Relationships between rhizoplane and rhizosphere bacteria and verticillium wilt resistance in potato

Six cultivars and breeding lines of potato (Solanum tuberosum) differing in susceptibility to verticillium wilt caused by Verticillium dahliae were studied with respect to quantitative and qualitative differences in the bacterial flora of their soil and rhizosphere-rhizoplane. Although, no association was observed between the types of bacteria that inhabited the soil or roots of wilt resistant and susceptible cultivars, quantitative differences were evident. These differences provide the first direct evidence that potato genotypes can influence bacterial populations. Bacterial populations were 9–25-fold higher on roots than in the adjacent soil. As the plants aged, the total number of rootcolonizing bacteria increased between 15 and 245%. Pseudomonas spp. were the most abundant microbes in the soil and rhizosphere-rhizoplane. The bacteria antagonistic to V. dahliae in vitro were identified as members of the genera Bacillus, Pseudomonas, Flavobacterium, and Gluconobacter. A statistically significant trend was evident toward the association of antagonistic bacteria with the more resistant potato cultivars.     

The Gossypium hirsutum TIR-NBS-LRR gene GhDSC1 mediates resistance against Verticillium wilt

Improving genetic resistance is a preferred method to manage Verticillium wilt of cotton and other hosts. Identifying host resistance is difficult because of the dearth of resistance genes against this pathogen. Previously, a novel candidate gene involved in Verticillium wilt resistance was identified by a genome-wide association study using a panel of Gossypium hirsutum accessions. In this study, we cloned the candidate resistance gene from cotton that encodes a protein sharing homology with the TIR-NBS-LRR receptor-like defence protein DSC1 in Arabidopsis thaliana (hereafter named GhDSC1). GhDSC1 expressed at higher levels in response to Verticillium wilt and jasmonic acid (JA) treatment in resistant cotton cultivars as compared to susceptible cultivars and its product was localized to nucleus. The transfer of GhDSC1 to Arabidopsis conferred Verticillium resistance in an A. thaliana dsc1 mutant. This resistance response was associated with reactive oxygen species (ROS) accumulation and increased expression of JA-signalling-related genes. Furthermore, the expression of GhDSC1 in response to Verticillium wilt and JA signalling in A. thaliana displayed expression patterns similar to GhCAMTA3 in cotton under identical conditions, suggesting a coordinated DSC1 and CAMTA3 response in A. thaliana to Verticillium wilt. Analyses of GhDSC1 sequence polymorphism revealed a single nucleotide polymorphism (SNP) difference between resistant and susceptible cotton accessions, within the P-loop motif encoded by GhDSC1. This SNP difference causes ineffective activation of defence response in susceptible cultivars. These results demonstrated that GhDSC1 confers Verticillium resistance in the model plant system of A. thaliana, and therefore represents a suitable candidate for the genetic engineering of Verticillium wilt resistance in cotton.     

Cellophane surface-induced gene,VdCSIN1, regulates hyphopodium formation and pathogenesis via cAMP-mediated signalling in Verticillium dahliae

The soil-borne vascular pathogen Verticillium dahliae infects many dicotyledonous plants to cause devastating wilt diseases. During colonization, V. dahliae spores develop hyphae surrounding the roots. Only a few hyphae that adhere tightly to the root surface form hyphopodia at the infection site, which further differentiate into penetration pegs to facilitate infection. The molecular mechanisms controlling hyphopodium formation in V. dahliae remain unclear. Here, we uncovered a cellophane surface-induced gene (VdCSIN1) as a regulator of V. dahliae hyphopodium formation and pathogenesis. Deletion of VdCSIN1 compromises hyphopodium formation, hyphal development and pathogenesis. Exogenous application of cyclic adenosine monophosphate (cAMP) degradation inhibitor or disruption of the cAMP phosphodiesterase gene (VdPDEH) partially restores hyphopodium formation in the VdΔcsin1 mutant. Moreover, deletion of VdPDEH partially restores the pathogenesis of the VdΔcsin1 mutant. These findings indicate that VdCSIN1 regulates hyphopodium formation via cAMP-mediated signalling to promote host colonization by V. dahliae.     

Reflexions on some aspects of the interactions among ROS,RNS, and Ca2+ in response to a mycorrhizal or a pathogenic fungus

In vivo redox activities in the apoplast of axenically cultured intact seedling roots (superoxide anion generation, and superoxide dismutase and peroxidase activities) in contact with the compatible arbuscular mycorrhizal fungus (AMF) were clearly attenuated in comparison with those in contact with the pathogenic fungus (PF) or treated with MeJA, even at the early stages of treatment. Contact of roots with the AMF did not enhance the biosynthesis of phenolic compounds (total phenolics, flavonoids, and phenylpropanoid glycosides), while contact with the PF significantly enhanced the biosynthesis of all phenolic fractions. Reactive oxygen and nitrogen species both seemed to be involved in these responses from the first moments of contact, but the fluorescence imaging of roots showed that ROS were mainly accumulated in the apoplast while NO was mainly stored in the cytosol. In conclusion, intact olive seedling roots clearly differentiated between AMF and PF.     

蜡蚧轮枝菌毒素Ⅷ对烟粉虱的忌避与拒食活性

蜡蚧轮枝菌毒素Ⅷ和化学合成的吡啶-2,6-二羧酸(DPA)对烟粉虱成虫的忌避作用均表现出忌避与拒食活性。利用罩叶选择法测定了蜡蚧轮枝菌毒素Ⅷ和DPA对烟粉虱成虫的忌避与拒食活性。试验表明蜡蚧轮枝菌毒素Ⅷ和DPA对烟粉虱具忌避与拒食作用活性,100mg·mL-1毒素Ⅷ的忌避率与拒食率分别为41%和23%。     

黄萎病菌胁迫下野生茄子托鲁巴姆防卫反应的生理生化分析

以野生茄子托鲁巴姆(Solanum torvum Swartz)和苏崎茄(Solanum melongena L.)为主要材料,通过比较两种茄子在大丽轮枝菌(Verticillium dahliae Kleb)侵染过程中体内的生理生化指标变化,分析托鲁巴姆对黄萎病的抗性响应机制。结果表明:(1)与苏崎茄植株相比,托鲁巴姆植株表现出很强的自我防御和自我修复能力。(2)托鲁巴姆体内存在的活性氧清除系统(如SOD、POD、CAT等酶的活性)高于苏崎茄;在侵染后,托鲁巴姆体内各种酶的活性快速增加,其幅度高于苏崎茄;MDA的变化则恰恰相反。这个结果提示,黄萎病菌胁迫可能激活了托鲁巴姆体内活性氧清除系统,从而加快了某些防御物质(如木质素和抗菌物质绿原酸等)的形成,同时减缓或降低MDA等有害物质在植株体内的积累。(3)黄萎病菌侵染后各生理指标的响应时间表现出差异。托鲁巴姆中POD、PAL的活性和可溶性蛋白含量在侵染后的12h内就迅速作出响应(POD增加、PAL和可溶性蛋白减少);而SOD、PPO、CAT的活性和MDA的含量则在处理后初始阶段(至少12h)进行了一些调整,随后才进入持续性的增加或减少阶段。由此可见,托鲁巴姆对黄萎病病菌侵染的响应具有时序性,其体内POD酶、PAL酶和可溶性蛋白首先参与植物的防卫反应以应对黄萎病的胁迫,其它酶随后参与响应,它们的共同作用形成对黄萎病菌的有效防卫。     

土壤中棉花黄萎病菌SYBR Green Ⅰ荧光RT-PCR定量检测技术研究

为实现田间土壤棉花黄萎病菌的早期检测,建立了土壤中棉花黄萎病菌的SYBR Green I荧光定量PCR检测方法。以含342bp PCR扩增产物的阳性质粒为参考,构建了标准曲线,并对该曲线的特异性、敏感性、可重复性进行了评价。结果表明,该方法具有快速、特异性强、敏感度高等特点。检测范围在3.8×103-3.8×108copies/μL之间有良好的线性关系,相关系数R2为0.996,扩增效率为101.5%,灵敏度比常规PCR方法高102倍。     

Functional analyses of small secreted cysteine-rich proteins identified candidate effectors in Verticillium dahliae

Secreted small cysteine-rich proteins (SCPs) play a critical role in modulating host immunity in plant–pathogen interactions. Bioinformatic analyses showed that the fungal pathogen Verticillium dahliae encodes more than 100 VdSCPs, but their roles in host–pathogen interactions have not been fully characterized. Transient expression of 123 VdSCP-encoding genes in Nicotiana benthamiana identified three candidate genes involved in host–pathogen interactions. The expression of these three proteins, VdSCP27, VdSCP113, and VdSCP126, in N. benthamiana resulted in cell death accompanied by a reactive oxygen species burst, callose deposition, and induction of defence genes. The three VdSCPs mainly localized to the periphery of the cell. BAK1 and SOBIR1 (associated with receptor-like protein) were required for the immunity triggered by these three VdSCPs in N. benthamiana. Site-directed mutagenesis showed that cysteine residues that form disulphide bonds are essential for the functioning of VdSCP126, but not VdSCP27 and VdSCP113. VdSCP27, VdSCP113, and VdSCP126 individually are not essential for V. dahliae infection of N. benthamiana and Gossypium hirsutum, although there was a significant reduction of virulence on N. benthamiana and G. hirsutum when inoculated with the VdSCP27/VdSCP126 double deletion strain. These results illustrate that the SCPs play a critical role in the V. dahliae–plant interaction via an intrinsic virulence function and suppress immunity following infection.