Purification and characterization of a lipopeptide produced by Bacillus thuringiensis CMB26

AIMS: To isolate an antagonist for use in the biological control of phytopathogenic fungi including Colletotrichum gloeosporioides, then to purify and characterize the biocontrol agent produced by the antagonist. METHODS AND RESULTS: Bacteria that exhibited antifungal activity against the causative agent pepper anthracnose were isolated from soil, with Bacillus thuringiensis CMB26 showing the strongest activity. A lipopeptide produced by B. thuringiensis CMB26 was precipitated by adjusting the pH 2 with 3 n HCl and extracted using chloroform/methanol (2:1, v/v) and reversed-phase HPLC. The molecular weight was estimated as 1447 Da by MALDI-TOF mass spectrometry. Scanning electron and optical microscopies showed that the lipopeptide has activity against Escherichia coli O157:ac88, larvae of the cabbage white butterfly (Pieris rapae crucivora) and phytopathogenic fungi. The lipopeptide had cyclic structure and the amino acid composition was L-Glu, D-Orn, L-Tyr, D-allo-Thr, D-Ala, D-Val, L-Pro, and L-Ile in a molar ratio of 3:1:2:1:1:2:1:1. The purified lipopeptide showed the same amino acid composition as fengycin, but differed slightly in fatty acid composition, in which the double bond was at carbons 13-14 (m/z 303, 316) and there was no methyl group. CONCLUSION: A lipopeptide was purified and characterized from B. thuringiensis CMB26 and found to be similar to the lipopeptide fengycin. This lipopeptide can function as a biocontrol agent, and exhibits fungicidal, bactericidal, and insecticidal activity. SIGNIFICANCE AND IMPACT OF THE STUDY: Compared with surfactin and iturin, the lipopeptide from B. thuringiensis CMB26 showed stronger antifungal activity against phytopathogenic fungi. This lipopeptide is a candidate for the biocontrol of pathogens in agriculture.     

A survey of black dot and other fungal tuber blemishing diseases in British potato crops at harvest

Black dot and other fungal blemishing diseases were assessed on tubers from crops in eastern England in 1987 and 1988, and in 1989 and 1990 from crops throughout Great Britain. Black dot was found on tubers in crops from all areas of the country, and was most common in those from eastern and southern England. The disease was scarce on most Scottish crops, but occasionally some were severely affected. In 1988 there was no consistent relationship between the amounts of disease on the seed and subsequent ware crop. On average, the disease was more prevalent in irrigated crops and where the interval between successive potato crops was short. In 1990 treating seed tubers with fungicide had no effect on subsequent levels of disease. Few crops were unaffected by silver scurf, and it was most common in crops from southern and eastern England. In the national survey common scab was the most prevalent disease, black scurf was less common and skin spot and powdery scab uncommon except in some Scottish crops.     

Development of black dot disease (Colletotrichum coccodes (Wallr.) Hughes) and its effects on the growth and yield of potato plants

Seed tubers of cvs Désirée and Pentland Crown with different severities of black dot were planted in 1988 and 1989 at Rothamsted in fields in 4– or 7-course rotations, respectively. Tubers treated with prochloraz (1988) or imazalil (1989) were planted in some plots, and in others Colletotrichum coccodes inoculum was added to the soil at planting. In further experiments at Mepal, Cambridgeshire in 1989 and 1990 and at Rothamsted in 1990 on sites where potatoes had not been grown for more than 15 years, large amounts of inoculum were added to the soil around disease-free seed tubers of two (1989) or three (1990) cultivars at planting. In all experiments plants were sampled during the season and the effects of treatments on disease development, growth and yield were recorded. Disease on roots, stem bases and tubers was found early in the season and was more severe on Désirée than on Pentland Crown plants from fields in 4– or 7-course rotations. Severity increased throughout the season and with increasing amounts of disease on the seed tubers, especially with Desiree. Disease was also found on plants from disease-free tubers and was more severe in 1988 than 1989. At harvest, black dot on tubers was significantly more severe from severely affected than from disease-free seed, and was most severe where inoculum, especially large amounts, had been added at planting. Fungicide treatment decreased disease early in the season but had no effect on tuber infection at harvest. In 1989 the weight loss of seed tubers during sprouting increased with increasing amounts of black dot, but the disease had little effect on plant size through the season. At harvest the yield of ware tubers (>50 mm) decreased with severe disease but total tuber yields were not significantly affected. However, at harvest in 1988 severely affected seed yielded significantly less than healthy seed. Plants grown from mini-tubers were free from disease on sites where potatoes had not been grown for at least 15 years. Inoculum applied at planting caused severe disease on all cultivars in both years, whereas disease was slight on uninoculated plants. Inoculated plants senesced early at Mepal in 1990, but there were no significant differences in total tuber yield in any experiment. However, yields of ware tubers (>50 mm) were sometimes decreased and the total tuber number per plant increased.     

营养物质对两种炭疽菌潜伏侵染的影响

通过不同浓度的几种糖类对潜伏侵染在青香蕉果实中的colletotrichum musae和芒果果实中的colletotichum gloeosporioides的菌体的影响进行了测定,结果表明,高浓度淀粉可极显著地提高两种炭疽菌的孢子萌发率,并有利于附着胞和分生孢子的形成。单糖和二糖在较低浓度时有利于孢子萌发和产孢,不利于附着胞形成。未成熟果实的坚硬结构和高淀粉含量为病菌提供了以附着胞形式潜伏侵染在寄主中的条件。     

Twin-screw extrusion of ‘Pesta’-encapsulated biocontrol agents

World Journal of Microbiology and Biotechnology -     

Postharvest disease control in mangoes using high humidity hot air and fungicide treatments

The disease control efficacy of quarantine heat treatments developed for fruit fly disinfestation in mangoes cv. Kensington Pride was evaluated in this study. Heat was applied using high humidity (>95% r.h.) hot air (HHHA) at temperatures ranging from 47–49°C. Anthracnose, caused by Colletotrichum gloeosporioides, was well controlled in mangoes heated to a core temperature of 46°C, 47°C or 48°C for 24, 10 or 8 min respectively, prior to ripening at 23°C for 16 days. Stem end rot, caused by Dothiorella dominicana and Lasiodiplodia theobromae, was not satisfactorily controlled by these treatments. In a subsequent experiment, fruit were immersed in a hot benomyl (0.5 g a.i. litre“¹ at 52°C for 5 min) or unheated prochloraz (0.25 ml a.i. litre¹ at 28°C for 30 s) dip before or after the application of HHHA (core temperature of 47°C for 10 min). During storage at 23°C for 15 days, the incidence of stem end rot was reduced by HHHA alone, although immersion in hot benomyl either before or after HHHA treatment greatly improved stem end rot control. HHHA treatment (core temperature of 46.5°C for 10 min) alone reduced the incidence of anthracnose in mangoes stored at 13°C for 14 days prior to ripening at 22°C, although a combination treatment consisting of HHHA and either hot benomyl or unheated prochloraz gave complete control of anthracnose under these storage conditions. HHHA treatment alone gave no control of stem end rot in mangoes stored at 13°C prior to ripening at 22°C. A supplementary hot benomyl treatment was required for acceptable control of this disease in cool-stored mangoes. The development of yellow skin colour in fruit was accelerated by HHHA treatment.     

瓜类刺盘孢诱导物对新疆甜瓜抗病相关酶活性的影响

近年来,应用病毒、细菌、真菌以及病原菌体及其代谢物作为诱导因子,已在多种作物上获得对病害的整体免疫,有些已开始在田间应用。我们也曾应用人工诱导免疫的方法使新疆甜瓜获得对瓜类疫霉病的抗性,但有关植物人工诱导免疫机理的研究,目前国外报道不多,国内尚未见报道。本文以瓜类刺盘孢(Colletotrichum langenarium)培养滤液和菌丝细胞壁作为诱导物,研究了免疫植株相关酶的活性以及可溶性蛋白的变化,探讨了人工诱导免疫的机理。     

More than 400 million years of evolution and some plants still can't make it on their own: plant stress tolerance via fungal symbiosis

All plants in natural ecosystems are thought to be symbioticwith mycorrhizal and/or endophytic fungi. Collectively, thesefungi express different symbiotic lifestyles ranging from parasitismto mutualism. Analysis of Colletotrichum species indicates thatindividual isolates can express either parasitic or mutualisticlifestyles depending on the host genotype colonized. The endophytecolonization pattern and lifestyle expression indicate thatplants can be discerned as either disease, non-disease, or non-hosts.Fitness benefits conferred by fungi expressing mutualistic lifestylesinclude biotic and abiotic stress tolerance, growth enhancement,and increased reproductive success. Analysis of plant–endophyteassociations in high stress habitats revealed that at leastsome fungal endophytes confer habitat-specific stress toleranceto host plants. Without the habitat-adapted fungal endophytes,the plants are unable to survive in their native habitats. Moreover,the endophytes have a broad host range encompassing both monocotsand eudicots, and confer habitat-specific stress tolerance toboth plant groups. Key words: Colletotrichum, fungal endophytes, stress tolerance, symbiosis, symbiotic lifestyle Received 19 June 2007; Revised 25 November 2007 Accepted 30 November 2007     

云南芒果采后炭疽病病原菌的鉴定及室内生防菌的筛选

随着对云南芒果需求量的增加,其种植面积日趋扩大,并不断引入新品种,这也导致云南省芒果炭疽病害发生日趋加重。为了有针对性地开展芒果炭疽病的生物防治,采用组织块分离法分离芒果采后炭疽病病原菌,通过形态学观察初步鉴定,并遵循柯赫氏法则对病原菌进行验证。随后,利用rDNA-ITS序列和系统发育树分析明确病原菌的分类学地位。最后,采用5种生防细菌对病原菌进行拮抗试验。通过对芒果采后炭疽病病原菌进行分离鉴定,确定胶孢炭疽菌(Colletotrichum gloeosporioides)是引起云南芒果采后炭疽病的病原菌,该菌株内转录间隔区(internal transcribed spacer,ITS)序列长度为536 bp,登录号为MH744668;5株生防细菌对芒果炭疽病病原菌都有一定的抑菌作用,具有较好的生防开发潜能,其中抗生素溶杆菌L-44的抑菌效果最好,抑制率达53.7%。研究结果为云南省芒果采后病害的生物防治提供了新的思路。