Design, semisynthesis and insecticidal activity of novel podophyllotoxin derivatives against Brontispa longissima in vivo

In an attempt to find the effective phytopesticides, a series of novel podophyllotoxin derivatives were firstly synthesized and preliminarily tested for their antifeedant and insecticidal effects against the fifth-instar larvae of Brontispa longissima. The different antifeedant and insecticidal activity ranges of compounds 3a-l showed that variations of NR₁R₂ groups in the 4-position of podophyllotoxin skeleton markedly affected the activity profiles of this compound class, and some important SAR information has been revealed from it. To clarify their mode of action of insecticidal activity, the docking models as well as tubulin inhibitory effect of representative compound 3i were also investigated, and the result indicated that the insecticidal activity of these compounds was due to the tubulin inhibitory effect of these derivatives, thereby possibly providing some useful information for rational designs of novel podophyllotoxin-based insecticides.     

Synthesis and insecticidal activities of pyridine ring derivatives of podophyllotoxin

In an attempt to find the biorational pesticides, we synthesized 12 pyridinyl derivatives of podophyllotoxin (PPT) and 4′-demthylepipodophylltoxin (4′-DMEP) in this study. Their structures and the α/β substitution at C-4 were confirmed by ¹H NMR, IR, MS spectral analyses and elemental analysis. The insecticidal activities were tested against fifth-instar larvae of Pieris rapae and the third-instar larvae of Cullex pipiens pallens at concentrations of 250 and 10 μg ml⁻¹. Four derivatives of PPT, 4.1, 4.2, 4.3 and 4.5, showed higher insecticidal activities against P. rapae than PPT, while three derivatives of PPT, 4.4, 4.5 and 4.6, displayed higher mosquito larvicidal activity than PPT, with LC₅₀ values of 1.66, 3.96 and 1.54 mg l⁻¹, respectively. Interestingly, we also found that the pyridine ring derivatives of PPT showed delayed insecticidal activity, which is different from traditional neurotic insecticides. The results suggest that 4′-OCH₃ in the PPT derivatives is essential to keep the insecticidal activity and the insecticidal activities of pyridine ring derivatives of PPT are higher than that of the derivatives of 4′-DMEP, supporting PPT has the potential to be a lead structure of semi-synthetic insecticides.     

Synthesis and insecticidal activities of novel derivatives of podophyllotoxin: Part XII

In order to find the biorational pesticides, eight novel 4β-substituted phenoxyaniline derivatives of podophyllotoxin have been synthesized with significant stereoselectivity and improved yields by employing the BF₃·Et₂O/NaI reagent system and evaluated for their antifeedant effect against 5th instar larvae of Pieris rapae and effect on the development of 5th instar larvae of P. rapae as well as for insecticidal activity against P. rapae. The antifeedant activities showed that these compounds exhibited less potent than podophyllotoxin. Though these derivatives showed less potent antifeedant activities than podophyllotoxin, they acted as growth development inhibitor to 5th instar larvae of P. rapae, which were found that the animals treated with podophyllotoxin and its derivatives showed moulting disturbances and/or deformities. Also, the insecticidal activity results show that all of these derivatives of PPT showed delayed insecticidal activity, which differs from traditional neurotoxic insecticides. Among them, compounds possessing a 4β-phenoxyaniline moiety substituted (CO₂C₂H₅, Cl and OH) at para position exhibited greater insecticidal activity against P. rapae than podophyllotoxin.     

Biological activity of the essential oil from the leaves of Piper sarmentosum Roxb. (Piperaceae) and its chemical constituents on Brontispa longissima (Gestro) (Coleoptera: Hispidae)

The results showed that the essential oil from Piper sarmentosum has strong antifeedant and toxicity effects on Brontispa longissima. The best antifeedant and contact toxicity effects were observed in the 1st-2nd instar larvae. The essential oil also displayed a notable fumigation effect on the eggs and pupae of B. longissima. It took the control-treated insects 43.34 d to complete one generation, while the insects treated with 2000 mg/L essential oil needed 73.58 d. The essential oil was analyzed by gas chromatography/mass spectrometry and 41 components were identified. Myristicin (65.22%) and trans-caryophyllene (13.89%) were the major components. Myristicin exhibited strong antifeedant and contact toxicity effects on both the 3rd instar larvae and the imagoes of B. longissima, when it showed a significant fumigation effect on the eggs and pupae of B. longissima. Another P. sarmentosum essential oil and myristicin all showed a strong inhibiting effect on the growth and development of B. longissima along with the activity of AChE, CarE, GSTs and Na⁺, K⁺-ATPase in B. longissima larvae.     

入侵害虫椰心叶甲在中国的风险性分析

为确定椰心叶甲科学的检疫管理措施,根据WTO的<卫生与植物卫生措施协定>(SPS)要求,对其进行风险性分析.依据国际植物检疫措施标准(ISPM)规定的有害生物风险分析(PRA)程序,利用相关模型,对各项分析指标进行分析并赋值运算.各分析指标及运算值为:(1)国内分布情况,风险值为2;(2)潜在的经济危害性,风险值为2;(3)寄主植物的经济重要性,风险值为3;(4)传播扩散的可能性,风险值为2.05;(5)风险管理难度,风险值为1.66.椰心叶甲的综合风险性值为2.09,符合高风险检疫性害虫条件,据此提出2条相关风险管理备选方案,并分别进行效率和影响评估,确定最终风险管理方案.     

椰心叶甲人工饲料的研制及应用效果评价

为了获得入侵害虫椰心叶甲Brontispa longissima(Gestro)的人工饲料,在分析其天然寄主叶片水分和营养成分基础上,借鉴其它鞘翅目昆虫人工饲料配方,进行椰心叶甲人工饲料的配制和筛选,并评价人工饲料饲养的椰心叶甲对其寄生蜂椰甲截脉姬小蜂Asecodes hispinarum和椰心叶甲啮小蜂Tetrastichus brontispae的适合性。人工饲料配方的成分:蔗糖4%、椰子粉2%、大豆粉2%、椰树叶粉10%、酵母2%、维生素E 0.3%、抗坏血酸0.2%、对羟基苯甲酸甲酯0.2%、琼脂4%、链霉素0.03%和水75%。用所配制的人工饲料饲养椰心叶甲初孵幼虫,其蛹成活率可达36%,蛹羽化率与对照无显著差异,均在90%以上;雌成虫占52.4%,与对照无显著差异。表明人工饲料饲养所得的椰心叶甲可用于其幼虫寄生蜂椰甲截脉姬小蜂和蛹寄生蜂椰心叶甲啮小蜂的扩繁。     

Ultra-structural changes in the integument induced by the use of three of six forms of allylisothiocyanate tested against Plutella xylostella and Pieris rapae larvae

The insecticidal activity of four forms of Hong Jing (HJ) allylisothiocyanate (AITC), AITC + cypermethrin (HJ_A, HJ_B, and HJ_C) with ratio of (1:1, 4:1, and 2:1), pure AITC (HJ_D), and two forms of Hong Du (HD) AITC, AITC + chlorpyrifos (HD_A and HD_B) with ratio of (2:1 and 2:1), respectively, were studied on the major cruciferous insect larvae Plutella xylostella (L.) and Pieris rapae (L.) by combining both spraying and dipping methods. The P. rapae was more susceptible than P. xylostella larvae. The LC₅₀ values 72 h after treatment of AITC forms (HJ_B, HJ_A, HJ_C, HJ_D, HD_B, and HD_A) on the P. rapae were; 0.07, 0.08, 0.16, 0.83, 0.26, 1.08 gL⁻¹, and 0.69, 0.26, 5.45, 0.93, 3.01, 5.98 gL⁻¹ on the P. xylostella, respectively. The toxicity of some of the AITC forms was very close to or better than that of the commercial contact insecticides such as chlorpyrifos (LC₅₀ = 0.03 and 0.04 gL⁻¹ on P. rapae and P. xylostella, respectively), and cypermethrin (0.65 and 0.78 gL⁻¹, respectively, against P. rapae and P. xylostella). The ultrastructural studies on the integument of the third larval instar of P. xylostella treated by sub-lethal concentration (LC₂₀) of HJ_B, HJ_D, and HD_B were carried out by using transmission electron microscope. The more pronounced alterations in the hypodermis and mitochondria cells. They exhibited changes in all treated samples. The hypodermis was almost completely destroyed, and the mitochondria exhibited morphological alterations, represented by enlargement, matrix rarefaction and vacuolization of the mitochondria matrix, quantity of cristae reduced, and density electron matrix lessened. These AITC forms have potential as contact insecticides, and the ultra structural observations confirm the insecticidal efficiency of different AITC forms on P. rapae and P. xylostella.     

Mosquito larvicidal activity of alkaloids from Zanthoxylum lemairei against the malaria vector Anopheles gambiae

Four alkaloids, 10-O-demethyl-17-O-methylisoarnottianamide 1, 6-acetonyl-N-methyl-dihydrodecarine 2, nitidine 3, and chelerythrine 4 were isolated from the plant Zanthoxylum lemairei (Rutaceae) and evaluated for mosquito larvicidal activity against the malaria vector Anopheles gambiae. The mortalities of the larvae were determined after 24 h. The results of the larvicidal tests demonstrated that compounds 1 and 2 were the most potent with mortality rates of 96.7% and 98.3% at a concentration of 250 mg/L, respectively. Compound 3 was less potent with a mortality of 28.3% at the same concentration. The percent mortality of 100% was observed at a concentration of 500 mg/L. The least potent of the four alkaloids was compound 4, which achieved 100% mortality at 1000 mg/L. These findings could be useful in the research for newer more selective, biodegradable and natural larvicidal compounds or can be used as lead compounds for the development of larvicides.     

Construction of the recombinant baculovirus AcMNPV with cathepsin B-like proteinase and its insecticidal activity against Helicoverpa armigera

The Helicoverpa armigera cathepsin B-like proteinase (HCB) has been shown to have a wide spectrum of substrates. It has been involved in the degradation of yolk protein during embryonic development and also in the decomposition of the adult fat body. To study the possibility of using HCB to improve the insecticidal activity of bioinsecticides, it was inserted into the pFASTBACDUAL-green fluorescent protein (GFP) donor plasmid under the strong polyhedrin promoter, and the polyhedrin gene was retained behind the HCB gene so that the virus could orally infect the host and survive in the natural environment. After the recombinant plasmid transfected the Sf21 cells, the recombinant baculovirus Autographa californica multiple nucleocapsid nucleopolyhedrovirus (AcMNPV)-GFP-HCB-Polh⁺ was produced. A 37-kDa recombinant procathepsin B was expressed in AcMNPV-GFP-HCB-Polh⁺-infected Sf21 cells and processed into 29/25-kDa mature forms. Gelatin zymography revealed that the proteolytic activities of the recombinant HCB and other proteases were activated or enhanced by HCB in AcMNPV-GFP-HCB-Polh⁺-infected larvae. Green fluorescence was observed earlier and was more intense in AcMNPV-GFP-HCB-Polh⁺-infected larvae than in HCB-free AcMNPV-GFP-Polh⁺-infected ones that were infected at the same time; this indicated that the AcMNPV-GFP-HCB-Polh⁺ virus spreaded faster in larvae than the AcMNPV-GFP-Polh⁺ virus. A bioassay revealed that infection with the AcMNPV-GFP-HCB-Polh⁺ virus shortened the median survival time of H. armigera larvae by 12 h as compared with those infected with the AcMNPV-GFP-Polh⁺ or the wild-type AcMNPV virus. These results suggest that HCB may possibly play a role in the recombinant virus in improving the rate of killing larvae.     

Antifungal activity of oligochitosan against Phytophthora capsici and other plant pathogenic fungi in vitro

Antifungal activity of oligochitosan against nine phytopathogens was investigated in vitro. Oligochitosan was more effective than chitosan in inhibiting mycelial growth of Phytophthora capsici and its inhibition on different stages in life cycle of P. capsici was observed. Rupture of released zoospores induced by oligochitosan was reduced by addition of 100 mM glucose. The effects of oligochitosan on mycelial growth and zoospore release, but not zoospore rupture, were reduced largely when pH value was above 7. The ultrastructural study showed that oligochitosan caused distortion and disruption of most vacuoles, thickening of plasmalemma, and appearance of unique tubular materials. Plasmalemmasomes in hyphal tip cells were not found in the presence of oligochitosan. These results suggest polycationic nature of oligochitosan contributes only partly to its antifungal activity and multiple modes of action of oligochitosan exist including the disruption of endomembrane system.     

枸骨内生菌No.2的鉴定及其对黄瓜立枯病的生防作用

从枸骨植物中分离到一株对立枯丝核菌Rhizoctonia solani具有抑制作用的内生真菌(编号为No.2),借助Biolog微生物自动分析仪对该真菌进行了分类鉴定,根据菌株的生理生化和形态特征,初步确定该菌株为哈茨木霉菌Trichoderma harzianum Rifai。双重培养试验表明,该哈茨木霉菌对黄瓜立枯病菌存在竞争和寄生作用;盆栽试验测定结果表明,喷施哈茨木霉菌胞外代谢产物木霉素的1 000倍液7 d后,其对黄瓜立枯病的保护和治疗效果分别为88.89%和95.83%。施药后各处理对黄瓜的株形、叶色均未产生药害症状,表明其安全性较好。     

四种熏蒸剂对辣椒疫霉和南方根结线虫的毒力

为明确不同熏蒸剂对土传病原菌辣椒疫霉Phytophthora capsici和南方根结线虫Meloidogyne incognita的毒力,采用密闭熏蒸法测定了4种土壤熏蒸剂氯化苦、棉隆、威百亩、1,3-二氯丙烯的毒力及1,3-二氯丙烯和氯化苦复配的联合毒力。结果表明:氯化苦、棉隆、威百亩、1,3-二氯丙烯对辣椒疫霉的EC₅₀分别为0.12、2.44、8.30、8.45 mg/L; 对南方根结线虫的EC₅₀ 分别为1.23、0.22、0.30、0.18 mg/L。1,3-二氯丙烯和氯化苦以2:1、1:1、1:2比例复配时对辣椒疫霉的EC₅₀ 分别为1.13、0.24、0.14 mg/L;对南方根结线虫的EC₅₀分别为0.19、0.32、0.61 mg/L。结合经济效益和多种病害兼治的原则,推荐使用1,3-二氯丙烯与氯化苦1:1配比,可兼治辣椒疫病和根结线虫病。     

两株棒束孢菌的鉴定及其对茶卷叶蛾和茶小卷叶蛾的致病力

为明确分离自茶卷叶蛾和中华大刀螳僵虫的2株虫生真菌的归属及生防潜力,采用形态学特征和rDNA-ITS序列分析其分类地位,测定了其对茶卷叶蛾和茶小卷叶蛾幼虫的致病力,并初步筛选了其最适培养条件.结果表明:2株真菌均为环链棒束孢Isaria cateniannulata,分别命名为ICBS918和ICTL911.在1.0×10⁸孢子/mL浓度下,ICBS918和ICTL911对茶卷叶蛾幼虫的累计校正死亡率均达100%,LT₅₀分别为3.13 d和3.15 d;对茶小卷叶蛾幼虫的累计校正死亡率分别为100%和95%,LT₅₀分别为3.25 d和3.31 d.接菌后8 d,2菌株对茶卷叶蛾幼虫的LC₅₀分别为0.47×10⁵孢子/mL和1.01×10⁵孢子/mL;对茶小卷叶蛾幼虫的LC₅₀分别为2.20×10⁵孢子/mL和1.34×10⁵孢子/mL.2菌株23 ℃下最适生长培养基均为综合马铃薯培养基,最适产孢培养基均为蛋白胨马铃薯葡萄糖琼脂培养基;在萨氏培养基上,菌株ICBS918和ICTL911分别在24~30 ℃及24 ℃时生长速度最快,在27 ℃和21 ℃时产孢量最大.表明2株环链棒束孢对茶卷叶蛾和茶小卷叶蛾幼虫均具有较好的生防潜力,可作为生防菌进行开发和应用.     

Design, synthesis and antifungal/insecticidal evaluation of novel nicotinamide derivatives

A series of nicotinamide derivatives based on Boscalid were prepared and tested for their activities against seven plant pathogenic fungi and two insects. The preliminary bioassays indicated that almost all of the synthesized target compounds displayed the antifungal activities and some of them also had certain insecticidal activities. And, compound 12 showed the strongest activity of all against Rhizoctonia solani (EC₅₀ = 0.010 mg L⁻¹) and Sphaceloma ampelimum (EC₅₀ = 0.040 mg L⁻¹), even stronger than Boscalid, a new nicotinamide fungicide. Additionally, both compounds 1 and 2 showed strong activities against Plutella xylostella (90% and 80%, respectively, at 1000 mg L⁻¹).     

渐狭蜡蚧菌Lecanicillium attenuatum产几丁质酶的活性及对南方根结线虫卵孵化的抑制作用

为探究卵寄生真菌产生的几丁质酶对南方根结线虫卵孵化的抑制作用,采用分离自黑龙江大豆孢囊线虫孢囊上的菌株CGMCC5328,利用NAG和pNP法测定其几丁质降解酶系和外切酶的活性,分析几丁质酶粗酶液对南方根结线虫卵的孵化抑制效果.结果表明,菌株CGMCC5328经形态学和ITS序列分析鉴定为渐狭蜡蚧菌Lecanicillium attenuatum;在几丁质酶诱导培养基中培养第4天时其分泌的几丁质降解酶系达酶活高峰,为16.90 U/mL;第6天外切酶达酶活高峰,为0.59 U/mL,之后酶活趋于稳定持续至第14天;几丁质酶分子量分别为79.6、65.6、54.1、42.1和32.9 kDa;其中培养第7天的几丁质酶粗酶原液对南方根结线虫卵孵化的抑制率为83.17%;第6天菌株CGMCC5328对卵的寄生率为85.10%.表明高效产几丁质酶的卵寄生真菌渐狭蜡蚧菌菌株CGMCC5328对南方根结线虫卵孵化具有明显的抑制效果.     

杠柳杀虫活性成分的分离

采用柱层析分离、高效液相色谱切分和生物活性追踪法,从杠柳 Periploca sepium 根皮甲醇提取物中分离出2个具有杀虫活性的化合物(G1和G2),经鉴定其分别为已知物杠柳新苷D和F。生物活性测定结果表明,化合物G1和G2 对3龄粘虫 Mythimna separata 48 h的胃毒致死中浓度(LC50)分别为0.39和0.34 mg/mL, 对小菜蛾 Plutella xyllostella 48 h的胃毒LC50值分别为1.21和1.39 mg/mL。     

Effect of Melia azedarach extract on the activity of NADPH-cytochrome c reductase and cholinesterase in insects

The effect of Melia azedarach extract on the activity of NADPH-cytochrome c reductase and cholinesterase was studied in Spodoptera frugiperda. Larvae were fed an artificial diet containing fruit extract and their midgut was used for enzyme determination. As compared to the control, consumption of the extract containing diet resulted in a 31% inhibition of the cholinesterase activity and a 34% activation of the NADPH-cytochrome c reductase activity. In the cockroach Leucophaea maderae, the effect on reductase activity was even more pronounced (43%).     

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。     

哈茨木霉Th-30几丁质酶的生产条件及对灰霉病菌的拮抗作用

病菌生长的抑制率为67.61%～71.31%,对灰霉病菌孢子产生及萌发的抑制率分别为80.95%和81.35%,对病菌孢子致病性的抑制率为65.22%～70.51%.