amounts of differentially expressed genes involved in hormone signalling, cell wall modification, oxidative stress and pathogen resistance. Collectively, these data demonstrate that pectate lyase plays an important role in both fruit softening and pathogen resistance. This may advance knowledge of postharvest fruit preservation in tomato and other fleshy fruit.     

Deregulated microRNAs in CD4+ T cells from individuals with latent tuberculosis versus active tuberculosis

The mechanisms of latent tuberculosis (TB) infection remain elusive. Roles of microRNA (miRNA) have been highlighted in pathogen–host interactions recently. To identify miRNAs involved in the immune response to TB, expression profiles of miRNAs in CD4⁺ T cells from patients with latent TB, active TB and healthy controls were investigated by microarray assay and validated by RT‐qPCR. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were used to analyse the significant functions and involvement in signalling pathways of the differentially expressed miRNAs. To identify potential target genes for miR‐29, interferon‐γ (IFN‐γ) mRNA expression was measured by RT‐qPCR. Our results showed that 27 miRNAs were deregulated among the three groups. RT‐qPCR results were generally consistent with the microarray data. We observed an inverse correlation between miR‐29 level and IFN‐γ mRNA expression in CD4⁺ T cells. GO and KEGG pathway analysis showed that the possible target genes of deregulated miRNAs were significantly enriched in mitogen‐activated protein kinase signalling pathway, focal adhesion and extracellular matrix receptor interaction, which might be involved in the transition from latent to active TB. In all, for the first time, our study revealed that some miRNAs in CD4⁺ T cells were altered in latent and active TB. Function and pathway analysis highlighted the possible involvement of miRNA‐deregulated mRNAs in TB. The study might help to improve understanding of the relationship between miRNAs in CD4⁺ T cells and TB, and laid an important foundation for further identification of the underlying mechanisms of latent TB infection and its reactivation.     

Facing warm temperatures during migration: cardiac mRNA responses of two adult Oncorhynchus nerka populations to warming and swimming challenges

The main findings of the current study were that exposing adult sockeye salmon Onchorhynchus nerka to a warm temperature that they regularly encounter during their river migration induced a heat shock response at an mRNA level, and this response was exacerbated with forced swimming. Similar to the heat shock response, increased immune defence‐related responses were also observed after warm temperature treatment and with a swimming challenge in two different populations (Chilko and Nechako), but with some important differences. Microarray analyses revealed that 347 genes were differentially expressed between the cold (12–13° C) and warm (18–19° C) treated fish, with stress response (GO:0006950) and response to fungus (GO:0009620) elevated with warm treatment, while expression for genes involved in oxidative phosphorylation (GO:0006119) and electron transport chain (GO:0022900) elevated for cold‐treated fish. Analysis of single genes with real‐time quantitative PCR revealed that temperature had the most significant effect on mRNA expression levels, with swimming and population having secondary influences. Warm temperature treatment for the Chilko population induced expression of heat shock protein (hsp) 90α, hsp90β and hsp30 as well as interferon‐inducible protein. The Nechako population, which is known to have a narrower thermal tolerance window than the Chilko population, showed even more pronounced stress responses to the warm treatment and there was significant interaction between population and temperature treatment for hsp90β expression. Moreover, significant interactions were noted between temperature treatment and swimming challenge for hsp90α and hsp30, and while swimming challenge alone increased expression of these hsps, the expression levels were significantly elevated in warm‐treated fish swum to exhaustion. In conclusion, it seems that adult O. nerka currently encounter conditions that induce several cellular defence mechanisms during their once‐in‐the‐lifetime migration. As river temperatures continue to increase, it remains to be seen whether or not these cellular defences provide sufficient protection for all O. nerka populations.     

Priming of protein expression in the defence response of Zantedeschia aethiopica to Pectobacterium carotovorum

The defence response of Zantedeschia aethiopica, a natural rhizomatous host of the soft rot bacterium Pectobacterium carotovorum, was studied following the activation of common induced resistance pathways—systemic acquired resistance and induced systemic resistance. Proteomic tools were used, together with in vitro quantification and in situ localization of selected oxidizing enzymes. In total, 527 proteins were analysed by label‐free mass spectrometry (MS) and annotated against the National Center for Biotechnology Information (NCBI) nonredundant (nr) protein database of rice (Oryza sativa). Of these, the fore most differentially expressed group comprised 215 proteins that were primed following application of methyl jasmonate (MJ) and subsequent infection with the pathogen. Sixty‐five proteins were down‐regulated following MJ treatments. The application of benzothiadiazole (BTH) increased the expression of 23 proteins; however, subsequent infection with the pathogen repressed their expression and did not induce priming. The sorting of primed proteins by Gene Ontology protein function category revealed that the primed proteins included nucleic acid‐binding proteins, cofactor‐binding proteins, ion‐binding proteins, transferases, hydrolases and oxidoreductases. In line with the highlighted involvement of oxidoreductases in the defence response, we determined their activities, priming pattern and localization in planta. Increased activities were confined to the area surrounding the pathogen penetration site, associating these enzymes with the induced systemic resistance afforded by the jasmonic acid signalling pathway. The results presented here demonstrate the concerted priming of protein expression following MJ treatment, making it a prominent part of the defence response of Z. aethiopica to P. carotovorum.