万寿菊根提取物对西瓜枯萎病菌的抑菌活性成分及作用机理研究

本文研究了万寿菊根提取物中西瓜枯萎病菌的抑菌活性成分及其抑菌作用机理。结果表明,在4类主要化学物质中,精油类含量最高(59.8%),且抑菌效果最佳;10mg/mL精油提取物的抑制率在24、36和48h分别为62.83%,58.31%和56.30%。在西瓜枯萎病的3个主要发病期(苗期、伸蔓开花期和座果期)施用精油提取液,能有效抑制西瓜枯萎病菌,促进植株生长,同时提高西瓜植株的POD和SOD酶活性,并维持CAT活性,有效减轻了西瓜枯萎病菌对植株的毒害作用。     

西瓜枯萎病菌镰刀菌酸对西瓜苗作用机制的初步探讨

 镰刀菌酸是由镰刀菌属中那些能引起多种植物萎蔫的病原菌所产生的一种非特异性毒素,从发现至今已有60多年了,但关于它在病程中作用问题的研究还不多,而且意见也不一致。     

芽孢杆菌B11拮抗蛋白性质及其对西瓜枯萎病菌的作用机理

芽孢杆菌(Bacillus sp.)B11分离自土壤,其拮抗蛋白粗提液在温度60、80℃各处理30 min后仍能保持抗菌活性,对紫外线、蛋白酶K、RNase处理稳定,对胃蛋白酶、胰蛋白酶、氯仿部分敏感,作用活性pH(pH4～10)范围较宽.B11拮抗蛋白对西瓜枯萎病菌具有较强的拮抗作用,其抑菌机制主要使病菌菌丝畸形,细胞质浓缩,孢子不萌发或萌发异常.     

拮抗链霉菌B28 的分类鉴定及其生防作用研究

 An antagonistic Streptomyces strain B28, isolated from the soil collected in Tianmu mountain of Zhejiang Province, was identified as Streptomyces diastatochromogenes by morphological , physiological & biochemical characteristics and 16S rDNA sequence alignment. Antagonistic study indicated that its secondary metabolite toyocamycin had an antifungal activity on the mycelial growth of seven species of plant pathogenic fungi. The results showed that toyocamycin possessed marked inhibitory activity against Rhizoctonia solani and the EC50 value was 0. 58 μg / mL. The effective of toyocamycin against Botrytis cinerea and Fusarium oxysporum f. sp. cucumerinum was much higher than that against Colletotrichum gloeosporioides, the EC50 value of
toyocamycin in inhibiting B. cinerea, F. oxysporum and C. gloeosporioides were 13. 65 μg / mL, 12. 33 μg / mL, and 30. 15 μg / mL respectively.     

枯草芽孢杆菌SB1的抑菌活性及其对番茄青枯病的防治作用

 Bacillus subtilis strain SB1 isolated from tomato roots was evaluated for its ability to control tomato bacterial wilt under greenhouse conditions.Application of strain SB1 culture suspension suppressed tomato wilt disease both in sterilized and non-sterilized soil,providing 76.03% and 77.78% protection,respectively.Wilt symptom was significantly reduced when strain SB1 was applied before pathogen inoculation,while less suppression was conferred by strain SB1 application after pathogen inoculati...     

芽孢杆菌B11拮抗蛋白性质及其对西瓜枯萎病菌的作用机理

芽孢杆菌(Bacillus sp．)B11分离自土壤，其拮抗蛋白粗提液在温度60、80℃各处理30min后仍能保持抗菌活性，对紫外线、蛋白酶K、RNase处理稳定，对胃蛋白酶、胰蛋白酶、氯仿部分敏感，作用活性pH(pH4～10)范围较宽。B11拮抗蛋白对西瓜枯萎病菌具有较强的拮抗作用，其抑菌机制主要使病菌菌丝畸形，细胞质浓缩，孢子不萌发或萌发异常。     

山东西瓜枯萎病菌致病性分化及品种抗性的初步研究

1989—1991年在自然病圃中观察了25个西瓜品种对西瓜枯萎病的抗病性。结果表明,国内只中8602F₁,是唯一保持高抗,中8601F₁,新澄1号和中育号等表现出中度抗性外,其他均属轻抗或感病;而来自国外的抗性品种显示出与原来一致的或更高的抗性。根据在国外鉴别寄主上的反应,致病性可分为2个不同的类型,类型Ⅰ占总分离物的74.4％,致病力中等;类型Ⅱ致病力较强,但仅分布于少数老瓜区。     

草莓枯萎病菌的生物学特性及7种杀菌剂对其抑制作用

采用十字交叉法和血球计数板法测定草莓枯萎病菌的生物学特性。结果表明,PSA培养基最适合草莓枯萎病菌的生长和产孢,菌落生长的适温为5~40℃(最适温度为28℃),适宜pH值为5,菌落产孢的适宜pH值为6,且光暗交替可促进菌落的生长和产孢,同时明确了不同碳氮源对草莓菌落生长及产孢的影响。采用菌落生长速率法,测定草莓枯萎病菌对7种杀菌剂的敏感性,烯唑醇和戊唑醇对菌落生长的抑制毒力最强,EC₅₀分别为0.2072μg·mL^-1和0.2130μg·mL^-1,分别是多菌灵的238.85倍和232.35倍;腈菌唑和苯醚甲环唑的毒力次之,EC₅₀分别为0.7929μg·mL^-1和0.8513μg·mL^-1,分别是多菌灵的62.42倍和58.13倍;三唑酮和霜霉威的毒力最小。     

多杀菌素的作用机理及其抗药性的研究进展

多杀菌素是一类新的杀虫剂,具有高效、低毒、选择性强、对环境安全的特点。其作用机制是通过激活烟碱型乙酰胆碱受体(nAChR),使正常昆虫神经细胞去极化,也可通过抑制γ-氨基丁酸受体(GABAR)使神经细胞超极化。本文综述了多杀菌素作用机理及其抗药性的研究进展。     

多杀菌素的作用机理及其执药性的研究进展

多杀菌素是一类新的杀虫剂,具有高效、低毒、选择性强、对环境安全的特点.其作用机制是通过激活烟碱型乙酰胆碱受体(nAChR),使正常昆虫神经细胞去极化,也可通过抑制γ-氨基丁酸受体(GABAR)使神经细胞超极化.本文综述了多杀菌素作用机理及其抗药性的研究进展.     

农杆菌介导的西瓜枯萎病菌遗传转化

为获得带GFP标记的西瓜枯萎病菌转化株,用于后期观察病原菌侵染过程,采用农杆菌介导的方法,对西瓜枯萎病菌1号生理小种进行了遗传转化。结果表明:共培养时间为36h,枯萎病菌孢子和农杆菌AGL1比例为1∶1时该菌株的遗传转化效率最高,可以达到117.33个转化子/107个孢子。转化株的孢子、菌丝体及萌发的孢子均能发出稳定而强的绿色荧光。转化株的致病力检测显示其致病力与转化前的野生菌株致病力无明显差异。结果表明本研究获得的带GFP标记的西瓜枯萎病菌转化株可用于观察病菌在西瓜根系的侵染过程。     

Mitochondrial dna restriction fragment length polymorphisms infusarium oxysporum f. sp.niveum

Mitochondrial DNA (mtDNA) extracts from 13 isolatesof Fusarium oxysporum f. sp.niveum, including 12 from widely separated geographic regions within the United States and representing the three races, and one race 2 isolate from Israel, were examined for the presence of plasmid DNA and were also subjected to restriction endonucleases analysis. None of the mtDNA from any isolate had a copurifying plasmid. The estimated size of mtDNA fromF. o. f. sp.niveum race 0, calculated as the average of the sum of restriction fragment sizes, was 45.1 ± 2.2 kb. The restriction enzymesBamHI, EcoRI, Hpal, HindIII andMbol resolved 2, 4, 9, 21, and more than 40 fragments, respectively, but no polymorphisms were observed among the 13 isolates with any of these endonucleases. However, PstI digestion showed three distinct polymorphic patterns among the isolates. Each appeared to derive from point mutations that resulted in a change of one or more restriction sites. The most common pattern was present in nine of the isolates (three of race 0, four of race 1, and two of race 2) and included a 1.5 kb fragment. A second polymorphic pattern occurred in three USA isolates (one each of race 0, race 1 and race 2) and was characterized by an apparent replacement of the 1.5 kb fragment by 0.6 and 0.9 kb fragments. The Israeli isolate [ISL-59(73) race 2| had a unique pattern lacking the 1.5 and 2.0 kb fragments present in the common pattern and, instead, had 0.6, 0.9 and 3.0 kb fragments. The mtDNA polymorphisms observed among the USA isolates were not correlated with either pathological race or geographic region of origin.     

主要环境及营养因素对西瓜枯萎病菌生长的影响

采用菌丝生长速率法和产孢量测定研究了不同温度、光照、pH及碳氮源对来源于海南、河北和上海的3株西瓜枯萎病菌Fo-HN-46、Fo-HB-12和Fo-SH-1生长速率和产孢量的影响。结果表明:西瓜枯萎病菌菌株的适宜生长温度为20~30℃,Fo-HN-46最适生长温度为28℃,Fo-HB-12和Fo-SH-1最适生长温度为25℃,产孢的最适温度为28~30℃;光照对西瓜枯萎病菌生长速率无显著影响,对产孢影响显著,Fo-HN-46在黑暗条件下单位面积产孢量最高,Fo-HB-12和Fo-SH-1在半光照条件下产孢量最高;pH对西瓜枯萎病菌菌株的生长速率和产孢量有显著影响,pH 7~9时菌丝生长速率快,pH 8~11时产孢量高;葡萄糖、淀粉和乳糖作为碳源能够显著增加西瓜枯萎病菌的产孢量;蛋白胨作为氮源能显著促进菌丝生长,硝酸钠、酵母粉和蛋白胨均能显著增加西瓜枯萎病菌的产孢量。用最适碳源、氮源培养,Fo-HN-46生长速率大于Fo-HB-12和Fo-SH-1。Fo-SH-1产孢量高于Fo-HN-46和Fo-HB-12。     

西瓜枯萎菌拮抗细菌的筛选、鉴定及防效测定

从内蒙古、新疆、安徽、河南、江苏、江西6省的西瓜、黄瓜和茄子3种作物根际土壤和病健组织中,共获得样本64份,分离得到细菌1050株。室内抑菌活性测定结果表明：9株细菌分离物对西瓜枯萎病菌Fusariumoxysporumf.sp.niveum、甘蓝黑斑病菌Alternaria brassicicola、辣椒疫霉病菌Phytophthora capsici和番茄灰霉病菌Botrytis cinerea均具有很强的拮抗作用,其中菌株NBT-15和PL-82对西瓜枯萎病菌菌丝生长的抑制作用比较明显,抑制率分别为60.5%和55.3%。盆栽防病试验结果表明：NBT-15和PL-82对西瓜枯萎病的防治效果分别为80.4%和75.6%。形态特征、生理生化特性及16S rDNA序列分析结果确定菌株NBT-15为枯草芽孢杆菌Bacillus subtilis,PL-82为铜绿假单胞菌Pseudomonas aeruginosa。     

AM真菌和镰刀菌对西瓜根系膜脂过氧化作用和膜透性的影响

 在温室盆栽条件下研究了丛枝菌根(arbuscular mycorrhiza,AM)真菌Glomus versiforme (Karsten) Berch和西瓜枯萎镰刀菌(Fusarium oxysporum f. sp. niveum)对西瓜根系中膜脂过氧化产物丙二醛(MDA)含量和细胞膜透性的影响。结果表明,接种AM真菌的西瓜根系中MDA含量、组织自动氧化速率和膜透性均低于对照,先接种G. versiforme,后接种F. oxysporum f. sp. niveum处理的MDA含量、组织自动氧化速率和细胞膜透性均低于只接种F. oxysporum f. sp. niveum的处理。接种G. versiforme感枯萎病西瓜品种"郑杂5号"MDA含量、组织自动氧化速率和膜透性降低幅度大于抗病品种"京欣1号"的接种处理,说明G. versiforme可降低感病西瓜品种的膜透性和MDA的产生,从而有效地保护细胞膜系统,减轻F. oxysporum f. sp. niveum对西瓜的为害程度。     

Occurrence of Fusarium oxysporum f. sp. melonis Race 1 in Japan

In 1994, Fusarium wilt of melon cultivars which are resistant to races 0 and 2 of Fusarium oxysporum f. sp. melonis was observed in southern area of the Lake Biwa region, Shiga prefecture. In commercial fields, mature plants of cv. Amus which were grafted onto cv. Enken Daigi 2, and of cv. FR Amus showed yellowing, wilting and finally death before harvesting of fruits. Diseased plants had vascular and root discolorations, and their stem sections yielded typical colonies of F. oxysporum. When the Shiga strains were tested for their pathogenicity to 12 species of cucurbits, they caused wilts only on melon. Using race differential cultivars of melon, the Shiga strains were classified as race 1 of F. oxysporum f. sp. melonis, which has not been reported in Japan. To further characterize their pathogenicity, the strains were used to inoculate 46 additional cultivars of melon, oriental melon and oriental pickling melon. All the race 1 strains were pathogenic to the cultivars tested, and their host range was apparently different from those of strains belonging to other races (races 0, 2 and 1,2y). DNA fingerprinting with a repetitive DNA sequence, FOLR3, differentiated race 1 strains from strains of races 0 and 2, but not from race 1,2y strains. Received 2 July 1999/ Accepted in revised form 30 September 1999     

西瓜枯萎病菌遗传多样性的相关序列扩增多态性分析

对30个西瓜枯萎病菌Fusarium oxysporum f.sp.niveum菌株基因组DNA进行相关序列扩增多态性(SRAP)分子标记分析,以探究其遗传多样性与地理来源的关系。采用尖孢镰刀菌西瓜专化型Fusarium oxysporumf.sp.niveum0、1、2号生理小种的基因组DNA为模板,对225对SRAP引物进行筛选,筛选出20对多态性、重复性较好且条带清晰的引物,对30个菌株进行PCR扩增,共扩增出386条带,其中多态性条带有371条,多态性比率为96.11%,平均每对引物扩增出19.3个位点和18.55个多态性位点。UPGMA法聚类分析结果显示,供试菌株两两之间的遗传相似系数范围为0.69~0.90,平均为0.79,说明尖孢镰刀菌西瓜专化型的遗传多样性较为丰富。基于SRAP标记聚类分析表明,30个菌株在遗传相似系数为0.70处被划分为3个类群,I类群包含24个菌株,其中18个来自湖南省,Ⅱ类群只包含1个来自黑龙江省哈尔滨市的菌株,它和另一个来自黑龙江地区的菌株被划分到不同的类群,且遗传距离相对较远;Ⅲ类群包含了5个菌株,其中3个来自海南三亚,其余两个来自湖南省。根据菌株的分布情况来看,菌株的聚集与地理来源没有明显的相关性。     

葫芦砧木萎蔫病病原镰刀菌鉴定及其防治药剂的筛选

本研究对葫芦砧木萎蔫病病原菌进行了分离鉴定及室内杀菌剂筛选, 为葫芦萎蔫病的防控提供参考依据。从嫁接西瓜主产区和葫芦砧木育种大棚采集具有典型萎蔫症状的病样, 进行病原菌的分离和鉴定, 共得到6株分离物。形态学观察发现分离物具有茄病镰刀菌Fusarium solani的特征。基于病原菌的rDNA-ITS和TEF-1α序列分析, 6株分离物的序列与茄病镰刀菌的序列相似性为100%, 与形态学鉴定结果一致。采用柯赫氏法则对分离菌株进行致病力验证, 结果表明, 6株分离物都能使葫芦砧木发病, 且症状与田间发病症状一致, 其中LS-2致病力最强。选用11种杀菌剂对葫芦茄病镰刀菌进行室内毒力测定, 结果表明, 供试药剂中, 20%硅唑·咪鲜胺EW、70%甲基硫菌灵WP和25%多菌灵WP能很好地抑制茄病镰刀菌菌丝的生长, 抑菌率达到80%以上。20%硅唑·咪鲜胺EW的毒力最强, EC50为0.08 μg/mL。