Abstract:Arabidopsis thaliana (Arabidopsis) Col-0 was inoculated with Phytophthora cinnamomi to assess the interaction and defence responses involved. Pathogen ingress and asexual reproduction occurred on root tissue but not leaf tissue. The colonisation of root tissue did not cause disease symptoms or plant death, indicating that Arabidopsis Col-0 was tolerant of the infection. The induction of several plant defence responses including the expression of defence-related genes were found, with differences displayed between inoculated root and leaf tissue. Arabidopsis defence-related gene mutant/over-expressing lines were also inoculated with P. cinnamomi but none of the lines tested exhibited a marked increase in susceptibility to the pathogen.
Abstract:Theobroma cacao, is a tropical understorey tree that is a major economic resource to several tropical countries. However, the crop is under increased threat from several diseases that are responsible for 30% loss of harvest globally. Although QTL data related to the genetic determinism of disease resistance exist in cocoa, QTL mapping experiments are heterogeneous, thus making comparative QTL mapping essential for marker assisted selection (MAS). Sixteen QTL experiments were analysed, and the 76 QTLs detected were projected on a progressively established consensus map. Several hot spots, with QTLs related to different Phytophthora species and other diseases, were observed. The likely number of real QTLs was estimated by using a meta-analysis implemented in BioMercator software. There was a twofold reduction in average confidence interval observed when compared to the confidence interval of individual QTLs. This alternative approach confirms the existence of several sources of resistance to different diseases of cocoa which could be cumulated in new varieties to increase the sustainability of cocoa resistance using MAS strategies.
Abstract: To test the hypothesis that resistance in Phytophthora cinnamomi to control by the fungicide phosphite (phosphonate) would arise in sites with prolonged use of phosphite, 30 P. cinnamomi isolates were collected from a range of sites with different phosphite-use histories, including phosphite-treated and untreated avocado orchards, and phosphite-treated and untreated native vegetation sites. The colonizing ability of these isolates was tested by different inoculation methods against a range of host tissues, treated and untreated with phosphite, including mycelial stem inoculation on clonally
propagated Leucadendron sp., mycelial root inoculation of lupin seedlings and zoospore inoculation of Eucalyptus sieberi cotyledons. Isolates from avocado orchards with a long history of phosphite use were, on average, more extensive colonizers of the phosphite-treated Leucadendron sp., lupin seedling roots and Eucalyptus sieberi cotyledons. These isolates did not colonize untreated plant tissue (Leucadendron sp.) more extensively than isolates from sites with no history of phosphite use and no isolates were resistant to control by phosphite. Analysis of all isolates with microsatellite markers revealed the majority were from a single clonal lineage. Selection for decreased sensitivity to phosphite in planta has taken place within asexual clonal lineages of P. cinnamomi in sites with prolonged use of phosphite.
Abstract:Phytophthora infestans, the causal agent of late blight is the most devastating pathogen of cultivated potato worldwide. Utilizing map based cloning; a genomic region containing a cluster of six nucleotide binding site-leucine-rich repeat resistance gene analogs was isolated from the wild potato species Solanum bulbocastanum. Four genes were pseudogenes, with coding sequences interrupted by either frame shift mutations or premature stop codons. However, neither of the two uninterrupted genes conferred resistance to
P. infestans when introduced into susceptible potatoes. Specific primers for one of the pseudogenes were used to amplify an uninterrupted cDNA from P. infestans-infected S. bulbocastanum leaves. A corresponding gDNA was amplified from a late blight-resistant bulbocastanum–tuberosum
introgression line (Rpi-bt1). The Rpi-bt1 gene under transcriptional control of the constitutive potato Ubi3 promoter was found to confer resistance to P. infestans in several transgenic potato lines in a whole plant greenhouse assay.
Abstract: McCHIT1 chitinase (DQ407723), a class I secretory endochitinase from bitter melon (Momordica charantia), had been demonstrated to enhance resistance against Phytophthora nicotianae and Verticillium wilt in transgenic tobacco and cotton. In order to obtain disease-resistant transgenic rice, McCHIT1 was transformed into a restorer line JinHui35 (Oryza sativa subsp. indica) by using the herbicide-resistance gene Bar as the selection marker. Transgenic rice lines and their progenies overexpressing the McCHIT1 gene showed enhanced resistance to Magnaporthe grisea
(rice blast) and Rhizoctonia solani (sheath blight), two major fungal pathogens of rice. McCHIT1-transgenic rice confirmed the inheritance of the transgene and disease resistance to the subsequent generation. The T2
transformants exhibited significantly increased tolerance to M. grisea, with a 30.0 to 85.7 reduction in disease index, and R. solani, with a 25.0 to 43.0 reduction in disease index, based on that of the control as 100. These results indicated that over-expression of the McCHIT1 gene could lead to partial disease reduction against these two important pathogens in
transgenic rice.
Abstract: This paper gives an overview of the activities in Sub-project 4 of the BIOEXPLOIT programme. The European genebank collections of potato need to be narrowed down into (customized) core collections by focusing on loci associated with disease resistance. Therefore, new and more efficient molecular methods are being developed to identify these loci, extensive genotyping of resistance to
Phytophthora infestans and signalling loci is being performed, and genotyped selections of accessions are phenotypically analysed for their resistance specificities. The results of these activities are being assembled in an integrated database.
Abstract: For breeding potato varieties resistant to late blight, identification of resistance genes to Phytophthora infestans (Rpi genes) is essential. Introduction of Rpi genes from wild Solanum species
into cultivated potato is likely to be a good method to achieve durable resistance to P. infestans. In this study, we identified two Rpi genes (Rpi-ber1 and Rpi-ber2) derived from two different accessions of Solanum berthaultii. These two genes are closely linked on the long arm of chromosome 10. There are similarities between the predicted genetic locations of the previously identified Rpi-ber and Rpi-ber1, which given the common origin of these genes, may indicate that they are the same. However, the genetic positions of Rpi-ber1 and Rpi-ber2 are different. Rpi-ber1 is positioned between CT214 and TG63, whereas Rpi-ber2 is located below both of these two markers. In addition, the sequences of four linked markers to both R genes showed different polymorphisms indicating the two Rpi genes could be transmitted from different haplotypes (chromosomes).
Abstract: Plant recognition of elicitors derived from pathogens induces various resistant reactions, including production of reactive oxygen species, hypersensitive cell
death and accumulation of phytoalexins. Previously, we isolated a ceramide elicitor from Phytophthora infestans, which activates O2- production of potato suspension-cultured cells. In this study, we employed nine ceramide-related chemicals to test their elicitor activity. Although, none of the tested chemicals induced O2- production, N,N-dimethylsphingosine (DMS) induced accumulation of phytoalexin in potato tubers. In potato, tobacco and Nicotiana benthamiana, DMS also induced rapid cell death. DMS-treated potato cells stained with 40,6-diamidino-2-phenylindole (DAPI) showed chromatin condensation, and isolated DNA from DMS-treated cells had ladder pattern, confirming that DMS-induced plant cell death is a hypersensitive reaction-like programmed cell death. Involvement of ceramide signaling in induction of plant defense reactions is discussed.
Abstract: Sterol acquisition by soilborne plant pathogens of the genus Phytophthora is presumed to involve extracellular proteins belonging to class-I elicitins. However, little is known about the relationship between sterol availability and elicitin secretion. The objective of this study was to determine the expression of class-I elicitin genes in Phytophthora sojae when grown in a medium containing stigmasterol or cholesterol. P. sojae growth was stimulated by nanomolar concentrations of stigmasterol and cholesterol, which also resulted in the down-regulation of its elicitin genes over time when expression profiles were monitored using real time Reverse Transcription Polymerase Chain Reaction (RT-PCR). The down-regulation of elicitin genes in response to the two sterols also coincided with a reduction in the amount of elicitins detected in spent filtrates. Our study is the first to show the influence of sterols on elicitin gene expression in Phytophthora, which is important with respect to the ecology of elicitin secretion as sterol carrier proteins in the environment.
Abstract: Late blight, caused by the oomycete pathogen, Phytophthora infestans, is a devastating disease of potatoes and tomatoes. A key long-term management strategy for combating this disease is to develop potato cultivars with broad-spectrum and durable resistance through identification and integration of major resistance genes. In support of previous results, we have determined that some accessions of the wild potato species Solanum verrucosum contain strong foliar resistance to P. infestans while others are susceptible. In addition, whole- and wounded-tuber inoculations revealed that most accessions lacked resistance to tuber infection. However, one accession (PI 570643) contained little foliar resistance but expressed tuber resistance, suggesting that expression of these two phenotypes in this species is not correlated.
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