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1.
 大麦黄矮病毒(barley yellow dwarf viruses,BYDVs)引起的小麦黄矮病严重威胁我国麦类生产,造成严重经济损失。植物中的miRNA调控植物生长发育、信号转导及对外界压力的反应,通过调控植物抗性基因的表达影响植物与病原物的互作。本研究对感染BYDV-GAV 后3 d、7 d及健康对照的‘小偃6号'小麦样品进行miRNA测序,合并去冗余后分别得到99、96、95个已知的miRNA序列和806、809、1 024个新miRNA序列。对这些miRNA进行差异表达分析,BYDV-GAV侵染后3 d和7 d的小麦样品,与对照相比上调表达的miRNA数量分别为3个和7个,下调表达的为14个和12个。将差异表达的miRNA利用psRNATarget进行靶基因预测,共得到1 254个靶标基因。靶基因的KEGG和GO富集分析,进一步明确了其功能及作用通路。对14个病毒病症状相关的靶基因进行定量分析,结果表明随病毒侵染时间的延长,这些靶基因出现差异性表达,显示miRNA参与了寄主与病毒的互作。研究结果有助于揭示BYDV-GAV与寄主小麦的互作机理。  相似文献   

2.
 大麦黄矮病毒(barley yellow dwarf viruses,BYDVs)引起的小麦黄矮病严重威胁我国麦类生产,造成严重经济损失。植物中的miRNA调控植物生长发育、信号转导及对外界压力的反应,通过调控植物抗性基因的表达影响植物与病原物的互作。本研究对感染BYDV-GAV 后3 d、7 d及健康对照的‘小偃6号'小麦样品进行miRNA测序,合并去冗余后分别得到99、96、95个已知的miRNA序列和806、809、1 024个新miRNA序列。对这些miRNA进行差异表达分析,BYDV-GAV侵染后3 d和7 d的小麦样品,与对照相比上调表达的miRNA数量分别为3个和7个,下调表达的为14个和12个。将差异表达的miRNA利用psRNATarget进行靶基因预测,共得到1 254个靶标基因。靶基因的KEGG和GO富集分析,进一步明确了其功能及作用通路。对14个病毒病症状相关的靶基因进行定量分析,结果表明随病毒侵染时间的延长,这些靶基因出现差异性表达,显示miRNA参与了寄主与病毒的互作。研究结果有助于揭示BYDV-GAV与寄主小麦的互作机理。  相似文献   

3.
为明确靶标害虫对Bt蛋白的抗性及对不同类型Bt蛋白的交互抗性,采用生物测定法进行了二化螟Chilo suppressalis(Walker)敏感品系和Cry1Ac汰选品系在Cry1Ac毒饲料上的时间-死亡率反应、对不同Bt杀虫蛋白的敏感性及不同Bt蛋白复配组合对二化螟毒力效果的研究。结果表明,二化螟Cry1Ac汰选品系已对36.67μg/mL(LC_(50))的Cry1Ac杀虫蛋白产生了一定的适应性,但对308.69μg/mL(LC90)的Cry1Ac蛋白反应仍较敏感。Cry1Ac敏感品系和汰选品系对Cry1Ab蛋白的LC_(50)分别为1.40μg/mL和3.86μg/mL,二者差异显著,但对Cry1Ca(1.63μg/mL和1.73μg/mL)或Cry2Aa(127.48μg/mL和144.50μg/mL)的LC_(50)差异不显著,即Cry1Ac汰选品系与Cry1Ab存在明显的交互抗性,但与Cry1Ca和Cry2Aa不存在交互抗性。在不同Bt蛋白复配组合中(1∶1复配),Cry1Ab+Cry1Ca、Cry1Ab+Cry2Aa和Cry1Ca+Cry2Aa增效作用最为显著。表明抗虫基因cry1Ca和cry2Aa可作为双价转基因抗虫水稻研发的候选基因。  相似文献   

4.
筛选对靶标害虫高效、低毒的Bt蛋白对未来开发新型Bt生物杀虫剂、转基因抗虫植物新品种等具有重要意义。我们在室内测定了Cry2Ab4、Cry2Ah1、Cry1Ca7及Vip3Aa11四种Bt蛋白对棉铃虫、甜菜夜蛾、斜纹夜蛾、黏虫、玉米螟、二化螟的杀虫效果。结果表明,Cry2Ab4杀虫谱广、对六种鳞翅目的重要害虫都具有很高的杀虫效果,尤其是对棉铃虫、甜菜夜蛾、斜纹夜蛾和玉米螟的毒力效果显著;Cry2Ah1对斜纹夜蛾、甜菜夜蛾具有很好的致死作用,对棉铃虫、玉米螟和二化螟也具有较好的杀虫效果,但对黏虫只有明显的体重抑制作用、致死效果不明显;Cry1Ca7对甜菜夜蛾和斜纹夜蛾有明显的防治效果,其次对棉铃虫、二化螟也有较好的杀虫作用,但对玉米螟毒力效果较差,对黏虫没有毒力作用;Vip3Aa11对甜菜夜蛾和斜纹夜蛾的防治效果好,对玉米螟、棉铃虫和二化螟有较好的防治效果,但对黏虫没有毒力作用。  相似文献   

5.
长链非编码RNA (long non-coding RNA, lncRNA) 是指长度大于200 nt且不具有编码蛋白质功能的一类RNA分子。当生物体暴露于环境污染物时,其lncRNA通常会异常表达,并与癌症等疾病的发生相关。采用亚致死浓度的三唑磷处理斑马鱼后,利用人源基因芯片初步筛选出了异常表达的斑马鱼lncRNA,并通过生物信息学软件预测了与lncRNA相互作用的微RNA (micro RNA, miRNA)及其靶基因,最后通过实时荧光定量PCR验证了该miRNA及其靶基因在斑马鱼中的表达,从而构建起了长链非编码RNA-微RNA-信使RNA (lncRNA-miRNA-mRNA) 的调控网络。荧光定量PCR检测结果表明:与空白对照组相比,经三唑磷处理后,斑马鱼lncRNA Sox2OT基因的表达下调了35%;lncRNA H19基因的表达上调了1.41倍,而与其可能存在相互作用的dre-let-7c基因的表达则下调了78%,同时dre-let-7c的靶基因ddx18的表达上调了2.11倍;差异均达显著水平 (P < 0.05)。据此推测,在三唑磷的作用下,lncRNA H19可能作为靶基因ddx18的竞争性内源RNA而与dre-let-7c发生相互作用,从而调控靶基因ddx18的表达。因此,lncRNA H19、dre-let-7c及ddx18都有可能作为生物标志物用于监测三唑磷对环境和生物体的影响。  相似文献   

6.
为获得新的鳞翅目害虫杀虫基因,根据已知cry1I类基因编码区设计简并引物,采用直接克隆法,以苏云金芽胞杆菌(Bacillus thuringiensis,Bt)菌株BN23-5质粒DNA为模板进行扩增,并对得到的基因进行鉴定和分析。结果表明:克隆得到一个完整的cry1Ie基因,全长2 160 bp,由719个氨基酸组成,该氨基酸序列与已知的4种Cry1Ie蛋白不同,与Cry1Ie2和Cry1Ie3的氨基酸序列同源性最高,为95.4%,被国际Bt杀虫晶体蛋白基因命名委员会命名为Cry1Ie5(登录号为KJ710646)。将该基因插入表达载体p ET-28a,转化大肠杆菌BL21(DE3),IPTG低温诱导成功表达,SDS-PAGE电泳验证其大小为81 k D,与预测的分子量相符合。生物活性测定表明,Cry1Ie5表达的包涵体蛋白对小菜蛾和亚洲玉米螟具有杀虫活性,LC_(50)分别为0.43μg/m L和48.39μg/m L;对棉铃虫的致死率不高,但能明显抑制其生长;对甜菜夜蛾没有杀虫活性。  相似文献   

7.
为明确转cry1Ab+cry1C双价抗虫水稻对二化螟的抗性及其杀虫蛋白的时空表达,本研究采用离体水稻组织生测法系统评价了cry1Ab、cry1C、正交cry1Ab+cry1C和反交cry1C+cry1Ab 4种抗虫水稻品系对二化螟的杀虫效果,并用ELISA方法测定了Cry1Ab、Cry1C蛋白在各个抗虫水稻品系中的表达情况。结果显示,二化螟在不同生育期的4种转基因抗虫水稻上取食的叶面积显著低于非转基因对照亲本明恢63。4个转基因抗虫水稻品系在生长前期（苗期、分蘖和拔节期）对二化螟表现极高的杀虫效果,生长后期（孕穗期和成熟期）杀虫效果有所下降。两双价抗虫水稻杀虫效果最好,其次为转cry1Ab和cry1C水稻。Bt蛋白表达水平随着抗虫水稻生育期的变化而变化,且差异显著;Cry1Ab的蛋白表达量在水稻整个生长期均显著高于Cry1C。Cry1Ab蛋白在单价抗虫水稻叶片和茎杆中均表现出随生育期先升高后下降的趋势,但在双价抗虫水稻中表现出随生育期逐渐下降的趋势。Cry1C蛋白在单、双价抗虫水稻的叶片组织中均表现出逐渐升高的趋势。与单价抗虫水稻相比,双价抗虫水稻中的Cry1Ab、Cry1C的蛋白表达水平并没有表现出显著的降低。因此,双价抗虫水稻不仅能高效防治害虫,还能延缓害虫抗性,在生产上表现出良好的应用前景。该研究结果可望为转Bt基因抗虫作物的环境安全评价提供科学数据和理论依据。  相似文献   

8.
Bt棉有效控制了棉田主要害虫棉铃虫(Helicoverpa armigera),然而原来处于次要地位的刺吸式口器害虫盲蝽(Heteroptera:Miridae)为害逐年加重,目前对绿盲蝽(Apolygus lucorum Meyer-Dür)有效的抗虫基因未见报道。本研究以本实验室保存的Bt杀虫基因和菌株为材料,对绿盲蝽进行杀虫活性筛选。利用本实验室先前克隆的Mtx类杀虫基因cry15Aa表达产物进行绿盲蝽杀虫活性测定,研究结果显示Cry15Aa蛋白对绿盲蝽具有一定的杀虫活性。通过杀虫活性测定,获得对绿盲蝽有效的Bt菌株20株;对这些菌株表达蛋白进行质谱鉴定,LC-MS/MS质谱鉴定结果显示,这些菌株可能含有Cry1Aa、Cry1Ab、Cry1Ac、Cry1Ae、Cry1Af、Cry1Ag、Cry1Ah、Cry1Ba、Cry1Be、Cry8Ha等十余种对鳞翅目、鞘翅目害虫有杀虫活性的蛋白片段,并且有4株Bt菌株样品中检出了Mtx类杀虫蛋白Cry15Aa的片段。但是进一步基因鉴定结果表明,这4株菌株并不含有cry15Aa全长基因,推测这些菌株中含有Cry15类的新基因。上述研究结果表明这些菌株中可能存在对盲蝽有效的新型杀虫蛋白。本研究在发现了cry15Aa基因所表达的蛋白对绿盲蝽具有杀虫活性的同时,获得了对绿盲蝽具有杀虫活性的Bt新菌株,对绿盲蝽的生物防治具有重要的意义。  相似文献   

9.
评价转Bt基因玉米对靶标生物亚洲玉米螟的杀虫作用是转基因玉米研发的重要一环。本文采用室内生测法对3种转Bt基因抗虫玉米‘瑞丰125’(表达Cry1Ab/Cry2Aj杀虫蛋白),‘DBN9936’‘DBN9978’(表达Cry1Ab杀虫蛋白)对亚洲玉米螟敏感品系ACB-S及抗Cry1Ab品系ACB-AbR、抗Cry1Ac品系ACB-AcR、抗Cry1F品系ACB-FR、抗Cry1Ah品系ACB-AhR、抗Cry1Ie品系ACB-IeR的杀虫活性进行测定,同时采用心叶期和抽丝期人工接虫法进行田间抗虫效果鉴定。结果表明,取食3种Bt玉米的ACB-S幼虫, 3 d死亡率100%,而取食对照常规玉米3 d存活率100%。取食3种Bt玉米的5个抗性品系幼虫除ACB-AbR和ACB-AcR有2%~6%的个体存活4~5 d, 6 d死亡率也达到了100%,其余品系均在3 d全部死亡,而取食对照玉米5~6 d的死亡率仅为4%~14%,差异显著。田间心叶期食叶级别及穗期活虫数、雌穗被害和茎秆被蛀等为害等级说明3种Bt玉米高抗亚洲玉米螟。明确了‘瑞丰125’‘DBN9936’和‘DBN9978’对亚洲玉米螟有很高的杀虫活性和田间防治效果。5个Bt蛋白抗性亚洲玉米螟品系幼虫在常规玉米上显示一定的适合度劣势。  相似文献   

10.
苏云金芽孢杆菌BtC008对棉铃虫、小菜蛾和甜菜夜蛾等鳞翅目害虫具有高毒力。本文分析了其杀虫基因类型包括cry1Ab、cry1Ca、cry1Ia、cry2Ab和一种新的cry1类基因,SDS-PAGE分析其至少2种cry1类基因获得了表达,在分子水平说明了该菌株高毒力的原因。在此基础上克隆了1种cry1Ab类杀虫晶体蛋白(insecticidalcrystalproteins,ICPs)基因,并被Bt杀虫晶体蛋白基因命名委员会命名为cry1Ab20。序列分析显示该基因所编码蛋白与已知的Cry1Ab13有1个氨基酸不同。将cry1Ab基因毒性片段(1.95kb)插入表达载体pET-21b,转化EscherichiacoliRosetta(DE3)菌株,表达的包涵体蛋白对小菜蛾和棉铃虫幼虫具有杀虫活性,LC50分别为100.74μg/mL和29.87μg/mL,进一步明确了Cry1Ab20蛋白的杀虫活性区。  相似文献   

11.
Research on root rot pathogens of peas in the Netherlands has confirmed the prevalence ofFusarium solani, F. oxysporum, Pythium spp.,Mycosphaerella pinodes andPhoma medicaginis var.pinodella. Aphanomyces euteiches andThielaviopsis basicola were identified for the first time as pea pathogens in the Netherlands. Other pathogens such asRhizoctonia solani andCylindrocarpon destructans were also found on diseased parts of roots. F. solani existed in different degrees of pathogenicity, and was sometimes highly specific to pea, dwarf bean of field bean, depending on the cropping history of the field.A. euteiches was specific to peas, whereasT. basicola showed some degree of physiological specialization.  相似文献   

12.
Forty-five onion (Allium cepa) cultivars were evaluated for disease resistance to the fungal pathogen Pyrenochaeta terrestris (pink root disease). Forty-three Iranian cultivars and two commercially exotic resistant cultivars (Texas Early Grano and Yellow Sweet Spanish) were grown in a glasshouse for two successive years (2003 and 2004) in Isfahan, Iran. Susceptibility of the cultivars was determined using a scale of 0–3. Six cultivars had significantly lower susceptibility of pink root compared to the exotic ones. High susceptibility tended to be associated with high mean scores (2–3), and the highly resistant cultivars had the lowest scores (0–1). In addition, the 45 examined cultivars were ranked from 1 to 45 according to their markedly differing reactions to P. terrestris, which differed markedly. Of 32 cultivars that were less resistant, 30 with an intermediate status or tolerance (1–2) differed considerably in their rank order. No pink root symptoms were seen in the early stages, but on blotter paper or wheat-straw agar some roots turned pink with no pycnidia formation, whereas on potato dextrose agar medium (PDA) with a few fresh healthy and sterile onion roots on the surface, mycelia and pycnidia were recovered. Variance and cluster analysis showed similar results among the cultivars with various levels of resistance, tolerance and susceptibility as determined by scoring scales.  相似文献   

13.
Broad bean mottle virus (BBMV) was transmitted from infected to healthy faba-bean plants by the curculionid weevilsApion radiolus Kirby,Hypera variabilis Herbst,Pachytychius strumarius Gyll,Smicronyx cyaneus Gyll, andSitona lineatus L. The latter appeared to be an efficient vector: acquisition and inoculation occurred at the first bite, the rate of transmission was c. 41%, and virus retention lasted for at least seven days.S. lineatus transmitted the virus from faba bean to lentil and pea, but not to the three genotypes of chickpea tested. This is the first report on the generaHypera, Pachytychius, andSmicronyx as virus vectors, and onA. radiolus, H. variabilis, P. strumarius, andS. cyaneus as vectors of BBMV.Out of 351 samples of food legumes with symptoms suggestive of virus infection, 16, 11, 19, and 17% of the samples of chickpea, lentil, pea, and common bean, respectively, were found infected when tested for BBMV in DAS-ELISA. This is the first report on the natural occurrence of BBMV in chickpea, lentil, pea, and common bean. The virus should be regarded as a food-legume virus rather than a faba-bean virus solely, and is considered an actual threat to food legume improvement programmes.  相似文献   

14.
The epidemiology ofSitobion avenae and its natural enemies in winter wheat was studied in 1975, 1976 and 1977. Immigration was important until the end of flowering. The alate immigrants had apterous offspring. These became the driving force in population growth. Their offspring were mostly alatae which usually left the field. A model of the epidemic was developed. Quantitative relations between the aphids and their environment were obtained from literature or established in laboratory trials. The model simulated population development and population composition from the beginning of June till the population peak at the end of June or early in July. Because quantitative data on relations between aphids and their natural enemies and pathogens are scarce, and since the knowledge on wing formation is still limited, the population collapse could not be predicted. In the future, prognosis over a period of three weeks seems possible.Samenvatting De toenemende betekenis van graanbladluizen (vooralSitobion avenae) gepaard gaande met een sterke toename van het gebruik van insecticiden op granen maakte verbetering van de prognose over het schadelijk optreden wenselijk. Door gedetailleerde tellingen in het veld (Fig. 1–7) werden gegevens verkregen over het verloop van de epidemie en het optreden van natuurlijke vijanden in 1975, 1976 en 1977.Een immigratieperiode tot in de bloei kon worden vastgesteld. Daarna lijkt de aantrekkelijkheid van het gewas voor alate luizen te verminderen. De alate immigranten krijgen aptere nakomelingen. Deze vormen de stuwende kracht van de populatiegroei. De nakomelingen van apteren zijn merendeels alaat. Zij verlaten het gewas.Een model van de populatieontwikkeling gedurende de epidemie werd opgesteld. De relatiediagrammen Fig. 9 en 10 laten groei en ontwikkeling vanS. avenae en een predator (Syrphus corollae) zien. Kwantificering van de betrekkingen werd mogelijk door literatuurgegevens en laboratoriumexperimenten.Met het model kon de populatieontwikkeling vanS. avenae vanaf begin juni tot aan de populatiepiek in 1975, 1976 en 1977 vrij goed worden gesimuleerd (Fig. 12). Ook de populatieopbouw kon worden gesimuleerd (Fig. 14). De teruggang van de populatiedichtheid blijkt moeilijker te voorspellen door het ontbreken van gegevens over natuurlijke vijanden.Het lijkt waarschijnlijk dat in de toekomst met het model een prognose over de piek van de bladluispopulatie circa 3 weken tevoren mogelijk zal zijn.  相似文献   

15.
Corynebacterium fascians, which causes leafy gall, has been shown to be responsible for the unusual symptoms recently found in lilies. The symptoms are described and the different bacterial isolates compared by biochemical, serological and pathogenicity experiments. No evidence could be found for the existence of specialized strains even though considerable variation in virulence could be demonstrated. Although the role of variation in susceptibility of the different lily cultivars should not be underestimated, it would appear that high inoculum levels ofC. fascians in the soil may be largely responsible for these outbreaks.Also included in this study are the results of biochemical, serological and pathogenicity experiments ofC. fascians which has been isolated fromKalanchoe, Euphorbia, Brodiaea, Hebe andVerbascum.Samenvatting Reeds een aantal jaren was in lelies een woekering in stengelbollen bekend, die echter sporadisch voorkwam en economisch van weinig betekenis was. Tijdens de zomer van 1977 echter trad de ziekte op één enkel perceel zo ernstig op dat zeer aanzienlijke oogstderving het gevolg was. Behalve de stengelbol kan ook de hoofdbol misvormd zijn. Bovendien kunnen misvormde stengelbollen lange spruiten vormen, zgn. sabeltandvorming. Soms is het wortelstelsel gereduceerd; in de kas zijn verdikte stengels waargenomen. De ziekte staat nu bekend onder de naam woekerziekte.Door de PD te Wageningen en het LBO te Lisse werd de bacterieCorynebacterium fascians (Tilford) Dows. uit het zieke materiaal geïsoleerd. In gezamenlijk onderzoek werd nagegaan of de bacterie de veroorzaker is van bovengenoemde symptomen en of hier sprake is van een op lelie gespecialiseerde stam.Biochemische en serologische vergelijkingen tussen isolaten vanC. fascians uit lelie,Kalanchoe, Euphorbia, Brodiaeea, Hebe enVerbascum laten zien dat er zeer weinig verschillen bestaan. Alle isolaten reageren op een standaard antiserum met titers variërend tussen 1: 640–1: 5120.Een oriënterende inoculatieproef werd uitgevoerd op het LBO in 1977. Daarna werden op de PD 10 van de 25 isolaten uit lelie en 8 uit andere waardplanten getoetst op lelie, chrysant en lathyrus. Uit de inoculatieproeven blijkt, dat de bacterie de veroorzaker is van de waargenomen symptomen. Tussen de isolaten blijkt een groot verschil in pathogeniteit te bestaan (12–100% aantasting), zonder dat dit aan herkomst gebonden is. Het voorkomen van op lelie of andere waardplanten gespecialiseerde stammen is daarom niet aan te nemen.In hoeverre de hevige aantasting een gevolg is van een opbouw van de bacterie-populatie door een te nauwe vruchtwisseling en mechanische rooimethoden moet worden nagegaan.  相似文献   

16.
Resistance of Brassica napus (oilseed rape, canola) conferred by three different major resistance genes has been overcome by changes in virulence of Leptosphaeria maculans populations in France and Australia. In South Australia where B. napus cultivars with major gene resistance derived from Brassica rapa ssp. sylvestris were grown extensively, resistance was rendered ineffective within 3 years of commercial release of the cultivar. Disease severity was higher on cultivars with sylvestris-derived resistance than cultivars with polygenic resistance. This Australian situation is compared to that in France, where resistance conferred by the Rlm1 gene was overcome nation-wide in 5 years under commercial cropping practices, and also where a source of resistance introgressed into B. napus from B. juncea was rendered inefficient in 3 years in experimental field plots near Rennes.  相似文献   

17.
Laboratory and field studies on aphid damage in wheat are described. Both direct and indirect effects of aphids on the behaviour of the crop were studied. In the laboratory a clear effect of honeydew and yeasts on photosynthesis was found. This effect could not be demonstrated under field conditions, since even at yield loss levels of 700 kg ha–1 the size of this effect may be undetectable with crop enclosures reaching an accuracy of 10%. The effect of honeydew on ageing, measured in the laboratory trials was also found in the field and may form a major cause of the yield losses found in the field. Yield losses under field conditions reached 700 kg ha–1 and were for 72% due to direct sucking damage of the 35 aphids, found maximally per culm, and direct and indirect honeydew effects. Saprophytic and possibly also some necrotrophic fungi caused 28% of the yield losses. The exact contribution of each of the damage factors was not revealed but it was demonstrated that direct and indirect effects contribute to the final yield losses.Samenvatting Laboratorium- en veldproeven over de schade door bladluizen in tarwe worden beschreven. Een analyse van de directe en indirecte effecten van bladluizen op de groei van het gewas werd verricht. In het laboratorium werd een duidelijk effect van honingdauw en gisten op de fotosynthese gevonden. Dit effect kon niet worden gedemonstreerd in het veld, hetgeen waarschijnlijk wordt veroorzaakt door de orde van grootte van dit effect dat valt binnen de meetfout van de gewasfotosyntheseapparatuur.Het effect van honingdauw op veroudering, dat in het laboratorium werd vastgesteld, kon ook worden aangetoond in het veld en is waarschijnlijk een van de hoofdoorzaken van de oogstverliezen. Deze oogstverliezen in het veld bedroegen 700 kg ha–1 bij een maximale dichtheid van 35 bladluizen per halm en werden voor 72% veroorzaakt door direct zuigsschade en honingdauweffecten. Van de totale schade werd 28% veroorzaakt door saprofytische en wellicht ook door enige necrotrofe schimmels. De precieze bijdrage van ieder van de schadefactoren tot de schade werd niet vastgesteld, maar de wijze waarop directe en indirecte effecten hun bijdrage leverden aan de uiteindelijke oogstverliezen werd zichtbaar gemaakt.  相似文献   

18.
Agave tequilana is the raw material for the production of the alcoholic beverage tequila. A bacterial disease has affected the A. tequilana crop in recent years. Previous reports based on colony and cell morphology, Gram stain and potato rot indicated that Erwinia sp. is the main pathogen. We isolated a several bacterial isolates capable of producing soft-rot symptoms in greenhouse pathogenicity assays. An extensive characterisation involving pathogenicity tests, fatty acid profile, metabolic and physiological properties, ribosomal DNA sequence and intergenic transcribed spacer amplification (ITS-PCR) and restriction banding pattern (ITS-RFLP) was made of each isolate. Three different species: Erwinia cacticida, Pantoea agglomerans and Pseudomonas sp. were identified. Fatty acid and metabolic profiles gave low similarity values of identification but 16S rDNA sequence, ITS-PCR and ITS-RFLP confirmed the identification of E. cacticida. In the phylogenetic tree, E. cacticida from blue agave was grouped neither with E. cacticida type strains nor with Erwinia carotovora. This is the first report that associates E. cacticida with A. tequilana soft-rot symptoms.  相似文献   

19.
Ecological properties and stability of benomyl resistance of three benomyl-resistant mutants of nonpathogenicFusarium-isolates antagonistic to fusarium wilt in carnation, and three benomyl-resistant mutants of a pathogenic isolate ofFusarium oxysporum f.sp.dianthi were evaluatedin vitro and in glasshouse experiments. The benomyl resistance of the nonpathogenic mutants was stable under all conditions tested, also after a 1000-fold increase of the population in sterilized soil. Mutants of the pathogen were stable during allin vitro tests, but after proliferation in carnation stems only part of the population was benomyl resistant.Compared to the wild type, mutants of the pathogen were less pathogenic, also if thein vitro propeties were similar. Colonization of carnation by benomyl-resistant nonpathogenicFusarium in the presence of the pathogen showed that the antagonistic effect correlated with the presence of the nonpathogenic isolates within the carnation stem. The wild types and two of the mutant nonpathogenicFusarium-isolates controlled fusarium wilt in the susceptible cultivar Lena for 50% or more.UV-induced benomyl resistance appeared to be a valuable marker to distinguish between differentFusarium isolates and to study the population dynamics, but intensive screening of the mutants is a prerequisite since alterations in antagonism and pathogenicity can occur.  相似文献   

20.
Several pathovars of Pseudomonas syringae produce the phytotoxin coronatine (COR)in vitro, and the availability of purified toxin has facilitated development of immunological detection methods. A modified, indirect competitive ELISA using the COR-specific monoclonal antibody 11B8 was developed to detect COR in various host plants infected by P. syringae. The estimated detection limit for COR was 50 pg per well, and COR could be reliably quantified from 5 to 40 ng ml−1. The subcellular localization of COR within infected tomato tissue was investigated using the COR-specific antibody MAb 8H3G2. Immunofluorescence microscopy and immunogold labelling showed that COR was present inside tomato cells and was associated with chloroplasts and particles of proteinase inhibitor I. Localization studies indicated that COR is mobile in infected plant tissue and can be detected in healthy tissue adjacent to the bacterial lesions.  相似文献   

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