Laboratory selection of resistance by the Red Flour Beetle,Tribolium castaneum (Herbst), To a carbon dioxide-enriched atmosphere

Adult populations of the red flour beetle,Tribolium castaneum (Herbst), were exposed for 40 generations to an atmosphere containing 65% CO₂, 20% O₂ and 15% N₂ at 95% RH, in order to select a strain resistant to a high carbon dioxide content (HCC) atmosphere. Selection pressure was maintained at between 50% and 70% mortality. At the 40th generation, comparison of sensitivity between the selected strain and the original non-selected strain indicated a resistance factor at the 50% mortality level (LT₅₀) of x 9.2. Throughout the selection process, log-time against probit-mortality lines remained roughly parallel and the slope remained low and similar to that of the non-selected strain until the last generation. Implications are that at a high level of selection, multiple genetic factors continued to act together even at the 40th generation to select for adaptation to survival at high CO₂ concentrations. Removal of selection pressure from a sub-population of the selected strain from the 13th to 21st generation revealed that resistance was partially retained with a decrease of 17% in the resistance factor. The modified atmosphere chosen in this study does not represent real-life situations where O₂ concentrations are reduced by flushing with CO₂, and intergranular relative humidities are generally not above 70%.     

Flufenoxuron, an acylurea insect growth regulator, alters development of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) by modulating levels of chitin, soluble protein content, and HSP70 and p34 in the larval tissues

The effect of sublethal concentrations, 0.00141% (LC₂₀), 0.00251% (LC₃₀), and 0.0036% (LC₄₀) of a dispersible formulation of an acylurea insect growth regulator, flufenoxuron (Cascade) on certain biochemical parameters in the larvae of Tribolium castaneum was investigated. When neonates were fed on diet treated with sublethal concentrations for 24 h, it was observed that at all concentrations tested, there was a significant reduction in chitin content on the 15th day of development. Total soluble protein content at LC₂₀ and LC₃₀ decreased with increasing age of the larvae. At LC₂₀ and LC₄₀ concentrations there was a progressive increase in the protein: chitin ratio as a function of increase in age of the larvae. SDS-PAGE analysis of the larval tissue extracts indicated gross quantitative changes in some of the protein bands (MW 50-97 kDa). Western blot analysis revealed significant increase in the level of HSP70 in the extracts of larvae fed on LC₃₀ treated diet, on the 7th and 10th day of development in the decreasing order. Conversely, a significant decrease in the hyper-phosphorylated form of p34^cdc2 kinase due to flufenoxuron treatment indicating modulation of cell cycle regulation was observed. Thus, sublethal concentrations of flufenoxuron alter expression of developmentally regulated proteins, HSP70 and p34^cdc2 and chitin formation in a stage-specific manner thereby resulting developmental abnormalities in T. castaneum.     

Preliminary study of the fumigant toxicity of allyl alcohol to three stored-product insects

The toxicity of allyl alcohol as a fumigant was assessed for adults ofTribolium castaneum (Herbst.),Rhyzopertha dominica (F.) andSitophilus oryzae (L.). Fumigations were carried out in 3.4–1 glass containers at concentrations of 1–15 mg/l for 3 h. The most sensitive species wasS. oryzae and the most tolerant wasT. castaneum. A dose of 15 mg/l for 3 h was lethal to all three species tested. The feasibility of proposing allyl alcohol as a candidate fumigant is discussed. Publication of the Agricultural Research Organization.     

CO2气调胁迫对药材甲能源物质的影响

为明确CO_2气调胁迫对药材甲能源物质的影响,采用不同CO_2浓度(10%、30%、50%、70%和90%)对其分别胁迫2、4、6、8 h后,测定其体内多糖、可溶性蛋白质及脂肪等能源物质的含量,并计算其利用率,探讨药材甲对气调胁迫的生理生化响应。结果表明,相较于对照,随胁迫时间的延长及CO_2浓度的升高,药材甲能源物质的含量均逐渐减少,利用率则逐渐增加。10%CO_2浓度胁迫2 h下,单头药材甲多糖、蛋白质及脂肪的含量均最高,分别为13.94、162.87和35.14μg;其利用率最低,分别为4.85%、2.04%和11.02%。90%CO_2浓度胁迫8 h下,其含量最低,分别为11.02、145.69和19.08μg;利用率最高,分别为24.80%、12.37%和51.68%。不同处理下,药材甲对3种能源物质的利用率均差异显著,为脂肪多糖可溶性蛋白质。表明CO_2浓度和胁迫时间是导致药材甲能源物质消耗的重要因素,而其对不同能源物质不同的利用率,则可能与昆虫抗气性的形成密切相关。     

Local and systemic resistance to fungal pathogens triggered by an AVR9-mediated hypersensitive response in tomato and oilseed rape carrying the Cf-9 resistance gene

Tomato and transgenic oilseed rape plants expressing the Cf-9 resistance gene develop a hypersensitive response (HR) after injection of the corresponding Avr9 gene product. It was investigated whether induction of a HR conferred resistance to different fungal pathogens in tomato and oilseed rape. Induction of an AVR9 mediated HR at the pathogen infection site delayed the development of the biotrophs Oidium lycopersicum in tomato and Erysiphe polygoni in oilseed rape, but enhanced the development of the necrotrophs Botrytis cinerea and Alternaria solani in tomato and Sclerotinia sclerotiorum in oilseed rape. Interestingly, delayed fungal disease development was observed in plant tissues surrounding the HR lesion regardless of whether a necrotrophic or biotrophic pathogen was used. In tomato, AVR9 injection induced systemic expression of PR1, PR2 and PR3 defence genes but did not induce systemic resistance to O. lycopersicum, B. cinerea or A. solani. In oilseed rape, AVR9 injection temporarily induced systemic resistance to Leptosphaeria maculans and E. polygoni, but did not induce detectable systemic expression of PR1, PR2 or Cxc750. These results give new insights into the potential uses of an induced HR to engineer disease resistance.     

麦田抗性生物型荠菜对苯磺隆的抗性机制研究

为明确抗性生物型荠菜对苯磺隆的抗性机制,分别测定了苯磺隆对抗性和敏感生物型荠菜体内乙酰乳酸合成酶(ALS)、谷胱甘肽-S-转移酶(GSTs)、超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)的影响。结果表明:离体条件下,抗性生物型荠菜体内ALS对苯磺隆的敏感性明显降低,苯磺隆对荠菜抗性和敏感生物型ALS的抑制中浓度(I50)分别为0.722 8和0.052 1 μmol/L,抗性与敏感生物型I50的比值为13.87;活体条件下,施用苯磺隆后,抗性和敏感生物型荠菜ALS活性均受到一定程度的抑制,但抗性生物型ALS活性受到抑制后能逐渐恢复,而敏感生物型则不能恢复;经苯磺隆处理后,抗性生物型GSTs相对活力明显高于敏感生物型,而抗性和敏感生物型体内POD、SOD和CAT相对活力无明显差异。研究表明,抗性生物型荠菜体内ALS对苯磺隆敏感性降低是其抗药性产生的原因之一,而GSTs对苯磺隆代谢能力的差异也可能与荠菜对苯磺隆的抗性有关。     

Studies on red light-induced resistance of broad bean to Botrytis cinerea: I. Possible production of suppressor and elicitor by germinating spores of pathogen

When detached broad bean leaves were preinoculated with virulent strain B304 of Botrytis cinerea 24 h before a challenge inoculation with strain B304, lesion formation by B304 was significantly inhibited in red light but not in the dark. In leaves that were preinoculated with avirulent strain 021 and then challenged by B304, however, lesion formation was not inhibited even under red light. Such differences in lesion formation after the challenge inoculation with an avirulent strain were also observed with lesions caused by Alternaria alternata, a nonpathogen of broad bean and by avirulent strain 021R in the presence of germination fluid from spores of strains B304 and 021R. These results suggest the possibility that virulent B. cinerea produced a suppressor involved in induced susceptibility and an elicitor involved in resistance induced by red light during spore germination.     

苹果蠹蛾对高效氯氟氰菊酯的抗性现状、机制及治理策略

苹果蠹蛾Cydia pomonella属鳞翅目卷蛾科,是全球仁果类水果种植地区最重要的果树害虫之一,也是我国重大农业入侵物种,对全球水果生产造成严重威胁。当前主要采用高效氯氟氯氰菊酯等杀虫剂对苹果蠹蛾进行防治,杀虫剂的频繁使用导致其对高效氯氟氰菊酯产生了抗性。针对苹果蠹蛾对高效氯氟氰菊酯的抗性问题,该文综述了全球苹果蠹蛾种群对高效氯氟氰菊酯的抗性现状,指出由编码细胞色素P450（cytochrome P450,P450）、羧酸酯酶（carboxylesterase,CarE）和谷胱甘肽S-转移酶（glutathione S-transferase,GST）的解毒酶基因过表达导致代谢能力增强是其最普遍的抗性机制,提出合理的抗性治理策略,并对苹果蠹蛾的绿色防控进行展望。     

Colonization of cabbage by the western black flea beetle (Phyllotreta pusilla) as affected by mulch and time of day

A 3-year study was conducted in Colorado (USA) to evaluate the response of the western black flea beetle (WBFB),Phyllotreta pusilla Horn (Coleoptera: Chrysomelidae), to different colored mulches and to determine if there were diurnal differences in the effect of mulch on their colonization on plants. The plants surrounded by aluminum mulch had the highest number of WBFB compared with the rest of the treatments in 1999 and 2000, but not in 2002; the black mulch had the lowest number of WBFB in 3 years of sampling. The highest number of WBFB were observed in the late day (4 p.m.) counting in 1999, whereas none of the treatments resulted in significant differences among early, mid, and late day counts in 2000 and in 2002. http://www.phytoparasitica.org posting May 20, 2005.     

Enhancing resistance of transgenic carrot to fungal pathogens by the expression of Pseudomonas fluorescence microbial factor 3 (MF3) gene

Microbial factor 3 (MF3) gene from a plant-growth promoting rhizobacteria Pseudomonas fluorescence was introduced into carrot genome using Agrobacterium-mediated transformation. The obtained transgenic plants expressing MF3 protein were grown in a glasshouse and evaluated for their resistance to phytopathogens. The assays were performed by the assessment of disease symptoms developing on the detached leaflets and petioles inoculated by three fungal pathogens. The results showed that transgenic plants had significantly enhanced resistance to Alternaria dauci, Alternaria radicina and Botrytis cinerea, on average, by 20–40% in comparison to the non-transformed control plants. This is the first report of enhancing plant resistance by expressing the bacterial protein homologous to the family of FK506-binding proteins.     

Metabolic resistance mechanisms to phosalone in the common pistachio psyllid, Agonoscena pistaciae (Hem.: Psyllidae)

The common pistachio psyllid, Agonoscena pistaciae, is the most damaging pest of pistachio in Iran, and is generally controlled by insecticides belonging to various classes especially, phosalone. The toxicity of phosalone in nine populations of the pest was assayed using the residual contact vial and insect-dip methods. The bioassay results showed significant discrepancy in susceptibility to phosalone among the populations. Resistance ratio of the populations to the susceptible population ranged from 3.3 to 11.3. The synergistic effects of TPP, PBO and DEM were evaluated on the susceptible and the most resistant population to determine the involvement of esterases, mixed function oxidases and glutathione S-transferases in resistance mechanisms, respectively. The level of resistance to phosalone in the resistant population was suppressed by TPP, PBO and DEM, suggesting that the resistance to phosalone is mainly caused by esterase detoxification. Biochemical enzyme assays revealed that esterase, glutathione S-transferase and cytochrome P450 monooxygenase activities in the resistant population was higher than that in the susceptible. Glutathione-S-transferases play a minor role in the resistance of the pest to phosalone.     

Application of pathogen surveys,disease nurseries and varietal resistance characteristics in an IPM approach for the control of wheat yellow rust

The present paper presents the rationale for the use of pathogen surveys, inoculated and non-inoculated disease nurseries and varietal resistance characteristics in an integrated approach to control wheat yellow rust in Denmark. The non-inoculated disease observation plots, which gave valuable information about yellow rust at the year, site and variety level, served as the primary sample source for the pathogen survey revealing pathogen virulence dynamics. This survey was also the main source for isolates of new pathotypes, a prerequisite for the assessment of the resistance characteristics of varieties and breeding lines in inoculated nurseries, and the postulation of race-specific resistance genes. A simple grouping of varieties into four categories with respect to resistance to the current yellow rust population proved robust, and this grouping was used as a determinant in a web-based decision support system for pesticide applications in cereals, Crop Protection On-line (CPO). The interplay between the different research and survey activities in the integrated pest management (IPM) approach demonstrated the need for a coherent and long-term involvement at all stages from plant breeding to the official variety approval system, extension service and research in disease epidemiology and resistance genetics.     

Resistance Reaction of Conifer Species (European Larch, Norway Spruce, Scots Pine) to Infection by Selected Necrotrophic Damping-off Pathogens

Three conifer species (European larch, Norway spruce, Scots pine) were investigated for their resistance to five damping-off pathogens (Rhizoctonia solani, Fusarium solani, F. oxysporum, F. culmorum, F. avenaceum). Inoculation of the primary roots of seedlings with these pathogens caused host cell death which did not prevent the invasive growth of these fungi; seedlings that had formed secondary and tertiary roots could overcome the infection to a significant degree. Infections with R. solani caused significant mortality to all the conifer species. In contrast, the tree species expressed different levels of resistance when challenged with the Fusarium isolates, with Norway spruce being the most resistant compared to uninoculated controls. Some of the Fusarium isolates were more pathogenic to certain hosts than others; F. oxysporum for European larch, F. avenaceum for Scots pine, F. solani for European larch; only F. culmorum was significantly pathogenic to Norway spruce. No significant differences in disease severity were observed at different soil pH (4.3–7.5). Disease progression was delayed at lower (10–15°C) rather than higher temperatures (20–25°C).     

Biochemical and molecular diagnosis of insecticide resistance conferred by esterase, MACE, kdr and super-kdr based mechanisms in Italian strains of the peach potato aphid, Myzus persicae (Sulzer)

In this paper we analysed the basis of insecticide resistance in 59 Italian strains of the peach potato aphid Myzus persicae using both molecular and biochemical assays. Our data as a whole clearly indicate that most M. persicae strains (76.3%) have high or extremely high production of an esterase enzyme which sequester and detoxify insecticides with esteric group. Kdr genotypes conferring resistance towards pyrethoids are present in 57.7% of the analysed populations. Moreover, 26.5% of the kdr positive strains possess also the M918T mutation conferring super-kdr phenotype. Strains with modified AChE (MACE) are not so numerous (27.1%), although they can be found almost everywhere in Italy. Considering all the strains analysed, both MACE and kdr phenotypes are associated with high levels of esterase activity. In Central–Southern regions, kdr and MACE resistance mechanisms resulted in linkage disequilibrium. Bioassays performed in order to evaluate the efficacy of a pyrethroid insecticide against a strain possessing a F979S mutation within its para-type sodium channel gene suggests that this amino acid substitution could affect the sodium channel responsivity to pyrethroids.     

甜菜夜蛾对氯虫苯甲酰胺抗性种群选育及鱼尼丁受体基因表达特征

为明确甜菜夜蛾Spodoptera exigua对氯虫苯甲酰胺的抗性发展及抗性种群中鱼尼丁受体(ryanodine receptor,RyR)基因的表达量变化,室内采用饲料混毒法进行甜菜夜蛾对氯虫苯甲酰胺的抗性选育,荧光定量PCR技术研究了抗性和敏感种群之间RyR基因mRNA表达量的差异。结果表明,室内选育31代后获得一个抗性倍数为105.60倍的甜菜夜蛾抗性种群,其mRNA表达量在甜菜夜蛾不同发育阶段及抗性种群和敏感种群之间均存在差异,以表达量最低的卵期作为对照,抗性种群中1龄幼虫表达量最高,是卵期的154.58倍;其次是雄性成虫,其表达量是卵期的101.51倍;2~5龄幼虫分别是卵期的59.56、35.35、72.99和19.84倍。抗性种群中1、2和4龄幼虫mRNA表达量分别是敏感种群的5.99、2.79和2.14倍,其余阶段低于敏感种群。表明甜菜夜蛾对氯虫苯甲酰胺的抗药性可能主要表现在幼虫阶段,RyR基因的表达量变化与氯虫苯甲酰胺诱导有关。     

广聚萤叶甲雄虫生殖系统特异性精液蛋白基因的筛选及表达分析

为明确广聚萤叶甲Ophraella communa雄虫精液蛋白基因在性别间的表达差异并筛选出在雄虫生殖系统特异性表达的基因,从转录组及蛋白组数据库选取13个精液蛋白基因,采用实时荧光定量PCR技术检测其在广聚萤叶甲不同性别、组织及发育阶段的表达特征。结果显示,在广聚萤叶甲雄虫13个精液蛋白基因中,有9个精液蛋白的mRNA表达水平在雄虫中更高,有5个精液蛋白基因具有专一的雄虫生殖系统特异性,分别为PGK、sucA、ENO、SDH和PLA2b基因,在雄虫生殖系统中的表达量分别较其在头部的表达量高273.01倍、401.48倍、11.21倍、64.27倍和1.86倍。PGK、sucA和ENO基因均在广聚萤叶甲老熟蛹期急剧上调表达,在性成熟成虫中的表达量最高,分别是卵期表达量的43.48倍、31.03倍和11.89倍,三者具有明显的组织和发育阶段时空特异性,推测PGK、sucA和ENO基因参与广聚萤叶甲雄虫生殖的可能性较其余基因更高。     

抗精噁唑禾草灵的日本看麦娘ACCase基因突变

为明确日本看麦娘抗性种群对精噁唑禾草灵的抗性水平及抗性产生的分子机制,采用整株水平测定法测定了日本看麦娘对精噁唑禾草灵的抗性水平,扩增和比对了日本看麦娘抗性和敏感种群间乙酰辅酶A羧化酶(acetyl-Co A carboxylase,ACCase)基因的差异。结果显示,与敏感种群AH-7相比,抗性种群AH-25对精噁唑禾草灵的抗性倍数为33.82;AH-25种群ACCase基因CT区域2 078位氨基酸发生了突变,由天冬氨酸GAT突变为甘氨酸GGT;AH-25种群对炔草酯、烯草酮和烯禾啶产生了高水平的抗性,抗性倍数分别为35.66、38.64和29.14,对高效氟吡甲禾灵产生了低水平的抗性,抗性倍数为3.04,对精喹禾灵和唑啉草酯较敏感。表明ACCase基因2 078位氨基酸的突变可能是导致精噁唑禾草灵产生高水平抗性的重要原因。