The evolutionary origins of pesticide resistance

Durable crop protection is an essential component of current and future food security. However, the effectiveness of pesticides is threatened by the evolution of resistant pathogens, weeds and insect pests. Pesticides are mostly novel synthetic compounds, and yet target species are often able to evolve resistance soon after a new compound is introduced. Therefore, pesticide resistance provides an interesting case of rapid evolution under strong selective pressures, which can be used to address fundamental questions concerning the evolutionary origins of adaptations to novel conditions. We ask: (i) whether this adaptive potential originates mainly from de novo mutations or from standing variation; (ii) which pre‐existing traits could form the basis of resistance adaptations; and (iii) whether recurrence of resistance mechanisms among species results from interbreeding and horizontal gene transfer or from independent parallel evolution. We compare and contrast the three major pesticide groups: insecticides, herbicides and fungicides. Whilst resistance to these three agrochemical classes is to some extent united by the common evolutionary forces at play, there are also important differences. Fungicide resistance appears to evolve, in most cases, by de novo point mutations in the target‐site encoding genes; herbicide resistance often evolves through selection of polygenic metabolic resistance from standing variation; and insecticide resistance evolves through a combination of standing variation and de novo mutations in the target site or major metabolic resistance genes. This has practical implications for resistance risk assessment and management, and lessons learnt from pesticide resistance should be applied in the deployment of novel, non‐chemical pest‐control methods.     

Sequence diversity in the large subunit of RNA polymerase I contributes to Mefenoxam insensitivity in Phytophthora infestans

Phenylamide fungicides have been widely used for the control of oomycete‐incited plant diseases for over 30 years. Insensitivity to this chemical class of fungicide was recorded early in its usage history, but the precise protein(s) conditioning insensitivity has proven difficult to determine. To determine the genetic basis of insensitivity and to inform strategies for the cloning of the gene(s) responsible, genetic crosses were established between Mefenoxam sensitive and intermediate insensitive isolates of Phytophthora infestans, the potato late blight pathogen. F1 progeny showed the expected semi‐dominant phenotypes for Mefenoxam insensitivity and suggested the involvement of multiple loci, complicating the positional cloning of the gene(s) conditioning insensitivity to Mefenoxam. Instead, a candidate gene strategy was used, based on previous observations that the primary effect of phenylamide compounds is to inhibit ribosomal RNA synthesis. The subunits of RNA polymerase I (RNApolI) were sequenced from sensitive and insensitive isolates and F1 progeny. Single nucleotide polymorphisms (SNPs) specific to insensitive field isolates were identified in the gene encoding the large subunit of RNApolI. In a survey of field isolates, SNP T1145A (Y382F) showed an 86% association with Mefenoxam insensitivity. Isolates not showing this association belonged predominantly to one P. infestans genotype. The transfer of the ‘insensitive’ allele of RPA190 to a sensitive isolate yielded transgenic lines that were insensitive to Mefenoxam. These results demonstrate that sequence variation in RPA190 contributes to insensitivity to Mefenoxam in P. infestans.     

Tris(triazolyl)borate ligands of intermediate steric bulk for the synthesis of biomimetic structures with hydrogen bonding and solubility in hydrophilic solvents

Tris(triazolyl)borate ligands (Ttz) of intermediate steric bulk were synthesized to investigate their potential for hydrogen bonding and improved solubility in hydrophilic solvents as applied to biomimetic chemistry. The crystal structure of 3-phenyl-5-methyl-1,2,4-triazole (Htz^Ph,Me) revealed hydrogen bonding and π stacking interactions. The new ligand salt, potassium tris(3-phenyl-5-methyl-1,2,4-triazolyl)borate (KTtz^Ph,Me) was synthesized as the first example of a Ttz ligand of intermediate steric bulk. Metathesis between KTtz^Ph,Me and NaCl followed by recrystallization produced [NaTtz^Ph,Me] · 6CH₃OH in which the geometry around the sodium is octahedral with an unusual N₃O₃ donor set; this structure also shows that a hydrogen bonding network is formed by methanol molecules and triazole nitrogens. (Ttz^Ph,Me)ZnCl was synthesized and characterized crystallographically as [(Ttz^Ph,Me)ZnCl] · 0.5CH₃OH in which the zinc is tetrahedral and the triazole rings are within hydrogen bonding distance of CH₃OH. All of these new compounds are methanol soluble to varying degrees and Htz^Ph,Me and KTtz^Ph,Me are soluble in methanol/water mixtures.     

Synthesis,Characterization, Antimicrobial Activity,and Docking Studies of New Triazolic Tripodal Ligands

The synthesis and characterization of new N‐donor bitriazolic tripods were reported. The in vitro antibacterial and antifungal activities of these products were screened against fungal strain (Candida pelliculosa) and against four bacterial strains (Micrococcus luteus, Bacillus subtilis, Listeria innocua, and Escherichia coli). Biological data revealed the effect of the chemical structure on antimicrobial activity. Molecular docking studies of some compounds showed that they could act as inhibitors for the biotin carboxylase enzyme.     

Discovery of triazole-based uracil derivatives bearing amide moieties as novel dipeptidyl peptidase-IV inhibitors

Dipeptidyl peptidase-IV (DPP-4) is a validated target for T2DM treatment. We previously reported a novel series of triazole-based uracil derivatives bearing aliphatic carboxylic acids with potent DPP-4 inhibitory activities in vitro, but these compounds showed poor hypoglycemic effects in vivo. Herein we further optimized the triazole moiety by amidation of the carboxylic acid to improve in vivo activities. Two series of compounds 3a-f and 4a-g were designed and synthesized. By screening in DPP-4, compound 4c was identified as a potent DPP-4 inhibitor with the IC₅₀ value of 28.62 nM. Docking study revealed compound 4c has a favorable binding mode and interpreted the SAR of these analogs. DPP-8 and DPP-9 tests indicated compound 4c had excellent selectivity over DPP-8 and DPP-9. Further in vivo evaluations revealed that compound 4c showed more potent hypoglycemic activity than its corresponding carboxylic acid in ICR mice and dose-dependently reduced glucose levels in type 2 diabetic C57BL/6 mice. The overall results have shown that compound 4c could be a promising lead for further development of novel DPP-4 agents treating T2DM.     

新型杀菌剂啶氧菌酯对香蕉叶斑病的防治效果

【目的】香蕉叶斑病是香蕉产业的重要病害,化学防治仍然是当前最为有效的防治手段。于2015—2016年连续2年开展香蕉叶斑病的田间化学防治试验,为生产上推广应用新型杀菌剂啶氧菌酯提供依据。【方法】试验设22.5%啶氧菌酯悬浮剂125、150和187.5 mg·kg~(-1),对照药剂250 g·L-1吡唑醚菌酯乳油125 mg·kg~(-1),以及空白对照共5个处理,3次药后第12或13天调查正常叶数、病叶数及病级,计算平均病指及平均防效。【结果】22.5%啶氧菌酯悬浮剂(有效成分用量125、150和187.5 mg·kg~(-1))2015年的防治效果分别为64.70%、68.16%和71.29%,2016年防治效果分别为68.44%、72.36%和76.29%。此外,在试验期间香蕉嫩叶未见药害现象,叶片生长均正常。【结论】22.5%啶氧菌酯悬浮剂是防治香蕉叶斑病的优良药剂,对香蕉比较安全,值得在香蕉产区推广应用。     

Function of the genetic element ‘Mona’ associated with fungicide resistance in Monilinia fructicola

The genetic element ‘Mona’ has been shown previously to be associated with resistance to demethylation inhibitors (DMIs) in Monilinia fructicola. In this study, the promoter activity of the ‘Mona’ element was demonstrated genetically and the activity was narrowed down to a 20‐bp active region through a series of deletions. ‘Mona’ knockout transformants (ΔMona) were generated from DMI‐resistant isolate Bmpc7, and EC₅₀ values and expression of the MfCYP51 gene were found to be reduced in transformants compared with the parental isolate. When the ‘Mona’ element was inserted into the upstream region of the MfCYP51 gene of the DMI‐sensitive isolate HG3, the EC₅₀ values and expression of the MfCYP51 gene increased in the transformants compared with the parental sensitive isolate. These results indicate that the ‘Mona’ element determines the DMI fungicide resistance through the up‐regulation of the expression of the downstream MfCYP51 gene. No fitness penalty was observed in knockout and insertion transformants, i.e. transformants showed similar mycelial growth rate, sporulation and ability to cause lesions on fruit compared with their parental isolates, suggesting that the ‘Mona’ element does not affect basal life activities.     

Integration of soil solarization with chemical, biological and cultural control for the management of soilborne diseases of vegetables

The long-term effectiveness of soil solarization integrated with (integration of pest management [IPM]) a biological control agent (Trichoderma virens), chemical fungicide (pentachloronitrobenzene [PCNB]), organic amendment (chicken litter) or physical method (black agriplastic mulch) to reduce southern blight (Sclerotium rolfsii) and southern root-knot diseases (Meloidogyne incognita) were evaluated on vegetable production. Results showed that the long-term effectiveness of IPM plus soil solarization reduced soilborne diseases of vegetables more than two years following the termination of solarization. These disease management strategies in 1991 and 1992, following soil solarization in 1990, reduced the numbers of sclerotia in the soil, and the number of plants killed by southern blight and root-knot of tomatoes, compared to nonsolarized bare soil treatment. The integration of a reduced dosage level of PCNB or T. virens in field plots, reduced southern blight of tomatoes by 100% and 71%, respectively, in solarized soil, compared to nonsolarized bare soil two years following soil solarization. PCNB effectively controlled southern blight in nonsolarized bare soil both years. All solarized treatments, except PCNB plus solarized soil increased tomato yields compared to nonsolarized bare soil plots. In the second study (1992) following soil solarization in 1991, the effectiveness of solarized bare soil, and nonsolarized bare soil mulched with black agriplastic film, with or without Reemay spunbounded polyester row cover, were effective in reducing root-knot of tomatoes as indicated by the root-knot gall index. Following a one year fallow period in 1994 three years following soil solarization, the root-knot gall index for severity of tomato roots grown in solarized bare soil, nonsolarized bare soil, black agriplastic mulched bare nonsolarized soil and black agriplastic mulched solarized bare soil, were 1.0, 3.0, 3.0 and 2.0, respectively, on a 0–5 scale, where 0=0% and 5=100% root-knot galled. In the third study 1992 and 1993, different dosage levels of chicken litter were used to amend soil artificially infested with sclerotia of S. rolfsii at different depths following solarization, decreased the number of viable sclerotia by 85–100%. All solarized treatments and nonsolarized bare soil amended with 18.8 MT/ha of chicken litter, were effective in controlling southern root-knot damage, and postharvest storage root rots of sweetpotato storage roots (Fusarium root rot [Fusarium solani] and Java black rot [Diplodia tubericola]). Our study showed that all soil solarization treatments, and soils amended with chicken litter, stimulated a shift in the soil microbial population dynamics. Rhizobacteria of Bacillus spp. and fluorescent pseudomonads increased significantly in the rhizosphere, rhizoplane, and interior root tissues of tomatoes and sweetpoatoes, grown in solarized soil compared to nonsolarized soil. These microorganisms may have contributed to the increased growth response of vegetables and some were probably suppressive to soilborne diseases     

Effects of biocides and rotation breaks on soil organisms associated with the poor early growth of sugarcane in continuous monoculture

Glasshouse and field experiments were conducted to determine the effects of biocides and rotation breaks on deleterious soil organisms associated with the poor early growth and subsequent yield decline of sugarcane grown in continuous monoculture. Fumigation of a soil that had been under sugarcane monoculture with minimal breaks for more than 30 years markedly improved the health and growth of the sugarcane sett and shoot root systems, increased the growth of the primary shoot and stimulated more and larger secondary shoots. It also reduced populations of culturable fungi in the rhizosphere of the sett roots and reduced colonization of the sett and shoot roots by lesion nematode (Pratylenchus zeae). Exposure of the developing sett root system for 14 days to mono-cultured sugarcane soil was sufficient to significantly retard subsequent plant growth. In field experiments, fungicide and nematicide (mancozeb + aldicarb), when applied together to land under sugarcane monoculture, was as effective as fumigation in improving early sugarcane growth and increasing sugarcane yields. Rotation breaks (alternate crops, sown pasture, bare fallow) that were in place for 54 months, increased sugarcane establishment and increased sugarcane yields to levels similar to that obtained following fumigation of land under sugarcane monoculture. Fumigation of land that had been under the rotation breaks gave plant growth responses that were in addition to that achieved by the breaks alone. A mancozeb + aldicarb treatment was as effective as fumigation in increasing sugarcane yields after a bare fallow break but accounted for only a portion of the fumigation response following the crop and pasture breaks. Improved plant nutrition may be a factor in the fumigation response following the crop and pasture breaks. Plant growth responses to fumigation and the manocozeb + aldicarb treatments that were manifested in final sugarcane yields (after one years growth) were evident as plant growth responses (sett root, shoot root and primary shoot dry weight) measured 54 days after planting. The experimental results support the concept that when sugarcane is grown as a monoculture, deleterious fungi and nematodes retard plant establishment and early plant growth and that this leads to reduced sugarcane yields.