Expression and inheritance of nerve insensitivity resistance in larvae of Helicoverpa armigera (Lepidoptera: Noctuidae) from China

Nerve insensitivity resistance to synthetic pyrethroids was detected in a resistant field strain (JSFX-R) of the cotton bollworm, Helicoverpa armigera (Hübner), using a neurophysiological assay in which extracellular spontaneous neuronal activity was measured in response to cis-cypermethrin. The nerve insensitivity mechanism was selected using a combination of toxicological and neurophysiological methods. The third-instar larvae in selected strains of Family-37 and CTR strain expressed a very high resistance to fenvalerate (RF = 2060-fold and 805-fold, respectively) and high cross-resistance to DDT (RF = 1927-fold and 2384-fold, respectively) which was not affected by two metabolic synergists, PBO and DMC. The frequency of nerve-insensitive individuals detected in neurophysiological assays (54, 81 and 100% for JSFX-R strain, and the selected strains Family-43 and Family-37, respectively) was not only positively correlated (R² = 0.968) with the frequency of non-PBO-synergisable resistant individuals detected in toxicological tests (37.5, 62.5 and 90% for JSFX-R strain, Family-43 and Family-37, respectively), but also positively correlated (R² = 0.978) with the frequency of DDT-resistant individuals detected in toxicological tests (40, 67.5 and 93.3% for JSFX-R strain, Family-43 and Family-37, respectively). Analysis of dose–mortality lines to DDT and fenvalerate from F₁ hybrids (R♀ × S♂) indicated that nerve insensitivity resistance to DDT and fenvalerate in the CTR strain was inherited in an incompletely recessive pattern. Degree of dominance (D) was estimated to be −0.66 (± 0.06) (DDT) and −0.26 (± 0.04) (fenvalerate). The dose–mortality curves to DDT in back-cross progeny were strongly suggested, by chi-square analysis, to be fitted with those expected of a one-gene model. Evidence for the co-existence of nerve insensitivity and oxidative metabolic resistance mechanisms within individual H armigera and the effects of their interaction on the expression of resistance to fenvalerate are discussed. © 1999 Society of Chemical Industry     

Nerve Insensitivity to cis-Cypermethrin is Expressed in Adult Heliothis virescens

Adult and third-instar larval Heliothis virescens were examined for nerve insensitivity to cis-cypermethrin using a rapid and robust neurophysiological method. Three laboratory strains, BRC susceptible, PEG-87 resistant and MS2 metabolic resistant, were compared with a series of pyrethroid-resistant strains collected from a field site in Louisiana, USA, during the 1992 cotton season. Nerve insensitivity was present in a proportion of the individuals in both the PEG-87 and the field strains, at both life stages, minimal in the MS2 strain and absent in the BRC strain. Frequencies of nerve-sensitive and nerve-insensitive individuals in each strain were not significantly different at the two life stages examined.     

Nerve insensitivity resistance to cypermethrin in larvae of the tobacco budworm Heliothis virescens from USA cotton field populations

Spontaneous neuronal activity was recorded from the peripheral nerves of third-instar larvae of strains of Heliothis virescens (F.) obtained directly from cotton fields in the USA. Following a control period the preparations were exposed to increasing concentrations of cis-cypermethrin in a cumulative dose-response assay. A positive response was defined as an increase of at least five-fold in the rate of neuronal activity over that seen during the control period. Up to 35 individuals of each strain were assayed and the responses used to construct a phenotypic profile categorising the individuals from nerve-susceptible to highly nerve-insensitive. An EC₅₀ for the action of cis-cypermethrin was also obtained. There was a positive, significant correlation between non-synergisable resistance to cypermethrin and nerve insensitivity as defined in the neurophysiological assay. It was shown that nerve insensitivity to cypermethrin increased throughout the cotton-growing season.     

Pyrethroid resistance mechanisms in Australian Helicoverpa armigera

In 1983, at the onset of pyrethroid resistance in Australian Helicoverpa armigera, three resistance mechanisms were identified. They were: a strong nerve insensitivity (Super-Kdr), penetration resistance (Pen), and a factor which was overcome by piperonyl butoxide (PBO). Super-Kdr nerve insensitivity appeared to be a major cause of pyrethroid resistance and contributed to high-order resistance (> 100-fold). From 1987 to 1990, to monitor the effect of the Australian Helicoverpa insecticide resistance management strategy on insecticide resistance, we conducted a survey of the frequencies of these mechanisms in field-collected H. armigera. The relative importance of the Pen and Pbo mechanisms in resistant H. armigera have increased but Pen and Pbo confer only low order (～ 20-fold) resistance. We found no evidence of the 1983 Super-Kdr mechanism in the latest samples, but, instead, found another distinct Kdr-type mechanism. This mechanism was correlated with very low-order nerve insensitivity and was of little toxicological significance. The impact of the Helicoverpa insecticide resistance management strategy on pyrethroid resistance in H. armigera is discussed.     

Virulence and toxin synthesis of an azole insensitive Fusarium culmorum strain in wheat cultivars with different levels of resistance to fusarium head blight

Fusarium culmorum causes head blight, produces toxins and reduces yield and quality of cereals. To prevent damage caused by fusarium head blight (FHB), azole fungicides are mainly applied. The occurrence of insensitivity to azoles is a major problem in agriculture. The present study shows that a tebuconazole insensitive strain of F. culmorum can be readily produced in the laboratory, but that the resulting strain of the fungus is of lower fitness in vitro. Insensitivity was confirmed microscopically and by cell viability and metabolic activity. The tebuconazole insensitive strain shows cross insensitivity to nine important azoles. In addition, plants inoculated with the insensitive F. culmorum strain showed no reduction of FHB symptoms and deoxynivalenol (DON) content after tebuconazole treatment, compared to an inoculation with the sensitive strain. Use of wheat cultivars carrying a high resistance level (i.e. cv. Toras) was the most effective method for reducing symptoms and decreasing DON content, independent from the level of fungicide insensitivity of the F. culmorum strain. In conclusion, resistant cultivars and a fungicide mixture which combines different mechanisms of action in fungal metabolism should be applied to avoid fungicide insensitivity of Fusarium spp. in future.     

Cross‐resistance,inheritance and biochemical mechanisms of imidacloprid resistance in B‐biotype Bemisia tabaci

BACKGROUND: The B‐type Bemisia tabaci (Gennadius) has become established in many regions in China, and neonicotinoids are extensively used to control this pest. Imidacloprid resistance in a laboratory‐selected strain of B‐type B. tabaci was characterised in order to provide the basis for recommending resistance management tactics. RESULTS: The NJ‐Imi strain of B‐type B. tabaci was selected from the NJ strain with imidacloprid for 30 generations. The NJ‐Imi strain exhibited 490‐fold resistance to imidacloprid, high levels of cross‐resistance to three other neonicotinoids, low levels of cross‐resistance to monosultap, cartap and spinosad, but no cross‐resistance to abamectin and cypermethrin. Imidacloprid resistance in the NJ‐Imi strain was autosomal and semi‐dominant. It is shown that enhanced detoxification mediated by cytochrome‐P450‐dependent monooxygenases contributes to imidacloprid resistance to some extent in the NJ‐Imi strain. Results from synergist bioassays and cross‐resistance patterns indicated that target‐site insensitivity may be involved in imidacloprid resistance in the NJ‐Imi strain of B. tabaci. CONCLUSION: Although oxidative detoxification mediated by P450 monooxygenases is involved in imidacloprid resistance in the NJ‐Imi strain of B‐type B. tabaci, target‐site modification as an additional resistance mechanism cannot be ruled out. Considering the high risk of cross‐resistance, neonicotinoids should be regarded as a single group when implementing an insecticide rotation scheme in B. tabaci control. Copyright © 2009 Society of Chemical Industry     

An electrophysiological investigation of the susceptible (cooper) and resistant (kdr; super-kdr) strains of the adult housefly,Musca domestica L

The electrical activity of abdominal nerves of the housefly, Musca domestica L., was used as a bioassay to study nerve sensitivity to DDT and deltamethrin in susceptible (Cooper) and resistant (kdr, super-kdr) strains. By this technique the resistant strains were less sensitive (approximately 10 000-fold) than Cooper, but the bioassay could not distinguish between super-kdr and kdr in their responses to either compound and so could not account for the greater resistance shown by flies with super-kdr above kdr flies when these insecticides are applied topically. Although factors other than nerve insensitivity may be involved, the compounds were applied to the preparation in aqueous saline solutions at, or close to, their solubility limits and this could have masked differences in responses of nerves from the resistant strains.     

Molecular analysis of pyrethroid resistance conferred by target insensitivity and increased metabolic detoxification in Plutella xylostella

BACKGROUND: The pyrethroid resistance of the diamondback moth Plutella xylostella (L.) is conferred by increased gene expression of cytochrome P450 to detoxify the insecticide and/or through gene mutation of the sodium channel, which makes the individual insensitive to pyrethroids. However, no information is available about the correlation between the increased metabolic detoxification and the target insensitivity in pyrethroid resistance. RESULTS: Frequencies of pyrethroid‐resistant alleles (L1014F, T929I and M918I) and two resistance‐related mutations (A1101T and P1879S) at the sodium channel and expression levels of the cytochrome P450 gene CYP6BG1 were examined individually using laboratory and field strains of P. xylostella. Real‐time quantitative PCR analysis using the laboratory strains revealed that levels of larval expression of the resistant strain, homozygous for the pyrethroid‐resistant alleles other than the M918I, are significantly higher than those of the susceptible strain. In the field strains, the expression levels in insects having the same resistant alleles as those of the resistant strains varied greatly among individuals. The expression levels were not significantly higher than those in the heterozygotes. CONCLUSION: Significant correlation between the target insensitivity and the increased metabolic detoxification in pyrethroid resistance of P. xylostella was observed in the laboratory but not in the field. Copyright © 2010 Society of Chemical Industry     

棉花内生真菌CEF-373菌株对棉花黄萎病的防效及其作用机理

为明确棉花内生真菌CEF-373菌株对棉花黄萎病的防效及其作用机理,利用圆盘滤膜法和平板对扣培养法测定菌株CEF-373对大丽轮枝菌Verticillium dahliae菌丝生长的抑制作用,测定其对棉花黄萎病的温室和田间防效,并通过活性氧含量及防御基因表达情况来分析其作用机理。结果表明,菌株CEF-373的挥发性代谢产物和非挥发性代谢产物对大丽轮枝菌菌丝的生长均有显著抑制作用,抑制率最高分别可达37.75%和100.00%。用1×107CFU/mL的菌株CEF-373分生孢子悬浮液灌根后,对棉花黄萎病的温室防效可达71.12%,用质量比为3%的菌株CEF-373固体菌剂拌土栽培后,对棉花黄萎病的温室防效可达62.74%,防治作用显著。菌株CEF-373的发酵液滴灌和微生物肥料处理40 d后,对棉花黄萎病的田间防效达到最大,分别为36.23%和27.71%,而在后期有所降低。菌株CEF-373可以诱导棉花叶片中细胞活性氧的爆发;且菌株CEF-373成功激活了苯丙氨酸解氨酶、过氧化物酶、多酚氧化酶、几丁质酶和病程相关蛋白基因PR10的表达,对大丽轮枝菌的侵染具有抵御作用。表明棉花内生真菌CEF-373菌株通过抑制大丽轮枝菌生长以及诱导寄主系统抗病性来有效防治棉花黄萎病,具有较好的生物防治应用前景。     

Seasonal Dynamics of Metabolic Mechanisms Mediating Pyrethroid Resistance in Helicoverpa armigera in Central India

Very high cypermethrin and fenvalerate resistance frequencies were recorded in Helicoverpa armigera (Hübner) populations in central India during the 1993–94, 1994–95 and 1995–96 cropping seasons. Synergism assays and biochemical analyses of detoxification enzyme levels indicated that mono-oxygenases and esterases were important metabolic mechanisms mediating pyrethroid resistance. Piperonyl butoxide- (PBO) and profenofos-suppressible pyrethroid resistance were correlated with enhanced levels of cytochrome P450 and general esterases respectively. Enzyme assay data indicated that high cytochrome P450 levels generally coincided with low esterase activity and vice versa. Similarly, synergist bioassays showed that PBO-insensitive resistance was frequently associated with profenofos-sensitive resistance and vice versa. Oxidase- and esterase-mediated mechanisms evidently alternated in a reciprocal manner, with perceptible shifts in relative importance occurring during mid-October in all three seasons and in late January in 1995. Apart from metabolic mechanisms, a synergist-insensitive resistance mechanism (believed to be nerve insensitivity), accounted for an average of 51, 30 and 28% of cypermethrin resistance during the 1993–94, 1994–95 and 1995–96 seasons respectively. © 1997 SCI.     

Cross‐resistance and possible mechanisms of chlorpyrifos resistance in Laodelphax striatellus (Fallén)

BACKGROUND: Laodelphax striatellus (Fallén) is a major pest of cultivated rice and is commonly controlled in China with the organophosphate insecticides. To develop a better resistance management strategy, a chlorpyrifos‐resistant strain of L. striatellus was selected in the laboratory, and its cross‐resistance to other insecticides and possible mechanisms of the chlorpyrifos resistance were investigated. RESULTS: After 25 generations of selection with chlorpyrifos, the selected strain of L. striatellus developed 188‐fold resistance to chlorpyrifos in comparison with the susceptible strain, and showed 14‐ and 1.6‐fold cross‐resistance to dichlorvos and thiamethoxam respectively. There was no apparent cross‐resistance to abamectin. Chlorpyrifos was synergised by the inhibitor triphenyl phosphate; the carboxylesterase synergistic ratio was 3.8 for the selected strain, but only 0.92 for the susceptible strain. The carboxylesterase activity of the selected strain was approximately 4 times that of the susceptible strain, whereas there was no significant change in the activities of alkaline phosphatase, acid phosphatase, glutathione S‐transferase and cytochrome P450 monooxygenase between the strains. The Michaelis constant of acetylcholinesterase, maximum velocity of acetylcholinesterase and median inhibitory concentration of chlorpyrifos‐oxon on acetylcholinesterase were 1.7, 2.5 and 5 times higher respectively in the selected strain. CONCLUSION: The high cross‐resistance to the organophosphate dichlorvos in the chlorpyrifos‐resistant strain suggests that other non‐organophosphate insecticides would be necessary to counter resistance, should it arise in the field. Enhanced activities of carboxylesterase and the acetylcholinesterase insensitivity appear to be important mechanisms for chlorpyrifos resistance in L. striatellus. Copyright © 2010 Society of Chemical Industry     

Biochemical and molecular mechanisms of diafenthiuron resistance in the whitefly,Bemisia tabaci

B-biotype Bemisia tabaci has developed high levels of resistance to many insecticides. To investigate the risks and explore possible mechanisms of resistance to diafenthiuron in B. tabaci, a 32.8-fold diafenthiuron-resistant strain (R-DfWf) was established after selection for 36 generations compared with the susceptible strain (S-Lab). Biochemical assays showed that the activity of cytochrome P450 towards p-NA was significantly higher (4.37-fold higher) in the R-DfWf strain than in the S-Lab strain. Similarly, the carboxylesterase (COE) activity and glutathione S-transferase (GST) activity were also significantly higher (3.12- and 1.83-fold higher, respectively) in the R-DfWf strain than in the S-Lab strain. The expression of five of seven P450 genes was significantly higher (>3-fold) in the R-DfWf strain than in the S-Lab strain. The expression of COE2 was significantly higher (>2.5-fold) in the R-DfWf than in the S-Lab strain. The expression of GST and GST2 was significantly higher (>2.3-fold) in the R-DfWf than in the S-Lab. Thus, cytochrome P450, COE and GST may appear to be responsible for the resistance to diafenthiuron in B. tabaci. It is also valuable for usage of insecticides for resistance management and control of this species.     

葡萄生单轴霉重寄生菌F3的鉴定及防治效果测定

为明确从生长异常的葡萄霜霉层上分离获得的菌株F3对葡萄生单轴霉Plasmopara viticola的重寄生作用及对葡萄霜霉病的防治效果,通过光学和扫描电子显微镜观察菌株F3对葡萄生单轴霉的重寄生作用,并对菌株F3进行形态特征观察及28S rDNA序列分析,采用孢子囊萌发抑制和离体叶片点样法测定该菌株对葡萄霜霉病的防治效果。结果表明,菌株F3对葡萄生单轴霉表现出覆盖、缠绕的重寄生现象。结合形态特征与序列分析结果将该菌株最终鉴定为层生镰刀菌Fusarium proliferatum。菌株F3分生孢子悬浮液及其无菌发酵液对葡萄霜霉菌孢子囊萌发具有显著的抑制作用,抑制率分别为86.8%和83.1%;经菌株F3分生孢子悬浮液预防处理后的离体葡萄叶片发病率为10.9%,显著低于对照的发病率98.8%,防治效果达88.9%;菌株F3分生孢子悬浮液与葡萄霜霉病菌孢子囊混合后点样接种处理的叶片发病率为50.0%,也显著低于对照,防治效果为49.1%;而治疗处理的叶片发病率与对照间无显著差异。表明层生镰刀菌对葡萄生单轴霉具有重寄生作用,且对葡萄霜霉病具有较强的预防控制作用。     

Factors associated with the incidence of Aspergillus section Flavi and aflatoxin contamination of peanuts in the Busia and Homa bay districts of western Kenya

The aim of this study was to identify agronomic, ecological and sociocultural factors that could be modified to reduce the risk of aflatoxin contamination of peanuts from western Kenya. Presence of fungi within section Flavi of the genus Aspergillus and levels of total aflatoxin were determined for 436 peanut samples from the Busia and Homa bay districts. A total of 1458 cultures of Aspergillus flavus or A. parasiticus isolated from the samples were assayed for production of aflatoxin B₁, B₂, G₁ and G₂. Associations among the incidences of fungal species, incidences of samples with ≥10 μg kg^?1 aflatoxin, production of specific aflatoxin types and various agronomic, ecological and sociocultural factors were modelled with chi‐squared and logistic regression methods. The predominant species were A. flavus L‐strain (78% incidence), A. flavus S‐strain (68%) and A. niger (65%). Occurrence of A. caelatus, A. alliaceus and A. tamarii in Kenya was also documented. Samples from the Busia district were three times (odds ratio = 3·01) as likely to contain ≥10 μg kg^?1 of total aflatoxin as were samples from the Homa bay district, while samples containing A. flavus S‐strain were 96% more likely to exceed this threshold compared with samples from which this fungus was not isolated. Grading, planting improved cultivars and membership of a producer marketing group were negatively associated with the incidence of A. flavus, while crop rotation was negatively correlated with the incidence of B aflatoxins. These sociocultural factors can be modified to reduce the risk of peanut contamination with aflatoxin.     

辣椒炭疽病生防菌株的筛选、鉴定及其抑菌机理

为有效防控由胶孢炭疽菌Colletorichum gloeosporioides引起的辣椒炭疽病,自辣椒上分离得到内生细菌,通过平板拮抗和辣椒离体生防试验筛选对胶孢炭疽菌有抑制作用的拮抗菌株,通过形态学特征、生理生化特征以及分子生物学技术对其进行鉴定,并于室内测定其对胶孢炭疽菌菌丝生长的影响、对辣椒炭疽病的防效及接种后辣椒内抗病活性物质含量以及防御酶活性。结果显示,从辣椒上共分离纯化获得46株细菌,其中菌株SQ-6对胶孢炭疽菌有明显的抑制作用,抑制率为61.11%,显著高于其他45株。结合菌株SQ-6的形态学特征、生理生化特征以及分子生物学特征,将该菌株鉴定为解淀粉芽胞杆菌Bacillus amyloliquefaciens。SQ-6菌株的50%无细胞滤液可引起胶孢炭疽菌菌丝畸形、断裂等,对其抑制率为57.87%。SQ-6菌株的10%、50%发酵液和10%、50%无细胞滤液均能显著降低由胶孢炭疽菌引起的辣椒炭疽病的发病率和病情指数,其中50%无细胞滤液的防效最好。SQ-6菌株能够提高辣椒内Vc、酚类和黄酮类物质含量,诱导辣椒内过氧化物酶（peroxidase,POD）、丙氨酸解氨酶（p...     

Distribution and metabolism of abamectin in larval stages of Spodoptera littoralis boisduval and Heliothis armigera hübner (Lepidoptera: Noctuidae)

Comparisons were made between the amounts of abamectin and total radioactivity recovered from Spodoptera littoralis and Heliothis armigera larvae after topical application of [³H]abamectin. Penetration (as shown by wash-off experiments) did not differ significantly between the Spodoptera instars. Significantly more abamectin was recovered from ventral nerve cord samples of larvae showing symptoms of poisoning than from larvae not showing these symptoms. Fifth-instar S. littoralis larvae had a significantly lower proportion of radioactivity as abamectin in the ventral nerve cord than in sixth-instar S. littoralis or fifth- and sixth-instar H. armigera. The proportion of radioactivity present as abamectin (but not total radioactivity) was significantly increased when the fifth-instar S. littoralis larvae were pre-treated with piperonyl butoxide (PB) suggesting that the relative insensitivity of fifth-instar S. littoralis larvae to abamectin is due at least in part to greater metabolism, particularly by microsomal oxidases. Fat body samples consistently had a greater proportion of radioactivity as abamectin than the nerve cord and the former may act as storage sites for abamectin.     

Resistance mechanisms and associated mutations in acetylcholinesterase genes in Sitobion avenae (Fabricius)

Wheat aphid, Sitobion avenae (fabricius), is one of the most important wheat pests and has been reported to be resistant to commonly used insecticides in China. To determine the resistance mechanism, the resistant and susceptible strains were developed in laboratory and comparably studied. A bioassay revealed that the resistant strain showed high resistance to pirimicarb (RR: 161.8), moderate reistance to omethoate (32.5) and monocrotophos (33.5), and low resistance to deltamethrin (6.3) and thiodicarb (5.5). A biochemistry analysis showed that both strains had similar glutathione-S-transferase (GST) activity, but the resistant strain had 3.8-fold higher esterase activity, and its AChE was insensitive to this treatment. The I₅₀ increased by 25.8-, 10.7-, and 10.4-folds for pirimicarb, omethoate, and monocrotophos, respectively, demonstrating that GST had not been involved in the resistance of S. avenae. The enhanced esterase contributed to low level resistance to all the insecticides tested, whereas higher resistance to pirimicarb, omethoate, and monocrotophos mainly depended on AChE insensitivity. However, the AChE of the resistant strain was still sensitive to thiodicarb (1.7-fold). Thus, thiodicarb could be used as substitute for control of the resistant S. avenae in this case. Furthermore, the two different AChE genes cloned from different resistant and susceptible individuals were also compared. Two mutations, L436(336)S in Sa.Ace1 and W516(435)R in Sa.Ace2, were found consistently associated with the insensitivity of AChE. They were thought to be the possible resistance mutations, but further work is needed to confirm this hypothesis.     

MPK4甲基化调控水杨酸信号介导的拟南芥对齐整小核菌菌株SC64的抗性机制

为提升齐整小核菌Sclerotium rolfsii菌株SC64作为生物除草剂的应用潜力，解析植物对其的防御机制，通过比较6种生态型拟南芥Arabidopsis thaliana(Col-0、Ms-0、Gr-1、Se-0、Tu-0和Wa-1)被菌株SC64侵染后病理特征、茎秆木质素水平以及抗病相关基因表达的差异，筛选出抗性最高和抗性最弱的典型生态型拟南芥，通过施加0.1 mmol/L外源水杨酸来验证其在拟南芥防御菌株SC64侵染过程中对木质素代谢及病理的调节作用，并比较分析6种生态型拟南芥水杨酸上游调控通路基因MPK4启动子区域的甲基化差异。结果显示，6种生态拟南芥中Col-0的木质素化程度最高，被菌株SC64侵染时木质素代谢通路基因CAD4和PAL3表达上调最显著，与其抗病能力最强相吻合，而Ms-0的病理表型和抗病能力最弱。外源水杨酸预处理使具典型抗性的生态型拟南芥Col-0受菌株SC64侵染后发病程度加重，木质素化程度减弱，同时木质素代谢通路基因MYB46、LAC17、PAL3和CAD4被菌株SC64侵染后的表达上调程度均显著减弱。另外，6种生态型拟南芥MPK4基因启动子区域CHH...