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Protease inhibitor expression in soybean roots exhibiting susceptible and resistant interactions with soybean cyst nematode.

ABSTRACT: Protease inhibitors play a role in regulating proteases during cellular development and in plant defense. We cloned and sequenced cDNA encoding six protease inhibitors expressed in soybean roots infected with soybean cyst nematode (SCN) and determined their expression patterns. Four of these protease inhibitors are novel and have not been reported previously. Using RT-PCR, we measured the relative transcript levels of each protease inhibitor in roots of the soybean cv. Peking inoculated with either SCN TN8 to examine the expression of protease inhibitors during the susceptible interaction or with SCN NL1-RHg representing the resistant interaction. Within 12 to 24 hours, mRNA transcripts encoding five of the six protease inhibitors were more highly elevated in soybean roots exhibiting the susceptible interaction than the resistant interaction. Transcripts encoding two protease inhibitors possessing Kunitz trypsin inhibitor domains were induced 37- and 27-fold in the susceptible interaction within 1 dpi, but were induced only 5- to 7-fold in roots displaying the resistant interaction. Our results indicate that soybean roots recognize differences between these two SCN populations before the nematodes initiate a feeding site, and accordingly the roots express transcripts encoding soybean protease inhibitors differentially. These transcripts were generally less abundant in roots exhibiting the resistant interaction.

SUBMITTER: Rashed NA

PROVIDER: S-EPMC2586541 | biostudies-literature | 2008 Jun

REPOSITORIES: biostudies-literature

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Protease inhibitor expression in soybean roots exhibiting susceptible and resistant interactions with soybean cyst nematode.

Rashed Nahed A NA Macdonald Margaret H MH Matthews Benjamin F BF

Journal of nematology 20080601 2

Protease inhibitors play a role in regulating proteases during cellular development and in plant defense. We cloned and sequenced cDNA encoding six protease inhibitors expressed in soybean roots infected with soybean cyst nematode (SCN) and determined their expression patterns. Four of these protease inhibitors are novel and have not been reported previously. Using RT-PCR, we measured the relative transcript levels of each protease inhibitor in roots of the soybean cv. Peking inoculated with eit ...[more]

PMID: 19259530

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Project description:Soybean cyst nematode (SCN; Heterodera glycines Ichinohe) reproduces on the roots of common bean (Phaseolus vulgaris L.) and can cause reductions in plant growth and seed yield. The molecular changes in common bean roots caused by SCN infection are unknown. Identification of genetic factors associated with SCN resistance could help in development of improved bean varieties with high SCN resistance. Gene expression profiling was conducted on common bean roots infected by SCN HG type 0 using next generation RNA sequencing technology. Two pinto bean genotypes, PI533561 and GTS-900, resistant and susceptible to SCN infection, respectively, were used as RNA sources eight days post inoculation. Total reads generated ranged between ~ 3.2 and 5.7 million per library and were mapped to the common bean reference genome. Approximately 70-90% of filtered RNA-seq reads uniquely mapped to the reference genome. In the inoculated roots of resistant genotype PI533561, a total of 353 genes were differentially expressed with 154 up-regulated genes and 199 down-regulated genes when compared to the transcriptome of non- inoculated roots. On the other hand, 990 genes were differentially expressed in SCN-inoculated roots of susceptible genotype GTS-900 with 406 up-regulated and 584 down-regulated genes when compared to non-inoculated roots. Genes encoding nucleotide-binding site leucine-rich repeat resistance (NLR) proteins, WRKY transcription factors, pathogenesis-related (PR) proteins and heat shock proteins involved in diverse biological processes were differentially expressed in both resistant and susceptible genotypes. Overall, suppression of the photosystem was observed in both the responses. Furthermore, RNA-seq results were validated through quantitative real time PCR. This is the first report describing genes/transcripts involved in SCN-common bean interaction and the results will have important implications for further characterization of SCN resistance genes in common bean.

Project description:BackgroundSoybean cyst nematode (Heterodera glycines, SCN) is the most economically damaging pathogen of soybean (Glycine max) in the U.S. The Rhg1 locus is repeatedly observed as the quantitative trait locus with the greatest impact on SCN resistance. The Glyma18g02680.1 gene at the Rhg1 locus that encodes an apparent leucine-rich repeat transmembrane receptor-kinase (LRR-kinase) has been proposed to be the SCN resistance gene, but its function has not been confirmed. Generation of fertile transgenic soybean lines is difficult but methods have been published that test SCN resistance in transgenic roots generated with Agrobacterium rhizogenes.ResultsWe report use of artificial microRNA (amiRNA) for gene silencing in soybean, refinements to transgenic root SCN resistance assays, and functional tests of the Rhg1 locus LRR-kinase gene. A nematode demographics assay monitored infecting nematode populations for their progress through developmental stages two weeks after inoculation, as a metric for SCN resistance. Significant differences were observed between resistant and susceptible control genotypes. Introduction of the Rhg1 locus LRR-kinase gene (genomic promoter/coding region/terminator; Peking/PI 437654-derived SCN-resistant source), into rhg1- SCN-susceptible plant lines carrying the resistant-source Rhg4+ locus, provided no significant increases in SCN resistance. Use of amiRNA to reduce expression of the LRR-kinase gene from the Rhg1 locus of Fayette (PI 88788 source of Rhg1) also did not detectably alter resistance to SCN. However, silencing of the LRR-kinase gene did have impacts on root development.ConclusionThe nematode demographics assay can expedite testing of transgenic roots for SCN resistance. amiRNAs and the pSM103 vector that drives interchangeable amiRNA constructs through a soybean polyubiqutin promoter (Gmubi), with an intron-GFP marker for detection of transgenic roots, may have widespread use in legume biology. Studies in which expression of the Rhg1 locus LRR-kinase gene from different resistance sources was either reduced or complemented did not reveal significant impacts on SCN resistance.

Project description:BACKGROUND: Extensive studies using the model system Arabidopsis thaliana to elucidate plant defense signaling and pathway networks indicate that salicylic acid (SA) is the key hormone triggering the plant defense response against biotrophic and hemi-biotrophic pathogens, while jasmonic acid (JA) and derivatives are critical to the defense response against necrotrophic pathogens. Several reports demonstrate that SA limits nematode reproduction. RESULTS: Here we translate knowledge gained from studies using Arabidopsis to soybean. The ability of thirty-one Arabidopsis genes encoding important components of SA and JA synthesis and signaling in conferring resistance to soybean cyst nematode (SCN: Heterodera glycines) are investigated. We demonstrate that overexpression of three of thirty-one Arabidoposis genes in transgenic soybean roots of composite plants decreased the number of cysts formed by SCN to less than 50% of those found on control roots, namely AtNPR1(33%), AtTGA2 (38%), and AtPR-5 (38%). Three additional Arabidopsis genes decreased the number of SCN cysts by 40% or more: AtACBP3 (53% of the control value), AtACD2 (55%), and AtCM-3 (57%). Other genes having less or no effect included AtEDS5 (77%), AtNDR1 (82%), AtEDS1 (107%), and AtPR-1 (80%), as compared to control. Overexpression of AtDND1 greatly increased susceptibility as indicated by a large increase in the number of SCN cysts (175% of control). CONCLUSIONS: Knowledge of the pathogen defense system gained from studies of the model system, Arabidopsis, can be directly translated to soybean through direct overexpression of Arabidopsis genes. When the genes, AtNPR1, AtGA2, and AtPR-5, encoding specific components involved in SA regulation, synthesis, and signaling, are overexpressed in soybean roots, resistance to SCN is enhanced. This demonstrates functional compatibility of some Arabidopsis genes with soybean and identifies genes that may be used to engineer resistance to nematodes.

Dataset Information

Protease inhibitor expression in soybean roots exhibiting susceptible and resistant interactions with soybean cyst nematode.

Publications

Protease inhibitor expression in soybean roots exhibiting susceptible and resistant interactions with soybean cyst nematode.

Similar Datasets

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