Comparison of the effects of salicylic acid and ethephon with virus-induced hypersensitivity and acquired resistance in tobacco

The induction of a hypersensitive reaction in Samsun NN tobacco by tobacco mosaic virus (TMV) at 20°C leads to the development of both localized and systemic acquired resistance, and is associated with the appearance of pathogenesis-related proteins (PR's) and large increases in peroxidase activity and ethylene production. Salicylic acid (SA) induced a similar resistance in treated plant parts and occasionally also in untreated upper leaves of plants of which three lower leaves had been injected. SA also induced the same four PR's, but these were confined to the treated leaves. Thus, the connection between the presence of PR's and the reduction of TMV multiplication and spread may not be direct.In contrast to TMV, SA did not stimulate ethylene production and hardly increased peroxidase activity. Induction of acquired resistance and PR's by SA developed equally well at 20°C and at 32°C. However, pricking leaves with needles moistened with the ethylene-releasing compound ethephon mimicked TMV infection in inducing acquired resistance and PR's in both treated and untreated leaves at 20°C, but not at 32°C. Ethephon increased peroxidase activity at both temperatures, but only at 20°C dit it induce changes in both the anodic and the cathodic isoenzymes that were similar to those induced as a result of TMV infection. SA induced PR's and reduced TMV multiplication in Samsun tobacco, and inhibited virus spread in Samsun NN at 32°C.These observation indicate that neither the induction of PR's, nor the development of acquired resistance is temperature-sensitive. On the other hand, the effects of ethephon are temperature-sensitive in the same way as the hypersensitive response to TMV. It can thus be hypothesized that ethylene, produced naturally during the hypersensitive reaction of tobacco to TMV, leads to the temperature-sensitive synthesis or release of a presumably benzoic acid-type compound that functions as the natural inducer of PR's and acquired resistance. Although vanillic acid has been shown to accumulate in hypersensitively reacting tobacco leaves, it produced none of the effects of SA, and thus cannot be the natural inducer.Samenvatting Inductie van een overgevoeligheidsreactie in Samsun NN-tabak door tabaksmozaïekvirus (TMV) bij 20°C leidt tot de ontwikkeling van een verworven resistentie die zowel lokaal als systemisch werkzaam is, en gaat samen met het verschijnen van pathogenesis-related proteins (PR's) en sterke toename in de activitieit van peroxidase en de produktie van ethyleen. Salicylzuur (SA) induceerde een vergelijkbare resistentie in behandelde plantedelen en af en toe ook in niet behandelde bovenbladeren van planten waarvan drie onderbladeren waren ingespoten. SA induceerde ook dezelfde vier PR's, maar deze waren beperkt tot de behandelde bladeren. Er bestaat dus geen directe samenhang tussen de aanwezigheid van PR's en de remming van de vermeerdering en uitbreiding van TMV in de plant.In tegenstelling tot TMV stimuleerde SA de ethyleenproduktie niet en verhoogde het de peroxidaseactiviteit nauwelijks. Inductie van verworven resistentie en PR's door SA trad even goed op bij 32°C als bij 20°C. Net als infectie met TMV leidde aanprikken van bladeren met naalden die gedoopt waren in een oplossing van ethefon — waaruit in het blad ethyleen vrijkomt — echter tot inductie van verworven resistentie en PR's in zowel behandelde als onbehandelde bladeren bij 20°C, maar niet bij 32°C. Ethefon verhoogde de peroxidaseactiviteit bij beide temperaturen, maar alleen bij 20°C induceerde het veranderingen in zowel de anodische als de kathodische isoënzymen die vergelijkbaar waren met die welke geïnduceerd werden als gevolg van infectie met TMV. SA induceerde PR's en verminderde de vermenigvuldiging van TMV in Samsun tabak, en remde de uitbreiding van het virus in Samsun NN bij 32°C.Deze waarnemingen tonen dat noch de inductie van PR's, noch de ontwikkeling van verworven resistentie een temperatuurgevoelig proces is. Daarentegen zijn de effecten van ethefon op dezelfde wijze temperatuurgevoelig als de overgevoeligheidsreactie op TMV. Men kan daarom veronderstellen dat ethyleen, dat op natuurlijke wijze geproduceerd wordt tijdens de overgevoeligheidsreactie van tabak op TMV, aanleiding geeft tot een temperatuurgevoelig proces, namelijk de synthese of het vrijkomen van een verbinding, vermoedelijk een benzoëzuurderivaat, dat fungeert als de natuurlijke inductor van PR's en verworven resistentie. Hoewel is aangetoond dat vanillinezuur zich ophoopt in overgevoelig reagerende tabaksbladeren, veroorzaakte deze verbinding geen enkel van de effecten van SA. Vanillinezuur kan dus niet de natuurlijke inductor zijn.     

Systemic acquired resistance delays race shifts to major resistance genes in bell pepper

ABSTRACT The lack of durability of host plant disease resistance is a major problem in disease control. Genotype-specific resistance that involves major resistance (R) genes is especially prone to failure. The compatible (i.e., disease) host-pathogen interaction with systemic acquired resistance (SAR) has been studied extensively, but the incompatible (i.e., resistant) interaction less so. Using the pepper-bacterial spot (causal agent, Xanthomonas axonopodis pv. vesicatoria) pathosystem, we examined the effect of SAR in reducing the occurrence of race-change mutants that defeat R genes in laboratory, greenhouse, and field experiments. Pepper plants carrying one or more R genes were sprayed with the plant defense activator acibenzolar-S-methyl (ASM) and challenged with incompatible strains of the pathogen. In the greenhouse, disease lesions first were observed 3 weeks after inoculation. ASM-treated plants carrying a major R gene had significantly fewer lesions caused by both the incompatible (i.e., hypersensitive) and compatible (i.e., disease) responses than occurred on nonsprayed plants. Bacteria isolated from the disease lesions were confirmed to be race-change mutants. In field experiments, there was a delay in the detection of race-change mutants and a reduction in disease severity. Decreased disease severity was associated with a reduction in the number of race-change mutants and the suppression of disease caused by the race-change mutants. This suggests a possible mechanism related to a decrease in the pathogen population size, which subsequently reduces the number of race-change mutants for the selection pressure of R genes. Thus, inducers of SAR are potentially useful for increasing the durability of genotype-specific resistance conferred by major R genes.     

Possible involvement of salicylic acid in systemic acquired resistance ofCucumis sativus againstSphaerotheca fuliginea

The possible involvement of salicylic acid in systemic acquired resistance ofCucumis sativus againstSphaerotheca fuliginea was studied. Cucumber plants were inoculated with tobacco necrosis virus on the cotyledons and the level of endogenous salicylic acid in the first true leaf was determined by gas chromatography. Salicylic acid increased continously from the second day after virus inoculation to the fifth day, when the same leaf was inoculated withSphaerotheca fuliginea. In healthy plants, the efficiency of exogenous salicylic acid in inducing resistance was assayed by applying aqueous solutions at different times beforeSphaerotheca fuliginea inoculation. To evaluate the level of induced resistance, the following parameters were examined by light microscopy: percentage of conidial germination, length of the hyphae derived from single conidia, number of haustoria, percentage of epidermal cells with lignified walls and of necrotic cells underlying fungal hyphae. In treated plants conidial germination was reduced, the total length of the hyphae was shorter, the number of haustoria was lower and the haustorium-containing epidermal cells had more frequently lignified walls. Moreover, an evident increase in callose deposition was observed leading to the formation of oversized papillae around the penetration pegs. These results indicate that the application of salicylic acid before inoculation withSphaerotheca fuliginea reduces the intensity of the infectious process and that salicylic acid is involved in the expression of systemic resistance in cucumber challenged by the biotrophic pathogenSphaerotheca fuliginea.     

No role for bacterially produced salicylic Acid in rhizobacterial induction of systemic resistance in Arabidopsis

ABSTRACT The role of bacterially produced salicylic acid (SA) in the induction of systemic resistance in plants by rhizobacteria is far from clear. The strong SA producer Pseudomonas fluorescens WCS374r induces resistance in radish but not in Arabidopsis thaliana, whereas application of SA leads to induction of resistance in both plant species. In this study, we compared P. fluorescens WCS374r with three other SA-producing fluorescent Pseudomonas strains, P. fluorescens WCS417r and CHA0r, and P. aeruginosa 7NSK2 for their abilities to produce SA under different growth conditions and to induce systemic resistance in A. thaliana against bacterial speck, caused by P. syringae pv. tomato. All strains produced SA in vitro, varying from 5 fg cell(-1) for WCS417r to >25 fg cell(-1) for WCS374r. Addition of 200 muM FeCl(3) to standard succinate medium abolished SA production in all strains. Whereas the incubation temperature did not affect SA production by WCS417r and 7NSK2, strains WCS374r and CHA0r produced more SA when grown at 33 instead of 28 degrees C. WCS417r, CHA0r, and 7NSK2 induced systemic resistance apparently associated with their ability to produce SA, but WCS374r did not. Conversely, a mutant of 7NSK2 unable to produce SA still triggered induced systemic resistance (ISR). The possible involvement of SA in the induction of resistance was evaluated using SA-nonaccumulating transgenic NahG plants. Strains WCS417r, CHA0r, and 7NSK2 induced resistance in NahG Arabidopsis. Also, WCS374r, when grown at 33 or 36 degrees C, triggered ISR in these plants, but not in ethylene-insensitive ein2 or in non-plant pathogenesis- related protein-expressing npr1 mutant plants, irrespective of the growth temperature of the bacteria. These results demonstrate that, whereas WCS374r can be manipulated to trigger ISR in Arabidopsis, SA is not the primary determinant for the induction of systemic resistance against bacterial speck disease by this bacterium. Also, for the other SAproducing strains used in this study, bacterial determinants other than SA must be responsible for inducing resistance.     

小麦抗白粉病基因Pm6的微卫星标记鉴定

 Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici, is a prevalent disease worldwide. Breeding and planting resistance cultivars have been proved effective and environmental friendly for control of the disease. To develop easily used PCR-based markers in marker assisted selection (MAS) for Pm6, a dominant powdery mildew resistance gene in wheat, 25 microsatellite markers on chromosome 2BL in wheat were screened between susceptible parent Yumai13 and resistance parent Timgalen carrying Pm6. F₂ population derived from Yumai13 and Timgalen was further analyzed by the marker Xgwm501. The results indicated that Xgwm501 was a co-dominant marker linked to Pm6 gene at a distance of 14.8 cM. 29 Pm-carrying varieties were tested by the marker Xgwm501 and only those carrying Pm6 showed 117 bp resistance specific band. This marker is proved to have high practicability and can be used in MAS of Pm6 gene in wheat breeding programs.     

番茄黄萎病抗病基因Ve的AFLP和SSR分子标记

 本研究以番茄抗病品种05046与感病品种051355配制杂交组合,接种鉴定F1代及F2代分离群体的黄萎病发生情况,结果表明,番茄黄萎病属单基因显性遗传。用545对AFLP引物和101对SSR引物对两个亲本、抗感池及F2代分离群体进行AFLP和SSR分析,得到3个与番茄抗黄萎病基因Ve连锁的AFLP标记和1个SSR标记,分别是E66M84-A、E78M84-D、E66M40-A和SSR599,与抗病基因Ve的连锁遗传距离分别为10.3、14.2、30.5 和12.5 cM。     

茶树病程相关蛋白基因CsPR5的克隆及表达分析

为探究茶树中病程相关蛋白5(pathogenesis-related protein 5,PR5)在茶树中的功能,采用cDNA末端快速扩增(rapid amplification of cDNA end,RACE)技术克隆了PR5基因的全长;通过荧光定量PCR方法检测了CsPR5在龙井43茶树不同组织部位、植物激素(茉莉酸、乙烯利和水杨酸)处理、茶炭疽病菌Colletotrichum camelliae侵染和小贯小绿叶蝉Empoasca onukii为害后的表达特征。结果表明,CsPR5基因开放阅读框为843 bp,编码281个氨基酸。CsPR5基因在茶树根、茎、嫩叶、花和种子5个组织中的相对表达量分别为0.187、3.093、0.928、0.045和0.012,且五者之间差异显著,主要在茎和叶中表达,其分别为根中相对表达量的16.54倍与4.96倍。水杨酸处理6 h及乙烯利处理6、12 h后,茶树CsPR5基因的相对表达量分别为1.622、2.344和1.845,分别比对照显著上调2.20倍、3.18倍和2.35倍;但是外施茉莉酸对茶树CsPR5基因的相对表达量无影响。茶炭疽病菌侵染显著诱导了茶树CsPR5基因的相对表达量,处理后的相对表达量为1.977,是对照的1.90倍;小贯小绿叶蝉取食显著抑制了其为害部位CsPR5的相对表达量,为7.273,仅为对照的38.80%,但该诱导不具系统性。     

Pathogenesis-related gene expression in rice is correlated with developmentally controlled Xa21-mediated resistance against Xanthomonas oryzae pv. oryzae

Disease resistance mediated by the resistance gene Xa21 is developmentally controlled in rice. We examined the relationship between Pathogenesis Related (PR) defense gene expression and Xa21-mediated developmental disease resistance induced by Xanthomonas oryzae pv. oryzae (Xoo). OsPR1a, OsPR1b, and OsPR1c genes were cloned and their induction was analyzed, in addition to the OsPR10a gene, at the juvenile and adult stages in response to a wildtype Xoo strain that induces a resistance response (incompatible interaction) and an isogenic mutant Xoo strain that does not (compatible interaction). We found that the adult stage leaves are more competent to express these OsPR1 genes and that the Xa21 locus is required for the highest levels of induction.     

与番茄枯萎病抗病基因I-1连锁的AFLP和SSR分子标记

本研究以番茄枯萎病抗病品种‘05045’与感病品种‘051451’为亲本配制杂交组合。用870对AFLP引物及319对SSR引物对348个F2代分离群体进行连锁分析,得到4个与番茄枯萎病抗病基因I-1连锁的AFLP标记和2个SSR标记,分别是E41M60-D、E41M62-C、E86M36-B、E32M44-E和SSR108、SSR276,与抗病基因I-1的连锁遗传距离分别为4.7、5.3、8.9、11.5cM和6.1、9.3cM。     

亚麻品系9801-1抗白粉病基因的RAPD标记

 F2 populations were obtained from the cross between 9801-1 and DIANE.Bulked segregate and RAPD analyses were employed to identify molecules linked to the resistance to powdery mildew.OPP02 amplified about 792 bp polymorphic band in all individuals from 9801-1 and resistant bulk,but absent in all individuals from DIANE and susceptible bulk.By further analysis in F2 segregating population,the polymorphic band was found to be cosegregated with the resistant gene possibly.The fragment was sequenced,     

Jasmonic and salicylic acid-induced resistance in sorghum against the stem borer Chilo partellus

Induced resistance was studied in three sorghum genotypes (IS2205, ICSV1 and ICSV700) against Chilo partellus (Swinhoe) (Lepidoptera; Pyralidae) infestation and jasmonic acid (JA) and salicylic acid (SA) application. The activity of plant defensive enzymes [peroxidase (POD), polyphenol oxidase (PPO), superoxide dismutase (SOD), and catalase (CAT)], and the amounts of total phenols, hydrogen peroxide (H₂O₂), malondialdehyde (MDA), and proteins were recorded at 6 days after infestation. The induction of enzyme activities and the amounts of secondary metabolites varied among the genotypes and treatments. The genotype IS2205 showed a stronger effect than that of ICSV1 or ICSV 700. Treatment with JA followed by insect infestation induced greater levels of enzymes and secondary metabolites. The results suggest that JA induces greater levels of resistance components in sorghum plants against insect pests. Thus, pretreatment of plants with elicitors including JA and SA could provide a greater opportunity for plant defense against herbivores.     

水杨酸和乙烯在烟草抗黑胫病中的作用

5mmol/L水杨酸或1g/L乙烯外源处理可诱导烟草对黑胫病的抗性，推迟植株死亡。不能积累水杨酸的转nahG基因植株和不表现乙烯应答反应的转etr1-1基因植株对黑胫病的抗性显著弱于野生型植株。组成性积累高水平水杨酸的转entC和pmsB基因植株对黑胫病的抗性明显强于野生型植株。这些结果说明水杨酸和乙烯增强烟草对黑胫病的抗性，减缓黑胫病的扩展。0．5mmol/L KCN连续浇灌植株4天或浸溃叶片2，4h均不能恢复转nahG基因植株丧失的对黑胫病的抗病性。此结果进一步支持关于KCN处理不能恢复转nahG基因植株丧失的对真菌病害抗病性的假说。     

Pathogenesis-related gene expression in rice is correlated with developmentally controlled Xa21-mediated resistance against Xanthomonas oryzae pv. oryzae

Disease resistance mediated by the resistance gene Xa21 is developmentally controlled in rice. We examined the relationship between Pathogenesis Related (PR) defense gene expression and Xa21-mediated developmental disease resistance induced by Xanthomonas oryzae pv. oryzae (Xoo). OsPR1a, OsPR1b, and OsPR1c genes were cloned and their induction was analyzed, in addition to the OsPR10a gene, at the juvenile and adult stages in response to a wildtype Xoo strain that induces a resistance response (incompatible interaction) and an isogenic mutant Xoo strain that does not (compatible interaction). We found that the adult stage leaves are more competent to express these OsPR1 genes and that the Xa21 locus is required for the highest levels of induction.     

Systemic acquired resistance plays a major role in bacterial blight resistance in a progeny of somatic hybrids of cultivated rice (Oryza sativa L.) and wild rice (Oryza meyeriana L.)

Journal of Plant Diseases and Protection - As the number of people in the world increases, people's demand for food becomes greater. In order to reduce reliance on pesticides that maintain crop...     

西瓜抗小西葫芦黄花叶病毒基因的连锁分子标记研究

 小西葫芦黄花叶病毒中国株系(Zucchini yellow mosaic virus Chinese strain,ZYMV-CH)是危害我国西瓜的主要病毒。本实验以抗病毒病西瓜野生种质P.I.595203与感病的普通西瓜自交系98R为亲本,采用单粒传方式得到109个E代株系,分别对亲本、F₁及109个F₃代株系群体进行了苗期抗ZYMV-CH接种鉴定,通过F₃代群体的抗感分离情况,推测得到F₂代各单株的基因型,采用集团分离分析法(bulked segregant analysis,BSA)在F₂代建立抗感基因池,以亲本、F₁和抗感基因池为模板,对640条RAPD引物进行PCR扩增筛选,其中引物AK13在亲本、F₁和抗感基因池之间扩增出一条多态性片段(644bp),在F₂代群体上验证该多态性条带与ZYMV-CH的抗性基因呈现连锁关系,遗传连锁距离为8cM,定名为AK13_-644,该连锁标记在ZYMV-CH抗性转育后代自交系上得到了验证。最终将此RAPD标记成功转化成SCAR标记SCAK13_-644,该标记可以作为西瓜抗病毒病辅助选择的分子标记。     

Compost and compost water extract-induced systemic acquired resistance in cucumber and Arabidopsis

ABSTRACT A biocontrol agent-fortified compost mix, suppressive to several diseases caused by soilborne plant pathogens, induced systemic acquired resistance (SAR) in cucumber against anthracnose caused by Colletotrichum orbiculare and in Arabidopsis against bacterial speck caused by Pseudomonas syringae pv. maculicola KD4326. A peat mix conducive to soilborne diseases did not induce SAR. The population size of P. syringae pv. maculicola KD4326 was significantly lower in leaves of Arabidopsis plants grown in the compost mix compared to those grown in the peat mix. Autoclaving destroyed the SAR-inducing effect of the compost mix, and inoculation of the autoclaved mix with nonautoclaved compost mix or Pantoea agglomerans 278A restored the effect, suggesting the SAR-inducing activity of the compost mix was biological in nature. Topical sprays with water extract prepared from the compost mix reduced symptoms of bacterial speck and the population size of pathogenic KD4326 in Arabidopsis grown in the peat mix but not in the compost mix. The peat mix water extract applied as a spray did not control bacterial speck on plants grown in either mix. Topical sprays with salicylic acid (SA) reduced the severity of bacterial speck on plants in the peat mix but did not further reduce the severity of symptoms on plants in the compost mix. The activity of the compost water extract was heat-stable and passed through a 0.2-mum membrane filter. beta-1,3-Glucanase activity was low in cucumber plants grown in either mix, but when infected with C. orbiculare, this activity was induced to significantly higher levels in plants grown in the compost mix than in plants grown in the peat mix. Similar results were obtained for beta-D-glucuronidase (GUS) activity driven by a PR2 (beta-1,3-glucanase) gene promoter in transgenic Arabidopsis plants grown in the compost or peat mix. GUS activity was induced with topical sprays of the compost water extract or SA in plants not inoculated with the pathogen, suggesting that compost-induced disease suppression more than likely involved the potentiation of resistance responses rather than their activation and that compost-induced SAR differed from SAR induced by pathogens, SA, or compost water extract.     

小麦诱导抗性基因TaWIR1b的克隆及表达分析

全蚀病是小麦上一种重要的土传病害。选育和种植抗病品种是防治小麦全蚀病的根本途径,抗病基因研究是抗病育种的基础性工作。根据基因TaWIR1b(Accession no.M94959.1)的全长序列设计引物扩增‘新农19’的cDNA,获得了完整ORF,编码85个氨基酸残基,比对后发现与TaWIR1b序列同源性达100%。根据获得的TaWIR1b基因全长序列设计定量引物,分析TaWIR1b在全蚀菌胁迫条件下不同互作模式的表达特征。结果表明接种全蚀病菌后抗病小麦品种‘新农19’中TaWIR1b基因被诱导表达,接菌后3d达到峰值143.97,感病品种‘新麦19’中峰值出现在接菌后8d,表达量仅为对照的4.22倍,提示该基因可能参与小麦对全蚀病的抗病过程。