水稻稻瘟病拮抗细菌的筛选与防治初探

从水稻作物根际分离筛选得到的 2 0 8个细菌菌株对稻瘟病病菌具有较强的抑制能力 ;抑菌圈直径在 2 0mm以上的有 65株。选用室内拮抗性较稳定的 8株细菌菌株经两年的田间防病试验 ,不同菌株间防效差异显著 ,其中两年防效较好的菌株是Ma 3 2 (蜡状芽孢杆菌 )和Xi 55(枯草芽孢杆菌 )。一些菌株处理种子对秧苗生长有促进作用。

一株家蚕病原物拮抗细菌的分离与鉴定

从泰山土壤中分离获得 1株对多种家蚕病原细菌和真菌具有强烈拮抗作用的细菌。经培养特征观察、生理生化指标测定、DNAG +Cmo1%值测定、16SrDNA碱基序列测定和同源性分析 ,鉴定该菌株属于类芽孢杆菌 ,定名为PaenibacillusCp-S316。该菌株已在GeneBank注册 ,注册号为AY2 92 989。初步研究发现 ,该菌株对家蚕生长发育无不良影响 ,发酵性状优异 ,抗菌物质耐高温 ,对盐酸和硫酸稳定 ,易于提取分离 ,具有一定开发价值。

Screening for Siderophore-producing Endophytic Bacteria against Fusarium oxysporum

[Objective] With a strain of endophytic antagonistic bacteria with good disease control efficacy in pot as trial material,the paper was to explore the antagonistic mechanism of siderophore against Fusarium oxysporum.[Method] Whether the strain produced siderophore and its fluorescent property was judged by MSA（Modified Sugar-Aspartic acid） plate.The siderophore activity of strains in liquid MSA medium was determined through the absorbance values at particular wavelength.The inhibition effects of it siderophore against F.oxysporum under different concentrations of Fe3＋ were compared.Then the strain was preliminarily identified based on morphological,physiological and biochemical characteristics [Result] The fluorescent siderophore produced by the endophytic bacteria in MSA medium had a strong inhibition effect against F.oxysporum.With the increasing concentration of Fe3＋,the inhibition effect against the pathogen weakened.The preliminary analysis showed these strains belonged to Bacillus.[Conclusion] Bacillus could compete the absorption of Fe3＋ by secreting siderophore to inhibit the growth of F.oxysporum.

Antagonistical Mode of Pichia membranefaciens to Rhizopus stolonifer in Wounds of Peach Fruit

膜醭毕赤酵母 (PichiamembranefaciensHansen)是本实验室从果实上分离获得的一种能有效防治桃果实采后软腐病的拮抗菌。本文将P .membranefaciens与葡枝根霉 (Rhizopusstolonifer)在桃果实伤口部位共培养 2 4h后 ,用扫描电子显微镜观测了它们的拮抗作用。结果表明 ,在有病原菌的地方聚集了大量的酵母拮抗菌 ,而且拮抗菌紧密地吸附在病原菌的菌丝体上。结合以前的研究结果可以推断 ,P .membranefaciens主要通过与病原菌进行营养和空间的竞争 ,紧密地吸附在病原菌菌丝体上分泌能降解病原菌细胞壁的水解酶 (如几丁质酶和 β_1,3_葡聚糖酶 ) ,并可能诱导寄主产生抗性 ,从而抑制桃软腐病的发生。

Screening and Identification of Antagonistic Bacteria against Soil-borne Diseases of Tomato

[Objective] This study was conducted to screen resources for biological control of soil-borne diseases of tomato. [Method] Antagonistic bacteria against Ral- stonia solanacearum and Fusarium oxysporum were isolated and purified from soil samples by dilution plating and confronting incubation on PDA plates, then the antibacterial spectrum and metabolic secretions of the bacterial isolates were measured, and their species were identified by establishing phylogenetic trees with Neighbor-Joining method. [Result] From the 60 soil samples, 10 of 59 antagonistic bacteria isolates against R. solanacearum showed inhibition zone diameter 〉 25 mm, and 4 of 30 strains against F. oxysporum exhibited inhibition rate 〉 30%. The bacteriostatic substances and antibacterial spectrum analysis on above 14 isolates revealed that four strains N23-2, N58-2, NF59-3 and NF61-1 had good antago- nism against pathogenic bacteria and fungi; 12 strains produced cellulose, 11 strains produced proteases, and 6 strains produced siderophores. The molecular identification experiments indicated that four strains were members of Paenibacillus, three strains of Streptomyces, three strains of Pseudomonas strains, and four strains of Bacillus strains.

Purification of Antifungal Protein from Biocontrol Strain XG-1 against Fusarium Wilt of Watermelon

[Objective] This study aimed to isolate and purify the antifungal protein against Fusarium oxysporum f. sp niveum （FON） from Bacillus subtilis XG-I. [Method] The crude protein was obtained by using fractional precipitation with am- monium sulfate, then chromatography using both DEAE-Sepharose FF anion ex- change and Sephacryl S-100 gel filtration columns was adopted for further purifica- tion of the protein, followed by MALDI-TOF-MS analysis of the structure of the at- tained protein which was antagonistic to FON. [Result] The result from MALDI-TOF- MS analysis suggested that the antifungal protein extracted from strain XG-1 was highly homologous to the flagellin （gi114278900） deriving from B. subtilis, with a pro- tein score of 248 and a coverage rate of 63%. It was thus speculated that this an- tagonistic protein was a kind of flagellin with a molecular mass of 30.6 kD. [Conclusion] The results from this study provide theoretical basis for clarifying the mechanisms of the action of strain XG-1 as well as important references for the preven- tion and control of watermelon Fusarium wilt.

Proteomic study of three component interactions: plant, pathogens and antagonistic fungi

The molecular factors involved in the three-way interaction between plant, pathogenic fungi and antagonistic/biocontrol fungi, such as Trichoderma, are still poorly understood, even if they represent a matter of interest for improving crop management and developing new strategies for plant diseases control. The aim of this work is to investigate the components involved in this interaction and, for this purpose, a proteomic approach was used. 2-D maps of the protein extracts from the single components in various interactions between plants (potato, bean, tobacco or tomato), pathogens (Botrytis cinerea, Rhizoctonia solani or Pythium ultimum) and biocontrol fungi (Trichoderma atroviride strain P1 or Trichoderma harzianum strain T22) were obtained. The proteome of each partner was collected separately and extracted by acetone precipitation in presence of trichloroacetic acid and a reducing agent (DTT). The extracted proteins were separated by isoelectrofocusing (IEF), using IPG (Immobilized pH gradient) strips, followed by SDS-PAGE. In order to improve resolution the separations were performed both on wide than narrow pH range and on different gel lengths. Differential spots were noted in the proteome of the three-way interaction when compared to each single component. These were further characterized by mass spectrometry and in silico analysis with the aim of identifying and cloning the relative genes. During the in vitro interaction of T. harzianum strain T22 with tomato and the culture filtrate or cell walls of pathogens, the spot number was higher than in the presence of pathogen biomass. In terms of Trichoderma differential proteins displayed on 2D gels, the most important changes were obtained in the presence of P. ultimum . During the in vivo interaction with tomato, the antagonist proteome changed much more in presence of soilborne fungi R. solani and P. ultimum than with the foliar fungus B. cinerea, both in terms of total and increased or novel spots. In silico analysis of some of those spots revealed homology with intracellular enzymes (GTPases, hydrolases) and with stress-related proteins (heat shock proteins HSP70, bacteriocin cloacin). Specific proteins in the plant proteome, i.e. pathogenesis-related proteins, have been identified during the in vivo interaction of bean with R. solani and T. atroviride strain P1. This is in agreement with the demonstrated ability of these beneficial fungi to induce plant systemic disease resistance by activating expression of defence-related genes. Proteins extracted from T. atrovride strain P1 which were analysed by mass spectrometry, revealed some interesting homologies with a fungal hydrophobin of Pleurotus ostreatus and an ABC transporter of Ralstonia metallidurans. These could represent molecular factors involved in the antagonistic mechanisms of Trichoderma and play a role in the three-way interaction with the plant and other microbes.

The Antagonistic Activity of the Lactic Acid Bacteria （Streptococcus thermophilus, Bifidobacterium bifidum and Lactobacillus bulgaricus） against Helicobacter pylori Responsible for the Gastroduodenals Diseases

The lactic acid bacteria are used in fermentation and the bio conservation of food, because it produces organic acids and other antibacterial substances as the bacteriocin to inhibit certain pathogenic stocks. In order to demonstrate the inhibitory effect of these bacteria, we studied the antagonistic capacity of the two leavens of yoghourt （Streptococcus thermophilus and Lactobacillus bulgaricus） and Bifidobacterium bifidum with respect to Helicobacterpylori by the method of diffusion by discs. Their interactions led to the emergence of large zones of inhibition. Bifidobacterium bifidum led the greatest halo with a diameter of 0.8 cm by contribution to Lactobacillus bulgaricus （0.5 cm） and Streptococcus thermophilus （0.3 cm）. The additional tests were required to know the exact nature of the inhibitors. The results showed that the antibacterial peptides （bacteriocins） have a narrow spectrum of activity against the species pathogen compared to that caused by acidity.

Controlling of Fusarium Wilt of Cucumber by Antagonistic Bacteria

Studying about the effect of four pseudomonas and two serratia isolates on growth of Fusarium oxysporum showed that, Pseudomonasfluorescens No.2 ＆ No.3 and Serratia marcescens No.2 gave highest inhibition zones which were 37.33, 35,00 and 31.33 mm, respectively. Evaluating about the effect ofpseudomonas and serratia culture filtrates at three concentrations （10, 25, 50%） on the linear growth and spore germination ofFusarium oxysporum revealed that, all filtrates of the tested isolates reduced the mycelial growth and spore germination of F. oxysporum. All filtrates of the tested isolates at 50% concentration completely inhabited spore germination ofF. oxysporurn. Culture filtrates of Pseudomonasputida and Serratia marcensens No.2 at 50% concentration reduced the mycelial growth of F. oxysporurn by 80.74 and 80.37% respectively. In this respect, all tested isolates made lysis to mycelial of F. oxysporum except Pseudornonasfluorescens No.3. Effect of treating cucumber seeds with cell suspension of pseudomonas and serratia isolates on incidence of Fusarium wilt disease revealed that all isolates were effective in reducing disease incidence and disease severity and increasing the percentage of healthy plants compared to the control. Pseudornonasfluorescens No.3 and Serratia marcensens No.2 were the best isolates and completely prevented the disease incidence.