Biosynthesis of Depsipeptide Mycotoxins in Fusarium

The cyclic hexadepsipeptide enniatin is known as a phytopathogenic compound from Fusaria causing necrosis and wilt. The molecule consists of three alternating residues each of a branched chain amino acid and D-hydroxyisovaleric acid (D-Hiv). Enniatins are synthesized by a 347kDa multienzyme (enniatin synthetase) via a thiol template mechanism. The corresponding gene esyn1 has an open reading frame of 9393 nucleotides and harbours two modules, one responsible for D-hydroxy acid activation and one for L-amino acid activation with an integrated N-methyltransferase domain. Such methyltransferases build an homologous group among N-methyl peptide synthetases. Enniatins are synthesized by step-wise condensation of dipeptidol building blocks in an iterative manner resembling fatty acid synthesis. A key enzyme in enniatin biosynthesis is the NADPH-dependent D-2-hydroxyisovalerate dehydrogenase, that supplies enniatin synthetase with D-Hiv. Enniatins contribute to the wilt toxic character of Fusaria. Virulence was significantly reduced in F. avenaceum after disruption of the esyn1 gene.     

皂荚提取物对植物病原菌的抑制作用

 豆科植物皂荚是一种传统药用植物,为我国特有种。本文在室内测定了皂荚叶和棘刺乙醇提取物对枯草芽孢杆菌、胡萝卜软腐欧文氏杆菌、番茄疮痂病菌、番茄灰霉病菌、番茄叶霉病菌、棉花枯萎病菌、瓜果腐霉的抑制作用。叶提取物未检测出抗菌活性。棘刺提取物对枯草芽孢杆菌和番茄疮痂病菌都表现出一定的抗菌活性,最低抑菌浓度(MIC)分别为2.5 mg/mL和10 mg/mL。棘刺乙醇提取物在浓度为1 mg/mL时,对4种植物病原真菌的生长都表现出抑制活性,其中对瓜果腐霉生长的抑制率为28.4%。将棘刺乙醇提取物分为石油醚等5个极性不同的萃取部分,其中乙酸乙酯部分对枯草芽孢杆菌、番茄疮痂病菌、欧文氏杆菌都具有明显的抑制作用,且活性强于其它的极性段,对枯草芽孢杆菌和番茄疮痂病菌的MIC分别为1.25 mg/mL和5 mg/mL。乙酸乙酯部分对瓜果腐霉和棉花枯萎病菌的生长也有较强的抑制作用,在1mg/mL时抑制率分别为45.8%和23.9%。研究结果表明,皂英抗菌活性成分主要存在于棘刺中的乙酸乙酯极性段和正丁醇极性段中。     

辽宁省辣椒土传病害镰刀菌鉴定及rDNA-ITS序列分析

［目的］ 对分离的辣椒镰刀菌形态、基因序列进行研究,以确定病原菌的种类及这些菌的同源性,为进一步开展该病害诊断和防治研究奠定基础。［方法］ 对采自辽宁省各地辣椒土传病害的16株病菌进行形态观察及rDNA-ITS序列分析。［结果］ 引起辽宁省辣椒土传病害的主要镰刀菌有尖镰孢、茄镰孢、锐顶镰孢和串珠镰孢等4种,根据菌株分生孢子形态和rDNA ITS序列同源性,将ZW13、LY3、PJLJ、HC5、CY2、CY5、TLLJ、LY5、LY1菌株聚类为尖镰孢;FSLJ2和FSLJ3菌株聚类为茄镰孢;ZW和ZW18菌株聚类为锐顶镰孢;KPLJ、LZLJ、FSLJ1菌株聚类为串珠镰孢。［结论］ 从分子系统树和菌株间的遗传距离看出镰刀菌同种同源性都很高,不同种之间同源性相对较低,种间遗传差异较大。     

Antifungal properties of pristimerin and celastrol isolated from Celastrus hypoleucus

Pristimerin and celastrol isolated from the roots of Celastrus hypoleucus (Oliv) Warb f argutior Loes exhibited inhibitory effects against diverse phytopathogenic fungi. Pristimerin and celastrol were found to inhibit the mycelial growth of Rhizoctonia solani Kuhn and Glomerella cingulata (Stonem) Spauld & Schrenk in vitro by 83.6 and 62.6%, respectively, at 10 microg ml(-1). Pristimerin showed good preventive effect (96.7% at 100 microg ml(-1)) and curative effect (66.5% at 100 microg ml(-1)) against wheat powdery mildew in vivo. For celastrol, the preventive and curative effects against wheat powdery mildew were 80.5 and 45.4%, respectively, at 100 microg ml(-1).     

番茄早疫病菌抗感异菌脲菌株特性研究

测定不同营养条件、温度和pH值对番茄早疫病菌抗感异菌脲菌株生长的影响。结果表明,抗感菌株均在以蛋白胨为氮源的培养基中生长最快,敏感菌株在以可溶性淀粉为碳源的培养基中生长最快,突变体和抗性菌株在以乳糖为碳源的培养基上生长最快。突变体在pH值为7时生长最快而敏感菌株在pH值为6时生长最快,在不同pH值条件下,敏感菌株都比突变体生长速度快。在4～35℃菌丝均能生长,以20～25℃生长最快。测定抗感菌株对渗透压的敏感性和菌丝内的甘油、可溶性糖含量。结果发现,突变体比亲本菌株对渗透压具有较高的敏感性,所有抗性菌株在含高于10g/LNaCl的PDA平板上菌落直径随着NaCl质量浓度的升高而减小,在含高于40g/LNaCl的平板上停止生长。含药培养液中敏感菌株菌丝内甘油合成大量增加和积累,突变体和抗性菌株菌丝内甘油含量增加较少或有所下降,菌丝内可溶性糖含量变化不明显。     

Screening for Differential Gene Expression in Atropa belladonna Leafy Gall Induced Following Rhodococcus fascians Infection

To better characterise at the molecular level the nature of plant responses to infection by Rhodococcus fascians PCR-based differential display patterns of Atropa belladonna leafy gall (LG) and non-infected plant tissues were compared. Six differentially expressed genes were identified and their altered expression was confirmed by RT-PCR. Three of them corresponded to up-regulated genes which encode proteins involved in plant defence. The three remaining cDNA fragments which correspond to down-regulated genes in LG, encoded proteins with similarity to a multicystatin, a miraculin and a methallothionein-like protein, respectively. Upon elimination of the bacteria from infected plant tissue, the expression of up-regulated genes was maintained, whereas expression of down-regulated genes resumed suggesting a potential role of these up-regulated genes in plant growth and development.     

白藜芦醇及其衍生物对植物病原真菌的抑菌活性

为了明确植物源活性成分芪类化合物在农业病害防治中的前景,在离体条件下分别测定了白藜芦醇(Ⅰ)及其衍生物(Ⅱ~Ⅴ)对植物病原真菌菌丝生长的抑制作用以及对菌丝形态和孢子萌发的影响,采用叶片法和温室盆栽法研究了白藜芦醇的防病作用原理。结果表明:5个供试化合物对6种供试病原菌的菌丝生长均有不同程度的抑制作用,但均对番茄早疫病菌 Alternaria solani 的抑制活性最高,其中又以3,5-二羟基-4'-甲氧基二苯乙烯(Ⅲ)的抑制活性最高;白藜芦醇可造成番茄早疫病菌菌丝体畸形,并可抑制该病菌孢子的萌发,但未引起孢子形态改变。叶片法和温室盆栽法试验的结论一致,即白藜芦醇能够抑制病原菌的侵染,对植株起到一定的保护作用。     

螺旋毛壳ND35抗生素的产生及其在病害生物防治中的作用

内生菌螺旋毛壳Chaetomium spirale ND35是一株广谱拮抗性生防因子。为了阐明菌株ND35抗生素在植物病害生物防治中的作用机制,系统研究了抗生素的产生、提取纯化及其在离体和温室条件下的抑菌防病作用。菌株ND35的发酵液经乙酸乙酯萃取,浓缩得黄褐色抗生素粗提液。平皿抑菌试验表明,抗生素粗提液对20余种植物病原真菌有抑制作用,浓度为400μg/mL时对病菌菌丝生长的抑制率范围从14．4%到72．6%。浓度为500μg/mL和1000μg/mL的抗生素粗提液对新月弯孢分生孢子萌发的抑制率分别是54．2%和75．9%。发酵时间也影响抗生素的产量,马铃著葡萄糖液体培养基中发酵15天时,抗生素产量最高。经薄层层析和高效液相色谱检测,证明有两个活性组分。温室试验表明,该抗生素粗提液稀释50倍液对玉米弯孢叶斑病的防治效果可达85．5%,稀释250倍液防治效果达63。3%。试验结果表明,抗生素是菌株ND35重要的生防决定因子之一,抗生素粗提液也展示了对玉米弯孢叶斑病具有较好的生防潜能。     

Specific Oligonucleotide Primers for the Rapid Identification and Detection of the Agent of Tomato Pith Necrosis, Pseudomonas corrugata, by PCR Amplification: Evidence for two Distinct Genomic Groups

Unique DNA bands from strains representative of two groups of Pseudomonas corrugata, as shown by amplification of their genomic DNA by polymerase chain reaction using short random sequence oligonucleotide primers (RAPD-PCR), were isolated, cloned and sequenced. Two pairs of specific primer sequences, based on the ends of the cloned unique DNA bands from strains IPVCT10.3 and IPVCT8.1, were used in multiplex PCR with a range of P. corrugata strains. All strains produced one of the two specific bands, 1100bp (from the IPVCT10.3-based primers) and 600bp (from the IPVCT8.1-based primers), representing groups designated I and II, respectively. The primers were also tested on a wider range of Pseudomonas species, including the closely-related fluorescent Pseudomonas genomospecies FP1, FP2 and FP3: none of these bacteria produced any bands following amplification by PCR with these primers. The primer sets detected P. corrugata in tomato pith necrosis-infected plants providing a useful tool for rapid identification and epidemiological studies.