Isolation and characterization of novel soilborne lytic bacteriophages infecting Dickeya spp. biovar 3 (‘D. solani’)

Nine bacteriophages infecting Dickeya spp. biovar 3 (‘Dickeya solani’) were isolated from soil samples collected in different regions in Poland. The phages have a typical morphology of the members of the order Caudovirales, family Myoviridae, with a head diameter of c. 90–100 nm and tail length of c. 120–140 nm. In host range experiments, phage ?D5 expressed the broadest host range, infecting members of all Dickeya spp., and phage ?D7 showed the narrowest host range, infecting isolates of Dickeya dadantii and ‘D. solani’ only. None of the phages was able to infect Pectobacterium spp. isolates. All phages were prone to inactivation by pH 2, temperature of 85°C and by UV illumination for 10 min (50 mJ cm^?2). Additionally, phages ?D1, ?D10 and ?D11 were inactivated by 5 m NaCl and phage ?D2 was inactivated by chloroform. Phages ?D1, ?D5, ?D7 and ?D10 were characterized for optimal multiplicity of infection and the rate of adsorption to the bacterial cells. The latent period was 30 min for ?D1, 40 min for ?D5, 20–30 min for ?D7 and 40 min for ?D10. The estimated burst size was c. 100 plaque‐forming units per infected cell. The bacteriophages were able to completely stop the growth of ‘D. solani’ in vitro and to protect potato tuber tissue from maceration caused by the bacteria. The potential use of bacteriophages for the biocontrol of biovar 3 Dickeya spp. in potato is discussed.     

抑制向日葵核盘菌菌核形成的生防菌S-16的鉴定及其生防作用研究

本研究从内蒙古包头市萨拉齐向日葵根围土壤中分离得到1株生防细菌 S-16，拮抗试验表明，菌株S-16能够显著抑制向日葵核盘菌的菌丝生长。显微镜观察发现，核盘菌菌丝生长点附近出现明显的异常分枝和囊泡状畸形，并且在距菌株S-16一定范围内核盘菌不能形成菌核。培养基内添加1%的菌株S-16发酵滤液能够有效抑制向日葵核盘菌菌核的形成。室内生测表明，2＆#215;106 cfu/mL浓度的菌株S-16菌液在离体叶片及幼苗上对向日葵菌核病的防效分别为94.62%和94.21%。通过细菌形态特征和生理生化反应，并结合API鉴定和16S rDNA序列分析，将菌株S-16鉴定为枯草芽孢杆菌Bacillus subtilis。     

利用枯草芽孢杆菌和荧光假单胞杆菌防治石榴枯萎病的初步研究

 石榴枯萎病是一种具有毁灭性的病害，生物防治是植物病害防治的可持续措施。本文在室内研究了枯草芽孢杆菌（Bacillus subtilis）菌株YZS B15和HB及荧光假单杆菌（Pseudomonas fluorescence）菌株YZS Ph02对石榴枯萎病的抑制作用，并进行了盆栽试验。结果表明，在室内，YZS B15，HB和YZS Ph02对石榴枯萎病菌甘薯长喙壳（Ceratocystis fimbriata Ellis & Halsted）具有很强的抑制作用，当它们的CFU为1×103时对甘薯长喙壳抑制效果就可达96%以上。盆栽试验的结果，YZS B15，HB及YZS B15与YZS Ph02的组合对石榴枯萎病有很好的防治效果，单用YZS Ph02的防治效果较差。随着接种后施用生物菌时间间隔的增加，防治效果逐渐下降。     

蚕桑多元化发展新方向——生物防治

蚕桑多元化发展是产业发展的方向，现主要集中于蚕桑资源综合利用，产业附加值低，产业水平亟待提高。生物防治的安全性、高技术含量和高附加值为产业的壮大提供了广阔市场前景。柞蚕卵繁育寄生蜂技术较为成熟，已经大量生产和推广。利用蚕桑副产物生产生物农药，开展生物防治，可成为蚕桑多元化发展的新方向。     

Alternative treatments for late blight control in organic potato: Antagonistic micro-organisms and compost extracts for activity againstPhytophthora infestans

Summary The effects of various fungi, bacteria and different compost extracts on foliar infection of potato (Solatium tuberosum) withPhytophthora infestans were tested in detached leaf bioassays in 2001–2003. Application of microorganism inocula and compost extracts as well as copper oxychloride to excised leaves resulted in different degrees of blight control. In general, suppression of blight lesion growth was not improved by applying the antagonists before the leaves were inoculated withPhytophthora spores. There was some evidence that extracts made from different compost feed stocks of different ages suppressed leaflet infection with blight. However, results were very limited and inconsistent. Moreover, the effects were much smaller than where copper oxychloride was used. Improved efficacy of acceptable alternatives to copper fungicides especially in organic farming is required. No promising effective alternative to the use of copper fungicides to reduce late blight infection in organic potato production systems was identified in the experiments.     

Lipid metabolism in tomato and bean as a sensitive monitor for biocontrol of wilt diseases

The lipids metabolism of tomato and bean plants during biological control of wilt pathogens (Fusarium oxysporum f.sp.lycopersici andF. oxysporum f.sp.phaseoli, respectively) byBacillus subtilis was investigated. The interaction of wilt pathogens with both tomato and bean caused an imbalance and drastic reduction in total lipids, triacylglycerol, sterol and all phospholipd fractions except phosphatidic acid. The application of a formulated biocontrol agent,B. subtilis, eliminated the detrimental effect of both wilt pathogens and consequently prevented catabolism of lipid fractions in both tomato and bean. Moreover, the changes in the lipid fractions as a sensitive monitor for biocontrol of wilt diseases suggest a positive correlation between the application ofB. subtilis and improvement in the host metabolism towards anabolism. http://www.phytoparasitica.org posting Sept. 20, 2006.     

Antagonistic Activities of Volatiles from Four Strains of Bacillus spp. and Paenibacillus spp. Against Soil-Borne Plant Pathogens

The four effective antagonistic Bacillus strains, isolated from the rhizosphere soil of cucumber in a greenhouse, produced antifungal volatiles. These volatiles strongly inhibited the growth of the most tested pathogenic fungi with wide host plants, induced the mycelial morphological abnormalities, and decreased the sclerotoid production of Sclerotinia sclerotiorum in sealed plates. Spores of Botrytis cinerea exposed to these volatiles for 24-48 h in cavity slides cracked and the sporaceous inclusion became brown and effused to the suspension. An interesting phenomenon observed was that all the bacterial volatiles exhibited intense inhibitory activities against the pigment formation of tested pathogenic fungi, including Ascochyta citrullina, Alternaria solani, Alternaria brassicae, and so on. Interactions mediated by microbial volatiles could be widespread in soils, and volatiles may play an important role in reducing disease levels. A phylogenetic analysis based on 16S rDNA sequence placed the four bacteria in three species Paenibacillus polymyxa （BMP-11）, Bacillus subtilis （BL02）, and Bacillus pumilus （BSH-4 and ZB 13）. Through headspace sampling and GC-MS analysis, a rich profile was found from B. subtilis and overlapping volatile patterns could be found among the different species. Studies are under the way to find the possible action mechanisms and to seek the effective application of bacterial volatiles in greenhouse.     

大唼蜡甲人工大量饲养的主要影响因素及其控制

通过对从比利时引进的红脂大小蠹Dendroctonus valens捕食性天敌-大唼蜡甲Rhizophagus grandis Gyll.5年的室内大量饲养,对影响该天敌的产卵量、生长发育和羽化的主要生物因子和环境条件等进行了深入研究。对发现的影响其正常繁殖和生长发育的主要有害生物进行了外部形态、为害特点及控制进行了初步的研究,成虫平均羽化率从2003年的29.61%上升到了2004年的76.03%。研究表明,大唼蜡甲室内大量饲养的主要影响因子有:两种寄生螨(Proclaelapssp.和Histiostomasp.)、球孢白僵菌Beauveria bassiana、种群退化和温湿度条件。严格的卫生条件、病原生物的有效控制和种群复壮是保证大唼蜡甲群体饲养成功的关键。     

Biocontrol potential of Pseudomonas azotoformans,Serratia marcescens and Trichoderma virens against Fusarium wilt of Dalbergia sissoo

Wilt disease caused by Fusarium solani is a serious constraint to Dalbergia sissoo (shisham) plantations in northern India. In this study, the antagonistic potential of 40 bacterial isolates recovered from rhizophere soil of healthy shisham trees, and a well‐characterized Trichoderma species (Trichoderma virens) were tested for their possibility as biocontrol agents for F. solani. Two promising isolates (S1 and S15) were identified which inhibited pathogen growth, caused chitin degradation, produced siderophores and solubilized phosphate in vitro. Isolate S15 scored highest for hydrogen cyanide (HCN) production while isolate S1 was a non‐HCN producer. These two isolates were identified as Serratia marcescens (S1) and Pseudomonas azotoformans (S15) following sequence analysis of 16S rDNA. In dual culture assays, T. virens caused 80% inhibition of mycelial growth of the test fungus. The three selected antagonists when tested in planta in the glasshouse completely suppressed production of wilt symptoms on 12‐month‐old shisham plants. Further work is needed to ascertain the potential of these isolates to be used as biocontrol agents to manage shisham wilt under field conditions.