Plant N-acylethanolamines play a crucial role in defense and its variation in response to elevated CO_(2) and temperature in tomato

The ubiquitous lipid-derived molecules N-acylethanolamines(NAEs)have multiple immune functions in mammals,but their roles and mechanisms in plant defense response during changing environment remain largely unclear.Here,we found that exogenous NAE18:0 and NAE18:2 promoted defense against the necrotrophic pathogen Botrytis cinerea but suppressed defense to the hemi-biotrophic pathogen Pseudomonas syringae pv.tomato(Pst)DC3000 in tomato.The knocking-down and overexpression function analysis of the pathogen-responsive NAE synthetic gene PHOSPHOLIPASE Dγ(PLDγ)and hydrolytic gene FATTY ACID AMID HYDROLASE 1(FAAH1)revealed that the NAE pathway is crucial for plant defense response.Using exogenous applications and SA-abolished NahG plants,we unveiled the antagonistic relationship between NAE and SA in plant defense response.Elevated CO_(2) and temperature significantly changed the NAE pathway in response to pathogens,while inhibition of the NAE pathway led to the alternation of environment-mediated defense variations against Pst DC3000 in tomato,indicating that NAE pathway is associated with plant defense variations in response to elevated CO_(2) and temperature.The results herein reveal a new function of NAE in plant defense,and its involvement in environment-mediated defense variation in tomato.These findings shed light on the NAE-based plant defense,which may have relevance to crop disease management in future changing climate.

A chorismate mutase from Radopholus similis plays an essential role in pathogenicity

In the process of infecting plants, plant parasitic nematodes release a series of proteins that play an essential role in the successful infection and pathogenesis of plant cells and tissues through stylets or body walls. In this study,based on transcriptome data, a chorismate mutase gene of Radopholus similis(RsCM) was identified and cloned,which is a single copy gene specifically expressed in the oesophageal gland and highly expressed in juveniles and females. Transient expression of RsCM in tobacco leaves showed that it was localised in the cytoplasm and nucleus of tobacco leaf cells, which inhibited the pattern-triggered immunity(PTI) induced by flg22, including callose deposition and defence gene expression, and cell death induced by immune elicitors BAX, but could not inhibit cell death induced by immune elicitors Gpa2/RBP-1. The RNA interference(RNAi) transgenic tomato of RsCM obviously inhibited the infection, pathogenicity, and reproduction of R. similis. However, the resistance of the overexpression transgenic tomato of RsCM to R. similis infection was significantly reduced, and the expression levels of two salicylic acid(SA) pathway genes(PR1 and PR5) in roots infected by the nematode were significantly down-regulated,which indicated that RsCM might be involved in the inhibition of SA pathway. The results of this study demonstrate that RsCM suppresses the host immune system and might be a new target for the control of R. similis, which also provides new data for the function and mechanism of CM genes of migratory parasitic plant nematodes.

Effect of plant extracts on activity of some defense enzymes of apple fruit in interaction with Botrytis cinerea

The efficacy of seven plant extracts(neem,fennel,lavender,thyme,pennyroyal,salvia and asafetida) in controlling postharvest of apple(caused by Botrytis cinerea) was evaluated in vitro and in vivo.In vitro,all plant extracts treatments inhibited spore germination.Inhibitory rates of pore germination was 17.41 and 20.83% for neem extract treatment(methanolic and aqueous extracts,respectively) with significant difference compared to control(73.6 and 85.33%) for aqueous and methanol extracts.In the storage conditions,the application of aqueous extract of neem(at concentration of 25%) resulted in 89.11% reduction of disease severity compared with the untreated control.Results of enzymes activity showed the plant extracts can increase the activity of peroxidase,phenylalanine ammonia-lyase,β-1,3-glucanase and polyphenol oxidase in the presence of pathogens,in apple fruits.However,the results of this research revealed that application of neem extracts was more effective than the application of other plant extracts.According to this study,it could be concluded that plant extracts may be useful to control postharvest disease as a safe alternative option to chemical fungicides.

In Vitro Antifungal Activity of the Extract and Compound from Acorus tatarinowii Against Seven Plant Pathogenic Fungi

Acorus tatarinowii Schott is a traditional Chinese medicine plant and has multiple bioactivities in medicine and pesticide field. In this study, the antifungal compound 1,2-dimethoxy-4（2-propenyl） benzene was isolated from A. tatarinowii Schott by activity-directed isolation method, and the inhibitory activity of the extract and 1,2-dimethoxy-4（2-propenyl） benzene against seven plant pathogenic fungi was evaluated. The results showed that the extract and 1,2-dimethoxy-4（2- propenyl） benzene had high inhibitory activity against hyphal growth of Thielaviopsis paradoxa （de Seynes） V. Hohnel, Pestalotia mangiferae P. Henn., Fusarium oxysporum f. sp. niveum （E. F. Smith） Syn. et Hans., Alternaria alternate Tanaka, Colletotrichum musae （Berk et Curt） V. Arx, Sphaceloma fawcettii Jenk., and Mycosphaerella sentina （Fr.） Schroter. The EC50 values of extract were 1.6162, 1.6811, 1.1253, 3.5771, 1.7024, 2.2284, and 2.2221 g L^-1, respectively, and the EC50 values of 1,2-dimethoxy-4（2-propenyl） benzene were 0.1021, 0.0997, 0.0805, 0.1742, 0.1503, 0.1853, and 0.1924 g L^-1, respectively. 1,2-Dimethoxy-4（2-propenyl） benzene also inhibited spores germination of T. paradoxa （de Seynes） V. Hohnel and F. oxysporum f. sp. niveum （E. F. Smith） Syn. et Hans., with the inhibitory rates of 98.81 and 100% at a concentration of 0.4 g L^-1 after 8 h, respectively. 1,2-Dimethoxy-4（2-propenyl） benzene is a potential botanical antifungal agent for controling of plant fungal diseases.

Functional identification of phenazine biosynthesis genes in plant pathogenic bacteria Pseudomonas syringae pv. tomato and Xanthomonas oryzae pv. oryzae

Phenazines are secondary metabolites with broad spectrum antibiotic activity and thus show high potential in biological control of pathogens. In this study, we identified phenazine biosynthesis （phz） genes in two genome-completed plant pathogenic bacteria Pseudomonas syringae pv. tomato （Pst） DC3000 and Xanthomonas oryzae pv. oryzae （Xoo） PXO99A. Unlike the phz genes in typical phenazine-producing pseudomonads, phz homologs in Pst DC3000 and Xoo PXO99A consisted of phzC/D/E/F/G and phzC/E1/E2/F/G, respectively, and the both were not organized into an operon. Detection experiments demonstrated that phenazine-l-carboxylic acid （PCA） of Pst DC3000 accumulated to 13.4 IJg L-1, while that of Xoo PXO99A was almost undetectable. Moreover, Pst DC3000 was resistant to 1 mg mL-1 PCA, while Xoo PXO99A was sensitive to 50 IJg mL ~ PCA. Furthermore, mutation of phzF blocked the PCA production and significantly reduced the pathogenicity of Pst DC3000 in tomato, while the complementary strains restored these phenotypes. These results revealed that Pst DC3000 produces low level of and is resistant to phenazines and thus is unable to be biologically controlled by phenazines. Additionally, phz-mediated PCA production is required for full pathogenicity of Pst DC3000. To our knowledge, this is the first report of PCA production and its function in pathogenicity of a plant pathogenic P. syringae strain.

Preliminary studies on differential defense responses induced during plant communication

We compared the expression patterns of three representative genes in undamaged tomato and tobacco plants in response to exposure to either tomato or tobacco fed on by Helicoverpa armigera (cotton bollworm). When tomato and tobacco, two species of one family, were incubated in the chambers with the tomato plants damaged by the cotton bollworm, the expression of the PR1, BGL2, and PAL genes was up-regulated in leaves of both plants. However, the levels of gene expression were significantly higher in the tomato than that in the tobacco. In addition, the activities of enzymes, peroxidase, polyphenol oxidase, and lipoxygenase were found to be higher in the tomato than those in the tobacco. Similar results were obtained when the damaged plants were replaced by the tobacco.

Pathogenesis-related protein genes involved in race-specific allstage resistance and non-race specific high-temperature adultplant resistance to Puccinia striiformis f. sp. tritici in wheat

Interactions of the stripe rust pathogen （Puccinia striiformis f. sp responses. Among various genes involved in the plant-pathogen related （PR） protein genes determine different defense responses tritici） with wheat plants activate a w^ae range OT nost nteractions, the expressions of particular pathogenesis-Different types of resistance have been recognized and utilized for developing wheat cultivars for resistance to stripe rust. All-stage resistance can be detected in seedling stage and remains at high levels throughout the plant growth stages. This type of resistance is race-specific and not durable. In contrast, plants with only high-temperature adult-plant （HTAP） resistance are susceptible in seedling stage, but become resistant when plants grow older and the weather becomes warmer. HTAP resistance controlled by a single gene is partial, but usually non-race specific and durable. The objective of this study was to analyze the expression of PR protein genes involved in different types of wheat resistance to stripe rust. The expression levels of 8 PR protein genes （PR1, PRI.2, PR2, PR3, PR4, PR5, PR9 and PRIO） were quantitatively evaluated at 0, 1, 2, 7 and 14 days after inoculation in single resistance gene lines of wheat with all-stage resistance genes YrTrl, Yr76, YrSP and YrExp2 and lines carrying HTAP resistance genes Yr52, Yr59, Yr62 and Yr7B. Races PSTv-4 and PSTv-37 for compatible and incompatible interactions were used in evaluation of PR protein gene expression in wheat lines carrying all-stage resistance genes in the seedling- stage experiment while PSTv-37 was used in the HTAP experiment. Analysis of quantitative real-time polymerase chain reaction （qRT-PCR） revealed that all of the PR protein genes were involved in the different types of resistance controlled by different Yr genes. However, these genes were upregulated at different time points and at different levels during the infection process among the wheat lines with different Yr genes for either all-stage resistance or HTAP resistance. Some of the genes were also induced in compatible interactions, but the levels were almost always higher in the incompatible interaction than in the compatible interaction at the same time point for each Yr gene. These results indicate that both salicylic acid and jasmonate signaling pathways are involved in both race-specific all-stage resistance and non-race specific HTAP resistance. Although expressing at different stages of infection and at different levels, these PR protein genes work in concert for contribution to different types of resistance controlled by different Yr genes.

Proteomics: A Successful Approach to Understand the Molecular Mechanism of Plant-Pathogen Interaction

In recent years, proteomics has played a key role in identifying changes in protein levels in plant hosts upon infection by pathogenic organisms and in characterizing cellular and extracellular virulence and pathogenicity factors produced by pathogens. Proteomics offers a constantly evolving set of novel techniques to study all aspects of protein structure and function. Proteomics aims to find out the identity and amount of each and every protein present in a cell and actual function mediating specific cellular processes. Structural proteomics elucidates the development and application of experimental approaches to define the primary, secondary and tertiary structures of proteins, while functional proteomics refers to the development and application of global (proteome wide or system-wide) experimental approaches to assess protein function. A detail understanding of plant defense response using successful combination of proteomic techniques and other high throughput techniques of cell biology, biochemistry as well as genomics is needed for practical application to secure and stabilize yield of many crop plants. This review starts with a brief introduction to gel- and non gel-based proteomic techniques followed by the basics of plant-pathogen interaction, the use of proteomics in recent pasts to decipher the mysteries of plant-pathogen interaction, and ends with the future prospects of this technology.

Antimicrobial Activity of Wild Plant Seed Extracts against Human Bacterial and Plant Fungal Pathogens

Five wild plant species belonging to different families (Chenopodium album, Plantago major, Elytrigia elongata, Filipendula ulmaria and Nigella sativa) widely spread in Russian Federation and the former USSR were evaluated for their ability to inhibit growth of two important human food-borne pathogens (Escherichia coli O157:H7 and Listeria monocytogenes strain EGD-e) and eight plant pathogens (Alternaria alternata, Alternaria tenuissima, Bipolaris sorokiniana, Stagonospora nodorum, Fusarium solani, Fusarium oxysporum, Fusarium culmorum and Phytophtora infestans). To isolate biologically active compounds from seeds, a step-wise procedure including extraction with hexane, ethyl acetate, ethanol, and 10% acetic acid followed by reversed-phase HPLC was developed. Using disc-diffusion assay, the highest activity against E. coli O157:H7 was observed with extracts from F. ulmaria (hexane and ethyl acetate extracts and the unbound RP-HPLC fraction) and P. major (ethyl acetate extract and the unbound RP-HPLC fraction);E. elongate (the unbound RP-HPLC fraction) was less active. The extracts from P. major and E. elongate (the unbound RP-HPLC fractions) were equally highly active against L. monocytogenes, while those of F. ulmaria (the unbound RP-HPLC fraction) and N. sativa (hexane and ethyl acetate extracts) were less active against this pathogen. The dynamics of L. monocytogenes EGD-е and E. coli O157:H7 growth in the presence of two most potent extracts (RP-HPLC-unbound fractions of P. major and E. elongate and the hexane extract of F. ulmaria) was studied.

NBS-LRR Proteins and Their Partners:Molecular Switches of Plant Defense

Specificity of the plant innate immune system is often conferred by resistance（R）proteins.Most plant disease resistance （R）proteins contain a series of leucine-rich repeats（LRRs）,a nucleotide-binding site（NBS）,and a putative amino-terminal signaling domain.They are termed NBS-LRR proteins.The LRRs are mainly involved in recognition,and the amino-terminal domain determines signaling specificity,whereas the NBS domain presumably functions as a molecular switch.During the past years,the most important discoveries are the role of partners in NBS-LRR gene mediated defenses,mounting support for the so-called＂guard hypothesis＂of R gene function,and providing evidence for intramolecular interactions and intermolecular interactions within NBS- LRR proteins as a mode of signaling regulation.The outcome of these interactions determines whether a plant activates its defense responses.

Effectiveness of Ringworm Cassia and Turmeric Plant Extracts on Growth Inhibition against Some Important Plant Pathogenic Fungi

Crude plant extracts of ringworm cassia, Cassia alata L. and turmeric, Curcuma longa L. were prepared by either hot water or organic solvents such as ethanol and ether. Various concentrations of the crude extract were then subjected to an in vitro test for their effectiveness on mycelia growth inhibition against some important plant pathogenic fungi such as Alternaria alternata, Colletotrichum gloeosporioides, Fusarium oxysporum fsp. lycopersici, Sclerotium rolfsii, Phytophthora infestans and Pythium sp. in comparison to commercial fungicides such as copper oxychloride and mancozeb. Reduction of the fungal growth was significantly obtained with C. longa extracts and the best median effective inhibitory concentration (IC50) value of 6.07, 6.50 and 7.13 mg/ml was from the ethanol extract for S. rolfsii, C. gloeosporioides and F. oxysporum fsp. lycopersici respectively. While all extracts from C. alata were almost the least effective against these fungi. The efficacy of C. longa extracts therefore, provided an alternative regime for the control of the fungal diseases and a promising appreciable choice for a replacement of chemical fungicides.

Exogenously applied methyl Jasmonate induces early defense related genes in response to Phytophthora infestans infection in potato plants

In order to elucidate the contributions of JA in orchestrating disease resistance in potato plants,the potato genotype‘SD20’,which exhibits strong resistance against the highly virulent Phytophthora infestans isolate CN152,while infected by the super virulent isolate 2013-18-306,was treated with exogenous JA and then challenged by inoculation with 2013-18-306.The results showed that exogenously applied JA significantly delayed the onset and alleviated the symptoms of late blight,indicating exogenous JA could induce resistance to P.infestans in the early biotrophic stage of infection in‘SD20’plants.To further clarify the role of JA in the early defense response and identify key genes involved in JA signal transduction,gene expression profiling via RNA sequencing(RNA-seq)in‘SD20’plants treated with exogenously applied JA was performed.A total of 2927 differentially expressed genes were specifically induced,the majority encoded transcription factors,protein kinases,secondary metabolites,defense enzymes and disease resistance related proteins.GO functional annotation and KEGG metabolic pathway analysis showed that exogenously applied JA rapidly induced the expression of genes related to immune response regulation,pathogen defense,and other biological processes,and stimulated endogenous JA synthesis and signal transduction,and the overall early pathogen defense response in‘SD20’.These results provide useful information in understanding the JA’s function involved in pathogen defense responses and a theoretical basis for the application of JA in potato production.

Advances in the Role of DNA Methylation and miRNAs in Plant Defense

DNA methylation and miRNAs are two important epigenetic molecular mechmxisms in plants. RNA-directed DNA methylation （RdDM） pathway is con- sidered as the most important mechanism of DNA methylation. A microRNA （miRNA） is processed from a 70-90-nucleotide-long single-stranded RNA precursor that forms a hairpin structure by Dicer enzyme, which plays an important role in plant development, regulation of plant gene expression and silencing of trans- posons. In recent years, a large number of studies have confirmed that DNA methylation and miRNAs play an important role in disease resistance mechanisms of plants. In this paper, recent progress on the role of DNA methylation and miRNAs in plant defense was summarized, which provided reference for revealing the re- lationship between disease resistance factors and epigenctic mechanisms in plants.

Antagonistic Potential of Bacterial Species against Fungal Plant Pathogens(FPP)and Their Role in Plant Growth Promotion(PGP):A Review

Since the 19th century to date,the fungal pathogens have been involved in causing devastating diseases in plants.All types of fungal pathogens have been observed in important agricultural crops that lead to significant pre and postharvest losses.The application of synthetic fungicide against the fungal plant pathogens(FPP)is a traditional management practice but at the same time these fungicides kill other beneficial microbes,insects,animal,and humans and are harmful to environment.The antagonistic microorganism such as bacteria are being used as an alternate strategy to control the FPP.These antagonistic species are cost-effective and eco-friendly in nature.These biocontrol bacteria have a broad mechanism against fungal pathogens present in the phyllosphere and rhizosphere of the plant.The antagonistic bacteria have different strategies against the FPP,by producing siderophore,biofilm,volatile organic compounds(VOCs),through parasitism,antibiosis,competition for limited resources and induce systemic resistance(ISR)in the host plant by activating the immune systems.The commercial bio-products synthesized by the major bacterial species Pseudomonas syringae,Burkholderia cepacia,Streptomyces griseoviridis,Pseudomonas fluorescens and Bacillus subtilis are used to control Fusarium,Pythium,Rhizoctonia,Penicillium,Alternaria,and Geotrichum.The commercial bio-formulations of bacteria act as both antifungal and plant growth regulators.The Plant growth-promoting rhizobacteria(PGPR)played a significant role in improving plant health by nitrogen-fixing,phosphorus solubilization,phytohormones production,minimizing soil metal contamination,and by ACC deaminase antifungal activities.Different articles are available on the specific antifungal activity of bacteria in plant diseases.Therefore,this review article has summarized the information on biocontrol activity of bacteria against the FPP and the role of PGPR in plant growth promotion.This review also provided a complete picture of scattered information regarding antifungal activities of bacteria and the role of PGPR.

Antibacterial activity of plant extracts in different solvents against pathogenic bacteria: An in vitro experiment

Objective:To assess the antibacterial activity of 5 selected plants against 4 pathogenic bacteria.Methods:Three solvents with different polarities were used to extract antimicrobial agents from the plants via maceration technique.The agar-disc diffusion technique was adopted to primarily screen antibacterial activities.Broth-dilution assay was employed to determine the minimum inhibitory concentration(MIC)and the minimum bactericidal concentration(MBC).Results:Among all extracts,the ethanol extract of Piper betle Linn showed the highest antibacterial activity against Gram-positive and the negative bacteria.MIC and MBC of the ethanol extract of Piper betle Linn against Salmonella typhimurium were the same(1?562.50 mg/L);while it showed the highest MIC and MBC against Pseudomonas aeruginosa of 6?250 mg/L and 12?500 mg/L,respectively.Conclusions:Salmonella typhimurium is the most susceptible bacteria while Pseudomonas aeruginosa is the most resistant bacteria towards the ethanol extract of Piper betle Linn.Piper betle possesses compounds with potential antibacterial activity and might be useful as an alternative to control infectious diseases.

Phytochemical Determination and Antibacterial Activity of Punica granatum Peel Extracts against Plant Pathogenic Bacteria

Plant pathogenic bacteria are recognized to be harmful microbes able to decrease the quantity and quality of crop production in the world. Punica granatum peel was screened for its potential use as biological control agent for plant pathogenic bacteria. P. granatum peel was successfully extract using n-hexane, methanol and ethyl acetate by maceration. The highest yield obtained by ethyl acetate showed that ethyl acetate extracted more compounds that readily soluble to methanol and n-hexane. For in-vitro antibacterial activity, three different species of plant pathogenic bacteria were used namely Erwinia carotovorum subsp. Carotovorum, Ralstonia solanacearum, and Xanthomonas gardneri. For all crude extracts, four different concentrations 25, 50, 100 and 200 mg/ml were used in cup-plate agar diffusion method. Streptomycin sulfate at concentration 30 μg/ml was used as positive control while each respective solvent used for peel extraction was used as negative control. The results obtained from in vitro studies showed only ethyl acetate extract possessed antibacterial activity tested on the plant pathogenic bacteria. Methanol and n-hexane did not show any antibacterial activity against plant pathogenic bacteria selected where no inhibition zones were recorded. R. solanacearum recorded the highest diameter of inhibition zones for all range of concentrations introduced followed by E. carotovorum subsp. Carotovorum and X. gardneri. For the minimum inhbitory concentration (MIC) and minimum bactericidal concentration (MBC), only the ethyl acetate extract was subjected to the assay as only ethyl acetate extract exhibited antibacterial activity. The minimum concentration of ethyl acetate extract that was able to inhibit plant pathogenic bacteria was recorded at a concentration of 3.12 mg/ml which inhibited R. solancearum and E. carotovorum subsp. Carotovorum, followed by X. gardneri at concentration 6.25 mg/ml. For the minimum bactericidal concentration (MBC), the results showed that at the concentration of 12.5 mg/ml, the extract was still capable of killing the pathogenic bacteria, R. solanacearum, and P. caratovora sub.sp. caratovora while for the bacteria X. gardneri, the concentration that was able to kill the bacteria was 25 mg/ml. The qualitative estimation of phytochemical constituents within P. granatum L. ethyl acetate peel extracts had revealed the presence of tannins, flavonoids, phenols alkaloid, Saponins, and terpenoids. This study has demonstrated that Ethyl Acetate peel extracts of P. granatum has significant antibacterial activity against pathogenic plant bacterial, and it could be of high agricultural value.

Viral pathogens and plant parasitic nematode

This special focus is dedicated to three parts：i）One of the most ubiquitous viral pathogens of stone fruit tree,Plum pox virus（PPV）;ii）a re-emerging pathogen,Wheat streak mosaic virus（WSMV）of cereal crops in Central Europe;and iii）a less studied plant parasitic,cyst-forming nematode in cereal crops Heterodera avenae.

Characterization of volatile organic compounds in grafted tomato plants upon potyvirus necrotic infection

A headspace solid-phase microextraction-gas chromatography-mass spectrometry(HS-SPME/GC-MS)method was used to study the volatile organic compounds(VOCs)associated with the differential immune response of tomato plants infected with the recombinant strain of potato virus Y(PVY^(C)-to),necrogenic to tomato.Analysis was carried out in UC82(UC),a virus susceptible tomato variety,comparing the same UC plants grafted or not onto a virus tolerant tomato ecotype,Manduria(Ma);the three types of samples used for the GC-MS analysis were mock-inoculated UC/Ma plants,UC/Ma+PVY^(C)-to and UC+PVY^(C)-to plants;the VOCs obtained were 111.Results from symptomatic PVY^(C)-to-infected UC plants showed a VOCs composition enriched in alcohols,fatty acid derivates,benzenoids,and salicylic acid derivatives,while in mock-inoculated UC/Ma plants VOCs were mainly characterized by methyl ester compounds.The VOC profile was in line with RNAseq data analyses,denoting that PVY^(C)-to viral RNA accumulation and disease symptoms induce the specific transcriptional activation of genes involved in VOCs biosynthesis.Furthermore,principal component analysis highlighted that VOCs of PVY^(C)-to-infected and mock-inoculated grafted plants were much closer each other than that of symptomatic PVY^(C)-to-infected non-grafted UC plants.These results suggest that VOCs profiles of tomato plants are related to the viral RNA accumulation,disease intensity and graft-derived tolerance to PVY^(C)-to infection.

A Meloidogyne incognita effector Minc03329 suppresses plant immunity and promotes parasitism

Meloidogyne incognita is a devastating plant-parasitic nematode.Effectors play important roles during the stages of nematodes infection and parasitism,but their molecular functions remain largely unknown.In this study,we characterized a new effector,Minc03329,which contains signal peptide for secretion and a C-type lectin domain.The yeast signal sequence trap experiments indicated that the signal peptide of Minc03329 is functional.In situ hybridization showed that Minc03329 was specifically expressed in the subventral esophageal gland.Real-time qPCR confirmed that the expression level of Minc03329 transcript was significantly increased in pre-parasitic and parasitic second-stage juveniles(pre-J2s and par-J2s).Tobacco rattle virus(TRV)-mediated gene silencing of Minc03329 in host plants largely reduced the pathogenicity of nematodes.On the contrary,ectopic expression of Minc03329 in Arabidopsis thaliana significantly increased plant susceptibility to nematodes.Transient expression of Minc03329 in Nicotiana benthamiana leaves suppressed the programmed cell death triggered by the pro-apoptotic protein BAX.Moreover,the transcriptome analysis of Minc03329-transgenic Arabidopsis and wild type revealed that many defense-related genes were significantly down-regulated.Interestingly,some different expressed genes were involved in the formation of nematode feeding sites.These results revealed that Minc03329 is an important effector for M.incognita,suppressing host defense response and promoting pathogenicity.

Gene Expression:A Review on Methods for the Study of Defense-Related Gene Differential Expression in Plants

The plant genes involved in cellular signaling and metabolism have not been fully identified, while the function(s) of many of those which have are as yet incompletely characterized. Gene expression analysis allows the identification of genes and the study of their relationship with cellular processes. There are several options available for studying gene expression, including the use of cDNA and microarray libraries and techniques such as suppression subtractive hybridization (SSH), differential display (DD), RNA fingerprinting by arbitrary primed PCR (RAP), expressed sequence tags (EST), serial analysis of gene expression (SAGE), representational difference analysis (RDA), cDNA-amplified fragment length polymorphism (cDNA-AFLP) and RNA sequencing (RNA-Seq). Focusing on defense-related processes in plants, we present a brief review and examples of each of these methodologies and their advantages and limitations regarding the study of plant gene expression.