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Co-CRISPR: A valuable toolkit for mutation enrichment in the gene editing of Spodoptera frugiperda

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insect Science 2023年第3期30卷 625-636页

作者： Wei-Kang Han Feng-Xian Tang Hao-Li Gao Yan Wang Na Yu Jian-Jun Jiang Ze-Wen Liu key laboratory of Integrated Management of crop diseases and pests(Ministry of Education) College of Plant ProtectionNanjing Agricultural UniversityNanjingChina Plant Protection Research Institute Guangxi Academy of Agricultural ScienceGuangxi Key Laboratory of Biology for Crop Diseases and Insect PestsNanningChina

The CRISPR/Cas9 system has been successfully applied in dozens of diverse species;although the screening of successful CRISPR/Cas9 editing events remains particularly laborious,especially for those that occur at relatively low ***,a co-CRISPR strategy was proved to enrich the desired CRISPR ***,the co-CRISPR strategy was developed in the Fall armyworm,Spodoptera frugiperda,with kynurenine 3-monooxygenase gene(kmo)as a *** kmo mosaics induced by single-guide RNAs(sgRNAs)/Cas9 displayed the darker green color phenotype in larvae,compared with wild type(brown),and mosaic-eye adults were significantly acquired from the mosaic larvae *** the kmo knockout strain,no significant difference was observed in larval development and adult *** 2(ace2)and Wntl were selected as target genes to construct the co-CRISPR strategy using kmo *** coinjection of kmo and ace2 sgRNAs,the mutant efficiency of ace2 was significantly increased in the kmo mosaic(larvae or adults)***,more malformed pupae with Wntl mutations were observed in the darker green larvae *** together,these results demonstrated that kmo was a suitable visible marker gene for the application and extension of co-CRISPR strategy in Fall *** darker green color in larvae or mosaic-eye in adults from kmo knockout as a marker,the mutant efficiency of a target gene could be enriched in a Fall armyworm group consisting of marked *** co-CRISPR strategy is helpful for gene function studies by the knockout technique with no or lethal phenotypes.

关键词： co-CRISPR kmo mutation enrichment Spodoptera frugiperda Wnt1

Synthesis and insecticidal Activity of Novel Camptothecin Derivatives Containing Analogs of Chrysanthemic Acid Moieties

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Journal of Integrative Agriculture 2014年第6期13卷 1320-1330页

作者： DENG Li ZHANG Lan CAO Li-dong XIE Ru-liang ZHANG Yan-ning HE Wei-zhi JIANG Hong-yun Hong-yun State key laboratory for biology of Plant diseases and insect pests Institute of Plant Protection Chinese Academy of Agricultural Sciences State key laboratory for biology of Plant diseases and insect pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

Creating high-efifcient and environment-friendly pesticides is very important to produce the pollution free agriculture food and maintain the balance of the survival environmental of the human being. According to reports, camptothecin （CPT） and its derivatives are now being explored as a class of botanical insecticide in agriculture due to its novel mode of action. In order to improve the insecticidal activity of CPT, ten novel camptothecin （1） and 10-hydroxycamptothecin （2） derivatives （1a, 1b, 1c, 1d, 1e;2a, 2b, 2c, 2d, 2e） were designed and synthesized via esteriifcation with analogs of chrysanthemic acid, which have outstanding insecticidal activity. The results showed that compound 2a exhibited potent antifeeding effect and the best contact toxicity among the target compounds against the third-instar larvae of beet armyworm, Spodoptera exigua H＆#252;bner. Compound 2a was also found to be the most effective cytotoxic compound to the tested insect cell lines, IOZCAS-Spex-II, which were established from the fat bodies of S. exigua. It was proposed that the 10-hydroxyl group in the camptothecin derivatives is a key factor for the antifeeding activity of a compound. The nature of the substituents was considered the major factor in determining the insecticidal activity of these compounds.

关键词： camptothecin analogs of chrysanthemic acid synthesis antifeeding activity contact toxicity cytotoxicity

Alanine-substituted mutant on Gly^(373) and Asn^(375) of Cry1Ai-h-loop 2 causes reduction in both toxicity and binding against Helicoverpa armigera

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Journal of Integrative Agriculture 2019年第5期18卷 1064-1071页

作者： LIU Yu-xiao ZHOU Zi-shan LIANG Ge-mei SONG Fu-ping ZHANG Jie State key laboratory for biology of Plant diseases and insect pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

Cry1Ai-h-loop 2 is a mutant of Cry1Ai constructed by exchanging loop 2 from Cry1Ah protein and shows insecticidal activity against Helicoverpa armigera. The toxicity of Cry1 Ai-h-loop 2, in contrast to the very low toxicity of Cry1Ai, is closely associated with the eleven residues in the loop 2 region. To characterize the key sites of loop 2 in Cry1Ai-h-loop 2, alaninesubstituted mutants were generated. The toxicity of these mutants against H. armigera indicated that dual-mutant on Gly373 and Asn375 caused a significant decrease in toxic activity. ELISA binding and competition binding assays demonstrated that the reduction of toxicity in the mutant of interest was correlated with decreased binding affinity.

关键词： Bacillus thuringiensis Cry1Ai Domain Ⅱ-loop2 Helicoverpa armigera binding affinity

Effects of intercropping vines with tobacco and root extracts of tobacco on grape phylloxera, Daktulosphaira vitifoliae Fitch

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Journal of Integrative Agriculture 2015年第7期14卷 1367-1375页

作者： WANG Zhong-yue SU Jun-ping LIU Wei-wei GUO Yu-yuan State key laboratory for biology of Plant diseases and insect pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

The effects of grape-tobacco intercropping patterns on populations of grape phylloxera, Daktulosphaira vitifoliae Fitch, as well as on the growth and development of the infested vines were evaluated in the field and the impact of an aqueous tobacco root extract on grape phylloxera was evaluated using a laboratory bioassay. The aqueous tobacco root extract exhibited biological activity against this pest. The egg mortality, nymph mortality, development period, life span and female fecundity were significantly affected. In the field trial, grape phylloxera populations were clearly lower as compared to the monoculture pattern. However, the rates of newly developed roots and newly infested grape roots were significantly higher and lower, in intercropping patterns than in the vine monoculture, respectively. The grape phylloxera population number on the grape roots decreased each year, and the vine trees gradually renewed upon continuous intercropping with tobacco over three years. These results confirmed that intercropping grapes with tobacco can effectively control grape phylloxera in an infested vineyard. The results also indicated that additional crops could be intercropped with grapes and are effective against grape phylloxera, which should be explored as an integrated approach for controlling the pest.

关键词： grape phylloxera tobacco intercropping developmental duration secondary metabolites control method

Comparing Immature Development and Life History Traits in Two Coexisting Host-Feeding Parasitoids, Diglyphus isaea and Neochrysocharis formosa(Hymenoptera: Eulophidae)

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Journal of Integrative Agriculture 2014年第12期13卷 2690-2700页

作者： ZHANG Yi-bo LU Shu-long LIU Wan-xue WANG Wen-xia WANG Wei WAN Fang-hao State key laboratory for biology of Plant diseases and insect pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

Coexisting natural enemies that share a common host resource in the same guild usually exhibit variation in their life history traits, due to their need to share a similar ecological niche. In this study, we compared the immature development times and adult life history traits of two coexisting, host-feeding parasitoids, Diglyphus isaea Walker and Neochrysocharis formosa Westwood （Hymenoptera：Eulophidae）, of which both attack larvae of the same agromyzid leafminers. These two species are both synovigenic, idiobiont parasitoids, whose adults consume host lfuids （“host feeding”） and lay anhydropic eggs. Of the two, D. isaea has a larger body but little or no initial egg load, and engages in similar lifetime host-feeding events. However, it achieves higher fecundity, longer adult longevity, and higher host suppression ability than N. formosa, which has a smaller body and higher initial egg load. Although D. isaea engages in similar lifetime host-feeding events with N. formosa, all of its gains in life history traits per host-feeding event of D. isaea were larger than those of N. formosa. The age-speciifc fecundity and host mortality curves of N. formosa were more skewed in early life than those of D. isaea. In addition, the ovigeny index of N. formosa was negatively correlated to body size. Our results conifrmed that two coexisting parasitoids, which share the same host resource, show different immature development patterns and life history traits, suggesting that different resource allocation mode could be a general rule of coexisting species sharing the same habitat or host.

关键词： parasitoid host feeding initial egg load ovigeny index fecundity longevity

More stressful event does not always depress subsequent life performance

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Journal of Integrative Agriculture 2019年第10期18卷 2321-2329页

作者： CHEN Ying-ying ZHANG Wei MA Gang MA Chun-sen State key laboratory for biology of Plant diseases and insect pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

Climate change has led to a substantial increase in intensity and duration of heat waves *** the ecological impacts of hot events should incorporate both immediate and potential carry-over effects in different intensities of heat waves. Previous studies suggested that higher heat dose in early life stage of insect generally decreased immediate survival and depressed adult reproduction through carry-over effects, or unchanged adult performance through recovery effects. However, our previous study showed a different pattern, in which longer heat exposures in larval stage did not always decrease but sometimes increase the subsequent adult maturation success in the diamondback moth. We speculated that it might be another important pattern in the carry-over effects vs. heat dose, and conducted experiments using a global pest, Plutella xylostella. Our present results suggested that heat exposures in early life stage reduced the immediate survival and produced general declines with significant zigzag fluctuating patterns in subsequent body size and reproduction as exposure durations increased. The similar patterns were also validated in other insect taxa and other stresses by reanalyzing the experiment data from literatures. The finding highlights the importance for differentiating the biological effects and consequences of changes in heat dose at fine scales;daily exposure hours of a hot day should be considered to predict population dynamic under climate change.

关键词： climate change Plutella xylostella extreme temperature reproduction carry-over effect

Effects of Droplet Distribution on insecticide Toxicity to Asian Corn Borers(Ostrinia furnaealis) and Spiders(Xysticus ephippiatus)

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Journal of Integrative Agriculture 2014年第1期13卷 124-133页

作者： YANG Dai-bin ZHANG Lin-na YAN Xiao-jing WANG Zhen-ying YUAN Hui-zhu State key laboratory for biology of Plant diseases and insect pests/Institute of Plant Protection Chinese Academy of Agricultural Sciences

Distribution of horizontal boom produced droplets downwards into maize canopies at flowering period and its effects on the efficacies of emamectin benzoate, lambda-cyhalothrin and chlorantraniliprole against the second generation of Asian corn borer （ACB） larvae and their toxicity to spiders were studied. When insecticides were sprayed downwards into the maize canopies, randomly filtering out droplets by upper leaves led to great variations of droplet coverage and density within the canopies. Consequently, the efficacies of lambda-cyhalothrin and emamectin benzoate against ACB larvae were decreased because of randomly filtering out droplets by upper leaves. But field investigation showed that lambda-cyhalothrin was extremely toxic to hunting spiders, Xysticus ephippiatus, and not suitable to IPM programs in regulation of the second generation of ACB. Therefore, randomly filtering out droplets by upper leaves decreased lambda-cyhalothrin＇s efficacy against ACB larvae, but did little to decrease its toxicity to X. ephippiatus. Amamectin benzoate can reduce the populations of X. ephippiatus by 58.1-61.4%, but the populations can recover at the end of the experiment. Chlorantraniliprole was relatively safe to X. ephippiatus. It only reduced the populations of X. ephippiatus by 22.3-33.0%, and the populations can totally recover 9 d after application.

关键词： droplet distribution insecticide toxicity Asian corn borer spider

Performance of Three Adelphocoris spp.(Hemiptera: Miridae) on Flowering and Non-flowering Cotton and Alfalfa

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Journal of Integrative Agriculture 2014年第8期13卷 1727-1735页

作者： GAO Zhen PAN Hong-sheng LIU Bing LU Yan-hui LIANG Ge-mei State key laboratory for biology of Plant diseases and insect pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

The genus Adelphocoris（Hemiptera： Miridae） is a group of key insect pests in cotton fields in China that includes three dominant species： A. suturalis（Jakovlev）, A. lineolatus（Goeze） and A. fasciaticollis（Reuter）. Previous field surveys have found that adults of these Adelphocoris species usually move onto specific host plants when the plants enter the flowering stage. To determine the potential trade-offs for this host-plant preference behavior, the performance of these three Adelphocoris spp. on flowering and non-flowering cotton and alfalfa were compared in the laboratory. The results showed that Adelphocoris spp. had significantly higher nymphal developmental and survival rates, along with increased adult longevity and fecundity on flowering cotton and alfalfa than on non-flowering plants of either species. In addition, compared with cotton plants, alfalfa generally promoted better performance of these three Adelphocoris species, especially for A. lineolatus. Simple correlation analysis indicated that female adult longevity was positively correlated to male adult longevity and female fecundity, and female fecundity was positively correlated to nymphal development and survival rates. This study demonstrated a positive correlation between adult preference and offspring/adult performance for all three Adelphocoris species, with no evidence of any trade-offs for this preference for flowering host plants. These findings support the hypothesis that hemimetabolous insects such as mirid bugs have positive adult preference-adult/nymphal performance relationships, which is likely due to the similar feeding habits and nutritional requirements of adults and nymphs.

关键词： Adelphocoris lineolatus Adelphocoris suturalis Adelphocoris fasciaticollis preference-performance relationship cotton alfalfa flowering stage

Relationship between copulation and cold hardiness in Ophraella communa(Coleoptera: Chrysomelidae)

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Journal of Integrative Agriculture 2019年第4期18卷 900-906页

作者： ZHAO Chen-chen YUE Lei WANG Yao GUO Jian-ying ZHOU Zhong-shi WAN Fang-hao State key laboratory for biology of Plant diseases and insect pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

Ophraella communa(Coleoptera: Chrysomelidae), the ragweed leaf beetle, is a biological control agent of the invasive common ragweed, Ambrosia artemisiifolia(Asterales: Asteraceae). Adults can survive cold conditions that occur during winter. The adults mate before entering overwintering. Understanding the connection between copulation and overwintering will be useful for determining O. communa seasonality. Determining the relationship between overwintering and copulation required comparison of mated and unmated beetles at mean lethal temperature(LTemp_(50)) exposures for 2 h. Cold-related physiological indices, including the water ratio, super cooling point(SCP), cryoprotectant levels, and energy reserve levels, were also measured. Mating treatment decreased the LTemp_(50) of both sexes by reducing their mean SCP and water ratios. Although the changes of cryoprotectant levels in mated adults were not precisely consistent in between the genders, they increased greatly in both males and females. Body sugar may play a role in copulation and may also elevate cold hardiness in O. communa.

关键词： Ophraella communa mating status cold tolerance hibernation super cooling point energy reserves