The <Emphasis Type="Italic">cyv-2</Emphasis> resistance to <Emphasis Type="Italic">Clover yellow vein virus</Emphasis> in pea is controlled by the eukaryotic initiation factor 4E

The same mutant allele of eukaryotic initiation factor 4E (eIF4E) that confers resistance to Pea seed-borne mosaic virus (sbm-1) and the white lupine strain of Bean yellow mosaic virus (wlv) also confers resistance to Clover yellow vein virus (ClYVV) in pea. The eIF4E genes from several pea lines were isolated and sequenced. Analysis of the eIF4E amino acid sequences from several resistant lines revealed that some lines, including PI 378159, have the same sequence as reported for sbm-1 and wlv. When eIF4E from a susceptible pea line was expressed from a ClYVV vector after mechanical inoculation of resistant PI 378159, the virus caused systemic infection, similar to its effects in susceptible line PI 250438. The resistance to ClYVV in line PI 378159 was characterized through a cross with PI 193835, which reportedly carries cyv-2. Mechanical inoculation of the F1 progeny with ClYVV resulted in no infection, indicating that the resistance gene in PI 378159 is identical to cyv-2 in PI 193835. Furthermore, particle bombardment of pea line PI 193835 with infectious cDNA of ClYVV (pClYVV/C3-S65T) resulted in the same resistance mode as that described for PI 378159. These results demonstrate that the resistance to ClYVV conferred by cyv-2 is mediated by eIF4E and that cyv-2 is identical to sbm-1 and wlv.     

Genetic analysis of lethal tip necrosis induced by <Emphasis Type="Italic">Clover yellow vein virus</Emphasis> infection in pea

Clover yellow vein virus (ClYVV) elicits lethal tip necrosis in the pea line PI 118501. Pea line PI 118501 develops necrotic lesions and veinal necrosis on inoculated leaves, followed by systemic necrosis, leading to plant death. To understand the genetic basis of this lethal tip necrosis, we crossed lines PI 226564 and PI 250438, which develop mosaic symptoms in response to ClYVV inoculation. In reciprocal crosses of PI 118501 with PI 226564, all F₁ plants had mosaic symptoms with slight stem necrosis and early yellowing of upper leaves. Essentially the same symptom was manifested in PI 118501 × PI 250438 F₁ plants. In F₂ populations from the cross between PI 118501 and PI 226564, the observed ratios of necrosis, mosaic with slight stem necrosis, and mosaic fit the expected 1 : 2 : 1 ratio. These results indicate that a single incompletely dominant gene confers the induction of necrosis in PI 118501. This locus in pea, conferring necrosis induction to ClYVV infection, was designated Cyn1 (Clover yellow vein virus-induced necrosis). A linkage analysis using 100 recombinant inbred lines derived from a cross of PI 118501 and PI 226564 demonstrated that Cyn1 was located 7.5 cM from the SSR marker AD174 on linkage group III.     

Turnip mosaic virus VPg does not disrupt the translation initiation complex but interferes with cap binding

Plant potyvirus RNAs have a virus protein covalently attached at their 5′-end: the VPg (virus protein genome-linked). In part because of its position at the 5′end of the virus RNA, the VPg has been speculated to play a role in virus translation and replication. We report that the VPg of Turnip mosaic virus (TuMV) interacts with, and does not disrupt, the translation initiation complex eIF(iso)4E-eIF(iso)4G. The purification of VPgPro or 6K2VPgPro by affinity chromatography from infected plant tissue also led to the recovery of eIF(iso)4G. The cap analogue m⁷GTP competed with the binding of VPg to both eIF(iso)4E and the eIF(iso)4E-eIF(iso)4G complex. This suggested that the viral VPg bound to the translation initiation complex but that interaction did not result in disassociation of the subunits but rather in competition for the binding of the 5′ cap structure. This would result in decreased affinity of the translation initiation machinery for capped cellular mRNAs.     

Quantitative resistance to Plum pox virus in Prunus davidiana P1908 linked to components of the eukaryotic translation initiation complex

A complex, polygenic resistance to Plum pox virus (PPV) was previously described in a wild peach-related species, Prunus davidiana clone P1908. In the current study, an analysis of quantitative trait loci (QTL) was performed on an F₂ population comprising 99 individuals obtained by selfing the F₁ individual #40 of an interspecific cross between susceptible nectarine cv. Summergrand and the resistant P. davidiana clone P1908. Six QTL were identified using both parametric and non-parametric methods of detection, individually explaining 5–28% of the phenotypic variance. The total phenotypic variation explained ranged from 29 to 58%. Alignment of the genetic map of the F₂ cross with the P. davidiana parent map showed consistency of QTL over generations, with three of the six QTL co-localizing at the 1-LOD interval and another one at the 2-LOD interval. Two of the QTL were mapped onto linkage group one, where resistance to PPV was previously mapped in apricot. Development and mapping of new microsatellite markers linked to candidate genes revealed a striking co-localization of three of the detected QTL with gene copies coding for eukaryotic translation initiation factors eIF4E and eIF(iso)4G. As co-localization of one QTL with candidate gene eIF(iso)4E was previously reported in the F₁ population, the results reported here strongly reinforce the idea that components of the eukaryotic translation initiation complex are correlated with resistance to PPV in P. davidiana P1908.     

Resistance to <Emphasis Type="Italic">Rice yellow mottle virus</Emphasis> in rice germplasm in Madagascar

Rice yellow mottle virus (RYMV) is a recent and major threat to rice production in Madagascar. A large scale screening of resistance to RYMV in rice germplasm in Madagascar was conducted by visual symptom scoring and virus-assessment through ELISA tests. The response to virus inoculation of 503 local or introduced rice accessions was assessed. Most of them were susceptible to RYMV. A few cultivars expressed partial resistance at a level similar to the partially resistant Oryza sativa japonica cv. Azucena. Only one O. sativa cultivar expressed high resistance characterised by a lack of symptoms and an undetectable level of virus. It was a Malagasy traditional indica cultivar, named Bekarosaka, which originated from the northwest of the country. It was selected by farmers for its field resistance to RYMV. The response of cv. Bekarosaka to four pathotypes of RYMV was similar to that of cv. Gigante, the only other highly resistant indica cultivar. The sequence of the middle domain of the eIF(iso)4G, the genetic determinant of Rymv1 resistance on chromosome 4, of cv. Bekarosaka was similar to that of cv. Gigante. Subsequently, cvs Bekarosaka and Gigante probably carried the same resistance allele Rymv1-2. Rymv1-2 resistance was efficient against isolates of the major strains of RYMV, but was readily overcome by a pathotype from the northwest of Madagascar.     

Erysiphe trifolii is able to overcome er1 and Er3, but not er2, resistance genes in pea

Until recently, Erysiphe pisi was thought to be the only causal agent of powdery mildew in pea, but recent studies showed that other species such as Erysiphe trifolii and Erysiphe baeumleri can also cause this disease. Three genes, er1, er2 and Er3, conferring resistance to E. pisi have been reported so far in pea. Previous studies showed that E. trifolii and E. baeumleri were able to overcome er1 resistance, but whether er2 and Er3 were effective against E. trifolii was not known. In this study, pea accessions carrying these three genes were evaluated for resistance to E. trifolii under controlled conditions at 20 and 25 °C. In addition, these accessions were also evaluated under field conditions in Spain and in India. Analysis of the ribosomal DNA (rDNA) internal transcribed spacer (ITS) sequences showed that E. trifolii was the causal agent of powdery mildew symptoms in lines carrying er1 in Spain and that this pathogen was also present in India. Our results showed that E. trifolii was able to overcome er1 and shows that this pathogen can also overcome Er3 resistance in some conditions. In contrast, er2 provided high level of resistance against E. trifolii in all conditions and locations studied. Temperature affected the expression of Er3 against E. trifolii, but not of er1 or er2. The pea accession JI2480, containing er2, was highly resistant and JI2302 containing er1 was susceptible to E. trifolii at both temperatures, whereas P660-4 containing Er3 was resistant at 20 °C but susceptible at 25 °C. The present study also identified sources of resistance effective against both E. pisi and E. trifolii.     

Pathogenic diversity of soybean rust in Argentina, Brazil, and Paraguay

Phakopsora pachyrhizi, the cause of soybean rust, is an economically important pathogen of soybean in South America. Understanding the pathogenicity of indigenous fungal populations is useful for identifying resistant plant genotypes and targeting effective cultivars against certain populations. Fifty-nine rust populations from Argentina, Brazil, and Paraguay were evaluated for pathogenicity in three cropping seasons, 2007/2008–2009/2010, using 16 soybean differentials. Only two pairs of P. pachyrhizi populations displayed identical pathogenicity profiles, indicating substantial pathogenic variation in the rust populations. Comparative analysis of 59 South American and five Japanese samples revealed that pathogenic differences were not only detected within South America but also distinct between the P. pachyrhizi populations from South America and Japan. In addition, seasonal changes in rust pathogenicity were detected during the sampling period. The differentials containing resistance genes (Rpp: resistance to P. p achyrhizi) Rpp1, Rpp2, Rpp3, and Rpp4, except for Plant Introduction (PI) 587880A, displayed a resistant reaction to only 1.8–14, 24–28, 22, and 36 % of South American P. pachyrhizi populations, respectively. In contrast, PI 587880A (Rpp1), Shiranui (Rpp5), and 3 Rpp-unknown differentials (PI 587855, PI 587905, and PI 594767A) showed a resistant reaction to 78–96 % of all populations. This study demonstrated that P. pachyrhizi populations from South America vary geographically and temporally in pathogenicity and that the known Rpp genes other than Rpp1 in PI 587880A and Rpp5 have been less effective against recent pathogen populations in the countries studied.     

Differential tolerance of peas to prometryne and terbutryn

The phytotoxicity of 2,4-bis(isopropyl)-6-(methylthio)-s-triazine (prometryne) and 2-(tert-butylamino)-4-(ethylamino)-6-(methylthio)-s-triazine (terbutryn) to peas (Pisum sativum L. var. Perfection 3040) was studied. No differences were found when the herbicides were applied to the roots of intact plants in nutrient solution or directly to leaf discs. However, prometryne was much more toxic when uptake was from soil. Absorption and translocation of ¹⁴C-labeled prometryne and terbutryn showed that the majority of terbutryn accumulated in the roots, whereas prometryne was uniformly distributed between the roots and the shoot. Thin layer chromatography of extracts from prometryne-treated peas showed that only 20% of the absorbed compound was metabolized to produce one breakdown product. Extracts of terbutryn-treated plants contained three different metabolites. After 120 hr of exposure to terbutryn, about half of the absorbed herbicide was metabolized. The results show that the main factors responsible for the differential toxicity of the herbicides to peas were availability from the soil, translocation pattern and initial detoxication.     

甜玉米小斑病抗性的遗传分析与主效QTL定位

为培育抗病品种,利用抗小斑病甜玉米自交系T14和感小斑病自交系T18为亲本配制杂交组合,对玉米抗小斑病性状进行遗传分析和抗病基因分子标记定位,用主基因＋多基因混合遗传模型和P₁、P₂、F₁、B₁、B₂、F₂ 6世代联合分析的方法对单位叶面积病斑数量进行遗传分析,并应用复合区间作图法检测抗小斑病QTL。结果表明,单位叶面积病斑数量受2对加性-显性-上位性主基因控制,自交系T14的抗病性在各个分离世代都以主基因遗传为主。在第4染色体上检测到4个相互连锁的小斑病抗性QTL,解释表型变异的7.7%、30.9%、14.8%和11.5%;在第6染色体上定位了1个抗病QTL,可解释表型变异的37.7%。检测到的小斑病抗性主效QTL位于第4和第6染色体的特征与2对主基因的遗传模型相吻合。     

Expression patterns of defence genes and antioxidant defence responses in a rice variety that is resistant to leaf blast but susceptible to neck blast

The rice stage resistance variety Jiajing3768 is resistant to leaf blast but susceptible to neck blast. This variety was used to analyze the expression patterns of defence genes and antioxidant defence responses in the leaves at the seedling stage (LSS) and necks at the preliminary heading stage (NPHS), after inoculation with Magnaporthe oryzae. We found that defence genes PR1b (pathogenesis-related class 1b), PR4, PR10a, JIOsPR10 (jasmonic acid induced rice PR 10), Gns1 (1,3; 1,4-β-glucanase), Cht-1 (chitinase), and LOX (lipoxygenase) may play roles in the stage resistance in Jiajing3768; PAL (phenylalanine ammonia-lyase) and CHS (chalcone synthase) could participate in defending Jiajing3768 against neck blast but not leaf blast. The antioxidant enzymes superoxide dismutase, peroxidase, catalase, glutathione reductase, and malondialdehyde coordinately participate in the stage resistance to blast in Jiajing3768, and that oxidative damage is less in the LSS than in the NPHS.     

Identification of a constitutive 45 kDa soluble protein associated with resistance to downy mildew in muskmelon (Cucumis melo L.), line PI 124111 F

Analysis by sodium dodecylsulphate-polyacrylamide gel electrophoresis of the soluble proteins extracted from muskmelon (Cucumis melo L.), inbred line PI 124111F (PI) with resistance to Pseudoperonospora cubensis (Berk, et Curt.) Rost, and two susceptible cvs, Hemed and AnanasYokneam revealed the presence of a unique 45 kDa protein (P45) in the resistant but not in the susceptible plants. Subcellular fractionation indicated that P45 is a cytoplasmic soluble protein. It was detected in leaves and cotyledons but not in stems or roots. F₁ hybrid plants (Hemed × PI) that displayed only a partial resistance against P. cubensis expressed P45 at an intermediate level, whereas plants of the inbred generations F₅, F₇ and F₁₀ that displayed a high degree of resistance, expressed P45 at a level similar to the parental PI. Back-cross progeny plants [(Hemed × PI) × Hemed] segregate 1:2:1 partially resistant: susceptible: highly susceptible to the disease. The partially resistant plants showed an intermediate level of P45 (similar to F₁) whereas the highly susceptible plants had no P45, thus indicating cosegregation of P45 with resistance. The resistance of PI to P. cubensis was found to decrease at a colonization temperature of 12 °C. ³⁵S-methionine in vivo protein labelling revealed a reduction in the intensity of the P45 band in plants incubated for 11 h at 12 °C. The application of P45 in breeding programs of muskmelon for downy mildew resistance and its possible involvement in resistance to P. cubensis are discussed.     

Use of real-time PCR to examine the relationship between disease severity in pea and <Emphasis Type="Italic">Aphanomyces euteiches</Emphasis> DNA content in roots

Aphanomyces euteiches causes severe root rot of peas. Resistance is limited in commercial pea cultivars. Real-time fluorescent PCR assay specific for A. euteiches was used to study the relationship between disease severity and pathogen DNA content in infected peas. Five pea genotypes ranging in levels of resistance were inoculated with five isolates of A. euteiches. Plants were visually rated for disease development and the amount of pathogen DNA in roots was determined using the PCR assay. The susceptible genotypes Genie, DSP and Bolero tended to have significantly more disease and more pathogen DNA than the resistant genotypes 90-2079 and PI 180693. PI 180693 consistently had less disease, while 90-2079 had the lowest amount of pathogen DNA. The Spearman correlation between pathogen DNA quantity and disease development was positive and significant (P < 0.05) for three isolates, but was not significant for two other isolates. This suggests that the real-time PCR assay may have limited application as a selection tool for resistance in pea to A. euteiches. Its utility as a selection tool would be dependent on the correlation between disease development and pathogen DNA content for a given pathogen isolate. The accuracy and specificity of the real-time PCR assay suggests considerable application for the assay in the study of mechanisms of disease resistance and the study of microbial population dynamics in plants.     

Validation of a QTL for resistance to ascochyta blight linked to resistance to fusarium wilt race 5 in chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.)

Ascochyta blight caused by Ascochyta rabiei and fusarium wilt caused by Fusarium oxysporum. f. sp. ciceris are the two most serious diseases of chickpea (Cicer arietinum). Quantitative trait loci (QTL) or genes for ascochyta blight resistance and a cluster of resistance genes for several fusarium wilt races (foc1, foc3, foc4 and foc5) located on LG2 of the chickpea map have been reported independently. In order to validate these results and study the linkage relationship between the loci that confer resistance to blight and wilt, an intraspecific chickpea recombinant inbred lines (RIL) population that segregates for resistance to both diseases was studied. A new LG2 was established using sequence tagged microsatellite sites (STMS) markers selected from other chickpea maps. Resistance to race 5 of F. oxysporum (foc5) was inherited as a single gene and mapped to LG2, flanked by the STMS markers TA110 (6.5 cM apart) and TA59 (8.9 cM apart). A QTL for resistance to ascochyta blight (QTL_AR3) was also detected on LG2 using evaluation data obtained separately in two cropping seasons. This genomic region, where QTL_AR3 is located, was highly saturated with STMS markers. STMS TA194 appeared tightly linked to QTL_AR3 and was flanked by the STMS markers TR58 and TS82 (6.5 cM apart). The genetic distance between foc5 and QTL_AR3 peak was around 24 cM including six markers within this interval. The markers linked to both loci could facilitate the pyramiding of resistance genes for both diseases through MAS.     

BTH处理对甜瓜叶片活性氧代谢的诱导作用

为了明确活性氧（reactive oxygen species,ROS）代谢在甜瓜抗病性诱导中的作用,以抗白粉病甜瓜品种Tam Dew和感病品种卡拉克赛幼苗为材料,通过盆栽试验研究了苯丙噻二唑（BTH）喷雾或白粉菌接种后甜瓜叶片超氧阴离子（O2.-）产生速率、过氧化氢（H2O2）含量及超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、苯丙氨酸解氨酶（PAL）活性的变化。BTH处理或白粉菌接种均可诱导甜瓜叶片SOD、PAL活性升高,抑制CAT活性,导致叶组织O2.-产生速率和H2O2含量增加,BTH喷雾＋白粉菌接种比二者单独处理效果更好。结果表明,BTH处理后叶片O2.-产生速率提高和H2O2积累是甜瓜抗白粉病能力提高的重要机制,BTH通过诱导ROS代谢酶活性调节H2O2含量,且BTH诱导的甜瓜抗病性与品种的基础抗性有关。     

High activities and mRNA expression of pyrophosphate-fructose-6-phosphate-phosphotransferase and 6-phosphofructokinase are induced as a response to Rhizoctonia solani infection in rice leaf sheaths

Rice sheath blight disease caused by Rhizoctonia solani Kuhn results in significant yield and quality losses in rice growing areas worldwide. The glycolytic pathway is important in the resistance response to R. solani infection in rice. This study examined one of the regulatory steps in this pathway catalyzed by pyrophosphate-fructose-6-phosphate-phosphotransferase (PFP) and 6-phosphofructokinase (PFK). PFP and PFK activity in R. solani-infected rice plants increased. The mRNA expression of PFP/PFK isozymes showed that PFK 1, 2, 4 and 5 in the resistant line at 1 dpi were high as compared to the gradual increase observed in the expression of all PFP isozymes. Also PFK 1, PFK 3, PFK 4, PFK 5, PFP 2 and PFP 5 were adaptive to sheath blight disease infection and linked to defence response while, the expressions of PFK 2, PFP 1, PFP 3 and PFP 4 although adaptive, were not specific to R. solani infection. These observations provide evidence that (a) both PFP and PFK have isozymes that play an adaptive role after R. solani infection but while those of PFK are expressed at higher levels within a short time after infection those of PFP are expressed gradually, (b) the adaptive activation of PFP in R. solani-infected rice plants is correlated with the paired expression of its α- and β-subunits as shown by PFP 2 and PFP 5, and (c) the expression of some α-subunits is not specific to R. solani infection as shown by PFP 1, PFP 3 and PFP 4.     

Tagging and mapping a second resistance gene for <Emphasis Type="Italic">Fusarium</Emphasis> wilt race 0 in chickpea

A second gene conferring resistance to the chickpea wilt pathogen, Fusarium oxysporum f. sp ciceris race 0, has been mapped to linkage group 2 (LG2) of the chickpea genetic map. Resistance to race 0 is controlled by two genes which segregate independently; one present in accession JG62 (Foc0 ₁ /foc0 ₁ ) and mapping to LG5 and the second present in accession CA2139 (Foc0 ₂ /foc0 ₂ ) but remaining unmapped. Both genes separately confer complete resistance to race 0 of the wilt pathogen. Using a Recombinant Inbred Line (RIL) population that segregated for both genes (CA2139 × JG62) and the genotypic information provided by two markers flanking Foc0 ₁ /foc0 ₁ ten resistant lines containing the resistant allele Foc0 ₂ /foc0 ₂ were selected. Genotypic analysis using these ten resistant lines paired with ten susceptible RILs, selected in the same population, revealed that sequence tagged microsatellite sites (STMS) markers sited on LG2 were strongly associated with Foc0 ₂ /foc0 ₂ . Linkage analysis, using data from two mapping populations (CA2139/JG62 and CA2156/JG62), located Foc0 ₂ /foc0 ₂ in a region where genes for resistance to wilt races 1, 2, 3, 4 and 5 have previously been reported and which is highly saturated with tightly-linked STMS markers that could be used in marker-assisted selection (MAS).     

Experimental evidence of a pathogenic change caused by homologous recombination between endogenous and introduced dysfunctional Avr-Pita genes in Pyricularia oryzae

To provide experimental evidence that somatic homologous recombination (HR) is involved in the instability and diversification of the avirulence gene Avr-Pita in Pyricularia oryzae, we generated a dysfunctional Avr-Pita homolog and integrated it into strain Hoku-1 containing Avr-Pita. In the transformants, the occurrence of somatic HR events between Avr-Pita and the dysfunctional homolog was confirmed by PCR–RFLP. Germlings from conidia from the HR-positive transformants had lost the avirulence function, which enabled it to infect rice cultivar Yashiromochi containing the corresponding resistant gene Pi-ta. These results suggested that genetic mutation caused by somatic HR is one of the mechanisms responsible for virulence diversity.     

The role of jasmonic acid signalling in wheat (Triticum aestivum L.) powdery mildew resistance reaction

Jasmonic acid (JA) signalling plays an important role in plant resistance to pathogens. Previously, JA has been found to play a role in induced disease resistance to necrotrophic pathogens in various plant species, but current researches showed that JA also enhanced resistance to biotrophic pathogens. However, its role in wheat (Triticum aestivum L.) powdery mildew (Blumeria graminis f. sp. tritici, Bgt) resistance reaction is largely unknown. To settle this issue, several typical powdery mildew resistant and susceptible wheat varieties were employed. The sensitivity to exogenous methyl jasmonate (MeJA) to wheat powdery mildew resistance, the concentration fluctuation of endogenous JAs after Bgt inoculation, and the expression profiles of nine pathogenesis-related protein genes (PR genes) after MeJA and Bgt treatments were studied systematically. Exogenous MeJA significantly enhanced the powdery mildew resistance of the susceptible varieties. After inoculation with Bgt, endogenous JAs accumulated rapidly, reached the maxima at 2 to 5 h post-inoculation (hpi), then decreased rapidly, and the concentration was almost the same as that of un-inoculated control at 96 hpi. The expression levels of the nine PRs were measured by real time quantitative RT-PCR (qRT-PCR) at different time points after MeJA application or Bgt inoculation respectively. The MeJA and Bgt strongly activated PR1, PR2, PR3, PR4, PR5, PR9, PR10 and Ta-JA2, but almost didn’t affect Ta-GLP2a. The induced powdery mildew resistance was positively correlated with the activated PR genes. JA plays a positive role in defence against Bgt. JA is a signalling molecule in wheat powdery mildew resistance and future manipulation of this pathway may improve powdery mildew resistance in wheat breeding.     

Insecticidal activity and mode of action of novel nicotinoids synthesized by new acylpyridinium salt chemistry and directed lithiation

Novel acylpyridinium salt chemistry and directed lithiation methodology was developed to add for the first time substitutions directly to the phenylpyridine heterocyclic ring of nicotine. A variety of 3-(1-methylpyrrolidin-2-yl)-4-(alkyl, aromatic, heterocyclic and silanyl) and -N-alkyl pyridines were synthesized (compounds 1-9). In vial tests with the green peach aphid, Myzus persicae, compounds 1-4 were 1.1, 1.8, 2.3 and 1.9×, respectively, more active than nicotine and 64, 40, 31 and 38×, respectively, less active than acetamiprid. Against the western flower thrips, Frankliniella occidentalis, 1-4 were 1.4, 2.1, 2.0 and 1.6×, respectively, more active than nicotine and 9, 6, 6 and 8×, respectively, less active than acetamiprid. For the cotton aphid, Aphis gossypii, the activity of 1-9 was similar to nicotine. Compounds 7 and 9 when incorporated into artificial diet produced low mortality for larvae of the beet armyworm, Spodoptera exigua, but were not active against the corn earworm, Helicoverpa zea. When 1-4 and 6-9 were injected into larvae of the beet armyworm, a variety of symptoms similar to acetamiprid were observed which included tremors, uncoordinated movement, diuresis, paralysis and death. In addition, imidacloprid-binding to membranes from the house fly head, Musca domestica, was inhibited by compounds 1-9, when using a concentration range of 1-100 μM. These studies demonstrate that our new chemistry enhances the insecticidal activity of nicotine with an apparent mode of action as an acetylcholine agonist.