基于聚氨酯海绵载体的棘孢木霉固体发酵培养基筛选

以棘孢木霉Trichoderma asperellum THGY-01(CGMCC No:22422)为试验菌株,对其在聚氨酯海绵载体上固体发酵产孢的培养基种类、浓度及促进产孢元素进行了筛选与优化.以聚氨酯海绵载体的单位质量产孢量为指标,通过单因素试验法对11种碳源、6种氮源和14种无机盐(8种含大量元素,6种含微量元...     

Effect of susceptible and resistant canola plants on Plasmodiophora brassicae resting spore populations in the soil

Clubroot, caused by Plasmodiophora brassicae, has become a serious threat to canola (Brassica napus) production in western Canada. Experiments were conducted to assess the effect of growing resistant and susceptible canola genotypes on P. brassicae soil resting spore populations under greenhouse, mini‐plot and field conditions. One crop of susceptible canola contributed 1·4 × 10⁸ spores mL^?1 soil in mini‐plot experiments, and 1 × 10¹⁰ spores g^?1 gall under field conditions. Repeated cropping of susceptible canola resulted in greater gall mass compared to resistant canola lines. It also resulted in reduced plant height, increased clubroot severity in susceptible canola, and increased numbers of resting spores in the soil mix.     

Direct evidence of surface infestation of seeds and tubers by Plasmodiophora brassicae and quantification of spore loads

Using quantitative PCR, DNA of Plasmodiophora brassicae, the causal agent of clubroot, was detected and quantified on canola, pea and wheat seeds, as well as on potato tubers, all harvested from clubroot‐infested fields in Alberta, Canada. Quantifiable levels of infestation were found on seven of the 46 samples analysed, and ranged from <1·0 × 10³ to 3·4 × 10⁴ resting spores per 10 g seeds; the vast majority (80–100%) of resting spores on these samples were viable, as determined by Evan’s blue vital staining. However, the levels of infestation found were generally lower than that required to cause consistent clubroot symptoms in greenhouse plant bioassays. While the occurrence of P. brassicae resting spores on seeds and tubers harvested from clubroot‐infested fields suggests that seedborne dissemination of this pathogen is possible, practices such as commercial seed cleaning may be sufficient to effectively mitigate this risk.     

亚蝗微粒子虫毒饵防治东亚飞蝗的研究

用亚蝗微粒子虫毒饵在室内和田间防治东亚飞蝗的试验结果表明，饵料含孢量约１×１０^１２个／ｋｇ，投放量２ｋｇ／６６７ｍ^２，于蝗蝻２龄期施用，１４ｄ内可获得８７．８％的杀蝻效果     

Studies into primary and secondary infection processes by Plasmodiophora brassicae on canola

The early stages of infection of canola roots by the clubroot pathogen Plasmodiophora brassicae were investigated. Inoculation with 1 × 10⁵ resting spores mL^?1 resulted in primary (root hair) infection at 12 h after inoculation (hai). Secondary (cortical) infection began to be observed at 72 hai. When inoculated onto plants at a concentration of 1 × 10⁴ mL^?1, secondary zoospores produced primary infections similar to those obtained with resting spores at a concentration of 1 × 10⁵ mL^?1. Secondary zoospores caused secondary infections earlier than resting spores. When the plants were inoculated with 1 × 10⁷ resting spores mL^?1, 2 days after being challenged with 1 × 10⁴ or 1 × 10⁵ resting spores mL^?1, secondary infections were observed on the very next day, which was earlier than the secondary infections resulting from inoculation with 1 × 10⁷ resting spores mL^?1 alone and more severe than those produced by inoculation with 1 × 10⁴ or 1 × 10⁵ resting spores mL^?1 alone. Compared with the single inoculations, secondary infections on plants that had received both inoculations remained at higher levels throughout a 7‐day time course. These data indicate that primary zoospores can directly cause secondary infection when the host is under primary infection, helping to understand the relationship and relative importance of the two infection stages of P. brassicae.     

Effects of dimehypo (disodium 2-methylaminotrimethylene di thiosulfonate) on growth and cocooning of the silkworm,Bombyx mori (Lepidoptera: Saturnidae)

Dimehypo (disodium 2-methylaminotrimethylene di thiosulfonate), is an insecticide used on rice and other crops in China. However, contamination of mulberry leaves with this has been implicated in a reduction of silk production. The acute and chronic toxicity of dimehypo to Bombyx mori L over the partial life cycle of the organism was determined based on survival, growth and cocooning of two strains of silkworm larvae. A change in the ultrastructure of the posterior silk gland cell was also observed in this study. The results showed that the growth and development of tested larvae was impeded and their life cycle was prolonged in both strains. It was also found that dimehypo was extremely harmful to the cocooning of B mori. Ultrastructural evidence suggests that adverse effects of dimehypo arise as a result of changes in the biosynthesis of fibroin and in the physiological activity of the posterior silk gland cell. The maximum acceptable daily dose of dimehypo based on growth and cocooning of B mori is less than 1.7 × 10⁻⁶ µg day⁻¹ in spring-reared larvae and less than 1.7 × 10⁻⁸ µg day⁻¹ in autumn-reared larvae. © 1999 Society of Chemical Industry     

Stability and controlled release properties of carboxymethylcellulose-encapsulated Bacillus thuringiensis var. Israelensis

The mosquito and blackfly pathogen, Bacillus thuringiensis Berl. var. israelensis (B.t.i.). containing spores and toxin crystal, was successfully encapsulated in an insolubilized carboxymethylcellulose (CMC)-aluminum matrix creating a controlled-release bead formulation. The release rate of B.t.i. from this formulation was sufficient to kill Culex sp. larvae during bioassays performed in the laboratory. The CMC concentration significantly affected the release rate of the bacterium. All types of beads tested exhibited a high initial release of B.t.i. in the first week of experiments, followed by a slower rate later. Levels of 10 g kg^?1 CMC solution and 0–05 m aluminium sulfate were found to be optimal in the formulation mixture for polysaccharide and gellant, respectively. This type of bead gave the fastest release of the bacterium and caused 100% larval mortality as from the second day of treatment. While acidic pH, high temperature and UV exposure drastically decreased spore viability of B.t.i., the CMC-encapsulated formulation was more stable to these conditions. Larvicidal activity of encapsulated B.t.i. was more resistant to high temperature (50°C) than the unformulated B.t.i. The high response of Culex larvae to the encapsulated B.t.i. suggests that the formulation could be employed effectively in the field.     

草地贪夜蛾病原微孢子虫的鉴定及其致病力分析

为明确从田间采集草地贪夜蛾 Spodoptera frugiperda幼虫体内发现的一种微孢子虫的分类地位和致病性,利用传统形态学观察和分子生物学技术对该微孢子虫进行鉴定,同时采用室内生物活性测定法对其致病性进行分析。结果显示,该微孢子虫的形态学特征与家蚕微孢子虫 Nosemabombycis相近,具有典型微孢子虫超微组成结构,孢子壁厚度为195.00~205.15 nm,极丝盘旋于孢子后极内侧10~12圈;其基因组的基因间隔区（intergenic spacer, ITS）和小亚基核糖体RNA（smallsubunit ribosomal RNA, SSU）序列与已报道的家蚕微孢子虫相关序列的相似度分别达94.34%和99.50%,系统发育树显示该微孢子虫属于微孢子虫属 Nosema,与家蚕微孢子虫亲缘关系最近。该微孢子虫侵染草地贪夜蛾1龄和2龄幼虫5 d时的LC50分别为2.51×10⁷孢子/mL和2.48×10⁷孢子/mL;侵染3龄幼虫10 d时的LC₅₀为3.79×10⁷孢子/mL;侵染4龄幼虫15 d时的LC50为3.98×10⁷孢子/mL;且当微孢子虫浓度为1.0×10⁸孢子/mL时,草地贪夜蛾1至4龄幼虫的LT₅₀分别为3.04、 3.86、 7.47和10.43 d。表明该微孢子虫隶属微孢子虫属,对草地贪夜蛾不同龄期幼虫均有较强的致病力,具有良好的开发应用潜力。     

生防菌Alternaria amaranthi-3对反枝苋的防治效果

反枝苋Amaranthus retroflexus是一种世界性恶性杂草。为了确定微生物除草剂候选菌Alternaria amaranthi-3防除反枝苋的潜力,通过盆栽试验研究了接种浓度、露期和水乳剂型对A.ama-ranthi-3菌株致病力的影响。结果显示,接种浓度显著影响菌株的致病力,在48h露期条件下,接种孢子浓度为105个/mL时,菌株水剂对反枝苋幼苗生长抑制率为35.55%;浓度为107个/mL时,生长抑制率达到75.25%。露期对该菌株的致病力也有较大影响,在不保湿条件下,菌株水剂对反枝苋的生长抑制率为26.43%,而保湿48h处理的生长抑制率达到77.96%。Span80∶Tween80=1∶3的复配乳化剂和大豆油制备的水乳剂可显著降低露期对菌株防效的影响和提高菌株的致病力,无露期时,菌株水乳剂对反枝苋的生长抑制率达到88.35%,显著高于水剂;48h露期条件下,菌株水乳剂处理的生长抑制率为90.59%,而菌株水剂处理为77.96%。表明通过剂型的改进菌株Alternaria amaranthi-3能有效防除反枝苋。     

菌株NDBJJ-BFG的鉴定及其对马铃薯甲虫不同虫态的毒力测定

为筛选出对马铃薯甲虫Leptinotarsa decemlineata(Say)具有防治潜力的菌株,本研究从马铃薯甲虫僵虫虫体上分离菌株NDBJJ-BFG,通过形态学特征和分子生物学方法明确其分类地位,采用喷雾法和覆土法测定了不同浓度下该菌株对马铃薯甲虫室内毒力的致死中浓度LC50及半致死时间LT50。结果表明,经形态特征与r DNA ITS序列分析最终确定菌株NDBJJ-BFG为球孢白僵菌Beauveria bassiana(Bals.)Vuill。该菌株对1、2、3和4龄马铃薯甲虫幼虫的LC50分别为0.91×10~6、1.51×10~6、5.09×10~6和6.84×10~6个/m L;采用喷雾法和覆土法处理蛹的LC50分别为1.43×10~7个/m L和8.15×10~6个/m L,成虫的LC50分别为5.08×10~7个/m L和2.97×10~7个/m L。在孢子悬浮液浓度相同时LT50随虫龄的增大而延长,其中成虫的LT50最长,其次是蛹;在相同龄期下LT50随着孢子悬浮液浓度的增大而缩短。表明菌株NDBJJ-BFG对马铃薯甲虫1龄和2龄幼虫具有高毒力,其在马铃薯甲虫的生物防治中将具有较大的应用潜力。     

Application of some bacterial varieties ofBacillus thuringiensis and its bioproduct Delfin onSynanthedon myopaeformis Borkh. (Lep. Aegeriidae) in apple orchards

Field experiments were conducted using the biological agentBacillus thuringiensis to control the clearwing moth,Synanthedon myopaeformis (Lep. Aegeriidae) in apple orchards at Alexandria governorate during 1995, 1996 and 1997. Three bacterial varieties ofB. thuringiensis showed significant influence on the reduction of borer infestation. Four applications with the sporic concentration 12×10⁷ spores/ml of each ofB. th. var.kurstaki, B. th. var.alesti andB. th. var.thuringiensis induced 82.5, 51.1 and 42.1% reduction of infestation, respectively. Methods of application showed highly significant effect on infestation level. Sparying after scraping was the best method (80.0%, reduction) followed by painting after scraping (71.5%, reduction) then spraying alone (59.7%, reduction) or painting alone (43.0%, reduction). A control programme was carried out using three applications of the biomaterial, Delfin at the rate of 200 g/100 I.w. during mid-March, early June and mid-August resulted in 88.5–91.4% reduction of infestation.     

Residues of the algicide endothal in water,soil and rice,after paddy field applications

In order to obtain residue data from the application of the algicide endothal in Italian rice paddy fields, two experiments were carried out using a 50 g kg^?1 granular formulation in a small pond and the same granular and two liquid formulations in actual paddy fields of the Italian rice growing area. Endothal decay in the pond water was very rapid, reaching residue levels of 0·01-1·02 mg litre^?1 in two days and 0·004-0·01 mg litre^?1 at the third day. The muddy soil of the pond was free from measurable endothal residues( <0·02 mg kg^?1). In the paddy-field waters, the endothal decay was slower, with an average half-life time of 3·3 days, independently of the type of formulation. The actual residues in water after 6 days ranged from 0·3 to 1·3 mg litre^?1 according to the initial amount of product applied, and, consequently, to the initial concentration in water. Rice samples collected at the normal harvest time from the two paddy fields, treated with three different formulations, showed no endothal residue at the minimum detectable level of 0·01 mg kg^?1.     

Assessment of production and dispersal of inoculum ofAlternaria Macrospora in various parts of the cotton canopy

Morning and evening counts of spores ofAlternaria macrospora in a cotton (Gossypium barbadense) field were assessed for the number of spores (a) produced on the last night, (b) dispersed during the day, and (c) retained in the spore reserve on leaves. Throughout the season, 1 m² of the field produced (P) and dispersed (D) approximately 163 x 10⁶ and 137 x 10⁶ spores, respectively. The 16% discrepancy between P and D values reflects experimental errors. Most spores were produced on shed leaves on the ground, fewer on leaves in the lower part of the canopy, and least on leaves in the upper canopy. The contribution to total D decreased from leaves in the lower part of the canopy, to shed leaves, and to leaves in the upper canopy. In tests with potted plants with spore-bearing lesions attached to various heights of the canopy under various conditions of weather, dispersal of spores at these heights was conditioned mainly by wind velocity, which decreased from the upper to the lower part of the canopy. In leaves close to the ground, approximately 29% of the released spores were removed by sedimentation to the ground. On any one day, the daily rate of D differed from the daily rate of P. On days in which D was higher than P, the difference was due to a reserve of spores remaining on lesions after dispersal during the previous days. In weather adverse for production of new spores, the reserve was depleted within 4 and 6 days in the top and bottom leaves, respectively, but remained relatively high in the shed leaves on the ground; a similar number of days was needed to restore the depleted reserve in sporulation-favorable weather.     

灭蚊幼球形芽孢杆菌漂浮颗粒剂研制与应用

根据蚊虫的生活习性和球形芽孢杆菌BS-10灭蚊特点,以玉米棒芯颗粒为载体、以球型芽胞杆菌BS-10为杀虫活性物质,加工制成生物灭蚊漂浮颗粒剂新剂型,经试验与示范应用,该颗粒剂具有高效,毒力效价达106.5 IU／mg,每平方米水面使用颗粒剂2 g,对蚊幼虫(孑孓)杀灭效果达95%以上;持效期长,药后12 d,灭蚊效果达96.8%;性质稳定,室内常温储存1 a,药效无变化;生产工艺简单,药剂使用方便,与环境相容性好。     

Formulation selection,and investigation of azadirachtin-A persistence in some terrestrial and aquatic components of a forest environment

Five commercial formulations of azadirachtin-A (AZ-A) Margosan-O^®, Azatin-EC^®, Neem-EC^®, RH-9999 and Neemix^® 4.5, were investigated for their volatilization and washoff potential in laboratory studies. Prior to the investigation, RH-9999 (a wettable powder) was mixed with water to provide an end-use formulation containing 35·6 g AZ-A kg^-1, while the remaining four formulations were investigated without dilution. Volatilization and washoff of AZ-A occurred more from white spruce foliage than from wax-coated glass plates. Neem-EC provided the lowest amount of loss, whereas Margosan-O provided the highest. Physical properties and atomization behaviour of the five formulations indicated that Azatin-EC was highly viscous and caused phase separation in droplets collected on glass plates after atomization in a rotary atomizer. RH-9999, despite its low viscosity, caused phase separation in droplets because of the heterogeneity of the wettable powder formulation. Based on the minimum loss of AZ-A due to volatilization and washoff from spruce foliage, and on the minimum potential for phase separation in droplets after atomization in a rotary atomizer, Neem-EC was considered to be the most appropriate choice for use in field studies to investigate environmental persistence and fate of AZ-A in terrestrial and aquatic matrices of a forest ecosystem. The Neem-EC formulation was sprayed at 40 and 80 g AI ha^-1 over single spruce trees and on litter and soil plots selected in a mixed-wood boreal forest in Ontario, Canada. In addition, outdoor aquaria containing stream water and sediment were also fortified with the formulation at 400 and 800 g AI ha^-1. Persistence of AZ-A was evaluated using one-year-old spruce needles, current-year shoots, spruce bark, litter, soil, stream water and sediment. The duration of persistence varied from 3 to 6 days in terrestrial matrices, whereas it ranged from 8 to 13 days in water, and 2 to 3 days in sediment. The half-life (DT₅₀) values ranged from 10·7 h (for soil) to 71·6 h (for spruce bark) at the lower dosage rate, and from 18·8 h (for litter) to 76·2 h (for bark) at the higher dosage rate. The DT₅₀ value for stream water was about 35 h regardless of the dosage rate applied. The data indicated that AZ-A was appreciably labile and short-lived in different forestry matrices, with low DT₅₀ values. © 1997 SCI     

三株球孢白僵菌对草地贪夜蛾的毒力比较

为筛选有效防治草地贪夜蛾Spodoptera frugiperda的微生物制剂,采用室内生物测定法测定从北京市、浙江省和海南省土壤样品中分离获得的3株球孢白僵菌Beauveria bassiana菌株bbbj、bbzj和bbhn对草地贪夜蛾3龄幼虫的毒力,并比较这3株菌株的产孢情况和合成白僵菌素的能力。结果显示,分离自北京市的菌株bbbj对草地贪夜蛾3龄幼虫的毒力最强,LC50为3.37×105个孢子/mL;其次为浙江省的菌株bbzj,毒力略逊于菌株bbbj;海南省的菌株bbhn毒力最弱,其在试验最高浓度108个孢子/mL处理7 d后对草地贪夜蛾3龄幼虫的致死率低于50%。菌株bbbj的产孢量远高于另外2株菌株,并且其菌丝结构上着生大量芽生孢子簇,而且菌株bbbj菌体中的白僵菌素含量最高,培养5 d后,分别为菌株bbzj和bbhn的40.08倍和65.85倍。虽然补充白僵菌素可以提高菌株bbhn的毒力,但是草地贪夜蛾幼虫对白僵菌素敏感度不高。表明球孢白僵菌菌株bbbj对草地贪夜蛾幼虫有较高的毒杀活性,具有作为草地贪夜蛾生防菌株的潜力。     

Cyt1Aa protein from Bacillus thuringiensis (Berliner) serovar israelensis is active against the Mediterranean fruit fly,Ceratitis capitata (Wiedemann)

BACKGROUND: Ceratitis capitata (Wiedemann) is one of the world's most destructive fruit pests. The aim of this study was to ascertain insecticidal activity of Bacillus thuringiensis (Berliner) δ‐endotoxins to C. capitata. RESULTS: Among 42 selected Bacillus strains, only B. thuringiensis serovar israelensis (Bti) solubilised protoxins showed biological activity against C. capitata neonate larvae, whereas Bti spore and crystal mixture was inactive. Insecticidal activity of Bti protoxins was significantly enhanced by incubation with Culex pipiens L. gut extracts. Overdigestion of Bti protoxins with Sesamia nonagrioides (Lefebvre) gut extracts suppressed biological activity against C. capitata, and this correlated with degradation of Cyt toxins. Cyt1Aa solubilised protoxin showed the highest toxicity, LC₅₀ after 7 days of 4.93 µg cm^?2, while proteolytical processing of Cyt1Aa protoxins by larval gut extracts did not enhance insecticidal activity. CONCLUSION: The present study provides evidence for the first time of the insecticidal activity of a B. thuringiensis strain against C. capitata and identifies a single δ‐endotoxin with potential for controlling this pest. Copyright © 2010 Society of Chemical Industry     

球孢白僵菌对松墨天牛的致病力及其与花绒寄甲的相容性

为探明昆虫病原真菌球孢白僵菌Beauveria bassiana与天敌昆虫花绒寄甲Dastarcus helophoroides的相容性,观察4株球孢白僵菌菌株的生长性状、产孢量以及孢子萌发率,并选用Bb3275和Bb202菌株测定其对松墨天牛Monochamus alternatus的致病力以及对花绒寄甲的毒力;同时在模拟环境条件下,分别用Bb3275和Bb202菌株的孢子悬浮液先侵染松墨天牛幼虫,再按1∶1比例将花绒寄甲与受侵染的松墨天牛置于同一环境下饲养,观察花绒寄甲的死亡率。结果显示,孢子悬浮液浓度为1×10⁶孢子/mL至1×10⁸孢子/mL时,Bb3275菌株和Bb202菌株对松墨天牛幼虫和成虫的致病力与对照组相比均有显著差异;经4种浓度Bb3275和Bb202菌株侵染花绒寄甲幼虫和成虫15 d后,仅在高浓度1×10⁸孢子/mL时表现出轻微致死作用,但花绒寄甲的存活率均高于85.00%;模拟条件下,松墨天牛幼虫校正死亡率均在80.00%以上,而同环境下花绒寄甲幼虫和成虫均表现出良好的活力,且侵染率为0。表明球孢白僵菌和花绒寄甲之间具有很好的相容性,可以同时应用于松墨天牛的生物防治。     

Effect of environmental conditions on spore production by Fusarium verticillioides, the causal agent of maize ear rot

Silk infection by Fusarium verticillioides is caused by conidia produced on maize crop residues and results in kernel infection and consequent accumulation of fumonisins. Studies were carried out in both controlled and field conditions to understand the dynamics of sporulation on maize residues. The effect of temperature (5°C to 45°C) and incubation time (3 to 41 days) on spore production on maize meal agar was described by a logistic model that accounted for 85% of variability. The rate parameter depended on the length of incubation and the asymptote on temperature. Maximum sporulation occurred at 27°C, with a progressive increase between 5°C and 27°C and then a rapid decline, with no sporulation at 45°C. Fusarium verticillioides strains from different geographic origins showed different sporulation capabilities, with similar optimum temperatures. Pieces of stalk residues inoculated with F. verticillioides and placed above the soil between rows of maize crops, in 2003 to 2005, produced conidia continuously and abundantly for some weeks, particularly during the period after silk emergence, with an average of 1.59 × 10⁷ conidia g⁻¹ of stalk, over a wide range of environmental conditions. Sixty-seven percent of variability of the spore numbers found on stalks was accounted for by a multiple regression model. Precipitation (rain or overhead irrigation) in the 14 days before stalk sampling decreased the number of spores, whilst the number of days with conducive conditions of moisture (i.e. days with rainfall, average relative humidity >85% or vapour pressure deficit <4 hPa) and greater degree-days (base 0°C) in the 14 and 3 days before sampling, respectively, increased sporulation.     

Effect of Adjuvants on the Therapeutic Activity of Dimethomorph in Controlling Vine Downy Mildew. II. Adjuvant Mixtures,Outdoor-Hardened Vines and One-Pack Formulations

Previous investigations have shown that certain C₁₂–C₁₈ alcohol ethoxylates with between 5 and 20 moles ethylene oxide gave large enhancements to the therapeutic activity of emulsifiable concentrate (EC) and wettable power (WP) formulations of dimethomorph in controlling two-day-old infections of downy mildew (Plasmopara viticola, Berl. & deT) on glasshouse propagated vines (Vitis vinifera, L.). In a continuation of that work it has been shown that similar enhancements could also be obtained on vines that had been grown under outdoor conditions for two to three weeks before treatment (outdoor-hardened vines) and it is concluded that the treatments would therefore be effective on field vines. Mixtures of either ammonium sulfate or the silicone ethoxylate, ‘Silwet’ L77, with the best alcohol ethoxylates, ‘Genapol’ C050 or ‘Genapol’ C080, produced slight antagonism of the action of the ‘Genapols’ and therefore were of no benefit. Trials with one-pack formulations prepared containing the ‘Genapol’ adjuvants produced good dose response curves from which estimated doses for 90% fungal control (ED₉₀ values) could be calculated by Logit analysis. These one-pack formulations were significantly more effective than the standard EC formulation (ED₉₀=335 g AI ha⁻¹, glasshouse vines and 445 g AI ha⁻¹, outdoor hardened vines) with optimum enhancements with those formulations containing an adjuvant: dimethomorph ratio of either 9:1 on glasshouse vines (ED₉₀= 13 g AI ha⁻¹) or 6:1 on outdoor-hardened vines (ED₉₀=30 g AI ha⁻¹). This last result was significantly better than that obtained with a commercial formulation of cymoxanil, ‘Fytospore’, (ED₉₀=86 g AI ha⁻¹) and encouraged the recommendation that these one-pack formulations should be examined in field trials.