三七主要病原菌对皂苷的敏感性分析

采用菌丝生长速率法测定了三七4种病原菌对三七皂苷粗提物及其含有的4种主要皂苷的敏感性。结果表明,三七皂苷粗提物对病原菌的抑制活性较低,在浓度1 000 μg/mL条件下对4种供试病原菌的抑制率均低于20%。在低浓度下对锈腐病菌和恶疫霉菌甚至表现出明显的促生作用。利用HPLC分析表明,三七皂苷粗提物中含有R1、Rb1、Rg1和Rd ?4种主要皂苷。在浓度为1 000 μg/mL条件下,R1、Rb1和Rg1对锈腐病菌均表现明显的促进生长作用;Rg1对恶疫霉D-1菌株表现出微弱的抑制作用,而R1、Rb1、Rd对恶疫霉菌均具有较低的促生长活性;R1和Rb1对腐皮镰刀菌PN-21具有较弱的促生长作用,而Rg1和Rd具有较弱的抑制作用;4种皂苷对侵染三七茎部的人参链格孢菌均具有抑菌活性,抑菌能力Rd>Rb1>R1>Rg1,Rd的抑制率可达23.43%。本试验结果表明,三七一些皂苷可以促进根腐病菌的生长,对根腐病的发生具有促进作用。     

人参锈腐病生防细菌的筛选及鉴定

从吉林省抚松地区采集到的人参根际土壤中分离、纯化获得97株细菌,采用平板对峙法筛选出13株对毁灭柱孢菌Cylindrocarpon destructans(Zinss.)Scholtan具有良好抑制作用的菌株。用生长速率法对抑菌效果最好的菌株SZ-22进行抑菌谱的测定,并进行盆栽试验验证其对人参锈腐病的防治效果。抑菌试验结果表明,菌株SZ-22发酵液对毁灭柱孢菌的抑制率达82.63%,具有较好的广谱抑菌作用。盆栽试验结果表明,菌株SZ-22对人参锈腐病的防治效果为63.02%,显著高于农药多菌灵的防效。经形态特征、生理生化特征鉴定及16S r DNA序列分析,确定菌株SZ-22为土壤短波单胞菌Brevundimonas terrae。     

人参黑斑病生防菌的分离鉴定及防效

本研究以人参链格孢为靶标菌,采用平板对峙法从人参根际土壤筛选出一株拮抗细菌YJ 3-7,结合细胞形态判断、16S rDNA和gyrA基因序列比对分析,鉴定菌株YJ 3-7为贝莱斯芽胞杆菌Bacillus velezensis。该菌株的发酵液、菌悬液和无菌发酵液对人参链格孢的生长均具有抑制作用,抑制率分别为98.76%、98.27%和70.62%。离体叶片和盆栽试验表明YJ 3-7发酵液可以较好地防治人参黑斑病,防效分别为63.24%和67.92%,盆栽防效略低于多菌灵(72.81%)。除此之外,菌株YJ 3-7还能有效抑制其他8种病原真菌的生长。研究表明贝莱斯芽胞杆菌YJ 3-7抑菌谱广,对人参黑斑病具有显著的防治效果,在植物病害防治中具有一定的应用潜力,可为人参黑斑病生物防治药剂的开发奠定基础。     

25%氟吗啉·唑菌酯悬浮剂在人参土壤中的残留动态

建立25%氟吗啉·唑菌酯悬浮剂在人参土壤上残留分析方法。样品经过乙腈提取,弗罗里硅土层析柱净化,高效液相色谱法进行定量分析。此条件下氟吗啉和唑菌酯的平均回收率分别是87.9%~99.0%和89.2%~99.9%,相对标准偏差为2.7%~7.3%和1.9%~13.4%。氟吗啉在人参土壤中的降解半衰期为2.7~10.7d,唑菌酯的半衰期为0.3~18.4d。该方法简单,快速,适用于人参土壤中氟吗啉和唑菌酯的残留分析。     

真菌甲硫氨酸生物合成途径研究进展

 甲硫氨酸是蛋白质的重要组分,同时还可通过生成S-腺苷甲硫氨酸（SAM）调控细胞内多种生理过程。本文根据酿酒酵母、粗糙脉孢菌和构巢曲霉3种模式真菌中甲硫氨酸生物合成的最新研究进展,勾画出真菌的甲硫氨酸合成途径,综述了该途径中关键酶的生物学功能,探讨了该途径的调控机制,为病原真菌甲硫氨酸合成的研究提供理论基础,同时为新杀菌剂的研发提供新思路。     

进境林木种子携带真菌的检测

以5种进境林木种子为材料进行种子带菌分析,采用保湿培养检验法和分离培养检验法分别对种子表面粘附真菌及内部寄藏真菌进行检测和鉴定.在供试的5个样品上共分离得到18属真菌,分别为链格孢属、枝孢属、镰孢属、加氏菌属、小丛壳属、毛壳菌属、拟茎点霉属、根霉属、附球菌属、节菱孢属、弯孢属、德氏霉属、拟盘多毛孢属、木霉属、散囊菌属、青霉属、毛霉属及曲霉属.种子表面附带真菌以曲霉菌和根霉菌分离频率最高;链格孢菌、镰孢菌、毛壳菌是种子寄藏真菌的优势菌群.     

皂苷与植物病原真菌相互关系研究进展

皂苷是一类植物次生代谢物,具有抗真菌活性,被认为是植物抵抗真菌侵染的主要化学防御物质。其后的研究发现,许多侵染含皂苷植物的真菌能够通过产生皂苷解毒酶破坏这些植物防御物质。人们推测,这种皂苷去毒机制在含皂苷植物病原真菌中可能普遍存在,是病原真菌产生致病性的主要原因。本文总结了近年来皂苷与植物病原真菌相互关系研究的现状和进展,分析了今后的研究方向及应用前景。     

相关防御酶对人参锈腐病诱导抗性的响应

 为明确水杨酸(SA)对人参抗人参锈腐病的诱导作用,本研究首先采用琼脂平板法测定了SA对人参锈腐病菌(Cylindrocarpon destructans)生长的影响。然后用SA溶液处理二年生人参移栽苗,温室接种人参锈腐病菌,测定了人参根内防御酶系活性（PAL,CAT,PPO,POD）、β-1,3-葡聚糖酶和几丁质酶活性的动态变化。结果表明:浓度为0~200 mg·L^-1 的SA溶液对人参锈腐病菌无直接抑制作用。但SA溶液处理后,人参锈腐病发病率较直接接种处理的下降30%,人参根系PAL、CAT、PPO、POD活性较对照均表现上升趋势,β-1,3-葡聚糖酶和几丁质酶活性也较对照增强,并且经SA诱导后接种锈腐菌的人参体内上述酶活性比只诱导不接种处理上升速度快。这表明SA处理可以改变人参根部相关防御酶的活性,从而提高人参对人参锈腐病的抗性。     

人参灰霉病拮抗细菌的筛选及鉴定

为获取对灰葡萄孢Botrytis cinerea Pers.ex Fr.防效优良的菌株,采用滤纸片法和牛津杯法从105株菌株中筛选出7株拮抗性良好的菌株,采用平板对峙法对抑菌效果较好的菌株SZ-35进行抑菌谱测定,采用盆栽法研究其对人参灰霉病的防效及在人参体内的定殖能力,并通过形态学、生理生化特征、16S rDNA及gyrB基因测序确立其分类地位。结果显示,菌株SZ-35对灰葡萄孢的抑制作用最强,抑菌率和抑菌带宽分别为86.53%和12.20 mm,且具有广谱抑菌能力;菌株SZ-35可有效定殖于人参体内,对人参灰霉病的防效达76.42%,显著高于70%代森锰锌可湿性粉剂;通过形态特征、生理生化特征观察,以及16S rDNA及gyrB同源性序列对比分析,鉴定菌株SZ-35为解淀粉芽胞杆菌Bacillus amyloliquefaciens(GenBank登录号为KC511113)。研究表明,解淀粉芽胞杆菌SZ-35对灰葡萄孢具有良好的抑菌能力,且在人参中的定殖能力较好。     

十字花科黑腐病菌纤维素酶主效基因XC0639 CBD结构域功能的分析

十字花科黑腐病菌(Xanthomonas campestris pv. campestris,Xcc)几乎能引起所有十字花科植物产生黑腐病[1]。该菌是研究植物与病原微生物互作的模式菌株之一[2]。植物病原菌在侵染宿主植物时需要先降解植物细胞的细胞壁,纤维素和半纤维素是细胞壁的主要成分。纤维素酶是降解纤维素生成葡萄糖的一类复合酶,主要由内切葡聚糖酶、外切葡聚糖酶以及β-葡萄糖苷酶3类酶组成,三者作用方式不同,但能协同催化水解纤维素[3]。     

Molecular Differentiation and Detection of Ginseng-Adapted Isolates of the Root Rot Fungus Cylindrocarpon destructans

ABSTRACT The soilborne fungus Cylindrocarpon destructans (teleomorph: Neonectria radicicola) causes root rot in a wide range of plant hosts; the disease is of particular concern in ginseng production, and in conifer and fruit tree nurseries. beta-Tubulin gene and rRNA gene internal transcribed spacer (ITS) sequence data and pathogenicity assays were used to characterize isolates of C. destructans from ginseng and other hosts. The results of these studies demonstrated a high amount of sequence divergence among strains identified as C. destructans or N. radicicola, suggesting the existence of several phylogenetic species in this complex. Accordingly, we propose that the two varieties of N. radicicola be raised to species status. Certain highly aggressive ginseng isolates from Ontario, Korea, and Japan have identical ITS and beta-tubulin sequences, and form a monophyletic clade (designated "clade a"); these strains are identified as C. destructans f. sp. panacis. Other ginseng strains clustered in monophyletic groups with strains from angiosperm and conifers. A subtractive hybridization method was used to isolate genomic DNA sequences with diagnostic potential from the aggressive C. destructans Ontario ginseng isolate 1640. One of these sequences was similar to the rRNA gene intergenic spacer from a Fusarium oxysporum isolate from Pinus ponderosa, and hybridized to DNA from F. oxysporum and all C. destructans isolates tested. Primers were designed that could be used to amplify this sequence specifically from the highly aggressive, ginsengadapted C. destructans isolates from Ontario and Korea and other members of clade a.     

The fate of [3-14C]metamitron in sugar beets after preemergence application in a lysimeter study

In a lysimeter experiment, [3-¹⁴C]metamitron was sprayed in a preemergence treatment of sugar beets, corresponding to approx 4.9 kg metamitron (7 kg Goltix)/ha. After 6 months, the beets contained metamitron equivalents amounting to 0.1 mg/kg fresh wt, calculated on the basis of the specific radioactivity of the [3-¹⁴C]metamitron employed. Radioactivity was also detected in the pure sugar isolates. The ¹⁴C activity represented approx 0.2 mg metamitron equivalent/kg pure sugar. Since the specific radioactivities of the sugar fractions were too low to employ physicochemical methods, a microbial degradation was used to investigate whether the radiocarbon was incorporated in the sucrose molecule. Microorganisms (Proteus vulgaris) degraded [U-¹⁴C] sucrose and the sugar isolates at the same ¹⁴CO₂ release rates under strictly controlled experimental conditions. This result indicates that about one fourth of the carbon from the C-3 position of the triazine ring of the metamitron, found in the sugar beets at harvest time, is partly being used as a substrate in the production of sucrose possibly via assimilation of mineralized ¹⁴CO₂.     

Factors Influencing Development of Root Rot on Ginseng Caused by Cylindrocarpon destructans

ABSTRACT The fungus Cylindrocarpon destructans (Zins) Scholten is the cause of root rot (disappearing root rot) in many ginseng production areas in Canada. A total of 80 isolates of C. destructans were recovered from diseased roots in a survey of ginseng gardens in British Columbia from 2002-2004. Among these isolates, 49% were classified as highly virulent (causing lesions on unwounded mature roots) and 51% were weakly virulent (causing lesions only on previously wounded roots). Pectinase and polyphenoloxidase enzymes were produced in vitro by C. destructans isolates when they were grown on pectin and phenol as a substrate, respectively. However, highly virulent isolates produced significantly (P < 0.001) higher enzyme levels compared with weakly virulent isolates. Histopathological studies of ginseng roots inoculated with a highly virulent isolate revealed direct hyphal penetration through the epidermis, followed by intracellular hyphal growth in the cortex. Subsequent cell disintegration and accumulation of phenolic compounds was observed. Radial growth of highly and weakly virulent isolates on potato dextrose agar was highest at 18 and 21 degrees C, respectively and there was no growth at 35 degrees C. Mycelial mass production was significantly (P 相似文献   

真菌寄生大豆孢囊线虫的初步研究

从山东省高密县、胶州市及江苏省沛县5个采集点的大豆孢囊线虫中,共分离出110个真菌菌株。优势真菌是镰刀菌,其它为柱孢菌、弯孢霉菌和一种子囊菌。真菌的出现机率平均为44％。在洋菜平板上测定,菌株对该线虫卵,均有不同程度的重寄生能力,其中茄病镰刀菌高达49％。该菌和柱孢菌的分生孢子萌发后可直接穿透卵壁,卵的内含物完全被其粗短的菌丝体取代。     

Development of an Arabidopsis thaliana-based bioassay for investigating seed colonization by mycotoxigenic Aspergillus species

Seeds of Arabidopsis thaliana are hosts for three mycotoxin-producing Aspergillus species, A. flavus , A. nidulans and A. parasiticus, enabling an Aspergillus - Arabidopsis infection (AAI) assay to be developed. The AAI assay involved inoculation of 10- to 12-mg aliquots of uniformly cultivated, surface-sterilized A. thaliana seeds in microcentrifuge tubes. Use of microcentrifuge tubes facilitated qualitative and quantitative analyses of post-infection characteristics such as sporulation and mycotoxin production. Cultivation of A. thaliana seeds under uniform environmental conditions is necessary to limit genotype-independent seed-lot variability. Using the A. nidulans oxylipin mutant, Δ ppoABC , and two well-characterized A. thaliana pathogen-defence mutants, ein2-1 and pad4-1 , the AAI assay permitted genetic analysis of seed infection and mycotoxin production. Sporulation, but not mycotoxin production, was impaired in A. nidulans ΔppoABC , while A. thaliana ein2-1 and pad4-1 had a small but detectable influence on A. nidulans sporulation that appeared to be dependent on seed age.     

Prevalence and pathogenicity of spinach root pathogens of the genera Aphanomyces, Phytophthora, Fusarium, Cylindrocarpon, and Rhizoctonia in Sweden

In a 4-year disease survey in commercial spinach fields, pathogens were isolated from spinach root pieces placed on selective agar media. Aphanomyces cladogamus was the most abundant pathogen, followed by Phytophthora. cryptogea and Fusarium oxysporum. Rhizoctonia solani was found only occasionally. Other pathogens isolated were F. redolens, F. sambucinum and Cylindrocarpon destructans. P. cryptogea was the most severe pathogen, causing death of most plants, but A. cladogamus also caused severe root damage. Isolates of F. oxysporum ranged from highly pathogenic, i.e. P. oxysporum f.sp. spinaciae race 1. to moderately pathogenic and non-pathogenic, Rhizoctonia solani isolates also varied widely in their pathogenicity. Only a small number of the F. redotens and F. sambucinum isolates were pathogenic and most C. destructans isolates were weakly pathogenic. Isolation frequencies were relatively stable from year to year, but P. cryptogea was isolated more frequently in autumn than in spring. No clear relationships were found between pathogen prevalence and disease severity index of surveyed field plants, between pathogen prevalence and plant developmental stage, or between prevalence of the different pathogens isolated.     

Pathogenicity of fungi associated with winter loss and injury in white lupin

Fungi isolated from diseased white lupin plants were tested for pathogenicity in standardized glasshouse tests to assess their potential as causes of plant death in winter in autumn-sown crops. Pleiochaeta setosa and, to a slightly lesser extent, Fusarium avenaceum caused lesions or plant death when inoculated onto hypocotyls of lupin seedlings. Disease was more severe when the hypocotyls were wounded before inoculation. F. avenaceum also caused injury when inoculated onto roots; wounding the roots had no effect. Some isolates of F. solani also caused injury, especially after root inoculation, but isolates of F. oxysporum and Cylindrocarpon destructans were only slightly or non-pathogenic. Isolates of Fusarium and C. destructans , but not of P. setosa , differed in pathogenicity. There were differences in susceptibility to disease among three lupin cultivars.     

Influence of iron on cylindrocarpon root rot development on ginseng

ABSTRACT Cylindrocarpon root rot, caused by Cylindrocarpon destructans, is an important disease on ginseng (Panax quinquefolius) in Canada. We studied the effects of iron (Fe) on disease severity and pathogen growth. When Hoagland's solution was amended with Fe at 56 and 112 mug/ml compared with 0 mug/ml, disease initiation and final severity on hydroponically maintained ginseng roots was significantly (P<0.0001) enhanced. Under field conditions, wounding of roots with a fine needle followed by application of 0.05% FeNaEDTA to the rhizosphere of treated plants significantly enhanced Cylindrocarpon root rot in 2003 and 2004 compared with unwounded roots with Fe or wounded roots without Fe. Foliar applications of Fe (as FeNaEDTA) to ginseng plants three times during the 2002 and 2003 growing seasons significantly increased Fe levels in root tissues. These roots developed larger lesions following inoculation with C. destructans in vitro. When radioactive Fe ((59)Fe) was applied to the foliage of ginseng plants, it was detected in the secondary phloem and in cortical and epidermal tissues within 1 week. Artificially wounded areas on the roots accumulated more (59)Fe than healthy areas. Diseased tissue also had threefold higher levels of phenolic compounds and Fe compared with adjoining healthy tissues. High-performance liquid chromatography analysis revealed enhanced levels of protocatechuic acid, chlorogenic acid, caffeic acid, ferulic acid, cinnamic acid, phloridizin, and quercetin. Phenolic compounds produced in diseased and wounded tissues sequestered Fe in vitro. The effects of Fe on mycelial growth, conidial germ tube length, and secondary branching of germ tubes of C. destructans were examined in vitro. When grown on Chrome-azurol S medium, Fe also was sequestered by C. destructans through siderophore production, which was visualized as a clearing pigmented zone at the margin of colonies. Mycelial dry weight was significantly increased in glucose/ yeast broth containing Fe at 56 or 112 mug/ml. Conidial germ tube length and secondary branching of hyphae also were enhanced after 8 and 16 h by Fe. Colony growth of C. destructans was not enhanced by Fe, but significantly greater spore production was observed with Fe at 56 and 112 mug/ml compared with no Fe in the medium. Although these levels of Fe had no effect on fungal pectinase enzyme activity, polyphenoloxidase (PPO) activity was significantly (P <0.0001) enhanced. We conclude that Fe enhances Cylindrocarpon root rot through enhanced pathogen growth, sporulation, and PPO enzyme activity. Fe sequestered by phenolic compounds produced in wounded tissues can enhance Fe levels at the site of infection. The pathogen also has the ability to sequester Fe at these sites.     

Aesculus pavia foliar saponins: defensive role against the leafminer Cameraria ohridella

BACKGROUND: Recently, the leafminer Cameraria ohridella Deschka & Dimic has caused heavy damage to the white‐flowering horse chestnut in Europe. Among the Aesculus genus, A. pavia L. HBT genotype, characterised by red flowers, showed an atypical resistance towards this pest. Its leaves, shaken in water, originated a dense foam, indicating the presence of saponins, unlike the common horse chestnut tree. The aim was to isolate and identify these leaf saponins and test their possible defensive role against C. ohridella. RESULTS: Spectroscopic analyses showed that A. pavia HBT genotype leaves contained a mixture of saponins, four of which were based on the same structure as commercial escin saponins, the typical saponin mixture produced by A. hippocastanum and accumulated only within bark and fruit tissues. The mixture showed a repellent effect on C. ohridella moth. The number of mines detected on the leaves of A. hippocastanum plants treated with A. pavia HBT saponins through watering and stem brushing was significantly lower than the control, and in many cases no mines were ever observed. CONCLUSION: The results showed that the exogenous saponins were translocated from roots/stem to the leaf tissues, and their accumulation seemed to ensure an appreciable degree of protection against the leafminer. Copyright © 2010 Society of Chemical Industry