Time-series expression profiling of sugarcane leaves infected withPuccinia kuehniireveals an ineffective defense system leading to susceptibility (original) (raw)
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Plant cell reports, 2012
Brown rust caused by the fungus Puccinia melanocephala is a major disease of sugarcane (Saccharum spp.). A sugarcane mutant, obtained by chemical mutagenesis of the susceptible variety B4362, showed a post-haustorial hypersensitive response (HR)-mediated resistance to the pathogen and was used to identify genes differentially expressed in response to P. melanocephala via suppression subtractive hybridization (SSH). Tester cDNA was derived from the brown rust-resistant mutant after inoculation with P. melanocephala, while driver cDNAs were obtained from the non-inoculated resistant mutant and the inoculated susceptible donor variety B4362. Database comparisons of the sequences of the SSH recombinant clones revealed that, of a subset of 89 non-redundant sequences, 88% had similarity to known functional genes, while 12% were of unknown function. Thirteen genes were selected for transcript profiling in the resistant mutant and the susceptible donor variety. Genes involved in glycolysis ...
Journal of Genetics and Genomics, 2011
Puccinia striiformis f. sp. tritici (Pst) causes stripe rust, one of the most important diseases of wheat worldwide. cDNA libraries had been constructed from urediniospores, germinated urediniospores and haustoria. However, little is known about the expression patterns of the genes related to the infection process and sporulation of the pathogen. In this study, a custom oligonucleotide microarray was constructed using sequences of 442 gene transcripts selected from Pst cDNA libraries. The expression patterns of the genes were determined by hybridizing the microarray with cDNA from Pst in vitro and Pst-infected wheat leaves. The time course study identified 55 transcripts that were differentially expressed during the infection process in a compatible interaction. They were identified to have functions related to the following biological processes, including carbohydrate and lipid metabolism, energy, cell signaling, protein synthesis, cell structure and division. In an incompatible interaction, 17 transcripts of the pathogen were differentially expressed in resistant wheat leaves inoculated with an avirulent Pst race, ten of which had similar expression patterns to those in the compatible interaction. Several candidates for pathogenicity and virulence/avirulence related genes were also identified. The results of quantitative real-time PCR validated the expression patterns of some selected genes. The study demonstrates that the custom oligonucleotide microarray technology is useful to determine the expression patterns of the pathogen genes involved in different types of the hostepathogen interactions and stages of development.
Frontiers in Plant Science
Sugarcane smut caused by the basidiomycetous fungus Sporisorium scitamineum is one of the most devastating diseases that affect sugarcane production, globally. At present, the most practical and effective management strategy for the disease is the cultivation of resistant cultivars. In this connection, a detailed understanding of the host’s defense mechanism in response to smut isolates with varying degrees of virulence at the molecular level would facilitate the development of reliable and durable smut-resistant sugarcane varieties. Hence, in this study, a comparative whole transcriptome analysis was performed employing Illumina RNA-seq in the smut susceptible cultivar Co 97009 inoculated with two distinct S. scitamineum isolates, Ss97009 (high-virulent) and SsV89101 (low-virulent) during the early phases of infection (2 dpi and 5 dpi) and at the phase of sporogenesis (whip emergence) (60 dpi). Though the differential gene expression profiling identified significant transcriptional...
Sugarcane transcriptome analysis in response to infection caused by Acidovorax avenae subsp. avenae
PloS one, 2016
Sugarcane is an important tropical crop mainly cultivated to produce ethanol and sugar. Crop productivity is negatively affected by Acidovorax avenae subsp avenae (Aaa), which causes the red stripe disease. Little is known about the molecular mechanisms triggered in response to the infection. We have investigated the molecular mechanism activated in sugarcane using a RNA-seq approach. We have produced a de novo transcriptome assembly (TR7) from sugarcane RNA-seq libraries submitted to drought and infection with Aaa. Together, these libraries present 247 million of raw reads and resulted in 168,767 reference transcripts. Mapping in TR7 of reads obtained from infected libraries, revealed 798 differentially expressed transcripts, of which 723 were annotated, corresponding to 467 genes. GO and KEGG enrichment analysis showed that several metabolic pathways, such as code for proteins response to stress, metabolism of carbohydrates, processes of transcription and translation of proteins, ...
Front. Plant Sci., 2014
Wheat leaf rust, caused by the basidiomycete Puccinia triticina, can cause yield losses of up to 20% in wheat producing regions. During infection, the fungus forms haustoria that secrete proteins into the plant cell and effect changes in plant transcription, metabolism, and defense. It is hypothesized that new races emerge as a result of overcoming plant resistance via changes in the secreted effector proteins. To understand gene expression during infection and find genetic differences associated with races, RNA from wheat leaves infected with six different rust races, at 6 days post inoculation, was sequenced using Illumina. As P. triticina is an obligate biotroph, RNA from both the host and fungi were present and separated by alignment to the P. triticina genome and a wheat EST reference. A total of 222,571 rust contigs were assembled from 165 million reads. An examination of the resulting contigs revealed 532 predicted secreted proteins among the transcripts. Of these, 456 were found in all races. Fifteen genes were found with amino acid changes, corresponding to putative avirulence effectors potentially recognized by 11 different leaf rust resistance (Lr) genes. Twelve of the potential avirulence effectors have no homology to known genes. One gene had significant similarity to cerato-platanin, a known fungal elicitor, and another showed similarity to fungal tyrosinase, an enzyme involved in melanin synthesis. Temporal expression profiles were developed for these genes by qRT-PCR and show that the genes expression patterns were consistent between races from infection initiation to just prior to spore eruption.
Red rot, a stalk disease in sugarcane caused by Colletotrichum falcatum an ascomycete fungus is a serious production constraint in many Asian countries. However, very limited studies at molecular level exist of the mechanisms related to interaction between sugarcane and the fungal pathogen C. falcatum (Cf). In the conventional system of pathogen inoculation, disease development is influenced by prevailing environmental conditions in the field. Hence an attempt was made to standardize an in vitro system of using sugarcane suspension cells and crude elicitor of Cf for transcriptome analysis and identifying defense related genes. Suspension cells of sugarcane cv Co 93009 was treated with Cf-elicitor at 60 glucose equivalents and transcriptome profile was monitored 30 min and 3 h later by differential display RT-PCR. From the experiment 241 transcripts were found to be differentially expressed and finally 75 of them were cloned and sequenced. Among the up-regulated transcripts, about 37 % were found to be defense related and which was followed by transcription and post transcription (13 %), general metabolism (11 %), transport (9 %), cell structure/ growth/division (9 %) and signal transduction (5 %). The down regulated transcript group constituted *27 % of the differentially expressed transcripts and the grouping pattern was different. Overall, the results revealed up regulation of many potential defense related transcripts like putative chitinase, glycine rich protein, 14-3-3-like protein, xylanase inhibitor protein, calmodulin related protein, Myb-related transcription factor CBM2-like, basal layer antifungal peptide etc. Further by adopting RACE-PCR approach, complete gene sequences of 14-3-3-like protein and xylanase inhibitor were identified and the genes were characterized to domain level. Our results demonstrate that the transcript profile in in vitro system of sugarcane suspension cells and Cf-elicitor is close to the cane tissue challenged with the pathogen and useful to identify defense related traits in sugarcane against Cf.
International Journal of Engineering Research and Technology (IJERT), 2014
https://www.ijert.org/statistical-analysis-of-microarray-data-to-elucidate-the-differential-gene-expression-of-puccinia-striiformis-f.sp.-tritici https://www.ijert.org/research/statistical-analysis-of-microarray-data-to-elucidate-the-differential-gene-expression-of-puccinia-striiformis-f.sp.-tritici-IJERTV3IS080662.pdf The Yellow rust of wheat (Puccinia striiformis f.sp. tritici), is one of the most destructive diseases causing extensive yield loss throughout the world. The present study deals with transcript profiling using Affymetrix Wheat Genome Array GeneChip. Molecular function enrichment analysis suggested that the differentially regulated genes were mainly related to protein degradation and modification, cell signaling and stress related mechanisms. The knowledge and comprehension of currently applied methods is one of the central criteria for a successful work. This study would be helpful in identification of early induced genes in wheat pathogens by the information of the resistance genes.
Functional Plant Biology, 2007
Eucalyptus grandis Hill ex Maiden and its hybrids are commonly planted by the Brazilian pulp and paper industry, but they are the most susceptible to the neotropical rust disease caused by Puccinia psidii Winter. In an initial attempt to understand the mechanisms of resistance, we constructed two contrasting Serial Analysis of Gene Expression (SAGE) libraries using susceptible and resistant individuals from a segregating half-sibling E. grandis population. Using the Z-test we identified tags differentially expressed between the libraries, preferentially 239 in the susceptible and 232 in the resistant type individuals. Using public (Expressed Sequence Tags) EST databases, 40 of the susceptible and 70 of the resistant tags matched ESTs and were annotated. By comparing the type of genes and their expression levels, distinct differences between the libraries were observed. Susceptible plants showed gene expression linked to leaf senescence, generalised stress responses and detoxification, and are apparently incapable of inducing a competent host defence response. On the other hand, resistant plants showed genes upregulated for cellular polarisation, cytoskeleton restructuring, vesicle transport, and cellulose and lignin biosynthesis. In the resistant individuals, evidence for systemic resistance, anti-oxidative responses and a hypersensitive response was also observed, although no R gene was identified.
International Journal of Plant Genomics, 2007
In this study, we detail the construction of a custom cDNA spotted microarray containing 7728 wheat ESTs and the use of the array to identify host genes that are differentially expressed upon challenges with leaf rust fungal pathogens. Wheat cultivar RL6003 (Thatcher Lr1) was inoculated with Puccinia triticina virulence phenotypes BBB (incompatible) or TJB (7-2) (compatible) and sampled at four different time points (3, 6, 12, and 24 hours) after inoculation. Transcript expression levels relative to a mock treatment were measured. One hundred ninety two genes were found to have significantly altered expression between the compatible and incompatible reactions. Among those were genes involved in photosynthesis, the production of reactive oxygen species, ubiquitination, signal transduction, as well as in the shikimate/phenylpropanoid pathway. These data indicate that various metabolic pathways are affected, some of which might be used by RL6003 to mount a coordinated defense against an incompatible fungal pathogen.
Applied Biochemistry and Biotechnology, 2013
Red rot is a serious disease of sugarcane caused by the fungus Colletotrichum falcatum imposing a considerable economic loss annually in all sugarcane-producing countries. In this study, we analyzed the early resistance response of sugarcane to red rot fungus by comparing the differences between control and inoculated stalk tissues. Differential display reverse transcription polymerase chain reaction (DD-RT-PCR) was employed to identify altered expression of genes in disease-resistant cv Co 93009, in response to pathogen infection. DD-RT-PCR identified 300 differentially expressed transcripts of which 112 were selected for further analysis. Cloning and sequence analysis of the isolated cDNA fragments resulted in functional categorization of these clones into five categories, of which the defense/stress/signaling group was the largest, with clones homologous to genes known to be actively involved in various pathogenesis-related functions in plant species. This group showed overexpression of several transcripts related to ethylene-mediated and jasmonic acid pathway of plant defense mechanisms. Of the 112 expressed sequence tags, validation of expression was carried out for five important genes whose role in plant defense mechanisms is well established. This is the first report of Colletotrichum-mediated gene regulation in sugarcane which has provided a set of candidate genes for detailed molecular dissection of signaling and defense responses in tropical sugarcane during the onset of red rot resistance.