Identification of Mulberry Bacterial Blight Caused by Klebsiella oxytoca in Bazhong,Sichuan,China

To provide a scientific basis for controlling mulberry bacterial blight in Bazhong,Sichuan,China(BSC),this study aimed to isolate and purify pathogenic bacteria from diseased branches of mulberry trees in the region and to clarify their taxonomic status using morphological observation,physiological and biochemical detection,molecular-level identification,and the construction of a phylogenetic tree.A total of 218 bacterial strains were isolated from samples of diseased mulberry branches.Of these,7 strains were identified as pathogenic bacteria based on pathogenicity tests conducted in accordance with Koch’s postulates.Preliminary findings from the analysis of the 16S rRNA sequence indicated that the 7 pathogenic bacteria are members of Klebsiella spp.Morphological observation revealed that the pathogenic bacteria were oval-shaped and had capsules but no spores.They could secrete pectinase,cellulase,and protease and were able to utilize D-glucose,D-mannose,D-maltose,and D-Cellobiose.The 7 strains of pathogenic bacteria exhibited the highest homology with Klebsiella oxytoca.This study identifies Klebsiella oxytoca as the causative agent of mulberry bacterial blight in BSC,laying the foundation for the prevention and control of this pathogen and further investigation into its pathogenic mechanism.

Comparative Assessment of Genetic Diversity of Peanut (Arachis hypogaea L.)Genotypes with Various Levels of Resistance to Bacterial Wilt Through SSR and AFLP Analyses

Bacterial wilt （BW） caused by Ralstonia solanacearum is an important constraint to peanut （Arachis hypogaea L.） production in several Asian and African countries, and planting BW-resistant cultivars is the most feasible method for controlling the disease. Although several BW-resistant peanut germplasm accessions have been identified, the genetic diversity among these has not been properly investigated, which has impeded efficient utilization. In this study, the genetic relationships of 31 peanut genotypes with various levels of resistance to BW were assessed based on SSR and AFLP analyses. Twenty-nine of 78 SSR primers and 32 of 126 AFLP primer combinations employed in this study were polymorphic amongst the peanut genotypes tested. The SSR primers amplified 91 polymorphic loci in total with an average of 3.14 alleles per primer, and the AFLP primers amplified 72 polymorphic loci in total with an average of 2.25 alleles per primer. Four SSR primers （14H06, 7G02, 3A8, 16C6） and one AFLP primer （P1M62） were found to be most efficient in detecting diversity. The genetic distance between pairs of genotypes ranged from 0.12 to 0.94 with an average of 0.53 in the SSR data and from 0.06 to 0.57 with an average of 0.25 in the AFLP data. The SSR-based estimates of the genetic distance were generally larger than that based on the AFLP data. The genotypes belonging to subsp, fastigiata possessed wider diversity than that of subsp, hypogaea. The clustering of genotypes based on the SSR and AFLP data were similar but the SSR clustering was more consistent with morphological classification ofA. hypogaea. Optimum diverse genotypes of both subsp, hypogaea and subsp.fastigiata can be recommended based on this analysis for developing mapping populations and breeding for high yielding and resistant cultivars.

Effects of Seedling-cake Integrated with Arbuscular Mycorrhizal Fungi on Resistance of Bacterial Wilt in Pepper (Capsicum annuum L.)

[Objective] The aim was to investigated the effects of seedling-cake with AMF on resistance of pepper （Capsicum annuum L.） to bacterial wilt. [Methods] A pot experiment was conducted to evaluate the dynamics of mycorrhizal colonization, disease index, morbidity rate, control effect and several agronomic traits of mycorrhizal seedlings after inoculation with RS in two kinds of seeding-cakes with AMF. [Results] Two kinds of seedling-cakes formed steadily mycorrhizal colonization after inoculation, which enhanced disease-resistance and decreased morbidity rate and disease index in different degrees. What’s more, the performance of self-made seedling-cakes was obviously better than that of commercial seedling-cakes. [Conclusions] Self-made seedling-cakes with AMF have superior performance on bacterial wilt resistance of pepper, which should be made use of in other crops extensively.

Effect of Fertilization on Tomato Bacterial Wilt Biocontrolling and Soil Health Restoration Using FAME Analysis

The effect of applying biological organic fertilizer(BOF)on bacterial wilt incidence of tomato and soil microbial community under continuous cropping was studied. The results showed that all the tomatoes were infected by bacterial wilt in the control. The infection rates of tomatoes in the treatments with un-composted BOF and decomposed BOF were 55 and 50% respectively. Fatty acid methyl esters(FAME)analysis indicated soil microbial community changed sensitively after applying BOF. Soil FAME total content, relative fungi content and ratio of fungi to bacteria were significantly increased in both BOF treatments. The soil odd-number fatty acid proportion changed after applying BOF, aC15 : 0, iC17 : 0 decreased, while cyC17 : 0 increased in soil odd-number fatty acid proportion. BOF application would strengthen soil health and disease suppression. The content of C16 : 1 11c in soil microbial community was obviously increased after decomposed BOF application. It indicated that the growth of AM fungi could be enhanced with decomposed BOF application. FAME microbial biomarkers could be used for an indicator of soil health and disease suppression. Odd-number fatty acid proportion was a sensitive indicator of the effect of applying un-composted and decomposed BOF respectively on soil health regulating.

Control Effect of Different Microbial Agents on Tobacco Bacterial Wilt

[ Objectives] The paper was to study effects of different microbial agents on occurrence of tobacco bacterial wilt. [ Methods I Antagonistic strains against Ralstonia solanacearum were screened from in situ soil, and prepared into four different combinations. The number of soil microorganisms and incidence of bacterial wilt of four combinations were observed. [ Results] The incidence rate of bacterial wilt treated by microbial agent OR-1 was significantly lower than that in control, and the control efficacy against bacterial wilt reached 67%. [ Conclusions] Application of microbial agent OR-1 could effectively reduce the incidence rate and disease index of bacterial wilt, thus improving the quality of tobacco.

Effects of Bio-organic Fertilizer with Antagonistic Bacteria against Tobacco Bacterial Wilt on Soil Microbial Communities and Disease Resistance of Tobacco

In this study, the effects of bio-organic fertilizer with antagonistic bacteria against tobacco bacterial wilt on soil microbial communities and disease resist- ance of tobacco were investigated by field experiment. The results showed that the incidence of tobacco bacterial wilt in bio-organic fertilizer treatments （T3 and T4） decreased remarkably among four treatments in the field. Compared with the local conventional fertilization group, the incidence of tobacco bacterial wilt was re- duced by 21.9% and 25.0% in T3 and T4, respectively ; the yield of flue-cured tobacco was improved by 5.7% and 5.3%, respectively ; the proportion of mid- high grade tobacco leaves increased by 2.3% and 2.6%, respectively. After application of bio-organie fertilizer with antagonistic bacteria against tobacco bacterial wilt, rhizosphere soil microbial communities exhibited vast amount and abundant species ; the amount of rhizosphere soil bacteria of infected tobacco plants was im- proved by 218.5% with fewer species. It could be concluded that the application of bio-organic fertilizer with antagonistic bacteria against tobacco bacterial wilt could improve the ecological environment of tobacco field, inhibit the growth of pathogenic bacteria, decrease the incidence of tobacco bacterial wilt, and enhance the quality of flue-cured tobacco. This study laid the foundation for further ecological prevention and control of soil-borne diseases of tobacco.

Extraction of Flavonoids and Polyphenols from Coronatinetreated Mulberry Leaves and Evaluation of Their Antibacterial Activity against Pseudomonas syringae

Flavonoids and polyphenols are the main secondary metabelites in mulberry leaves, which have multiple biological functions such as antibacterial activity, free radical scavenging activity and antioxidant activity. In this paper, the parameters for the extraction of flavonoids and palyphenols from eoronatine （ COR）- treated mulberry leaves were optimized, and the components of the extracts were analyzed by high performance liquid chromatography （ HPLC）, and their antibacterial activity against Pseudomonas syringae, which causes mulberry bacterial blight, was evaluated in vitro to explore the mechanism how COR acted as a plant immune elicitor to improve the resistance of mulberry to bacterial blight. The results showed that ethanol concentration of 70%, extraction temperature of 80 ℃, extraction duration of 6 h and solid-liquid ratio of 1 ： 40 were the optimal parameters for the extraction of flavonoids and polyphenols from mulberry leaves. Flavonoids and polyphenols were mostly distributed in ethyl acetate and n-butyl alcohol fractions which showed better antibacterial activity than petroleum ether, dichloromethane and water fractions. Meanwhile, the antibacterial activity of ethyl acetate and n-butyl alcohol fractions of COR-treated mulberry leaves were more evident than that of water-treated mulberry leaves. Scanning electron microscopy showed that flavonoids and pelyphenols inhibited the growth of pathogenic bacteria by changing cell morphology, damaging cell wall structure and unbalancing cell osmotic pressure. HPLC analysis confirmed the contents of morin, quercetin, quereitrin, kaempferol, chlorogenic acid and rutin were increased to a different extent in COR-treated mulberry leaves. So it can be concluded that the increased contents of flavonoids and polyphenols induced by COR may be one of the reasons for the enhanced resistance to mulberry bacterial blight.

Effect of Integrated Use of Inorganic Fertilizer and Organic Manures on Bacterial Wilt Incidence (BWI) and Tuber Yield in Potato Production Systems on Hill Slopes of Central Kenya

Bacterial wilt (BW) caused by Ralstonia solanacearum is one of the most damaging diseases of potato (Solanum tuberosum L.) in Kenya and worldwide. In Kenya, potato tuber yield losses due to BW infection are estimated at 50 ~ 100%. Low soil fertility is also one of the most important constraints limiting potato production in central Kenya highlands. Farmers tackle this problem through use of inorganic fertilisers and organic manures, both of which amend the soil environment to influence bacterial wilt development. Undecomposed organic manures can also introduce the pathogen into a clean field. Between short rains 1999 and 2000, 10 on-farm extension-researcher- farmer-designed and farmer-managed trials were done at Kianjuki catchment in Embu District. The objective was to use farmers?participatory research approach and select the most suitable organic and inorganic fertiliser combination(s) with lowest BWI and acceptable usable tuber yields, and also to demonstrate use of some components of integrated disease management methods in reduction of disease incidence and spread. Seven treatments were proposed, presented to the farmers for discussion and the most relevant four were selected for evaluation. A newly released potato variety 慉sante?was planted during the short rains 1999 and long rains 2000. BWI didn抰 result in significant differences between treatments but the tuber yields were significantly different in short rains of 1999 and 2000. During short rains 2000, both BWI and tuber yields and unusable tubers differed significantly between treatments. The results confirmed that well- decomposed manures or manures from pathogen-free areas can be used in combination with inorganic fertilisers to improve soil fertility and potato tuber yields in smallholder farms without influencing BWI. Use of certified seed tubers in pathogen free field and following recommended field sanitation measures resulted in apparently bacterial wilt free crop. Considering the high cost of inorganic fertiliser and its negative effects on the environment, reduced usage at half the recommended rates combined with half rates of FYM is feasible option friendly to the farmer, soil and environment. The interviewed farmers also ranked the option as the most appropriate combination for soil fertility improvement for potato production in smallholder farms.

Integrated Biological Control of Tobacco Bacterial Wilt (<i>Ralstonia solanacearum</i>) and Its Effect on Rhizosphere Microbial Community

Biological control agents and soil amendments have been applied to control tobacco bacterial wilt, but the mechanism is not well-known. In the present study, a field experiment was performed to investigate the soil physicochemical properties, disease index (DI) and disease incidence of tobacco bacterial wilt, and rhizosphere microbial community. The results show that the control efficacy of single application of antagonistic bacteria and calcium cyanamide was 46.43% and 51.92%, respectively. While the combined control efficacy of antagonistic bacteria and calcium cyanamide was 65.79%. Besides, the combined application of antagonistic bacteria and calcium cyanamide could increase soil pH, total N alkaline N, and exchangeable Ca, which were negatively associated with the microbial diversity, soil-borne pathogenic microorganisms, and incidence of tobacco bacterial wilt. Additionally, the combination of antagonistic bacteria and calcium cyanamide can improve the proportion of some antagonistic microbial species, and these antagonistic microbial species were inversely associated with the DI of tobacco bacterial wilt. In conclusion: The integrated measure may influence soil microbial communities through enhancing soil physicochemical properties and rebuild healthy soil microbial community structure to mitigate tobacco bacterial wilt. The current study presented valuable insights into the mechanisms enhancing soil health in the integrated measure.

Testing of Bacterial Endophytes from Non-Host Sources as Potential Antagonistic Agents against Tomato Wilt Pathogen <i>Ralstonia solanacearum</i>

The study was taken up with the objective of testing whether the endophytic organisms isolated from crops that are normally non-hosts to the bacterial wilt pathogen Ralstonia solanacearum possessed pathogen-antagonistic activity and to evaluate the selected isolates for the alleviation of wilt disease in the target tomato crop through horizontal movement of promising organisms. Sixteen endophytic bacteria (EB) isolated from the micropropagated cultures of grape, watermelon and papaya were tested for potential antagonistic effects against R. solanacearum tomato isolate “NH-01” through agar-well diffusion assay. Enterobacter cloacae from papaya (EB-11) displayed the maximum antagonistic effect followed by Bacillus subtilis (EB-06) and B. flexus (EB-07) from watermelon and B. pumilus (EB-02) from grape. Testing the above organisms for crop protection through seed fortification of susceptible tomato cv. Arka Vikas at sowing in R. solanacearum inoculated (Ral+) organic cocopeat showed EB-02 and EB-11 promising (33% and 32% survival, respectively, four weeks after sowing against 15% in Ral+ control). A second trial showed 37%, 28%, 21% and 55% seedling survival 6 weeks after sowing for EB-02, EB-06, EB-07 and EB-11 respectively, compared to 2.5% in non-treated control. Assessing the four endophytes for crop protection in Ral+ sick-soil through seedling fortification at transplanting indicated less disease incidence in treated sets (40%, 40%, 20% and 20% survival, respectively, six weeks after transplanting) over non-fortified control (5%). Endophytic fortification of seedlings through hypocotyl inoculation showed some systemic resistance induction upon seedling transplanting to sick soil but not with petiole fortification. Seedling growth was enhanced by the isolates EB-06 and EB-07. The study thus identifies four endophytic organisms from crops unrelated to tomato possessing potential antagonistic activity against the wilt pathogen and prospects for exploitation as biocontrol agents coupled with seedling growth promotion effects.

Screening, Identification and Biocontrol Effect of Antagonistic Streptomyces Strain Tra69 against Tobacco Bacterial Wilt

[Objective] The paper was to screen the biocontrol agents with good control effects against tobacco bacterial wilt. [Method] The strains with antagonistic activity against tobacco bacterial wilt were isolated from 36 copies of rhizosphere soil sample. The strain with best inhibition effect was identified, and carried out pot test and growth-promoting experiment. [Result] The strain with best inhibtion effect was Tra69, which was identified to be Streptomyces flavogriseus. The fermentation liquid of Tra69 had good control effect against Ralstonia solanacarum, and also had good growth-promoting effect on tobacco. [Conclusion] Tra69 was the biocontrol agent with excellent control effect against R. solanacarum, which could be further developed and used in biological control against tobacco diseases.

Threshold of Soil pH for Occurrence of Tobacco Bacterial Wilt in Henan Province

[Objective]The paper was to study the relationship between spatial distribution characteristics of soil p H and bacterial wilt in tobacco-growing area in Henan Province.[Method]The 21 tobacco-growing fields infected by bacterial wilt and 91 tobacco-growing fields with no cases were selected from Henan Province,and the pH of fields was determined.[Result]Bacterial wilt mainly occurred in the tobacco-growing fields in Luoshan,Pingqiao,Queshan,Biyang,Suiping,and Xiping,and the soil pH mainly ranged from 5.5 to 6.5(accounted for 84.75%of total surveyed tobacco fields);bacterial wilt had not been found in tobacco-growing fields in Tanghe,Zhengyang,Shangcai,Sheqi,Wuyang and Chenqu,and the soil pH mainly ranged from 6.5 to 7.0(accounted for 81.25%of total surveyed tobacco fields).There were significant differences in soil pH between the fields with and without bacterial wilt,and soil pH 6.5 could be regarded as the threshold for the occurrence of tobacco bacterial wilt in Henan Province.There was extremely significantly positive linear correlation between soil p H and latitude(y=0.7375 x-17.799,R2=0.386,P<0.01).The geographical boundary of soil pH 6.5 was roughly 33°N,and bacterial wilt was less likely to occur in the north tobacco-growing areas of the boundary.[Conclusion]The study will provide a scientific basis for the prevention and control of tobacco bacterial wilt in Henan.

Genome-wide investigation of defensin genes in peanut(Arachis hypogaea L.)reveals AhDef2.2 conferring resistance to bacterial wilt

Peanut(Arachis hypogea L.)bacterial wilt(BW)is a devastating disease caused by Ralstonia solanacearum that results in severe yield and quality losses.Plant defensins are short cysteine-rich peptides with antimicrobial activity.The role of defensin genes(Ah Def)in peanut is unclear.A genome-wide investigation of Ah Def genes was undertaken,and 12 identified Ah Def genes were classified into two groups containing the gamma-thionin domain formed by four disulfide pairs:Cys1-Cys8,Cys2-Cys5,Cys3-Cys6,and Cys4-Cys7.In silico analysis revealed that Ah Def genes showed highly conserved architectural features and contained cis-elements associated with phytohormone signaling and defense responses.A highly resistant cultivar,H108(R)and a susceptible accession,H107(S)were tested by R.solanacearum inoculation.H108(R)showed fewer symptoms than H107(S)owing to inhibition of bacterial reproduction and spread in the vascular bundles of roots and stems.In a transcriptomic expression profile,Ah Def genes,particularly Ah Def1.6 and Ah Def2.2,were up-regulated in H108(R)compared with H107(S)under R.solanacearum infection and phytohormone treatment.Subcellular localization showed that the Ah Def1.6 and Ah Def2.2 proteins were both expressed specifically on the plasma membrane.Overexpression of protein fusion Ah Def2.2-YFP in Nicotiana benthamiana and peanut leaves increased resistance to R.solanacearum,suggesting its role in response to BW infection.Ah Def2.2 may be valuable for peanut resistance breeding.

Analysis of Meteorological Epidemic Factors and Soil Improvement Controlling Technology of Tobacco Bacterial Wilt (Ralstonia solanacarum)

[Objective] The paper was to analyze the meteorological epidemic factors for occurrence and prevalence of tobacco bacterial wilt （ Ralstonia solanaca- rum）, and to study control effects of different soil conditioners on the bacterial disease in Gacligongshan demonstration area of green, ecological, high quality tobac- co leaf production. [Method] The plots attacked by tobacco bacterial wilt over the years were selected and the incidence of the disease was periodically surveyed in tobacco growth period in 2012, 2103 and 2014, respectively. 10 d Effective accumulated temperature and rainfall were counted according to the meteorological data, and the relationship between meteorological factors and disease index was analyzed. The control effects of three kinds of soil conditioners ＂Zhuanggenfeng＂, refined fulvic acid and lime on tobacco bacterial wilt were tested. [ Result] The analysis results of meteorological factors showed that 10 d effective accumulated temperature and rainfall were positively correlated to disease index. The variation curve of 10 d effective accumulated temperature and rainfall reflected the change trend of disease index. The pH values were increased by 0.57, 0.50 and 0.72 respectively after applying ＂Zhuanggenfeng＂, refined fulvic acid and lime. The aver- age control effects on tobacco bacterial wilt were 60.74% -62. 18%, 53.05% -59.53%, and 48.59% -58.53%, respectively. [ Conclusion] 10 d Effective accumulated temperature and rainfall could be used as important reference for disease forecasting and controUing. The usage of soil conditioner has a certain preven- tion and control effect on tobacco bacterial wilt disease by forming soil conditions conducive to flue-cured tobacco growth but adverse to disease survival, which is an effective auxiliary method against the disease.

Effects of Integrated Technology of Medicine and Fertilizer on Bacterial Wilt of Tobacco

In order to validate the control effects of integrated technology of medicine and fertilizer on tobacco bacterial wilt,field experiments were conducted by using youmeili and tetracycline as the test agents.The results obtained were listed as follows:(1)The incidence rate of bacterial wilt ascended more quickly after July 10 th,the incidence of CK reached 78.30%on July 25 th.(2)The control effect of“youmeili 3 g+tetramycin 200 times liquid”reached 69.47%,significantly higher than other treatments.(3)The output value of“Youmeil 3 g+tetramycin 200 times liquid”served as 1635.66 yuan/667m^2,with the highest economic character in all treatments.In a word,integrated technology of medicine and fertilizer was a feasible approach to control tobacco bacterial wilt.

Improvement of Bio-Efficacy to Reduce Bacterial Wilt Complex Disease in Tomatoes through Bacillus amyloliquefaciens and Pseudomonas fluorescens under Field Conditions

Bacterial wilt complex disease of tomato(Solanum lycopersicum L.)was incited jointly by bacterial wilt pathogen Ralstonia solanacearum and Meloidogyne incognita worldwide.Bio-efficacy of bacterial antagonists i.e.B.amyloliquefaciens DSBA-11 and P.fluorescens DTPF-3 was studied against the wilt disease complex in tomato at National Phytotran facility Indian Agricultural Research Institute(IARI),New Delhi,at 26±2°C.Minimum wilt disease incidence(26.00%)with the highest bio-control efficacy(64.15%),less juvenile population(19.33 J2/g of soil)of M.incognita was recorded in the combined application of DTPF-3+DSBA-11 after 30 d of inoculation under glasshouse conditions.In a field study,minimum bacterial wilt disease incidences 19.0%and 20.4%were recorded in the bleaching powder treatment followed by mixed application of DSBA-11+DTPF-3,19.6%and 21.2%wilt incidence in 2014 and 2015 respectively.However,a reduction of root-knot gall index was recorded a maximum of 59.76%and 69.62%in DSBA-11+DTPF-3 treated plants followed by 54.88%and 60.13%over control in DTPF-3 treatment in 2014 and 2015 respectively.The yield of tomato fruit was increased over control by 17.48%and 16.97%in 2014 and 2015 respectively under field conditions.A combination of P.fluorescens DTPF-3+B.amyloliquefaciens DSBA-11 suppressed bacterial wilt and root-knot diseases and also increased the yield of the tomato fruit significantly(p<0.05)under field conditions.

Analysis of Rhizosphere Microbial Community Structure in Different Pathogenesis Stages of Watermelon Fusarium Wilt

High-throughput sequencing technique was applied to analyze the microbial community structure of rhizosphere soil at different stages of watermelon fusarium wilt to find out the difference of dominant microbial community in rhizosphere during the occurrence of watermelon fusarium wilt.Illumina-Hiseq high-throughput sequencing platform was used to sequence 16S and ITS rDNA in rhizosphere soil.The soil was named CK1 before planting,CK2 at peak stage and CK3 at stable stage.The results showed that the soil bacterial diversity was in the order of CK1>CK3>CK2,indicating no significant difference between CK1 and CK3 and a significant difference between CK2 and CK1,CK3.At the genus level,the dominant bacteria were Mizugakiibacter(21.9299%),Rhodanobacter(5.0933%),and Lactobacillus(3.1921%).The diversity of soil fungi was in the order of CK1>CK3>CK2,all showing significant differences.At the genus level,the dominant fungus was Lysurus(54.4601%),Papulaspora(12.4252%),Acrophialophora(3.1729%).The results showed that the diversity and abundance of bacteria and fungi in rhizosphere soil decreased during the peak period of watermelon fusarium wilt.With the gradual stabilization of the disease,the diversity and abundance of bacteria and fungi in rhizosphere soil recovered to a certain extent.

蚯蚓联合生防细菌Bacillus velezensis改善连作百合土壤细菌群落结构及防治枯萎病的效果

【目的】从土壤理化性状和微生物群落结构及多样性角度,研究蚯蚓联合生防细菌改善百合连作障碍的机理和效果,为湖南百合产业化种植提供技术措施。【方法】采用盆栽试验方法,在湖南长沙连续种植了两季百合(Lilium lancifolium Thunb.)。供试土壤为红壤,生防菌为Bacillales velezensis YFB3-1菌液(菌体浓度为10^(9) CFU/mL),蚯蚓为赤子爱胜蚓(Eisenia fetida)。试验设单施菌液(T1)、只接种蚯蚓(T2)、同时使用菌液和蚯蚓(T3) 3个处理,以不使用菌剂和蚯蚓为对照(CK)。调查了两茬百合产量和枯萎病病情指数,分析了第二茬百合根际土壤理化性质、细菌群落结构和多样性,以及连作百合产量与根际土壤理化性质、细菌群落和枯萎病的相关性。【结果】与第一茬相比,同一处理的第二茬百合产量都显著下降。两年连作百合产量都以同时使用菌液和蚯蚓(T3)处理为最高,分别为16464、15674 kg/hm^(2),都显著高于同年份其他处理。两年连作百合枯萎病发病率和病情指数都以T3为最低,都显著低于同年份其他处理,T3处理百合枯萎病病情指数分别为18.15(2019年)、25.00 (2020年),并且两年的防治效果(T3)都明显优于单施菌液(T1)和接种蚯蚓(T2)。相比于对照(CK),2019、2020年T3处理百合枯萎病防治效果分别为44.55%、37.66%,分别增产15.05%、14.78%。蚯蚓联合生防菌YFB3-1增加了根际土壤Rhizobiales、Flavobacteriales、Pseudomonadales、Bacillales等有益微生物的种群丰度,蚯蚓与生防菌YFB3-1在抑制连作百合枯萎病方面具有显著的互作效应,因而缓解连作百合产量下降的效果最佳。【结论】在施用芽孢杆菌生防菌剂防治连作百合枯萎病的同时,可以增施有机肥(牛粪)来提高土壤中蚯蚓数量,有效预防百合枯萎病发生,减少连作百合产量损失。

香葱伴生番茄对青枯病及根际微生态特征的影响

【目的】为明确香葱伴生对番茄青枯病及根际微生态特征的影响,揭示香葱伴生番茄降低青枯病发病率的机理。【方法】通过田间试验设置番茄单作和香葱伴生番茄2个处理,分别采集单作发病番茄(QK)、单作健康番茄(NBS)和香葱伴生番茄(BS)的根际土壤,应用16S rDNA高通量测序和液相色谱-质谱联用(LC-MS)技术,探究香葱伴生对番茄根际微生态特征的影响。【结果】田间试验结果表明,番茄单作处理和香葱伴生番茄处理的青枯病平均病株率分别为50.48、31.43%,香葱伴生番茄处理对番茄青枯病的防治效果为37.74%。16S rDNA高通量测序结果表明,BS、NBS根际土壤细菌群落相对丰度和多样性均显著高于QK;与QK相比,BS和NBS根际土壤细菌茄科雷尔氏菌属相对丰度显著降低,而芽孢杆菌属相对丰度升高;与NBS相比,BS根际土壤细菌茄科雷尔氏菌属相对丰度降低,芽孢杆菌属相对丰度显著提高。利用LC-MS技术共鉴定出番茄根际土壤代谢物586种,其中正离子模式代谢物320种,负离子模式代谢物266种,并将代谢物的功能注释至新陈代谢、遗传信息加工和环境信息加工等代谢通路;与NBS相比,BS根际土壤代谢物中的缬氨酸、L-天冬酰胺和柠檬酸的含量均显著降低,L-赖氨酸、谷氨酸、D-谷氨酰胺和异亮氨酸的含量也低于NBS,但差异未达到显著水平。【结论】香葱伴生番茄显著降低番茄青枯病田间发病率,改变了番茄根际土壤中细菌群落结构和番茄根际土壤代谢物含量。

河南省花生青枯病菌的分子鉴定及其生物学特性研究

为了明确河南省花生青枯病的病原菌分类属性,对采自河南省内不同市的30个青枯病菌株在16S rDNA序列、碳水化合物利用、致病性、演化型及序列变种等方面进行了鉴定。30个青枯病菌株的16S rDNA测序结果表明,引起河南花生青枯病的病原为茄科雷尔氏菌(Ralstonia solanacearum);所有青枯病菌株可侵染茄子、辣椒、马铃薯、烟草和番茄,但不侵染姜,属于生理小种1号;根据能否利用3种双糖和3种己醇的能力,可将其中4个菌株归于生化型Ⅱ,10个菌株归于生化型Ⅲ,16个菌株归于生化型V;所有青枯病菌株均能用多重PCR扩增得到144 bp演化型Ⅰ特异条带和280 bp茄科雷尔氏菌特异条带,表明青枯病原菌均属于茄科雷尔氏菌演化型Ⅰ,即亚洲组13;利用egl基因的系统发育进化分析表明,来自河南的30个菌株与Gx525、Ah-XnJn-12-6、HA2-1聚为一支,属于序列变种14。