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Jujube witches' broom (JWB) is a destructive disease for Chinese jujube caused by phytoplasma. A suppression subtractive hybridization library of resistant cultivar ‘Xingguang’ was constructed under phytoplasma stress to identify genes related to JWB resistance. 77 of 200 unique expressed sequence tags had significant sequence homologies and were classified into 10 functional groups. The most abundant group was disease/defense (20.8%), which was consistent with the phytoplasma stress. These differentially expressed genes provide the groundwork for addressing the plant–phytoplasma interaction. Meanwhile, the expression of five selected genes (TLP, PR10, HSP70, ERF, kinase-related protein) was confirmed to upregulate at different infection periods. 相似文献
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西瓜与枯萎病菌非亲和互作相关基因的分离及表达分析 总被引：1，自引：0，他引：1
从整体水平阐明西瓜对西瓜枯萎病菌1号生理小种的抗病分子机制和抗病相关基因的表达特征,以高抗枯萎病菌1号生理小种的西瓜品种“卡红(Calhoun Gray)”为试材,接种西瓜枯萎病菌和蒸馏水的根尖组织作为测验方(Tester)和驱动方(Driver),构建西瓜枯萎病菌胁迫的SSH-cDNA正向文库。利用反向 Northern 斑点杂交技术对文库中克隆进行杂交筛选。随机测序300个阳性克隆,序列比对分析,利用RT-PCR技术分析抗病相关基因的表达特性。259条EST成功测序,167条与已知基因具有较高的同源性,占全部ESTs的65.5%,其中与抗病和防卫相关的有64条23种,占24.7%;卡红对枯萎病菌1号生理小种的抗性相关基因主要涉及抗病信号传导、抗病防卫、转录因子、次生代谢合成和细胞保护等方面;Aquaporin和Peroxidase基因在接种后表达量均增加。Calhoun Gray对枯萎病菌侵染作出的反应是全方位多方面的,抗病相关基因主要集中在系统获得性抗性反应中,获得了一些Calhoun Gray与野生西瓜PI296341在与枯萎病生理小种1互作中差异表达的基因,为深入研究西瓜与枯萎病菌互作的分子机制以及关键基因的功能分析奠定基础。 相似文献
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In an ongoing effort to investigate the mechanism of auxinic herbicide resistance in Kochia scoparia (kochia), polymerase chain reaction‐based cDNA suppression subtractive hybridization was used to identify genes that are differentially expressed between dicamba‐resistant (HRd) and dicamba‐susceptible (S1) kochia biotypes in response to herbicide treatment. Both the HRd and S1 adaptor‐ligated cDNAs were used in separate hybridizations in order to generate biotype‐specific clones. A total of 710 cDNAs, representing putative differentially expressed mRNAs, were isolated and subjected to further screening. The false‐positive cDNAs were removed by conducting two colony hybridizations and at least one Northern hybridization. Differential or biotype‐specific expression was confirmed for six clones each from the HRd and S1 plants. The S1‐related genes were constitutively expressed at higher levels than in the HRd plants, but none had significant sequence similarity to known genes. Among the HRd‐related genes, HRd‐88 had 42% amino acid sequence identity to a conserved domain within thiol peptidases, which might be involved in auxin‐regulated gene expression. The constitutively expressed and inducible (by the dicamba treatment) HRd‐39 had 40% identity and 60% similarity to a domain from the Fe(II)/α‐ketoglutarate‐dependent hydroxylase superfamily. The HRd‐39 gene product had the characteristics of an enzyme that is able to detoxify dicamba via oxidative hydroxylation and thus its overexpression might confer the dicamba resistance phenotype. 相似文献
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Inducible responses in plants against pathogen attack play a major role in resistance to disease. The defense responses are
mostly associated with the expression of various kinds of inducible genes. We employed differential hybridization to isolate
elicitor-inducible genes (EIGs) of tobacco (Nicotiana tabacum cv. Samsun NN) using the tobacco-fungal elicitor system. A cDNA library was constructed from tobacco leaves treated for 12
hr with hyphal wall components (HWC) prepared from Phytophthora infestans, and six EIGs were identified. Expression of all EIGs was induced after inoculation with the soybean pathogen Pseudomonas syringae pv. glycinea (nonpathogenic on tobacco) or treatment with salicylic acid, and a variety of expression patterns of EIG mRNAs was observed.
Sequence analysis of EIG cDNAs revealed similarities to genes for SAR8.2 (EIG-B39 and EIG-D14), glycine-rich protein (EIG-G7), extensin (EIG-I30), acyltransferase (EIG-I24) and unknown protein (EIG-J7). Possible roles of EIG products in disease resistance are discussed.
Received 30 August 2000/ Accepted in revised form 30 November 2000 相似文献
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A. Aryan G. Brader J. Mörtel M. Pastar M. Riedle-Bauer 《European journal of plant pathology / European Foundation for Plant Pathology》2014,140(2):213-227
Bois noir (BN) associated with ‘Candidatus Phytoplasma solani’ (Stolbur) is regularly found in Austrian vine growing regions. Investigations between 2003 and 2008 indicated sporadic presence of the confirmed disease vector Hyalesthes obsoletus and frequent infections of bindweed and grapevine. Infections of nettles were rare. In contrast present investigations revealed a mass occurrence of H. obsoletus almost exclusively on stinging nettle. The high population densities of H. obsoletus on Urtica dioica were accompanied by frequent occurrence of ‘Ca. P. solani’ in nettles and planthoppers. Sequence analysis of the molecular markers secY, stamp, tuf and vmp1 of stolbur revealed a single genotype named CPsM4_At1 in stinging nettles and more than 64 and 90 % abundance in grapevine and H. obsoletus, respectively. Interestingly, this genotype showed tuf b type restriction pattern previously attributed to bindweed associated ‘Ca. P. solani’ strains, but a different sequence assigned as tuf b2 compared to reference tuf b strains. All other marker genes of CPsM4_At1 clustered with tuf a and nettle derived genotypes verifying distinct nettle phytoplasma genotypes. Transmission experiments with H. obsoletus and Anaceratagallia ribauti resulted in successful transmission of five different strains including the major genotype to Catharanthus roseus and in transmission of the major genotype to U. dioica. Altogether, five nettle and nine bindweed associated genotypes were described. Bindweed types were verified in 34 % of grapevine samples, in few positive Reptalus panzeri, rarely in bindweeds and occasionally in Catharanthus roseus infected by H. obsoletus or A. ribauti. ‘Candidatus Phytoplasma convolvuli‘(bindweed yellows) was ascertained in nettle and bindweed samples. 相似文献
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Thomas K. Walsh 《Pesticide biochemistry and physiology》2007,88(1):26-30
Lepeophtheirus salmonis is a copepod ectoparasite of wild and farmed salmonids and can cause considerable damage to infected fish. We have examined the effect of the organophosphate trichlorphon, which was one of the early treatments for sea lice as Neguvon® on L. salmonis. Suppression subtractive hybridisation was used to prepare a cDNA library enriched for cDNAs expressed in response to the organophosphate trichlorophon and using this technique a total of 33 cDNAs were found to be differentially regulated in response to organophosphate exposure. Sequence analysis of the cDNAs revealed that many were involved in cellular stress responses and or neurotoxicity. The expression of two of these cDNAs was confirmed to be up-regulated in response to trichlorophon. 相似文献
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《Physiological and Molecular Plant Pathology》2006,68(1-3):51-60
In order to understand the defense machinery in the model cereal crop rice, we performed a large-scale analysis of rice gene expression in response to rice blast Magnaporthe grisea (M. grisea) or Magnaporthe oryzae and bacterial blight Xanthomonas oryzae pv. oryzae (Xoo) during the early incompatible and compatible interactions. Using a gene chip containing 10 254 rice cDNAs representing 9240 unique genes, we identified 794 and 612 genes differentially expressed in the incompatible and compatible rice–M. grisea interactions, respectively, with 274 genes co-regulated during both interactions. In the rice–Xoo pathosystem, 454 and 498 differentially expressed genes were identified in the incompatible and compatible interactions, respectively, including 237 co-regulated genes in the both interactions. By clustering differentially regulated genes from all these interactions, we identified 29 co-regulated genes in the all four interactions, and 86 and 74 co-regulated genes in the two incompatible and two compatible interactions, respectively. These differentially expressed genes could be classified into three categories, including M. grisea- and Xoo-regulated, M. grisea-specific, and Xoo-specific. The expression patterns of representative defense-related genes were further confirmed by RT-PCR. The large-scale expression data from our microarray analysis indicated the existence of distinctive as well as shared defense pathways between the rice–M. grisea and rice–Xoo interactions. 相似文献
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Litao Yang Hong Lin Yuri Takahashi Feng Chen M. Andrew Walker Edwin L. Civerolo 《Physiological and Molecular Plant Pathology》2011,75(3):90-99
Xylella fastidiosa (Xf) is the bacterial causal agent of Pierce’s disease (PD) as well as other economically important diseases in a number of agronomic, horticultural and ornamental plants. The objective of this research was to tentatively identify proteins that are differentially expressed in grapevines and involved in disease development or defense responses to Xf-inoculation. We comparatively analyzed proteins differentially expressed in Xf-inoculated grape stems using a pair of siblings of 9621-67 (highly susceptible) and 9621-94 (highly resistant) from a cross of Vitis rupestris × Vitis arizonica. Total proteins were extracted from the stems of uninoculated controls and Xf-inoculated plants at 1, 6, and 12 weeks after inoculation, separated by a 2D-PAGE system, and spots representing differentially expressed proteins were analyzed and tentatively identified using LC/MS/MS. Protein identification was performed using BLASTp and tBLASTn against NCBI non-redundant protein databases and EST databases, respectively. Ten tentatively identified proteins were differentially expressed at different time points after inoculation. A thaumatin-like protein and the pathogenesis-related protein 10 from both genotypes, and the 40S ribosomal protein S25 from the susceptible genotype were up-regulated in response to Xf-inoculation. Furthermore, the expression of the thaumatin-like protein increased sharply 12 weeks post-inoculation in the PD-resistant genotype only. Three heat shock proteins, 17.9 kDa class II, protein 18 and 21 were highly expressed in healthy tissues compared with those in tissues infected with Xf, and heat shock protein 21 was not detectable in the Xf-inoculated PD-susceptible genotype. In addition, a down-regulated putative ripening related protein was found in the Xf-inoculated PD-susceptible genotype. Glycoprotein and formate dehydrogenase were identified in the PD-resistant genotype and their expression was constant during plant development. A putative GTP-binding protein was down-regulated in the PD-susceptible genotype. Our results revealed that differential expression of proteins in response to Xf-inoculation was genotype and tissue development stage dependent. The specific roles of these candidate proteins in alleviation or aggravation of this disease are under investigation. The information obtained in this study will aid in the understanding of the mechanisms related to the host–pathogen interactions involved in PD. 相似文献
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ABSTRACT Magnaporthe grisea (anamorph = Pyricularia grisea) causes blast on rice (Oryza sativa) and gray leaf spot on turfgrass. Gray leaf spot is a serious disease on St. Augustinegrass (Stenotaphrum secundatum), perennial ryegrass (Lolium perenne), and tall fescue (Festuca arundinacea). Virulence assays performed in this study revealed that M. grisea collected from rice could also cause disease on St. Augustinegrass and tall fescue. One rice isolate, Che86061, caused similar disease reactions on susceptible cultivars of rice and St. Augustinegrass and an incompatible interaction on resistant cultivars of both species. To explore whether similar defense-related genes are expressed in rice and St. Augustinegrass, a rice cDNA library was screened using pooled cDNAs derived from M. grisea-infected St. Augustinegrass. Thirty rice EST (expressed sequence tag) clones showing differential expression in St. Augustinegrass following M. grisea inoculation were identified and classified into six putative functional groups. Northern blot analyses of seven EST clones that collectively represented each putative functional group confirmed that the expression of five out of seven EST clones was similar in both rice and St. Augustinegrass. This study represents one of the first attempts to use a broad-scale genomic approach and resources of a model monocot system to study defense gene expression in St. Augustinegrass following M. grisea infection. 相似文献
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为克隆和研究链孢粘帚霉Gliocladium catenulatum寄生核盘菌菌核的相关基因,应用抑制消减杂交技术构建了cDNA消减文库并进行了筛选。通过PCR技术从文库中共筛选到1315个阳性克隆,克隆中插入片段大小主要集中于300～600bp之间。随机挑取120个克隆,经测序和同源性分析,获得60条有效序列,其中部分序列所编码的血红素加氧酶、核糖体蛋白L11、细胞色素P450及热激蛋白等均参与机体对胁迫条件的应答反应。11条序列在NCBI数据库中未找到显著匹配的序列,可能为新基因片段。分别将寄生于核盘菌菌核上的粘帚霉cDNA和粘帚霉与核盘菌纯培养的cDNA混合物经RasⅠ酶切后进行标记作为探针,利用反向Northern杂交技术验证了所选取的25条序列全部为差异表达基因片段。 相似文献
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利用cDNA-AFLP技术, 对甜瓜抗白粉病品种‘云甜-930’在接种Podosphaera xanthii生理小种2F.后的基因表达谱进行分析。256对引物共产生188个具良好多态性的转录本(TDF), 其中109个上调表达, 79个下调表达。经过对差异片段的回收、克隆、测序分析, 最终得到60个EST。Blastx比对和功能分类分析表明, 参与物质合成与代谢的属第一大类, 占48%, 其他主要涉及物质运输(12%)、防御系统(12%)、转录调控(8%)、能量代谢(8%)、信号转导(4%)等, 7条EST(8％) 与未知功能蛋白同源性较高。选取与代谢、抗病防御、信号转导及蛋白转运等相关的4个差异基因TDF12(SEH)、TDF67(SAMDC)、TDF76(CDPK)和TDF82(PDR8)进行qRT-PCR验证, 结果显示其表达模式符合cDNA-AFLP表达谱, 同时表明这些基因可能参与了甜瓜与白粉病菌的互作过程。 相似文献
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Geleta Dugassa Barka Eveline Teixeira Caixeta Robson Ferreira de Almeida Samuel Mazzinghy Alvarenga Laércio Zambolim 《European journal of plant pathology / European Foundation for Plant Pathology》2017,149(3):543-561
Countering the economic hurdle caused by coffee leaf rust disease is most appealing at this time as it has posed a major threat to coffee production around the world. Establishing differential expression profiles at different times following pathogen invasion in both innate and acquired immunities unlocks the molecular components of resistance and susceptibility. Suppression subtractive hybridization (SSH) was used to identify genes differentially over-expressed and repressed during incompatible and compatible interactions between Coffea arabica and Hemileia vastatrix. From 433 clones of expressed sequence tags (ESTs) sequenced, 352 were annotated and categorized of which the proportion of genes expressed during compatible interaction were relatively smaller. The result showed upregulation and downregulation of various genes at 12 and 24 h after pathogen inoculation in both interactions. The use of four different databases in searching for gene homology resulted in different number of similar sequences. BLASTx against EMBL-EBI (European Molecular Biology Laboratory-European Bioinformatics Institute) database being with the maximum (100%) hits for all the annotated sequences. RT-qPCR analysis of seven resistance-signaling genes showed similar expression patterns for most of the genes in both interactions, indicating these genes are involved in basal (non-specific) defense during which immune reactions are similar. Using SSH, we identified different types of resistance related genes that could be used for further studies towards resistant cultivar development. The potential role of some of the resistance related proteins found were also discussed. 相似文献