Induction of programmed death and cytoskeletal damage on Trichoplusia ni BTI-Tn-5B1-4 cells by azadirachtin

Plenty of insect cells are sensitive to azadirachtin, an ideal biopesticide. However, the mode of action of azadirachtin on insect cells still remains unexplained. In this study, Trichoplusia ni BTI-Tn-5B1-4 cell line was used as the model system to evaluate induction of programmed death and cytoskeletal damage by azadirachtin. The result of colorimetric [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] (MTT) assay indicated that the effect of azadirachtin on cells growth was time-dependent with different concentrations (0.75, 1.5 and 3 μg/mL). As autophagic vacuoles were observed obviously by transmission electron microscopy, azadirachtin could induce autophagy in cells. Furthermore, lysosomal function was affected after treatment with azadirachtin in time-dependent manner. Aberrant morphological figures appeared in treated cells, such as contraction and turning round, especially the appearance of apoptotic bodies and membrane blebbing, which was the evidence of azadirachtin-induced apoptosis, with apoptosis rate of 13.98% at 1.5 μg/mL for 120 h. As the stress fibers changed after treatment, the cytoskeletal function could be influenced. The effect on cell cycle was also analyzed by flow cytometry, with the result that azadirachtin enhanced cell cycle arrest at the G2/M phase gradually.     

几种杀虫剂对亚非玛草蛉的毒力

采用药膜法测定了高毒农药替代品的6种药剂对柑桔害虫重要捕食性天敌亚非玛草蛉Mallada desjardinsi的毒力作用。结果表明,6种药剂对亚非玛草蛉的毒力大小依次为毒死蜱〉氧化乐果〉丁硫克百威〉高效氯氟氰菊酯〉阿维菌素〉印楝素。其中前5种药剂的LC95分别为18.67、324.48、1694.80、11520.83、642592.76mg/L;LC50分别为8.20、41.03、134.85、527.89、7799.57mg/L。印楝素对亚非玛草蛉的毒性极小,其LC50和LC95因毒力低而未测出。     

保护地常用农药对蜡蚧轮枝菌菌丝生长影响研究

6种杀虫剂和 6种杀菌剂对昆虫病原真菌蜡蚧轮枝菌菌丝生长影响的研究结果表明 ,杀虫剂对蜡蚧轮枝菌菌丝生长有程度不同的抑制作用。在田间常规使用浓度下 ,5种杀虫剂对菌丝生长的抑制率在 10 %以上 ,最高者为甲氰菊酯达 17.6%。田间常规使用浓度稀释 10倍下 ,杀虫剂万灵、快灭净和锐劲特对菌丝生长抑制率低于 5% ,锐劲特在3.3～33.3mg/L时 ,菌丝生长与对照无显著差异 ,表现出与蜡蚧轮枝菌有极好的相容性 ,是菌药混用较好的候选杀虫剂。杀菌剂对蜡蚧轮枝菌菌丝生长有强烈的抑制作用 ,浓度愈高 ,抑制程度愈强。因此 ,在利用蜡蚧轮枝菌防治保护地蔬菜害虫时 ,须与杀菌剂使用保持一定的间隔期     

Azadirachtin-A and tetrahydroazadirachtin-A concentrates: preparation, LC-MS characterization and insect antifeedant/IGR activity against Helicoverpa armigera (Hübner)

A 60% azadirachtin-A concentrate has been obtained through repeated precipitation with hexane from a methanolic solution of a 20% concentrate. Azadirachtin-A (90%) has been obtained by medium-pressure liquid chromatography of the 60% concentrate with an RP-18 column and a methanol + water (1 + 1 by volume) solvent system. Catalytic hydrogenation of the 60 and 90% azadirachtin concentrates yielded the corresponding tetrahydroazadirachtin concentrates. Dihydroazadirachtin and tetrahydroazadirachtin formed during the first 5 h of hydrogenation were identified by LC-ESI-MS on the basis of their unique mass fragmentation pattern. The efficacy of tetrahydroazadirachtin concentrates in inhibiting the feeding and growth of Helicoverpa armigera (Hübner) larvae has been compared with that of azadirachtin concentrates. They were in general more active and deterred larvae from feeding at all concentrations. Tetrahydroazadirachtin-A (90%) and azadirachtin-A (90%) with respective IC(50) values of 280 and 390 mg L(-1) were effective as insect growth regulators, while tetrahydroazadirachtin-A (90%) displayed higher antifeedant activity (AI(50) 14 mg L(-1)) against the test insect.     

The commercial use of azadirachtin and its integration into viable pest control programmes

With the continued robust growth of the global biopesticide market, azadirachtin is uniquely positioned to become a key insecticide to expand in this market segment. In the USA the actual or impending cancellation of some organophosphate and carbamate insecticides that have either lost patent protection or are not being re-registered in many markets because of the Food Quality Protection Act of 1996, has opened new opportunities for biopesticides and reduced-risk pesticides in general. The broad-spectrum activity of azadirachtin at low use rates (12·5 to 40 g AI ha^-1) coupled with the insect growth regulator activity (in all larval/nymphal instars including the pupal stage) and unique mode of action (ecdysone disruptor), make azadirachtin an ideal candidate for insecticide resistance, integrated pest control and organic pest control programmes. Azadirachtin has been exempted from residue tolerance requirements by the US Environmental Protection Agency for food crop applications. Azadirachtin exhibits good efficacy against key pests such as whiteflies, leafminers, fungus gnats, thrips, aphids and many leaf-eating caterpillars. Azadirachtin has minimal to no impact on non-target organisms, is compatible with other biological control agents and has a good fit into classical Integrated Pest Management programmes. The world's largest azadirachtin extraction facility has been fully commissioned in India to process over 10,000 tonnes neem seeds per annum. This will ensure the wide availability of azadirachtin technical grade material in the future. © 1998 Society of Chemical Industry.     

Antiproliferative effect of azadirachtin A on Spodoptera litura Sl-1 cell line through cell cycle arrest and apoptosis induced by up-regulation of p53

In order to illustrate the mechanism of cell cycle arrest and apoptosis induced by botanical insecticide, the effects of cell proliferation inhibition and apoptosis induction by azadirachtin A were investigated in Spodoptera litura cell line Sl-1 in vitro. The cell proliferation, cell cycle division, mitotic and apoptotic figure of nucleus, and the expression of related proteins were detected by a variety of methods, including MTT assay, flow cytometry, fluorescence microscope observation, and Western blot technique. Sl-1 cells grew in a dose-dependent inhibition manner and the proliferation inhibition rates were 18.95%, 33.45% and 41.00%, after 48 h of treatment with azadirachtin A at the concentrations of 0.1, 0.5 and 1.0 μg/mL, respectively. Sl-1 cell cycle was arrested in G2/M phase with lower expression of cyclin B1. The apoptosis rates were found to be 19.71%, 23.40% and 24.91% after 48 h of treatment with 0.1, 0.5, and 1.0 μg/mL of azadirachtin A, which increased with the activities of caspase-3. The up-regulation of p53 was observed before apoptosis and cell cycle arrest occurred. These findings suggest that p53 protein was involved in cell cycle arrest, apoptosis induction and cell proliferation inhibition when the insect cultured cells were treated with azadirachtin A.     

昆虫生长调节剂和绿僵菌对意大利蝗几丁质酶活力的影响

[目的]探明3种昆虫生长调节剂(苯氧威、氟虫脲、灭幼脲)和一种生物农药(绿僵菌189菌株)对意大利蝗3龄蝗蝻几丁质酶活力的影响.[方法]用药剂处理3龄意大利蝗蝗蝻,用分光光度法测定几丁质酶分解几丁质产物N-乙酰氨基葡萄糖的含量.[结果]3种昆虫生长调节剂浓度达到50mg/L时对几丁质酶的抑制作用超过200％;绿僵菌189菌株对意大利蝗几丁质酶活力也有一定程度的抑制,且抑制作用随着孢子浓度的增加而增大,当孢子浓度达到5×107时,意大利蝗几丁质酶的活力仅为3.8U.[结论]昆虫生长调节剂和绿僵菌均抑制意大利蝗几丁质酶活力.     

Inhibitory activity against plant pathogenic fungi of extracts from Myoporum bontioides A. Gray and indentification of active ingredients

BACKGROUND: In order to understand the bioactivity of Myoporum bontioides A. Gray against plant pathogens and determine its active ingredients, the inhibitory activities of methanol extracts from M. bontioides against Fusarium oxysporum f. sp. niveum (E. F. Smith) Snyder & Hansen, Pestalotia mangiferae P. Henn., Thielaviopsis paradoxa (De Seynes) v. Hohnel, Colletotrichum musae (Berk. & M. A. Curtis) v. Arx, Alternaria alternata (Fr.) Keissler, Mycosphaerella sentina (Fr.) Schroter and Sphaceloma fawcettii Jenk. were evaluated using a growth rate method, and the active ingredient was isolated by activity-directed isolation and identified by determination and analysis of IR, (1)H NMR, (13)C NMR and mass spectra and correlative physical constants. RESULTS: The results showed that the extracts from stems and leaves of M. bontioides exhibited inhibitory activity against the seven fungi, with > 58% inhibition at 10 g L(-1) after 72 h. The active compound was isolated and identified as (-)-epingaione, and showed inhibitory activity against the above seven fungi. The inhibitory activity against P. mangiferae was the highest, with an EC(50) value of 77 mg L(-1). The EC(50) values against the other six fungi were 147-245 mg L(-1). (-)-Epingaione also inhibited spore germination of F. oxysporum f. sp. niveum, T. paradoxa and S. fawcettii.CONCLUSION: (-)-Epingaione demonstrated broad-spectrum inhibitory activity against plant pathogenic fungi and is promising for exploitation as a fungicide.     

常用杀虫剂和杀菌剂对褐飞虱类酵母共生菌生长的影响

本文研究了常用杀虫剂和杀菌剂对褐飞虱类酵母共生菌生长的影响。结果表明,10%吡虫啉可湿性粉剂、48%毒死蜱乳油、10%溴氰菊酯乳油、25%噻嗪酮可湿性粉剂、1.2%印楝素乳油、75%百菌清可湿性粉剂、20%三环唑可湿性粉剂的500、1 000、2 000倍液处理后共生菌的生长（菌落数量）显著减少,共生菌的假菌丝生长不舒展,有所萎缩,酵母多形成空泡。浓度越高、共生菌生长受到抑制越显著。低浓度（常规推荐剂量及以下）井冈霉素对共生菌生长影响较小,而高浓度（2倍常规推荐剂量）处理影响很大。这说明常用杀虫剂和杀菌剂农药对褐飞虱类酵母共生菌的生长均有较大的抑制作用。     

Laboratory and field evaluations of rhodojaponin-III against the imported cabbage worm Pieris rapae (L.) (Lepidoptera: Pieridae)

The activity of rhodojaponin-III (R-III), a grayanoid diterpene compound isolated from Rhododendron molle G. Don flowers, was determined under laboratory and field conditions as an antifeedant, stomach poison, contact toxicant and insect growth inhibitor against Pieris rapae (L.) larvae. The median antifeedant concentration (AFC(50)) values in no-choice leaf disc tests were 1.16 and 15.85 mg L(-1) at 24 h after treatment when tested against third and fifth instars respectively. The median lethal concentration (LC(50)) values in leaf disc tests were 2.84 and 9.53 mg L(-1) at 96 h after treatment against third and fifth instars respectively. R-III showed an almost 30 times higher contact toxicity against third instars than for fifth instars, and the median lethal dose (LD(50)) values for topical application were 1.18 and 34.09 mg kg(-1) at 72 h after treatment respectively. R-III disrupted the development of larvae to pupae or adults with median concentration for inhibiting growth (IC(50)) values of only 1.36 mg L(-1) for third instars and 11.28 mg L(-1) for fifth instars. In field trials, a greater than 80% reduction in the adjusted larval numbers was obtained against P. rapae 14 days after treatment when Rhodo 0.1% EC, a commercial botanical insecticide based on R-III, was applied at both 937.5 and 625 mL ha(-1). These results suggest that further research to develop R-III, and extracts from R. molle, as biorational pesticides or as lead compounds for integrated pest management deserve consideration.     

五种药剂对黏虫卵的室内触杀效果及胚胎发育的影响

为有效防控黏虫Mythimna separata,选用灭多威、苦参·印楝素、苏云金芽胞杆菌Bacillus thuringiensis、甲维盐和高效氯氟氰菊酯5种药剂对其卵进行室内触杀试验,比较这5种药剂对黏虫卵和初孵幼虫的触杀效果及对卵的毒力,测定不同药剂处理后黏虫卵初孵幼虫存活率、发育历期、化蛹率、蛹重和羽化率;并通过冷冻切片技术观察不同药剂处理后黏虫卵的外部形态和胚胎发育情况。室内触杀试验结果表明,苦参·印楝素浓度大于0.1 mg/L时,对产后不同时间黏虫卵的校正死亡率介于42.89%～97.34%之间,苏云金芽胞杆菌浓度为0.01 mg/L时对产后72 h黏虫卵的校正死亡率已达71.99%;各浓度苦参·印楝素和苏云金芽胞杆菌处理产后24、48和72 h黏虫卵后,其初孵幼虫校正死亡率介于39.02%～83.59%之间。苦参·印楝素对产后24、48 h黏虫卵的毒力最大,致死中浓度LC_(50)分别为0.018、0.109 mg/L;苏云金芽胞杆菌对产后72 h黏虫卵的毒力最大,LC_(50)为0.004 mg/L。黏虫卵经5种药剂处理后,幼虫化蛹率、蛹重和羽化率分别介于91.07%～93.64%、0.272～0.284 g和86.60%～87.78%之间,与对照差异不显著,幼虫存活率较对照显著下降43.02%～60.27%,幼虫发育历期较对照延长0.23～0.88 d,其中灭多威、苦参·印楝素、苏云金芽胞杆菌处理的幼虫发育历期较对照显著延长。苦参·印楝素、苏云金芽胞杆菌使胚胎无法形成正常胚带,同时会使已发育的虫体出现蜷缩、腹部皱缩;高效氯氟氰菊酯会延缓胚胎发育时间,使已形成的虫体畸形或使其肠道断裂;甲维盐影响虫体对营养物质的吸收,造成虫体萎缩,使其不能正常孵化。在黏虫卵防治中推荐使用苏云金芽胞杆菌和苦参·印楝素。     

5种杀虫剂和黄瓜新小绥螨对棉田朱砂叶螨的协调防治

为协调使用杀虫剂及害虫天敌,本文测定了阿维菌素、印楝素、苦参碱、除虫菊素和吡虫啉5种常用杀虫剂分别与天敌黄瓜新小绥螨(原名胡瓜钝绥螨)对棉田朱砂叶螨的联合防治效果。结果表明:1.8%阿维菌素EW(1∶8 000)处理6d后释放黄瓜新小绥螨对朱砂叶螨的防治效果最佳,20d后相对防治效果高达96.63%;其次是0.3%印楝素EC(1∶250)处理7d后释放黄瓜新小绥螨,1d和20d后防效分别为59.7%和90.16%;0.5%苦参碱AS(1∶2 000)处理6d后释放黄瓜新小绥螨,20d后相对防治效果达到82.65%。本研究为朱砂叶螨可持续防控提供了可选方案,为延缓抗药性产生提供了研究思路。     

Inhibitory effects of pesticides on growth and respiration of cultured cells

Effects of various drugs including pesticides on the growth and respiration of cultured cells were evaluated comparatively using cell lines derived from mosquito ovary and subcutaneous mouse tissues. The concentration producing 50% inhibition of cell growth, I₅₀ (M), was determined for each of 42 drugs. Inhibitors of respiration and nucleic acid and protein biosyntheses such as rotenone, piericidin A₁, actinomycin D, and puromycin had very high pI₅₀ values of approximately 8. Except for the compounds known to be uncouplers of oxidative phosphorylation, the drugs suppressed the respiration rate of the cultured cells to various degrees. The pI₅₀ value (and the pEC₁₅₀, 150% enhancement of the control, value for uncouplers) was determined for each compound. By examining the relation of pI₅₀ (and pEC₁₅₀) values between cell growth and respiration, the compounds could be classified into two groups according to their modes of inhibitory action against the cultured cells. One mode relates to the inhibition of energy synthesis and the other, perhaps, to interference with the biosynthesis of biomacromolecules.     

新型砜类化合物对辣椒枯萎病菌的抑菌作用初探

离体条件下采用生长速率法测得2-(苄基砜)-5-(2,4-二氯)苯基-1,3,4- NFDA1 二唑( 2008334 )对辣椒枯萎病菌Fusarium oxysporum菌丝生长具有较强的抑制作用,其EC50值为27.54 μ g/mL,与对照药剂 NFDA1 霉灵(EC50值为29.13 μ g/mL)相当;同时该化合物对辣椒枯萎病菌孢子萌发亦有一定的抑制作用,EC50值为41.37 μ g/mL。以质量浓度为100 μ g/mL的 2008334 处理辣椒枯萎病菌菌丝,在短时间内菌丝细胞膜通透性显著增强,菌体内还原糖及可溶性蛋白含量均出现先升高后下降的趋势。     

降香黄檀炭疽病药剂筛选及林间防效

为了筛选出防治降香黄檀炭疽病的环境友好型药剂,本文测定了10种杀菌剂的室内毒力,并从中筛选出毒力较强的药剂进行林间防效试验。结果表明:25%丙环唑EC,60%唑醚·代森联WG和40%氟硅唑EC对降香黄檀炭疽病菌的毒力较强,其EC50分别为0.12,0.15和1.12 mg/L。其他供试药剂的EC50分别为:75%百菌清WP231.42mg/L,10%苯醚甲环唑WG 3.60mg/L,80%醚菌酯WG 1.58mg/L,70%甲基硫菌灵WP 4.51mg/L,3%中生菌素WP 19.36mg/L,80%代森锰锌WP 79.76mg/L,80%多菌灵WP 11.47mg/L。考虑到氟硅唑、唑醚·代森联价格较高,建议将这两种药剂使用在降香黄檀苗木炭疽病的防治上。人工林炭疽病的防治采用25%丙环唑EC、80%醚菌酯WG、70%甲基硫菌灵WP和3%中生菌素WP,每隔15d依次施用,林间防治效果达到71.53%,明显高于其他防治方法。因此,在人工林降香黄檀炭疽病的防治中采用这4种药剂每隔15d依次施用可以有效控制降香黄檀炭疽病的暴发。喷雾时间应选择在阴天或晴天早晨露水干后8:00-10:00或者15:00-17:00。     

Field efficacy of azadirachtin-A, tetrahydroazadirachtin-A, NeemAzal and endosulfan against key pests of okra (Abelmoschus esculentus)

BACKGROUND Unlike synthetic pesticides, azadirachtin-based neem pesticides are environmentally friendly and are well known for their diverse pest control properties. Their use is, however, limited by the instability of azadirachtin, necessitating application at short intervals. The efficacy of relatively stable tetrahydroazadirachtin-A, therefore, needed to be established under field conditions. RESULTS: Azadirachtin-A (Aza-A), its stable derivative tetrahydroazadirachtin-A (THA) and other neem pesticides have been evaluated for their field efficacy against major insect pests of okra, Abelmoschus esculentus (L.) Moench., during summer (kharif) 2003 and 2004. The optimum doses of Aza-A and THA against the fruit borer, Earias vittella F., were also established. Reductions in population of whitefly, Bemisia tabaci (Genn.), and leafhopper (jassid), Amrasca biguttulla biguttulla Ishida, after application of THA or endosulfan was evident up to 10 days after treatment (DAT), whereas with Aza-A and NeemAzal (NZ) the effect was observed up to 7 DAT only. Endosulfan and THA also caused higher reduction in the larvae of shoot and fruit borer E. vittella and E. insulana Boisd., and recorded the highest yields of 4600 and 4180 kg ha(-1). The efficacy of THA (0.05 g L(-1) emulsion) was comparable with that of 0.5 g L(-1) endosulfan emulsion in reducing fruit borer infestation, the reduction over the control being 86.0 and 87.3%, 84.9 and 94.1% and 90.2 and 92.6% at first, second and third picking. THA 0.02 g L(-1) and Aza-A 0.05 g L(-1) were on a par. Laboratory-made neem oil emulsifiable concentrate was the least effective, but was superior to untreated check. CONCLUSION: Three consecutive sprays of THA, a neem-based biopesticide, and endosulfan have been found to be superior in controlling field pests of okra to Aza-A and NZ, which were on a par. THA thus holds potential as a component of pest management strategies against okra pests.     

Effect of uneven application of azadirachtin on reproductive and anti-feedant behaviour of Rhyzopertha dominica (Coleoptera: Bostrichidae)

The effects of uneven application of an azadirachtin-enriched neem extract to wheat grain on the reproduction and feeding behaviour in Rhyzopertha dominica (F) were investigated. The evenness of distribution amongst individual wheat grains during grain treatment was not important in ensuring its effectiveness as an anti-feedant or as an insect growth regulator, provided that, overall, the grain was treated with an effective level of azadirachtin. The effect of azadirachtin on reproduction was not improved when the diluent application rates were increased from 0.5 ml kg⁻¹ to 5 ml kg⁻¹. Treating 10% of the grain was shown to provide the same level of protection as treating 100% of the grain. The anti-feedant effect of azadirachtin on the beetle was also unaffected by unevenness of treatment, provided that 50% of the grain was treated. The fact that azadirachtin remains effective even when application to grain is uneven may be advantageous in field situations where uniform distribution of insecticides is difficult to achieve. © 1999 Society of Chemical Industry     

四种羧酸酰胺类杀菌剂对辣椒疫霉菌三个不同发育阶段的毒力比较

室内离体条件下测定了4种羧酸酰胺类(CAAs)杀菌剂双炔酰菌胺、烯酰吗啉、丁吡吗啉和氟吗啉对辣椒疫霉3个不同生长发育阶段的抑制活性。结果表明:4种杀菌剂抑制辣椒疫霉菌丝生长的EC50值分别为1.95×10-2、1.41、1.85、2.31 μ g/mL;对病菌孢子囊形成的抑制效果最好,其EC50值分别为2.00×10-4、1.50×10-3、2.60×10-3、4.30×10-3 μ g/mL;抑制游动孢子萌发的EC50值分别为4.60×10-3、0.373、0.494、0.635 μ g/mL。4种CAAs杀菌剂对辣椒疫霉的抑制作用均高于对照药剂甲霜灵、吡唑醚菌酯及嘧菌酯。     

Increased activity and reduced sensitivity of glutamine synthetase in glufosinate‐resistant vetiver (Vetiveria zizanioides Nash) cells

Vetiver (Vetiveria zizanioides Nash) cells derived from an inflorescence were cultured in a modified N6 liquid medium supplemented with 10 µm 2,4‐D and 10 mm proline. Exponentially growing cell suspensions were subcultured with a selection medium containing glufosinate (ammonium dl ‐homoalanin‐4‐yl(methyl)phosphinate). The glufosinate‐resistant cells which can grow in a medium containing 5 × 10^?5 M glufosinate was selected by a stepwise selection, and its I₅₀ value was determined to be 4.2 × 10^?5 M. The growth of susceptible cells was inhibited by lower concentrations of glufosinate and its I₅₀ value was 2.5 × 10^?7 M. This indicated that the selected cells were 170‐fold resistant compared with the susceptible cells. Glutamine synthetase (GS) activity of the resistant cells was twice as high as that of the susceptible cells. The I₅₀ values of glufosinate were 3.2 × 10^?5 M and 9.0 × 10^?7 M for GS from the resistant and susceptible cells, respectively. The accumulation of ammonia caused by GS inhibition was higher in the susceptible cells. Absorption of [3,4–¹⁴C]glufosinate was not significantly different between the resistant and susceptible cells. Both cell types did not metabolize glufosinate. These results suggest that the resistance of the selected vetiver cell suspension to glufosinate is mainly due to increased GS activity and its decreased sensitivity to the herbicide.     

五种杀虫剂对苜蓿种子田中主要害虫和传粉昆虫的影响

为筛选高效、低毒的杀虫剂，测定1.5%除虫菊素、0.3%印楝素、4%阿维·啶虫脒、2.5%高效氯氟氰菊酯和70%吡虫啉5种杀虫剂对苜蓿种子田主要害虫牛角花齿蓟马Odontothrips loti和豌豆蚜Acyrthosiphon pisum的室内毒力和田间防效，并分析这5种药剂对主要传粉昆虫数量及多样性指数的影响。结果显示，0.3%印楝素对牛角花齿蓟马和豌豆蚜的毒力相对较高，LC₅₀分别为1.093 mg/L和2.864 mg/L。70%吡虫啉以及4%阿维·啶虫脒对牛角花齿蓟马和豌豆蚜均有较高的防效，最高防效均达到85.00%以上，但这2种药剂对传粉昆虫威胁较大，而0.3%印楝素则对传粉昆虫威胁较小。推荐花期苜蓿种子田使用0.3%印楝素1 000倍稀释液防控害虫，而非开花期可交替使用70%吡虫啉1 000倍稀释液和4%阿维·啶虫脒1 000倍稀释液防控害虫。