Evidence for the Preemptive Nature of Tomato Race 3 of Xanthomonas campestris pv. vesicatoria in Florida

ABSTRACT Until recently, tomato race 1 (T1) of Xanthomonas campestris pv. vesicatoria was the only race causing bacterial spot of tomato in Florida. In 1991, tomato race 3 (T3) was first identified in 3 of 13 tomato production fields surveyed. By 1994, T3 was observed in 21 of 28 fields and was the only race identified in 14 fields. In field studies, tomato genotypes with resistance to either T1 or T3 or susceptibility to both were co-inoculated with strains of both races. Lesions on 10 plants in each of three replications for each genotype were sampled three times during the experiment; bacterial isolations were made from each lesion, and tomato race identifications were made for each strain. At the third sampling date, T3 was isolated from 97% of the lesions on the susceptible genotype Walter and the T1-resistant genotype Hawaii 7998, while T3 was isolated from 23% of the lesions and T1 from the remaining 77% on the T3-resistant genotypes PI 128216 and PI 126932. In surface population studies done in growth rooms, suspensions of T1 and T3 were applied alone and in combination to the leaf surfaces of susceptible and resistant genotypes. T1 populations were reduced more than 10-fold when applied in combination with T3, compared with populations that developed when T1 was applied alone. T3 populations were not affected when applied in combination with a T1 strain. In greenhouse studies with the T3-resistant genotype Hawaii 7981, disease was significantly reduced in plants inoculated with T3 in combination with T1, compared with plants inoculated with T1 alone. These results clearly demonstrate the competitive nature of T3 in the presence of T1 and help explain the emergence of T3 as a prevalent race in Florida.     

Races of Xanthomonas campestris pv. vesicatoria overcoming the gene Bs2 for bacterial spot resistance in pepper, prevalent on Capsicum chinense in Barbados and Grenada and weakly pathogenic on bell pepper and tomato in the field

A total of 404 isolates of Xanthomonas campestris pv. vesicatoria , obtained from Capsicum chinense cv. West Indian Red grown in Barbados and Grenada, were differentiated into pathogenic races, and of these, 96 were tested also for selected taxonomic group phenotypes. The response of C. chinense to infection by several X. campestris pv. vesicatoria races and the contribution of races isolated from this cultivar to severity of bacterial spot of bell pepper and tomato were also investigated. P4T2, P5T2 and P6T2 were the predominant races of X. campestris pv. vesicatoria isolated from C. chinense grown in Grenada, whereas nine races (T1, P4, P6, P0T2, P1T2, P4T1, P4T2, P6T1 and P6T2) were isolated in Barbados. Race P4T2 comprised 46.0 and 71.4% of the isolates from Barbados and Grenada, respectively. The 96 isolates, all of which overcame resistance conferred by the gene Bs2 , shared taxonomic group B strain characteristics, including the presence of the β-protein band, positive amylolytic activity and inability to oxidize cis -aconitate. The C. chinense cv. West Indian Red was susceptible only to races of X. campestris pv. vesicatoria that can overcome Bs2 gene resistance. Of six such races identified in Barbados, only P4T1, P4T2 and P6T1 affected bacterial spot-susceptible bell pepper or tomato in the field, and they amounted to only 1.5–2.1% of each sample of isolates from these plant species. Moreover, they were confined to the smallest bacterial spot lesions. Bell pepper was most severely affected by combinations of races T1 with P3T2 and T2 with P0T1, and tomato by race T1 only and combinations of races P0T1 with P0T2 and P1T1 with P1T0, all of which prevailed in the field despite selection against them by C. chinense cv. West Indian Red.     

Bacterial spot of capsicum and tomato in Yugoslavia

The causal agent of bacterial spot of capsicum and tomato grown in different regions in Yugoslavia was investigated. Isolations were made from diseased material collected in recent years. The biochemical and physiological characteristics of isolated bacteria were studied by standard bacteriological tests. The race of the pathogen was determined on differential cultivars of capsicum and tomato. The causal agent of the disease was identified according to the concepts of the time as Xanthomonas campestris pv. vesicatoria. Strains isolated from diseased capsicum were non-pectolytic and non-amylolytic, and did not infect tomato plants. According to the reaction of capsicum cv. Early Calwonder and its isogenic lines, these strains belonged to'pepper races'1 and 3 of X. vesicatoria. Tomato strains showed pectolytic and amylolytic activity and were not pathogenic to capsicum. Accordingly, the capsicum strains could now be considered to be X. axonopodis pv. vesicatoria and the tomato strains X. vesicatoria.     

辣椒、番茄细菌性疮痂病及生理小种鉴定

 近3年,从北京、山西、内蒙、新疆和云南等地的辣椒和番茄病株上分离到19个菌株,经致病性测定和细菌学鉴定,确定这19个菌株为甘蓝黑腐黄单胞菌疮痂致病变种(Xanthomonas campestris pv.vesicatoria(Doidge) Dye,1978)。供试19个菌株在国内首次采用国际标准鉴别寄主进行了生理小种鉴定。其中,3个菌株为番茄小种1(XcvT race1),仅存在于北京地区,其它16个菌株均属于辣椒-番茄小种3(XcvPT race3),分布广,为我国优势小种。     

植物土传病原菌拮抗细菌的筛选与鉴定

从作物根际土壤中分离到1056株细菌，筛选出7个具有较强拮抗活性的菌株。室内测定对水稻纹枯病菌、辣椒疫病菌、瓜果腐霉、油菜菌核病菌、棉花枯萎病菌、茄根腐病菌、棉花黄萎病菌、番茄青枯病菌、甘蓝黑腐病菌等重要土传病原菌有较强拮抗作用；温室盆栽试验对番茄青枯病表现出较好防效，其中以BOH2和OH11效果较为明显，防效分别为90．9％和86．4％。通过形态观察、生理生化试验和16SrDNA序列分析，确定OH11为产酶溶杆菌。BOH3为荧光假单胞菌，其余5个菌株为不同芽孢杆菌。     

Identification of Isolates that Cause a Leaf Spot Disease of Brassicas as Xanthomonas campestris pv. raphani and Pathogenic and Genetic Comparison with Related Pathovars

ABSTRACT Twenty-five Xanthomonas isolates, including some isolates received as either X. campestris pv. armoraciae or pv. raphani, caused discrete leaf spot symptoms when spray-inoculated onto at least one Brassica oleracea cultivar. Twelve of these isolates and four other Xanthomonas isolates were spray- and pin-inoculated onto 21 different plant species/cultivars including horseradish (Armoracia rusticana), radish (Raphanus sativus), and tomato (Lycopersicon esculentum). The remaining 13 leaf spot isolates were spray-inoculated onto a subset of 10 plant species/cultivars. The leaf spot isolates were very aggressive on several Brassica spp., radish, and tomato causing leaf spots and dark sunken lesions on the middle vein, petiole, and stem. Based on the differential reactions of several Brassica spp. and radish cultivars, the leaf spot isolates were divided into three races, with races 1 and 3 predominating. A differential series was established to determine the race-type of isolates and a gene-for-gene model based on the interaction of two avirulence genes in the pathogen races and two matching resistance genes in the differential hosts is proposed. Repetitive-DNA polymerase chain reaction-based fingerprinting was used to assess the genetic diversity of the leaf spot isolates and isolates of closely related Xanthomonas pathovars. Although there was variability within each race, the leaf spot isolates were clustered separately from the X. campestris pv. campestris isolates. We propose that X. campestris isolates that cause a nonvascular leaf spot disease on Brassica spp. should be identified as pv. raphani and not pv. armoraciae. Race-type strains and a neopathotype strain for X. campestris pv. raphani are proposed.     

2株分泌型铁载体真菌对番茄青枯病的防效

为筛选对番茄青枯病防治效果好的天然产物,本文以分泌型铁载体产生菌云南木霉Trichoderma yunnanense 2-14F2和拟球孢白僵菌Beauveria pseudobassiana 2-8F2为材料,考察其铁载体活性物质对番茄青枯病的防效。采用CAS检测法及全波段紫外光吸收法判定铁载体化学结构类型及其活性。采用Sephadex-LH20凝胶划段法分离活性物质。采用平板扩散及96孔板倍半稀释法检测铁载体活性物质对青枯雷尔氏菌Ralstonia solanacearum的离体抑菌活性;采用盆栽法检测活性物质对番茄青枯病的防效。结果显示,菌株2-14F2和2-8F2产铁载体活性单位(SU)分别为62.02%和52.06%,铁载体化学结构类型均为异羟肟酸盐型(hydroxamates)。当铁载体活性物质浓度为0.15 mg/mL时,两菌株对青枯雷尔氏菌抑菌率分别为73.26%(2-14F2)和37.23%(2-8F2);对番茄青枯病的防治效果分别达到51.36%(2-14F2)与50.27%(2-8F2)。此外,当两菌株铁载体活性物质中含1 mol/L FeCl₃     

抗不同生化型青枯菌的生防菌筛选鉴定及其活性分析

为更好地利用生防菌控制青枯病危害,从不同地区的土壤中分离到569株细菌菌株,筛选到3株对5种不同生化型青枯劳尔氏菌Ralstonia solanacearum具有较强拮抗活性的菌株,其中菌株BS2004的拮抗活性最强。以BS2004的菌悬液为对照,分别测定无菌滤液、蛋白酶K及高温热处理后拮抗物质抑菌活性的变化。结果显示,蛋白酶K及高温热处理后,该菌的抑菌活性显著降低,表明其主要抑菌成分为蛋白类物质。在设施栽培条件下用生防菌BS2004菌悬液处理番茄植株,能有效控制番茄青枯病的发生,防治效果达66.75%,同时还发现,重新分离得到的青枯菌菌体数明显受到生防菌的抑制。通过对BS2004的形态、生理生化特征、脂肪酸鉴定、16S rDNA序列等进行分析,该菌株被鉴定为解淀粉芽孢杆菌Bacillus amyloliquefaciens。     

Pathogenicity assays restrict the species Xanthomonas campestris into three pathovars and reveal nine races within X. campestris pv. campestris

The species Xanthomonas campestris (Vauterin) groups bacteria associated with cruciferous plants. In order to clarify and refine the pathovar and race structures within X . campestris , 47 representative strains of six pathovars were characterized for their pathogenicity on a large host range of Brassicaceae, including all original hosts. Three diseases were observed on tested plants: (i) black rot disease on cruciferous plants; it was proposed that all strains causing black rot on at least one cruciferous plant be grouped in the single pathovar X . c . pv. campestris ; (ii) leaf spot disease caused by X . c . pv. raphani on hosts belonging to the Brassicaceae and Solanaceae; the sequenced strain 756C identified as X . c . pv. armoraciae was included in this pathovar and the existence of another leaf spot disease caused by X . c . pv. armoraciae was not supported; and (iii) bacterial blight of garden stocks caused by X . c . pv. incanae . No plants susceptible to X . c . pv. barbareae were found. Strains that did not induce any symptom on cruciferous plants tested, including their original hosts, were removed from the pathovar scheme and were named X . campestris only. Three new races were described in addition to the six races previously described within X . c . pv. campestris . The sequenced strains ATCC 33913 (CFBP 5241) and Xcc 8004 (CFBP 6650) belonged to race 3 and to race 9 (one of the new races described), respectively.     

The basis of host recognition in Fusarium oxysporum f. sp. lycopersici

Filtrates from shake-cultures of Fusarium oxysporum f. sp. lycopersici race 1, concentrated to 20% of the original volume, caused cell death in tomato leaf protoplasts from near-isogenic lines corresponding to the compatible cultivar/race reactions of whole plants. Maximum activity was found in late log phase cultures on Czapek-Dox supplemented with 2% casamino acids. Selective toxicity was associated only with the protein fraction of the culture filtrate. LD₅₀ values for susceptible Ace and Moneycross to F. oxysporum f. sp. lycopersici race 1 culture filtrates were 1·92 and 0·36 μg protein ml⁻¹. Corresponding values for cvs Royal Ace and MM161, each containing the I-gene conferring resistance to race 1, were >350. Culture filtrates from F. oxysporum f. sp. lycopersici race 2 gave LD₅₀ values of 2·34 and 2·08 μg protein ml⁻¹ on cvs Ace and Royal Ace, both susceptible to race 2. The LD⁵⁰ of cv. Ace to a non-pathogenic isolate of F. xysporum f. sp. lycopersici was > 350. Culture filtrates from non-host formae of F. oxysporum were 9–149-fold less toxic on cv. Ace. Protoplasts from Pisum sativum, Lactuca sativa, Zea mays, Gossypium barbadense and Solanum melongena, all non-hosts of F. oxysporum f. sp. lycopersici, were 6–175 times less sensitive to F. oxysporum f. sp. lycopersici filtrates than susceptible tomato. The putative toxins lycomarasmin and fusaric acid showed no differential toxicity to I⁺ and I⁻ tomato protoplasts. The results are discussed in the wider context of host-pathogen interaction in which specificity is considered as the recognition of susceptibility by a proteinaceous toxic metabolite of the pathogen. This hypothesis is further extended to include the specificity of F. oxysporum formae and races.     

Molecular mapping of hypersensitive resistance from tomato 'Hawaii 7981' to Xanthomonas perforans race T3

Bacterial spot of tomato (Solanum lycopersicum) is caused by four species of Xanthomonas. The disease causes significant yield losses and a reduction in fruit quality. Physiological races have been described with tomato race 3 (T3) corresponding to strains of Xanthomonas perforans. The breeding line Hawaii 7981 (hereafter H7981) shows a hypersensitive reaction (HR) to race T3 strains conditioned by the interaction of the host resistance locus Xv3 and the bacterial effector avrXv3. The Xv3 gene is required for H7981-derived resistance to be effective under field conditions, though its expression is subject to genetic background. The segregation of HR in F(2) populations derived from H7981 crossed to processing tomato parents OH88119 and OH7870 was studied in 331 progeny, with the two independent crosses providing validation. We screened 453 simple-sequence repeat, insertion/deletion, and single-nucleotide polymorphism markers and identified 44 polymorphic markers each for the OH88119 and OH7870 populations covering 84.6 and 73.3% of the genome, respectively, within 20 centimorgans (cM). Marker-trait analysis using all polymorphic markers demonstrated that Xv3-mediated resistance maps to chromosome 11 in the two independent crosses. Allelism tests were conducted in crosses between lines carrying Xv3 derived from H7981, Rx-4 derived from plant introduction (PI) 128216, and resistance derived from PI 126932. These allelism tests suggested that the loci conditioning HR to race T3 strains are linked within 0.1 cM, are allelic, or are the same gene.     

一株绿针假单胞菌桔黄亚种在防治番茄匍柄霉叶斑病中的应用

为筛选对番茄匍柄霉叶斑病具有良好拮抗效果的生防细菌,本研究采用稀释涂布法,从四川番茄根际土壤分离筛选到一株对番茄匍柄霉Stemphylium lycopersici（Enjoji）Yamamoto具有显著抑制效果的生防菌株YS05。经过形态学特征观察、生理生化特征、Biolog测定和多基因系统发育树分析,鉴定其为绿针假单胞菌桔黄亚种Pseudomonas chlororaphis subsp.aurantiaca。生防机理初步研究发现,菌株YS05在代谢过程中可产生蛋白酶、氢氰酸、吩嗪-1-羧酸（phenazine-1-carboxilic acid）和硝吡咯菌素（pyrrolnitrin）,具有广谱拮抗作用,能够有效抑制9种病原真菌和3种病原细菌的生长。采用二分皿法测定其挥发性物质对7种病原菌均有抑制效果。离体叶片和活体盆栽条件下,接种菌株YS05的番茄发病率和病情指数均显著降低,离体叶片防效可达71.52%,活体盆栽防效可达61.27%。综上所述,菌株YS05具有良好的生防潜力和应用前景。     

蓖麻枯萎病菌高效生防菌株LX1的筛选与鉴定

为筛选对蓖麻枯萎病菌Fusarium oxysporum Schl.具有抑制活性的生防菌株,本研究利用利福平选择性培养法,从花生根际土壤中筛选了一株对蓖麻枯萎病菌具有显著抑制活性的菌株 LX1。菌株 LX1在对峙培养中出现了明显的抑菌圈,孢子萌发试验显示孢子萌发抑制率为55%～78%,普通光学显微镜观察显示,菌株LX1可显著抑制F. oxysporum菌丝生长,明显造成菌丝分支增加,部分菌丝顶端和中央膨大,随着时间的延长,畸变结构和程度加剧。通过对菌株LX1的形态特征、染色反应、培养性状、生理生化测试以及16S rDNA分析,鉴定该菌株为枯草芽孢杆菌。本研究结果为菌株LX1应用于蓖麻枯萎病菌的生物防治及生物制剂的开发提供了依据。     

Multiphasic analysis of xanthomonads causing bacterial spot disease on tomato and pepper in the Caribbean and central america: evidence for common lineages within and between countries

ABSTRACT Four hundred thirty-three xanthomonad strains isolated from tomato or pepper plants from 32 different fields in four Caribbean and Central American countries were screened for the ability to hydrolyze starch and sodium polypectate and for resistance to copper and streptomycin. Of these, 95 representative strains were further characterized by various phnetic tests, and 63 of these strains were then analyzed by genomic fingerprinting. Most of the strains (>90%) were tolerant to copper. However, there was much more variability in sensitivity to streptomycin. All strains in Guadeloupe and 93% of the strains in Barbados were sensitive to streptomycin. The majority of strains were typical Xanthomonas campestris pv. vesicatoria group A strains. In Barbados, however, a unique group of strains was identified that was serologically similar to group A strains but was amylolytic. These strains were designated A1. The occurrence of X. campestris pv. vesicatoria group B strains in Central America was found to be limited to two fields in Costa Rica and one in Guatemala. No group B strains were identified in the Caribbean, in contrast to common occurrence in the central United States and in South America. T3 strains were not found in this study, despite the recent increase of such strains in Florida and Mexico. Unique strains from Costa Rica belonging to the X. gardneri group were identified. Little linkage was found among phenotypic and rep-polymerase chain reaction (rep-PCR) genomic fingerprinting profiles of the pathogens except at the species/pathovar level; strains displaying virtually identical fingerprint profiles were found to correspond to distinct races and vice versa. The rep-PCR genomic fingerprinting analyses suggest that certain lineages may have evolved or predominated in specific regions or specific countries.     

Diversity and distribution of Ralstonia solanacearum strains in Guatemala and rare occurrence of tomato fruit infection

Fifty-nine Ralstonia solanacearum isolates from diverse crops and regions were collected and characterized to determine the distribution and diversity of this soilborne pathogen in Guatemala. Three distinct types were present: a phylotype I, sequevar 14 strain, probably originating from Asia, infecting tomatoes and aubergines at moderate elevations; a phylotype II, sequevar 6 strain of American origin causing Moko disease in lowland banana plantations; and a phylotype II, sequevar 1 (race 3 biovar 2) strain causing brown rot on potatoes, Southern wilt of Pelargonium spp. and bacterial wilt of greenhouse tomatoes at high elevations. These data on strain diversity will inform effective regional efforts to breed for wilt resistance. A sensitive enrichment method did not detect the pathogen in fruits from naturally infected commercial tomato plants in Guatemalan fields and greenhouses, although it was detected in 6% of fruits from a wilt-resistant hybrid. Low numbers of R. solanacearum cells were also infrequently detected in fruits from plants artificially inoculated in the growth chamber with either race 3 biovar 2 or a phylotype II tomato strain.     

Resistance in Lycopersicon esculentum Intraspecific Crosses to Race T1 Strains of Xanthomonas campestris pv. vesicatoria Causing Bacterial Spot of Tomato

ABSTRACT We used molecular markers to identify quantitative trait loci (QTL) that confer resistance in the field to Xanthomonas campestris pv. vesicatoria race T1, a causal agent of bacterial spot of tomato. An F(2) population derived from a cross between Hawaii 7998 (H 7998) and an elite breeding line, Ohio 88119, was used for the initial identification of an association between molecular markers and resistance as measured by bacterial populations in individual plants in the greenhouse. Polymorphism in this cross between a Lycopersicon esculentum donor of resistance and an elite L. esculentum parent was limited. The targeted use of a core set of 148 polymerase chain reaction-based markers that were identified as polymorphic in L. esculentum x L. esculentum crosses resulted in the identification of 37 markers that were polymorphic for the cross of interest. Previous studies using an H 7998 x L. pennellii wide cross implicated three loci, Rx1, Rx2, and Rx3, in the hypersensitive response to T1 strains. Markers that we identified were linked to the Rx1 and Rx3 loci, but no markers were identified in the region of chromosome 1 where Rx2 is located. Single marker-trait analysis suggested that chromosome 5, near the Rx3 locus, contributed to reduced bacterial populations in lines carrying the locus from H 7998. The locus on chromosome 5 explained 25% of the phenotypic variation in bacterial populations developing in infected plants. An advanced backcross population and subsequent inbred backcross lines developed using Ohio 88119 as a recurrent parent were used to confirm QTL associations detected in the F(2) population. Markers on chromosome 5 explained 41% of the phenotypic variation for resistance in replicated field trials. In contrast, the Rx1 locus on chromosome 1 did not play a role in resistance to X. campestris pv. vesicatoria race T1 strains as measured by bacterial populations in the greenhouse or symptoms in the field. A locus from H 7998 on chromosome 4 was associated with susceptibility to disease and explained 11% of the total phenotypic variation. Additional variation in resistance was explained by plant maturity (6%), with early maturing families expressing lower levels of resistance, and plant habit (6%), with indeterminate plants displaying more resistance. The markers linked to Rx3 will be useful in selection for resistance in elite x elite crosses.     

防治南方根结线虫的根瘤内生细菌的筛选及鉴定

根结线虫的严重发生给农作物生产造成巨大经济损失,生物防治是控制该病的一种安全有效措施。本研究从448株根瘤内生细菌分离株中筛选出3株对南方根结线虫有较好防效的生防细菌Sneb1994、Sneb1995和Sneb2000,3个菌株发酵滤液对南方根结线虫2龄幼虫（J2）均有高毒杀活性,处理24 h校正死亡率达85%以上,同时对卵孵化具有抑制作用,并可促进番茄种子生长。盆栽试验结果表明,3个菌株发酵液灌根处理均能显著减少根结数和卵囊数,相对防效为37.95%~44.51%,同时促进番茄植株的生长。经形态特征、生理生化特性和16S rDNA序列分析,鉴定菌株Sneb1994、Sneb1995和Sneb2000分别为氧化微杆菌Microbacterium oxydans、油菜假单胞菌Pseudomonas brassicacearum和蜡样芽胞杆菌Bacillus cereus。氧化微杆菌为首次报道对根结线虫有效的生防菌株。这些根瘤内生细菌菌株的发掘,为植物线虫病害的生物防治提供了新的微生物资源。     

Comparisons of single versus multiple bacterial species on biological control of anthurium blight

ABSTRACT Effects of single versus multiple biological control agents (BCAs) on suppression of bacterial blight of anthurium were studied using a bioluminescent strain (V108LRUH1) of Xanthomonas campestris pv. dieffenbachiae. When five BCAs (GUT3, GUT4, GUT5, GUT6, and GUT9) were coinoculated in various combinations with V108LRUH1 into filter-sterilized guttation fluids of anthurium plants, a mixture of all five strains or four strains without GUT9 was most inhibitory to V108LRUH1. None of the individual BCAs inhibited V108LRUH1 in the guttation fluid. When BCAs were sprayed at congruent with10(8) CFU/ml on the foliage of a susceptible cultivar 1 day prior to inoculation with V108LRUH1, GUT6 alone and any mixtures containing GUT6 were highly effective in suppressing wound invasion and subsequent leaf infection by V108LRUH1. When tested on several cultivars that differed in susceptibility to the disease, the mixture of five strains or four strains without GUT9 consistently suppressed leaf infection regardless of the cultivars. In some cultivars, BCAs completely suppressed both wound and hydathode invasion by V108LRUH1, resulting in no infection in many leaves. These results indicate that application of bacterial mixtures provides anthurium cultivars with bacterial communities suppressive to X. campestris pv. dieffenbachiae. The results also suggest that selecting an effective mixture of BCAs first and then removing ineffective strains may be a better general approach to finding the most effective BCAs than finding individual strains and combining them.     

7种检疫性植物病原黄单胞菌III型分泌系统和效应蛋白编码基因的差异性分析

 γ-变形菌纲(γ-Proteobacteria)的黄单胞菌属(Xanthomonas)的大多数种类可引起植物病害,多数是我国检疫对象。与其他革兰氏阴性植物病原细菌一样,植物病原黄单胞菌可通过高度保守的III型分泌系统(type-III secretion system, T3SS)分泌效应蛋白(T3SS-secreted effectors, T3SEs)进入植物细胞,在非寄主植物和抗病寄主植物上产生过敏反应(hypersensitive response, HR)以及在感病寄主植物上具有致病性。尚不清楚哪些种类的黄单胞菌具有T3SS和缺少哪些T3SE是否可作为检疫的依据。搜集7种检疫性植物病原黄单胞菌,通过PCR和Southern杂交试验结果发现:香蕉细菌性青枯病菌(X. campestris pv. musacearum)的ICMP287和ATCC49084菌株、甘蔗流胶病菌(X. axonopodis pv. vasculorum)ATCC13901菌株、洋葱细菌性叶枯病菌(X. axonopodis pv. allii)的LMG576和LMG578菌株中不含有tale基因,并且ATCC13901菌株既不含有T3SS基因也不含有hpa1和xopQ基因;菜豆细菌性疫病菌(X. campestris pv. phaseoli)ATCC49119菌株不含有hpa1基因。相应地,推测含有2~12个tale基因的黄单胞菌有:大豆斑疹病菌(X. axonopodis pv. glycines)ICMP5732和ATCC43911菌株、豌豆细菌性疫病菌(X. axonopodis pv. vignicola)ATCC11648菌株、棉花细菌性角斑病菌(X. campestris pv. malvacearum)ATCC12131和(X. campestris pv. phaseoli)ATCC49119菌株。大豆细菌性斑疹病菌ATCC43911菌株尽管含有hpa1、xopQ和hrcC基因,但在非寄主烟草上不能激发HR反应;而甘蔗流胶病菌ATCC13901菌株不含有hpa1、xopQ和hrcC基因,却激发烟草产生HR反应。这些结果对于分析比较不同植物病原黄单胞菌的致病性因子和设计特定的植物检疫靶点提供了科学线索。     

番茄叶霉病菌拮抗菌的筛选及其发酵条件

从蔬菜地田间土壤中筛选出对番茄叶霉病具拮抗作用的22株菌株,其中14株细菌,8株放线菌。孢子萌发抑制试验表明,细菌X14菌株和放线菌F15菌株的发酵液及其离心后上清液对番茄叶霉病菌孢子萌发抑制率均达100%。菌丝生长抑制试验表明,X14菌株对番茄叶霉病菌的平均抑菌直径为22.2mm,初步鉴定该菌株为芽孢杆菌。利用离体叶片法测定对番茄叶霉病致病性的影响,结果表明,X14菌株对番茄叶霉病的防治效果为79.8%~89.0%。对X14菌株发酵条件研究,最佳碳源为淀粉,最佳氮源为NH4NO3,最适pH为6.0。