Evaluation of O,O-Dialkyl O-Cyanophenyl Phosphates and Phosphorothioates as Insecticides

Various O,O-dialkyl O-cyanophenyl phosphates and phosphorothioates were prepared and their biological activities were examined. Among them, O,O-dimethyl O- (4-chloro-2-cyanophenyl) phosphorothioate was found to have selective and high toxicity to houseflies. O,O-Dimethyl O- (4-cyanophenyl) phosphorothioate, O,O-diethyl O- (4-cyanophenyl) phosphorothioate and O,O-diethyl O- (2-chloro-4-cyanophenyl) phosphorothioate showed high insecticidal activty to American cockroaches, though the former two were not so effective to houseflies. The dimethyl esters of these series exhibited markedly lowered mammalian toxicity. Among the O-ethyl O-cyanophenyl phenylphosphonothioates, O-ethyl O- (2-chloro-4-cyanophenyl) phenylphosphonothioate was highly effective to mites, while less effective to insects.     

水稻螟虫神经肽PBAN及其受体序列的生物信息学分析

【目的】性信息素合成激活肽(PBAN)是控制昆虫产生性信息素的激素,本文旨在分析水稻螟虫神经肽PBAN及其受体的序列。【方法】通过t Blastn同源检索从水稻螟虫基因组和转录组数据库中鉴定水稻螟虫PBAN神经肽及其受体序列,在此基础上进行序列比对及系统发生分析。【结果】发现二化螟Chilo suppressalis、三化螟Tryporyza incertulas和大螟Sesamia inferens的PBAN成熟肽序列均含有33个氨基酸残基,其C端五肽序列完全相同,3种水稻螟虫PBAN多肽相似度为54.55%~63.64%;发现二化螟PBAN受体3个异构体全长氨基酸序列(PBANR-A、PBANR-B和PBANR-C),均含有7个跨膜区域。【结论】进化树分析发现不同昆虫PBAN神经肽及其受体存在一定的保守性和多样性,并且在进化树上的位置几乎与昆虫系统发育分类一致,推测PBAN神经肽和PBAN受体在昆虫系统进化过程中可能存在协同进化现象。本研究为水稻螟虫PBAN神经肽及其受体的结构和功能分析提供基础。     

Field evaluation of resistance of transgenic rice containing a synthetic cry1Ab gene from Bacillus thuringiensis Berliner to two stem borers

Two transgenic rice (Oryza sativa L.) lines, KMD1 and KMD2 at the R4 generation, transformed with a synthetic cry1Ab gene from Bacillus thuringiensis Berliner, were first evaluated for stem borer resistance in the field during the rice growing season of 1998 in two areas of Zhejiang Province, China. Both KMD1 and KMD2 were highly resistant to the stem borers Chilo suppressalis (Walker) and Scirpophaga incertulas (Walker), and were completely undamaged during the whole rice growing season. In contrast, damage to the plants of the untransformed parental control (Xiushui 11) was in the form of deadhearts or whiteheads. Under natural infestation by the C. suppressalis, the damage to control plants reached a peak of 88.7% of plants and 20.1% of tillers encountered with deadhearts. Under artificial and natural infestation of neonate striped stem borers at the vegetative stage and booting stage, 100% of plants and 25.6% of tillers, 78.9% of plants and 15.6% of productive tillers among artificially infested control plants were observed with the symptom of deadhearts and whiteheads, respectively. Damage to the control plants from artificial infestation by the S. incertulas reached a peak of 97.0% of plants and 22.9% of tillers damaged. The field research indicated that both KMD1 and KMD2 show great potential for protecting rice from attack by these two stem borers.     

Purification and characterization of three parathion hydrolases from gram-negative bacterial strains.

Three unique parathion hydrolases were purified from gram-negative bacterial isolates and characterized. All three purified enzymes had roughly comparable affinities for ethyl parathion and had broad temperature optima at ca. 40 degrees C. The membrane-bound hydrolase of Flavobacterium sp. strain ATCC 27551 was composed of a single subunit of approximately 35,000 daltons (Da) and was inhibited by sulfhydryl reagents such as dithiothreitol (DTT) and by metal salts such as CuCl2. The cytosolic hydrolase of strain B-1 was composed of a single subunit of approximately 43,000 Da and was stimulated by DTT and inhibited by CuCl2. The membrane-bound hydrolase of strain SC was composed of four identical subunits of 67,000 Da and was inhibited by DTT and stimulated by CuCl2. The substrate ranges of the three enzymes also differed, as evidenced by their relative affinities for parathion and the related organophosphate insecticide O-ethyl-O-4-nitrophenyl phenylphosphonothioate (EPN). The B-1 hydrolase displayed equal affinity for both compounds, the Flavobacterium enzyme showed twofold-lower affinity for EPN than for parathion, and the SC hydrolase displayed no activity toward EPN. The range in characteristics of these three enzymes can be exploited in different waste disposal strategies.     

Impact of insect-resistant transgenic rice on target insect pests and non-target arthropods in China

Progress on the research and development of insect-resistant transgenic rice, especially expressing insecticidal proteins from Bacillus thuringiensis （Bt）, in China has been rapid in recent years. A number of insect-resistant transgenic rice lines/varieties have passed restricted and enlarged field testing, and several have been approved for productive testing since 2002 in China, although none was approved for commercial use until 2006. Extensive laboratory and field trials have been conducted for evaluation of the efficiency of transgenic rice on target lepidoteran pests and potential ecological risks on non-target arthropods. The efficacy of a number of transgenic rice lines currently tested in China was excellent for control of the major target insect pests, the rice stem borers （Chilo suppressalis, Scirpophaga incertulas, Sesamia inferens） and leaffolder （ Cnaphalocrocis medinalis）, and was better than most insecticides extensively used by millions of farmers at present in China. No significantly negative or unintended effects of transgenic rice on non-target arthropods were found compared with non-transgenic rice. In contrast, most of the current insecticides used for the control of rice stem borers and leaffolders proved harmful to natural enemies, and some insecticides may directly induce resurgence of rice planthoppers. Studies for developing a proactive insect resistance management of transgenic rice in the future are discussed to ensure the sustainable use of transgenic rice.     

Differential fipronil susceptibility and metabolism in two rice stem borers from China

The susceptibilities of larvae of two rice stem borers, namely, Chilo suppressalis (Walker) (Lepidoptera: Crambidae) and Sesamia inferens (Walker) (Lepidoptera: Nocutidae) to fipronil and its metabolites were investigated, and then the activities of microsomal O-demethylase, and glutathione transferase (GST) in two species were measured. The metabolism of fipronil in both stem borers was determined in vivo and in vitro. The LD50 value of fipronil to S. inferens was 118.5-fold higher than that of C. suppressalis. The bioassay results offipronil metabolites showed that the toxicities of sulfone and sulfide were higher than fipronil for both species, and the differential toxicity between sulfone and fipronil was remarkable. Alternatively, the activities of microsomal O-demethylase and GST of C. suppressalis were 1.35- and 2.06-fold higher than S. inferens, respectively. The in vivo and in vitro studies on metabolism of fipronil showed that all of fipronil, sulfone, and sulfide were detected and the content of sulfone was higher than sulfide in both stem borers. The residue of sulfone in C. suppressalis was significantly higher than that in S. inferens. These results suggest that the higher activity of mixed function oxidases may cause the higher capacity of C. suppressalis to produce fipronil-sulfone, which is more toxic than fipronil leading to the higher susceptibility of this species.     

Purification and characterization of three parathion hydrolases from gram-negative bacterial strains

Three unique parathion hydrolases were purified from gram-negative bacterial isolates and characterized. All three purified enzymes had roughly comparable affinities for ethyl parathion and had broad temperature optima at ca. 40 degrees C. The membrane-bound hydrolase of Flavobacterium sp. strain ATCC 27551 was composed of a single subunit of approximately 35,000 daltons (Da) and was inhibited by sulfhydryl reagents such as dithiothreitol (DTT) and by metal salts such as CuCl2. The cytosolic hydrolase of strain B-1 was composed of a single subunit of approximately 43,000 Da and was stimulated by DTT and inhibited by CuCl2. The membrane-bound hydrolase of strain SC was composed of four identical subunits of 67,000 Da and was inhibited by DTT and stimulated by CuCl2. The substrate ranges of the three enzymes also differed, as evidenced by their relative affinities for parathion and the related organophosphate insecticide O-ethyl-O-4-nitrophenyl phenylphosphonothioate (EPN). The B-1 hydrolase displayed equal affinity for both compounds, the Flavobacterium enzyme showed twofold-lower affinity for EPN than for parathion, and the SC hydrolase displayed no activity toward EPN. The range in characteristics of these three enzymes can be exploited in different waste disposal strategies.     

二化螟水稻种群与茭白种群光周期反应的比较

本文报道了用水稻和茭白分别饲养二化螟水稻种群和茭白种群的光周期反应。光周期反应曲线显示,用水稻饲养茭白种群或用茭白饲养水稻种群,无论是在短光照还是在长光照条件下,绝大多数幼虫被诱导进入滞育,丧失了各自原有的光周期反应特性,表明这两个种群已分化到仅适应其本身寄主的程度。用水稻饲养的茭白种群仅有5%～6%的个体化蛹,且其幼虫期较用茭白饲养的延长了15～18天;而用茭白饲养的水稻种群有30%～40%的个体化蛹,其幼虫期与水稻饲养的仅相差3～8天。茭白种群用水稻饲养时32日龄幼虫体重仅为茭白饲养的53.1%,而水稻种群用水稻饲养时32日龄幼虫体重为茭白饲养的79.5%。这些结果表明,茭白种群不适应取食水稻,而水稻种群对取食茭白则有一定的适应能力。根据这些结果,我们认为：（1）这两个种群已出现种下分化的迹象;（2）茭白种植不会对水稻田二化螟的发生产生大的影响。     

我国稻螟灾害的现状及损失估计

我国稻螟为害的确切记载始于 1 0 2 2年。 2 0世纪 5 0年代中期以前 ,以二化螟Chilosuppressalis(Walker)为害为主。此后十几年中 ,三化螟Tryporyzaincertulas(Walker)普遍大发生。 70年代 ,我国螟害轻微。 1 993年 ,二化螟急剧回升 ,三化螟也随之回升。 1 996年 2种稻螟大发生 ,此后连年暴发成灾。目前我国螟灾的主要特点是虫口密度高、受害范围广、受害程度重、持续时间长、经济损失大。首要螟灾区是沿江稻区 ,二化螟和三化螟分别约占 2 3和 1 3。全国年发面积约 1 5 0 0万hm2 ,防治约 3 80 0万hm2 次。该文首次估计年防治代价约 45 7～ 60亿元 ,残虫造成作物损失近 65亿元 ,总经济损失约 1 1 5亿元。由此可见 ,当前稻螟已取代稻飞虱、棉铃虫成为影响我国国计民生的头号害虫。     

Seasonal infestations of two stem borers (Lepidoptera: Crambidae) in noncrop grasses of Gulf Coast rice agroecosystems

Infestations of two stem borers, Eoreuma loftini (Dyar) and Diatraea saccharalis (F.) (Lepidoptera: Crambidae), were compared in noncrop grasses adjacent to rice (Oryza sativa L.) fields. Three farms in the Texas rice Gulf Coast production area were surveyed every 6-8 wk between 2007 and 2009 using quadrat sampling along transects. Although D. saccharalis densities were relatively low, E. loftini average densities ranged from 0.3 to 5.7 immatures per m(2) throughout the 2-yr period. Early annual grasses including ryegrass, Lolium spp., and brome, Bromus spp., were infested during the spring, whereas the perennial johnsongrass, Sorghum halepense (L.) Pers., and Vasey's grass, Paspalum urvillei Steud., were infested throughout the year. Johnsongrass was the most prevalent host (41-78% relative abundance), but Vasey's grass (13-40% relative abundance) harbored as much as 62% of the recovered E. loftini immatures (during the winter). Young rice in newly planted fields did not host stem borers before June. April sampling in fallow rice fields showed that any available live grass material, volunteer rice or weed, can serve as a host during the spring. Our study suggests that noncrop grasses are year-round sources of E. loftini in Texas rice agroecosystems and may increase pest populations.     

烯效唑对水稻幼苗内源IAA含量的影响

研究了烯效唑对3~H-IAA色氨酸合成3~H-IAA的效率及对IAA氧化酶活性的影响,以探讨烯效唑延缓植物生长的作用机理。结果表明,烯效唑对水稻(Oryza sativa L.)幼苗生长的控制效应与其降低内源IAA含量有关,烯效唑浸种处理降低水稻幼苗内源IAA含量有两条途径,其一是提高水稻幼苗IAA氧化酶活性,增强内源IAA的氧化;其二是阻抑内源IAA的合成。烯效唑除阻抑内源赤霉素的生物合成而延缓作物生长外,通过降低内源IAA水平也可能是其延缓作物生长的一个原因。     

Constitutive expression of a cowpea trypsin inhibitor gene,CpTi, in transgenic rice plants confers resistance to two major rice insect pests

The gene encoding a cowpea trypsin inhibitor (CpTI), which confers insect resistance in trangenic tobacco, was introduced into rice. Expression of the CpTi gene driven by the constitutively active promoter of the rice actin 1 gene (Act1) leads to high-level accumulation of the CpTI protein in transgenic rice plants. Protein extracts from transgenic rice plants exhibit a strong inhibitory activity against bovine trypsin, suggesting that the proteinase inhibitor produced in transgenic rice is functionally active. Small-scale field tests showed that the transgenic rice plants expressing the CpTi gene had significantly increased resistance to two species of rice stem borers, which are major rice insect pests. Our results suggest that the cowpea trypsin inhibitor may be useful for the control of rice insect pests.     

Determination of Binding of Bacillus thuringiensis (delta)-Endotoxin Receptors to Rice Stem Borer Midguts

Insecticidal activity and receptor binding properties of Bacillus thuringiensis toxins to yellow and striped rice stem borers (Sciropophaga incertulas and Chilo suppresalis, respectively) were investigated. Yellow stem borer (YSB) was susceptible to Cry1Aa, Cry1Ac, Cry2A, and Cry1C toxins with similar toxicities. To striped stem borer (SSB), Cry1Ac, Cry2A, and Cry1C were more toxic than Cry1Aa toxin. Binding assays were performed with (sup125)I-labeled toxins (Cry1Aa, Cry1Ac, Cry2A, and Cry1C) and brush border membrane vesicles (BBMV) prepared from YSB and SSB midguts. Both Cry1Aa and Cry1Ac toxins showed saturable, high-affinity binding to YSB BBMV. Cry2A and Cry1C toxins bound to YSB BBMV with relatively low binding affinity but with high binding site concentration. To SSB, both Cry1Aa and Cry1Ac exhibited high binding affinity, although these toxins are less toxic than Cry1C and Cry2A. Cry1C and Cry2A toxins bound to SSB BBMV with relatively low binding affinity but with high binding site concentration. Heterologous competition binding assays were performed to investigate the binding site cross-reactivity. The results showed that Cry1Aa and Cry1Ac recognize the same binding site, which is different from the Cry2A or Cry1C binding site in YSB and SSB. These data suggest that development of multitoxin systems in transgenic rice with toxin combinations which recognize different binding sites may be useful in implementing deployment strategies that decrease the rate of pest adaptation to B. thuringiensis toxin-expressing rice varieties.     

Structure-activity relationship of S-benzylisothiourea derivatives to induce spherical cells in Escherichia coli

We have previously reported that a novel S-benzylisothiourea derivative, S-(3,4-dichlorobenzyl)isothiourea, tentatively named A22, induced spherical cells in Escherichia coli. To elucidate the structural element(s) required for inducing these spherical cells, the biological activity of S-benzylisothiourea derivatives and related compounds toward E. coli cells was investigated. S-(4-Chlorobenzyl)isothiourea revealed spherical cell-inducing activity, although being slightly less potent than A22, and S-benzylisothiourea itself showed much less activity. S-Cyclohexylmethylisothiourea did not show antibacterial activity and had little effect on the cell shape. S-Heptylisothiourea showed antibacterial activity and induced elongated cells rather than spherical cells. Benzylisothiocyanate inhibited cell growth but did not induce spherical cells. S-Ethylisothiourea, benzylthiocyanate, benzylisocyanate, and N-phenylthiourea did not show any activity under the present experimental conditions. These results indicate that the S-benzylisothiourea structure was necessary and sufficient for inducing spherical cells and that 3- and/or 4-chloro-substitution of the S-benzyl group enhanced this activity.     

Screen of Bacillus thuringiensis toxins for transgenic rice to control Sesamia inferens and Chilo suppressalis

Transgenic rice to control stem borer damage is under development in China. To assess the potential of Bacillus thuringiensis (Bt) transgenes in stem borer control, the toxicity of five Bt protoxins (Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba and Cry1Ca) against two rice stem borers, Sesamia inferens (pink stem borer) and Chilo suppressalis (striped stem borer), was evaluated in the laboratory by feeding neonate larvae on artificial diets containing Bt protoxins. The results indicated that Cry1Ca exhibited the highest level of toxicity to both stem borers, with an LC₅₀ of 0.24 and 0.30 μg/g for C. suppressalis and S. inferens, respectively. However, S. inferens was 4-fold lower in susceptibility to Cry1Aa, and 6- and 47-fold less susceptible to Cry1Ab and Cry1Ba, respectively, compared to C. suppressalis. To evaluate interactions among Bt protoxins in stem borer larvae, toxicity assays were performed with mixtures of Cry1Aa/Cry1Ab, Cry1Aa/Cry1Ca, Cry1Ac/Cry1Ca, Cry1Ac/Cry1Ba, Cry1Ab/Cry1Ac, Cry1Ab/Cry1Ba, and Cry1Ab/Cry1Ca at 1:1 (w/w) ratios. All protoxin mixtures demonstrated significant synergistic toxicity activity against C. suppressalis, with values of 1.6- to 11-fold higher toxicity than the theoretical additive effect. Surprisingly, all but one of the Bt protoxin mixtures were antagonistic in toxicity to S. inferens. In mortality-time response experiments, S. inferens demonstrated increased tolerance to Cry1Ab and Cry1Ac compared to C. suppressalis when treated with low or high protoxin concentrations. The data indicate the utility of Cry1Ca protoxin and a Cry1Ac/Cry1Ca mixture to control both stem borer populations.     

水稻二化螟和三化螟基因组ddRADseq文库的构建

本文介绍了构建水稻二化螟和三化螟"双酶切限制性酶切位点关联DNA测序"(Double digest restrictionsite associated DNA sequencing,ddRADseq)文库的方法。利用安捷伦2100生物分析仪对4种单酶切及2种双酶切的酶切产物片段大小及分布范围进行分析,筛选出Mlu C I和Nla III两种限制性内切酶组合对螟虫基因组DNA进行酶切。酶切后的DNA片段两端连接上特定的P1、P2接头后,用Pippin Prep回收大小为285-435 bp的DNA片段。通过PCR扩增进行文库的富集并引入index序列。构建好的ddRADseq文库用琼脂糖凝胶电泳和生物分析仪进行质量检测。本方法所构建的文库DNA片段长度、分布和摩尔浓度能够达到Illumina平台测序的技术要求。本研究证实了利用Mlu C I和Nla III组合酶切构建水稻螟虫基因组ddRADseq文库的可行性,为在水稻螟虫中利用ddRADseq技术开展生物地理学、种群遗传学和系统发育重建等方面的研究奠定基础。     

Synthesis and evaluation of phosphoramidate and phosphorothioamidate analogues of amiprophos methyl as potential antimalarial agents

A series of phosphoramidate and phosphorothioamidate compounds based on the lead antitubulin herbicidal agents amiprophos methyl (APM) and butamifos were synthesised and evaluated for antimalarial activity. Of these compounds, phosphorothioamidates were more active than their oxo congeners and the nature of both aryl and amido substituents influenced the desired activity. The most active compound was 46, O-ethyl-O-(2-methyl-4-nitrophenyl)-N-cyclopentyl phosphorothioamidate, which was more effective than the lead compound.     

人胎肝造血干细胞增殖刺激物的分离及鉴定

经过超滤、ＤＥＡＥ－Ｓｅｐｈａｃｅｌ、ＳｅｐｈａｃｒｙｌＳ－２００和Ｓｕｐｅｒｏｓｅ１２ＨＲ多步分离纯化,从人胎肝细胞原代培养上清中分离到一分子量为３５ｋＤ的单一活性组分,具有造血干细胞增殖刺激活性,定义为ＦＬＳ－４。ＦＬＳ－４可能是一种新型造血干细胞增殖刺激因子,与具有这类活性的ＩＬ－３、ＩＬ－６、ＧＭ－ＣＳＦ、ＦＬＴ３配基和ＳＣＦ等在理化特性或生物学性质上均有所差异,在胎肝造血活跃时期,是启动早期造血干细胞从Ｇ＿０期进入Ｓ期的主要候选活性物质。     

油菜甾醇内酯类化合物结构与活性的相关性

比较了19种油菜甾醇内酯类似物和有关甾体化合物在水稻叶片倾斜及萝卜幼苗生长试验中的生物活性。表油菜甾醇内酯(24—Epi—BR)在两个系统中都具有很强的生物活性。C_2位失去羟基(香蒲甾醇)仅在水稻试验中有高活性,改变C_22位侧链结构(2α,3α双羟基—6—酮—23,24—双失碳—β—高-5α—胆烷酸甲酯)在萝卜试验中仍有活性。     

越桔两种蛀干害虫的发生与防治

咖啡豹蠹蛾Zeuzera coffeae Nietner和樟泥色天牛Uraecha angusta(Pascoe)是江苏地区的美国兔眼越桔上2种主要蛀干害虫。前者在枝干中向顶端方向蛀食,排出圆粒状虫粪;后者向植株基部方向蛀食,排出粉末状虫粪,均造成枝干枯死。咖啡豹蠹蛾发生量较大,是主要虫种。防治这2种蛀干害虫,应采用保护利用天敌、改善果园生态环境、促使树体健壮生长等综合管理措施。