Identification of differentially expressed genes in tolerant and susceptible potato cultivars in response to Spongospora subterranea f. sp. subterranea tuber infection

Powdery scab caused by Spongospora subterranea f. sp. subterranea (Sss) has recently become one of the most devastating potato diseases of economic importance in South Africa. The use of resistant cultivars has long been considered the most effective and sustainable strategy to manage the pathogen. However, little is known about the molecular mechanisms underlying resistance of potato tubers to Sss. Using RNA-sequencing (RNA-seq), 2058 differentially expressed genes (DEGs) were identified from two potato cultivars (tolerant and susceptible) in response to Sss infection. Analysis of the expression patterns of 10 selected defence-response genes was carried out at two different stages of tuber growth using RT-qPCR to validate the RNA-seq data. Several defence-related genes showed contrasting expression patterns between the tolerant and susceptible cultivars, including marker genes involved in the salicylic acid hormonal response pathway (StMRNA, StUDP and StWRKY6). Induction of six defence-related genes (StWRKY6, StTOSB, StSN2, StLOX, StUDP and StSN1) persisted until harvest of the tubers, while three other genes (StNBS, StMRNA and StPRF) were highly up-regulated during the initial stages of disease development. The results of this preliminary study suggest that the tolerant potato cultivar employs quantitative resistance and salicylic acid pathway hormonal responses against tuber infection by Sss. The identified genes have the potential to be used in the development of molecular markers for selection of powdery scab resistant potato lines in marker-assisted breeding programmes.     

Insights on gene expression response of a characterized resistant genotype of Solanum commersonii Dun. against Ralstonia solanacearum

Solanum commersonii is a wild species related to the cultivated potato. Some S. commersonii genotypes have been proven to be resistant to the pathogenic bacteria Ralstonia solanacearum, which causes damage in potato and other economically important crops. Here an expression analysis of the response of a resistant S. commersonii genotype against R. solanacearum was performed using microarrays. The aims of this work were to elucidate the molecular processes involved in the interaction, establish the timing of the response, and contribute to identify genes related to the resistance. The response to the treatment was already initiated at 6 h post-inoculation (hpi) and was established at 24 hpi; during this period, a high number of genes was differentially expressed and several candidate genes for the resistance of S. commersonii to R. solanacearum were identified. At an early stage, the photosynthetic process was highly repressed and several genes encoding proteins related to reactive oxygen species (ROS) production were differentially expressed. The induction of ERF and ACC-oxidase genes related to the ethylene pathway and PR1 related to the salicylic acid pathway suggested the induction of both pathways, and back up the previously reported hemibiotrophic nature of the pathogen. Five genes related to plant defence and observed to be differentially expressed at the first two time points were validated by real time PCR. This work gives a glimpse to the molecular processes involved in S. commersonii resistance and identifies the species as a valuable genetic source for potato breeding against bacterial wilt.     

Tight regulation of allene oxide synthase (AOS) and allene oxide cyclase-3 (AOC3) promote <Emphasis Type="Italic">Arabidopsis</Emphasis> susceptibility to the root-knot nematode <Emphasis Type="Italic">Meloidogyne javanica</Emphasis>

Biosynthesis of the oxylipin jasmonic acid (JA) in Arabidopsis thaliana is catalyzed by a single allene oxide synthase (AOS)-encoding gene and four genes encoding four functional allene oxide cyclase (AOC) polypeptides (AOC1, AOC2, AOC3, and AOC4). To elucidate the biological activities of the JA pathway in regulating the plant defense response to plant-parasitic nematodes, transgenic lines carrying the GUS reporter gene under the control of individual AOC or AOS promoters were examined. Upon penetration by second-stage juveniles (J2 s), promoter activities of AOC1, AOC3 and AOC4 appeared in the root tip and root-elongation zone, with AOC3 demonstrating highest induction. At 5 days AOC3 activity continued to be highly pronounced in the stele and root cortex, associated with nematode invasion throughout gall initiation and maturation. AOS expression appeared 3 days postinfection and accompanied all later infection stages. Mutant lines were analyzed: disruption in AOS rendered plants more resistant to nematode infection, as reflected by the decreased number of females produced on this line; loss-of-function of AOC3 rendered plants more susceptible to nematode infection. Oxylipins derived from the 9- and 13-lipoxygenase pathways were assayed for their direct inhibitory activity toward M. javanica J2 s. Clear nematicidal activity of the bioactive 9- and 13-hydroperoxides was observed. Oxylipins produced by divinyl ether synthase, colneleic acid, colnelenic acid and ω5(Z)-etherolenic acid demonstrated strong inhibitory activity. These data, along with those of other assayed oxylipins, suggest that temporal and spatial fine tuning of the JA route allowing nematodes parasitism on plant host.     

Turnip mosaic virus induces expression of the LRR II subfamily genes and regulates the salicylic acid signaling pathway in non-heading Chinese cabbage

In order to study the defense response to turnip mosaic virus (TuMV) infection in non-heading Chinese cabbage (Brassica campestris ssp. chinensis Makino), we cloned the LRR II subfamily genes which comprises six members. They were high homologous to the function-known LRR II genes of Arabidopsis. We investigated their expression through quantitative real-time PCR analysis. TuMV infection induced the expression of these genes locally and systematically, and regulated the endogenous accumulation of salicylic acid (SA). Exogenous SA spraying was able to induce resistance to the susceptibility of the TuMV-infected plants, which might function via inhibiting the viral duplication. Though TuMV-induced SA accumulation was not the determinant in regulating gene expression, it mediated the reaction oxygen species (ROS) burst as a channel of defense.     

Jasmonic acid and abscisic acid play important roles in host–pathogen interaction between Fusarium graminearum and wheat during the early stages of fusarium head blight

The effect of phytohormones on the defense response of wheat against Fusarium graminearum infection was investigated. Infection of heads with F. graminearum induced accumulation of salicylic acid (SA), jasmonic acid (JA), abscisic acid (ABA), and indole acetic acid (IAA). Exogenous phytohormone treatments showed crosstalk between them and a complex effect on expression of the genes ATB2, ExpB6, LEA Td16, PR1, Pdf1.2, PR4. JA treatment reduced F. graminearum growth and fusarium head blight (FHB) symptoms while an increase in FHB was observed with ABA. Transient down-regulation of allene oxide synthase (AOS) supports a complex role for JA in wheat head.     

Molecular and biochemical mechanisms of defence induced in pea by Rhizobium leguminosarum against Orobanche crenata

Orobanche species (broomrapes) are parasitic weeds which dramatically decrease the yields of many economically important dicotyledonous crops, including pea (Pisum sativum), in Mediterranean areas. Previously, we identified some Rhizobium leguminosarum strains, including P.SOM, which could both promote pea development and significantly reduce infection by Orobanche crenata, notably through induction of necrosis of attached parasites. In the present study, induced resistance against broomrape in the nodulated pea was shown to be associated with significant changes in rates of oxidative lipoxygenase (Lox) and phenylpropanoid/isoflavonoid pathways and in accumulation of derived toxins, including phenolics and pisatin (pea phytoalexin). Changes were followed for 5 weeks after inoculation and attack by Orobanche. In contrast to non‐inoculated plants or Orobanche only infected plants, polyphenoloxidase (PPO) activity and hydrogen peroxide content increased in response to bacteria inoculation indicating the involvement of oxidative processes. In parallel, the nodulated roots displayed high Lox activity related to the overexpression of the lox1 gene. Similarly, the expression of phenylalanine ammonia lyase (PAL) and 6a‐hydroxymaackiain 3‐O‐methyltransferase (Hmm6a) genes were induced early during nodule development, suggesting the central role of the phenylpropanoid/isoflavonoid pathways in the elicited defence. As a consequence, the derived products, phenolics and pisatin, accumulated in response to rhizobacteria and conferred mechanical and chemical barriers to the invading parasite. These results highlight the likely role of signalling pathways in induced resistance and suggest these mechanisms should be enhanced through integrated Orobanche management practices.     

Germination stimulatory activity of bacterial butenolide hormones from Streptomyces albus J1074 on seeds of the root parasitic weed Orobanche minor

Damage caused by Orobanchaceae root parasitic weeds is a substantial agricultural problem for global food security. Many studies have been conducted to establish practical methods of control, but efforts are still required for successful management. Seed germination of root parasitic weeds requires host-derived germination stimulants including strigolactones (SLs). Studies on SLs have revealed that a butenolide ring is the essential moiety for SL activity as a germination stimulant. Interestingly, recent studies have revealed that butenolide hormones regulate the biosynthesis of secondary metabolites and mediate communication in actinomycete bacteria. Because of the structural similarity between SLs and the bacterial butenolides, we evaluated the germination stimulatory activity of butenolides isolated from Streptomyces albus J1074 on root parasitic weeds. These butenolides were found to specifically induce seed germination of Orobanche minor. Our findings contribute to understanding the molecular mechanisms of germination stimulant perception and to the development of a method for their biological control.     

Pepper 9- and 13-lipoxygenase genes are differentially activated by two tobamoviruses and by hormone treatments

Two novel pepper 13-lipoxygenase (LOX) genes were cloned and their expressions were compared with those of three 9-LOX genes in pepper leaves inoculated with two different tobamoviruses. Obuda pepper virus (ObPV) inoculation led to a massive induction of pathogenesis-related genes and to the development of hypersensitive reaction (incompatible interaction), while Pepper mild mottle virus (PMMoV) inoculation resulted in a compatible interaction. Both virus infections markedly activated the expression of the two novel 13-LOXs. The magnitudes of induction of 13-LOXs did not differ substantially between the ObPV- and PMMoV-inoculated leaves. The induction of three 9-LOXs was markedly more robust and rapid in ObPV-inoculated leaves than in PMMoV-inoculated ones. LOXs were very differentially activated in pepper leaves treated with defense hormones. A large number of hormone-related cis-regulatory elements were identified in the promoter regions of LOXs. ObPV inoculation resulted also in the substantial up-regulation of an omega-6-fatty acid desaturase gene. Our results suggest that 9-LOX-dependent pathways are more probably involved in the suppression of virus replication than 13-LOX-dependent plant responses.     

<Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> upregulates marker genes of the salicylic acid and ethylene signaling pathways but not those of the jasmonic acid pathway in leaflets of <Emphasis Type="Italic">Solanum</Emphasis> lines during early stage of infection

Real-Time PCR assay was used to quantify the expression of marker genes of the salicylic acid, jasmonic acid and ethylene signaling pathways in seven Solanum lines after inoculation with a Ralstonia solanacearum phylotype I strain, R008. Four Solanum lycopersicum lines (CRA 66, Hawaii 7996, MST 32/1, Quatre carrées), one S. tuberosum line (Spunta), the wild Lycopersicon cerasiforme and Solanum commersonii were used for this investigation. Results revealed very little activation of the jasmonic acid pathway marker genes, lipoxygenase A (LoxA) and protease inhibitor II (Pin2), with no significant difference (p > 0.05) in fold change expression among the Solanum lines. In contrast the salicylic acid pathway marker genes, glucanase A (GluA) and PR-1a, and the ethylene pathway marker genes, osmotin-like (Osm) and PR-1b, were expressed at higher levels with a statistically significant difference (p < 0.05) in fold change expression among the Solanum lines. The resistant lines L. cerasiforme, CRA 66, Hawaii 7996 and S. commersonii showed stronger activation of the salicylic acid and ethylene marker genes than the moderately resistant cultivar (MST 32/1) and the susceptible lines (Quatre carrées and Spunta). The marker genes reached their highest expression levels earlier (4 h.p.i) in the resistant and moderately resistant lines than in the susceptible lines (48 h.p.i.). These results indicate that salicylic acid and ethylene signaling pathways have a significant role in defense against R. solanacearum. The timing and magnitude of the upregulation of gene expression may determine the plant ability to put up a defense response against the pathogen.     

Pathogenesis-related gene expression in rice is correlated with developmentally controlled Xa21-mediated resistance against Xanthomonas oryzae pv. oryzae

Disease resistance mediated by the resistance gene Xa21 is developmentally controlled in rice. We examined the relationship between Pathogenesis Related (PR) defense gene expression and Xa21-mediated developmental disease resistance induced by Xanthomonas oryzae pv. oryzae (Xoo). OsPR1a, OsPR1b, and OsPR1c genes were cloned and their induction was analyzed, in addition to the OsPR10a gene, at the juvenile and adult stages in response to a wildtype Xoo strain that induces a resistance response (incompatible interaction) and an isogenic mutant Xoo strain that does not (compatible interaction). We found that the adult stage leaves are more competent to express these OsPR1 genes and that the Xa21 locus is required for the highest levels of induction.     

近年水稻主要线虫病害的研究进展

目前,以拟禾谷根结线虫Meloidogyne graminicola、水稻干尖线虫Aphelenchoide besseyi、水稻潜根线虫Hirschmanniella oryzae、水稻茎线虫Ditylenchus angustus和旱稻孢囊线虫Heterodera elachista为主的水稻寄生线虫侵染水稻以及其它禾本科作物引致的线虫病害,已造成了严重的经济损失。近年来,对拟禾谷根结线虫的致病机制和寄主的防御机制等方面的研究已取得了显著进展,但对其它水稻寄生线虫与寄主互作机制的研究还存在较大空白。水稻寄生线虫的防治多依赖化学农药,抗性品种和生物防治等措施的实施存在一定的局限性。本文对上述5种主要水稻寄生线虫病的发生为害、致病机制以及防治方法等进行了综述,并对水稻寄生线虫致病机理的研究以及抗性品种、生物防治和诱导化合物的应用进行了探讨。