新型杀虫剂Spinosad产生菌株的诱变选育

对spinosad产生菌Saccharopoly,Spora,Spinosad508,采用离子束辐照和亚硝基胍(NTG)诱变相交替的办法,选育得到一突变株,其发酵水平较原始菌株提高了140%。

48%Spinosad浓溶剂防治棉铃虫试验研究

通过室内和田间试验 ,全面评价新型生物农药 Spinosad防治抗性棉铃虫的效果及对棉田主要天敌龟纹瓢虫的安全性。结果表明 ,该农药每公顷有效用量 3 0 .2 4～ 40 .3 2 g控制抗性棉铃虫 ,尤其是大龄幼虫效果显著 ,持效期长 ( 8～ 1 0天 ) ,保顶、保蕾效果明显 ,且对棉田主要天敌安全 。

Spinosad对烟草甲生物活性的影响

用含不同剂量Spinosad的烟叶和面粉为食料,研究其对不同形态的烟草甲及幼虫解毒酶的影响;同时配制不同浓度的Spinosad溶液点滴处理烟草甲幼虫。结果表明:Spinosad对烟草甲5龄幼虫的触杀作用和胃杀作用均较弱,但在5、10 mg/kg两种用量下,能完全抑制烟草甲卵的有效孵化和烟草甲成虫的繁殖,在处理后3～18个月内,Spinosad对烟草甲的防效分别为87.11%～99.11%和96.89%～100%;同时,Spinosad对烟草甲幼虫体内的羧酸酯酶(CarE)、谷胱甘肽转移酶(GSTs)和O-脱甲基酶活性均有明显的诱导作用,并对羧酸酯酶的动力学常数Vmax和Km也有一定影响。

新型杀虫剂——Spinosad发酵工艺的研究

采用60 Co正交设计的方法 ,选择最优种子培养基配方和发酵培养基配方 ,在此基础上进行UV、60 Co等诱变 ,得突变株 ,其发酵水平较原始菌株提高了 1 5 0 %。同时 ,对菌株的发酵接种量、发酵液处理及发酵曲线也作了研究。

2.5%菜喜悬浮剂(spinosad)对三种储粮害虫的毒力测定

用药膜法室内测定了新型生物杀虫剂2.5%菜喜悬浮剂(主要成分为多杀菌素,spinosad)对储粮害虫嗜虫书虱、米象、锯谷盗的作用效果。结果表明菜喜对嗜虫书虱、锯谷盗和米象触杀毒力LC50分别为0.0022、0.0046、0.0059mg/m2。对书虱的防治效果明显高于对其他两种储粮害虫的防治效果。本试验结果对多杀菌素在储粮害虫防治方面的合理应用提供了基本依据。

新型微生物杀虫剂—Spinosad(多杀菌素)的毒理学研究进展

多杀菌素是一种大环内酯类化合物,由于其兼有生物农药的安全性和化学农药的速效性,而获得美国“总统绿色化学品挑战奖”。文章主要从多杀菌素对靶标昆虫的杀虫活性和对非靶标昆虫的毒性,多杀菌素的残留评价及分析检测方法,多杀菌素对昆虫的毒理机制等方面进行综述,并对多杀菌素的应用前景作了展望。

农用抗生素刺糖菌素(Spinosads)的研究进展

本文综述了农用抗生素刺糖菌素的研究进展，主要论述了它的理化性质、生理 特性、发酵生产、分离提取、分析检测、活性谱和ＬＤ５０，旨在为刺糖菌素的研究开发提 供参考。

Field resistance to spinosad in western flower thrips Frankliniella occidentalis(Thysanoptera:Thripidae)

The western flower thrips, Frankliniella occidentalis, is one of the most destructive sucking pests of vegetables, fruits and ornamental crops in China. Spinosad is one of the most commonly used insecticides to manage thrips. To assess the incidence of spinosad resistance in F. occidentalis field populations in eastern China, survival rates for 24 different populations were compared with those of a susceptible laboratory strain. All populations showed significantly higher resistance to spinosad compared with the control as determined by comparing median lethal concentrations. Two populations from Shouguang and Liaocheng in Shandong Province were classified as having moderate and high levels of resistance to spinosad with a mean resistance ratio of 17.0 and 89.2, respectively. Our research indicates a widespread reduction in spinosad efficacy for controlling F. occidentalis field populations, and that resistance management strategies should be implemented as soon as practicable, to reduce the potential of progressive resistance development and loss of efficacy.

Spinosad:天然产物研究开发的成功范例

多杀菌素是一类新型杀虫剂。在美国环保署(EPA)登记,能够控制多种害虫。这一独特有机物的发现和后续的研发给世界带来了一类全新的产品。多杀菌素具有独特的作用方式,对目标害虫有很高的活性,对非靶标有机体低毒。多杀菌素对于害虫的控制有较好的效果。

Effects of Glucose and Phosphate on Spinosad Fermentation by Saccharopolyspora spinosa

The effects of glucose and inorganic phosphate on mycelium growth and spinosad production with Saccharopolyspora spinosa were studied. The results showed that the increase of glucose concentration from 18.6g·L^-1 to 58.8g·L^-1 could promote the mycelium growth and spinosad production. And when the glucose con- centration increased from 58.8g·L^-1 to 79.6g·L^-1, no obvious change was detected but a slight drop in spinosad production was observed, whereas, when the glucose concentration increased from 79.6g·L^-1 to 115.3g·L^-1, substantial decrease in both mycelium growth and spinosad production occurred. The increase of phosphate concentra- tion from 3.68mmol·L^-1 to 29.41mmol·L^-1 rendered corresponding increase in mycelium growth and spinosad production. When phosphate concentration increased from 29.41mmol·L^-1 to 44.12mmol·L^-1, mycelium growth slightly increased and spinosad production dropped, while when phosphate concentration increased from 44.12mmol·L^-1 to 57.62mmol·L^-1, both mycelium growth and spinosad production decreased sharply. Conclusively, the optimal initial concentration of glucose and phosphate is 58.8g·L^-1 and 29.41mmol·L^-1, respectively. The spinosad fermentation in the production medium containing 58.8g·L^-1 glucose and 29.41mmol·L^-1 phosphate was scaled up in 5-L fermentation and the spinosad production reached 507mg·L^-1, which was 28% higher thar that in the flask fermentation.

Larvicidal and pupicidal activity of spinosad against the malarial vector Anopheles stephensi

Objective:To investigate the larvicidal and pupicidal activity of spinosad against Anopheles stephensi Listen.Methods:Spinosad from the actinomycete,Saccharopolyspora spinosa was tested against Anopheles stephensi at different concentrations（0.01,0.02,0.04,0.06 and 0.08 ppm.）, and against first to fourth instar larvae and pupae.Results:The larval mortality ranged from 36.1±1.7 in（0.01 ppm） to 79.3±1.8（0.08 ppm） the first instar larva.The LC50 and LC90 values of first, second,third and fourth instar larva were 0.001,0.031,0.034,0.036 and 0.0113,0.102,0.111,0.113, respectively.The pupal mortality ranged from 33.0±2.0（0.01 ppm） to 80.0±0.9（0.08 ppm）.The LC50 and LC90 values were 0.028 and 0.1020,respectively.The reduction percentage of Anopheles larvae was 82.7%,91.4%and 96.0%after 24,48,72 hours,respectively,while more than 80% reduction was observed after 3 weeks.Conclusions:In the present study spinosad effectively caused mortality of mosquito larvae in both the laboratory and field trial.It is predicted that spinosad is likely to be an effective larvicide for treatment of mosquito breeding sites.

Fed-Batch Fermentation for Spinosad Production in an Improved Reactor

As a kind of aerobic bacteria, Saccharopolyspora spinosa exhibits a high demand for oxygen. In the fermentation process, the methods of increasing ventilation and improving agitation speed are usually adopted to achieve higher values of dissolved oxygen. These methods decrease the efficiency of spinosad biosynthesis. In this study, an improved reactor was designed to solve these problems. The exhaust gas reflux device, impellers, and baffles were improved. Furthermore, we established the kinetic models for the cell growth, substrate consumption and spinosad generation in batch fermentation process. The simulation results were in good agreement with the experimental data. Spinosad production reached 583.86 mg/L after employing the suitable feeding strategy by fed-batch fermentation in the improved reactor, whereas it was only 157.01 mg/L before optimization. The method described can provide insight to strengthen spinosad production and can be extended to the culturing process of filamentous aerobic bacteria. © 2017 Tianjin University and Springer-Verlag Berlin Heidelberg

生物农药乙基多杀菌素的研究进展

多杀菌素类化合物是一类新型高效的生物源杀虫剂,对多种害虫具有显著的活性抑制作用。多杀霉素(spinosad)和乙基多杀菌素(spinetoram)为多杀菌素类化合物第1代和第2代商业化产品,具有杀虫活性高、降解速度快、对环境友好以及对非靶标生物毒性低等特点。本文着重对乙基多杀菌素的理化性质、杀虫机制、应用、合成方法以及衍生化研究进行了阐述,并对多杀菌素类化合物的研究进行了展望。

多杀霉素/羧甲基壳聚糖缓释悬浮剂的制备

基于羧甲基壳聚糖在等电点附近溶解度低的特性,采用pH调节法制备多杀霉素/羧甲基壳聚糖缓释悬浮剂,并初步探究其对小菜蛾幼虫的防治效果。结果表明:缓释悬浮剂适宜的制备条件为羧甲基壳聚糖盐酸溶液质量分数1.0%,多杀霉素甲醇溶液质量分数1.5%,羧甲基壳聚糖与多杀霉素的质量比2∶1,15000 r/min下剪切乳化3 min。缓释悬浮剂呈乳白色,载药量、包封率和中位粒径分别约为25%、74%和48μm,稀释施用时粒径降为约3μm。红外光谱分析表明,羧甲基壳聚糖成功包封多杀霉素,其相互间作用力主要为物理吸附。相对传统悬浮剂,多杀霉素/羧甲基壳聚缓释悬浮剂在防治小菜蛾方面具有优异的速效性和更长的持效期,在多杀霉素减施增效方面具有一定的应用潜力。

Mechanisms of resistance to spinosad in the western flower thrip, Frankliniella occidentalis （Pergande） （Thysanoptera： Thripidae）

Cross-resistance, resistance mechanisms, and mode of inheritance of spinosad resistance were studied in the western flower thrip, Frankliniella occidentalis （Pergande）. Spinosad （naturalyte insecticide） showed low cross-resistance to prothiophos （organophosphorus insecticide） and chlorphenapyr （respiratory inhibitor） showed some cross-resistance to thiocyclam （nereistoxin）. The synergists PBO （piperonyl butoxide）, DEM （diethyl maleate）, and DEF （s,s,s-tributyl phosphorotrithioate） did not show any synergism on the toxicity of spinosad in the resistant strain （ICS）, indicating that metabolic- mediated detoxification was not responsible for the spinosad resistance, suggesting that spinosad may reduce sensitivity of the target site： the nicotinic acetylcholine receptor and GABA receptor. Following reciprocal crosses, dose-response lines and dominance ratios indicated that spinosad resistance was incompletely dominant and there were no maternal effects. The results of backcross showed that spinosad resistance did not fit a single-gene hypothesis, suggesting that resistance was influenced by several genes.

13种杀虫剂对木橑尺蠖的室内毒力测定及田间防效

【目的】木橑尺蠖Culcula panterinaria是一种杂食性鳞翅目害虫,为了对浙江省临安区山核桃等多种果树木和经济作物进行有效防治。【方法】本研究分别采用浸液法、浸叶法测定了13种杀虫剂对木橑尺蠖卵与1龄、3龄、5龄3个龄期幼虫的室内毒力,并通过田间试验测定了13种杀虫剂对木撩尺蠖的田间防效。【结果】乙基多杀菌素、多杀菌素和氯虫苯甲酰胺对木橑尺蠖卵的致死中浓度(LC_(50))分别为0.322、0.514和0.656 mg/L;多杀菌素、氯虫苯甲酰胺和乙基多杀菌素对木橑尺蠖5龄幼虫的毒力较高,致死中浓度(LC_(50))分别为1.414、2.147和2.658 mg/L。多杀菌素、氯虫苯甲酰胺和乙基多杀菌素对木橑尺蠖的田间防效高于其他杀虫剂,其中多杀菌素的田间防效为94.56%。【结论】推荐生物源杀虫剂多杀菌素或乙基多杀菌素与氯虫苯甲酰胺轮换使用可防治木橑尺蠖。

多杀菌素生产菌的复合诱变选育及发酵优化

生物农药多杀菌素的发酵水平限制了其产业化发展。为了提高多杀菌素产量,以刺糖多孢菌T1(Saccharopolyspora spinose T1)为出发菌,通过常压室温等离子体(atmospheric and room temperature plasma,ARTP)诱变和硫酸二乙酯(diethyl sulfate,DES)诱变,并以链霉素和鼠李糖作为筛选因子结合微孔板培养进行筛选,获得高产菌株S.spinosa F5;发酵后其多杀菌素产量为572.3 mg/L,比出发菌提高35.9%;通过单因素实验和响应面实验确定柠檬酸钠、异亮氨酸的最适添加量分别为0.46、0.10 g/L,豆油的最适添加体积分数为1.77%,此时多杀菌素产量为707.2 mg/L。在5 L发酵罐中进行分批发酵,发酵240 h产量接近800 mg/L;进一步分批补料发酵264 h,多杀菌素产量达1175.5 mg/L。结果表明,ARTP和DES复合诱变育种及添加外源诱导物可以显著提高多杀菌素产量。

Promotion of spinosad biosynthesis by chromosomal integration of the Vitreoscilla hemoglobin gene in Saccharopolyspora spinosa

To promote spinosad biosynthesis by improving the limited oxygen supply during high-density fermentation of Saccharopolyspora spinosa, the open reading frame of the Vitreoscilla hemoglobin gene was placed under the control of the promoter for the erythromycin resistance gene by splicing using overlapping extension PCR. This was cloned into the integrating vector pSET152, yielding the Vitreoscilla hemoglobin gene expression plasmid pSET152EVHB. This was then introduced into S. spinosa SP06081 by conjugal transfer, and integrated into the chromosome by site-specific recombination at the integration site ФC31 on pSET152EVHB. The resultant conjugant, S. spinosa S078-1101, was genetically stable. The integration was further confirmed by PCR and Southern blotting analysis. A carbon monoxide differential spectrum assay showed that active Vitreoscilla hemoglobin was successfully expressed in S. spinosa S078-1101. Fermentation results revealed that expression of the Vitreoscilla hemoglobin gene significantly promoted spinosad biosynthesis under normal oxygen and moderately oxygen-limiting conditions （P〈0.01）. These findings demonstrate that integrating expression of the Vitreoscilla hemoglobin gene improves oxygen uptake and is an effective means for the genetic improvement of S. spinosa fermentation. Saccharopolyspora spinosa, spinosad, Vitreoscilla hemoglobin, integrating vector, homologous recombination