杀菌剂抗性监测研究进展

使用现代选择性杀菌剂防治植物病害是保证农作物高产、稳产的重要措施之一,但是现代杀菌剂很容易出现抗性而导致化学防治失败．使农业生产蒙受巨大损失。本文从杀菌剂的抗性检测和治理技术等方面概述杀菌剂的抗性研究进展。     

Genetic Control of Resistance to the Piperidine Fungicide Piperalin in Ustilago maydis

Mutants of Ustilago maydis resistant to the piperidine fungicide piperalin were isolated in a mutation frequency of 2.4 × 10^–5 after UV-irradiation and selection on media containing 50gml^–1 piperalin. Genetic analysis with 15 such mutant isolates resulted in the identifications of two unlinked chromosomal loci, the U/ppl-1 locus with two allelic genes (U/ppl-1A and U/ppl-1B) and the U/ppl-2 locus. The U/ppl-2 and U/ppl-1A mutations are responsible for two levels of moderate and high resistance to piperalin (resistance factor, Rf: 54 and 135, respectively, based on effective concentration causing a 50% reduction in the growth rate, EC₅₀), while the U/ppl-1B mutation gives only a small reduction (approximately 8-fold) in piperalin sensitivity. Cross-resistance studies with other SBIs shows that the major gene (U/ppl-2 and U/ppl-1A) mutants are resistant to fenpropidin (Rf: 43 and 68), fenpropimorph (Rf: 261 and 283) and tridemorph (Rf: 9 and 10), but not to the inhibitors of C-14 demethylase (DMIs) and squalene epoxidase. The minor gene mutation U/ppl-1B codes a low-level of resistance (approximately 5—12-fold) to the above morpholine-type fungicides, but in contrast with the major gene mutations it increases 2–10 times the sensitivity to triazoles: triadimefon, triadimenol, propiconazole and flusilazole. Crosses between mutants carrying the U/ppl-genes with compatible isolates carrying the U/fpd, U/fpm or U/tdm mutations, which have been identified in previous genetic works for resistance to morpholine-type fungicides, yielded, with the exception of U/ppl-2 ×U/fpm-2 cross, a large number of recombinants with wild-type sensitivity, indicating that the mutant genes involved in these crosses, were not allelic. An additive gene effect was observed only between nonallelic minor genes U/ppl-1B and U/fpm-1B or U/tdm-1,2. Studies of the fitness of piperalin-resistant isolates showed that the reduced sensitivity of major gene mutants was not associated with changes on the phytopathogenic fitness determining characteristics, such as growth in liquid culture and pathogenicity on young corn plants. Conversely, the minor gene mutation U/ppl-1B appeared to be pleiotropic, having significantly adverse effects on the phytopathogenic fitness.     

The Abrasive Action of a Copper-based Fungicide on Four Different Nozzle Tips

Abstract

The abrasive action of a copper fungicide on four nozzles commonly used in Nigeria when spraying cocoa for black pod control was evaluated. The spray solution flocculated and the nozzles frequently blocked indicating the need for an efficient filter. The single hollow cone type nozzle and the variable spray nozzle were the most highly abraded.     

多杀菌素的作用机理及其抗药性的研究进展

多杀菌素是一类新的杀虫剂,具有高效、低毒、选择性强、对环境安全的特点。其作用机制是通过激活烟碱型乙酰胆碱受体(nAChR),使正常昆虫神经细胞去极化,也可通过抑制γ-氨基丁酸受体(GABAR)使神经细胞超极化。本文综述了多杀菌素作用机理及其抗药性的研究进展。     

Fungicide resistance inBotrytis cinerea isolates from strawberry crops in Huelva (southwestern Spain)

The severity of disease caused byBotrytis cinerea in strawberries is very high and chemical control is common practice; low residue levels of chemical products are required. Thus, it is important to be aware of the development of fungicide resistance in order to choose the best strategies of chemical control. In the present study we evaluated the response of 36B. cinerea isolates against eight different fungicides. The isolates were sampled twice, at the beginning and the end of the season, in 11 commercial strawberry fields located in the area of Huelva (Spain). In addition, two reference isolates, SAS56 and SAS405, were evaluated. The proportion of isolates resistant to benomyl was very high (86%). Resistance to dicarboximides was detected in 44% of the isolates and resistance to pyrimethanil in 25% of the isolates. Different degrees of sensitivity to captan and dichlofluanid were recorded. No resistance was found to diethofencarb plus carbendazim. http://www.phytoparasitica.org posting Sept. 18, 2002.     

Selection for Fungicide Resistance Within a Growing Season in Field Populations of Phytophthora infestans at the Center of Origin

ABSTRACT The central highlands of Mexico should provide an optimal testing ground for evaluating the potential threat of selection for resistance to fungicides in the population of Phytophthora infestans. We evaluated the hypotheses that exposure to the fungicides azoxystrobin, cymoxanil, dimethomorph, fluazinam, mancozeb, metalaxyl, and propamocarb hydrochloride would lead to (i) a shift in the sensitivity distributions (i.e., selection) and (ii) a lower genotypic diversity of the population. We compared populations from unsprayed plots with populations that had been exposed to several applications of each of the fungicides within a single field season. This study provides novel baseline data and shows that the Toluca valley P. infestans population has a wide range of sensitivities to the fungicides fluazinam, cymoxanil, dimethomorph, metalaxyl, and propamocarb. Directional selection toward resistance combined with a reduction in genetic diversity of the P. infestans population was observed only for the fungicide metalaxyl. The results obtained provide direct experimental support for continuing vigilance regarding further introductions of exotic strains of P. infestans into the United States.     

The Relationship Between Pathogen-induced Systemic Resistance (ISR) and Multigenic (horizontal) Resistance in Plants

Plants have developed mechanisms to successfully co-exist in the presence of pathogenic organisms. Some interactions between plants and pathogens are based on recognition of specific elicitor molecules from avirulent pathogen races (avr gene products), which is described in the gene-for-gene resistance theory. Another type of resistance, multigenic (horizontal) resistance, is a less well-studied phenomenon that depends upon multiple genes in the plant host. All plants possess resistance mechamisms which can be induced upon pre-treatment of plants with a variety of organisms or compounds. This general phenomenon is known as induced systemic resistance (ISR). At least in some plant species, ISR depends on the timely accumulation of multiple gene products, such as hydrolytic enzymes, peroxidases or other gene products related to plant defences. The pre-treatment of plants with an inducing organism or compound appears to incite the plant to mount an effective defense response upon subsequent encounters with pathogens, converting what would have been a compatible interaction to an incompatible one. Our studies in three plant–pathogen systems clearly document that multigenic-resistant plants constitutively express specific isozymes of hydrolytic enzymes that release cell wall elicitors, which in turn may activate other defense mechanisms. ISR induces constitutive accumulation of these and other gene products prior to challenge. ISR is known to function against multiple organisms, and there is no specificity observed in the accumulation patterns of defense-related gene products when ISR is induced. It is therefore hypothesized that the constitutive accumulation of specific isozymes of hydrolytic enzymes, or other defense related gene products, is an integral part of both multigenic resistance and the phenomenon of ISR. Further, plants in which ISR has been activated appear to move from a latent resistance state to one in which a multigenic, non-specific form of resistance is active.     

Resistance Risk Assessment of Cereal Eyespot, Tapesia Yallundae and Tapesia Acuformis, to the Anilinopyrimidine Fungicide, Cyprodinil

Eyespot pathogens, Tapesia yallundae and Tapesia acuformis, were isolated from two trial sites in the UK over several years. Both sites were treated with 2 applications per year of cyprodinil (a new anilinopyrimidine fungicide), prochloraz and a mixture of cyprodinil with prochloraz. One trial site was exposed to cyprodinil for 3 years, and the second for a total of 11 years, including 5 years before the trial was initiated. Control of eyespot and sensitivity to cyprodinil were monitored. During the first 3 years of the trial, disease control with all fungicide treatments ranged from 43% to 82%. At the site, where the trial was extended for a further 3 years, control then began to decline but no practical resistance was detected. The decline in control by both fungicides suggests that factors other than reduced sensitivity might be involved. Field isolates of both T. yallundae and T. acuformis with reduced sensitivity to cyprodinil were found predominantly in plots treated with cyprodinil. A reduction in sensitivity to cyprodinil was identified in the population from cyprodinil-treated plots in two years out of six, and in the population from mixture plots in the final year. No obvious trends could be identified and in-vivo studies showed control of most isolates with reduced sensitivity could be regained by increasing the dose to one tenth of the recommended field rate. Analysis of progeny from sexual crosses involving a sensitive isolate and a field isolate with an ED₅₀ value higher than the baseline sensitivity range indicated that a single gene controlled the reduction in sensitivity to cyprodinil in one T. yallundae isolate. There is clearly a resistance risk in eyespot to cyprodinil. The reduction in sensitivity is monogenic in inheritance and at a significant level in some isolates, but any shift in sensitivity in field populations has so far been gradual.     

Inhibition of Methionine Biosynthesis in Botrytis cinerea by the Anilinopyrimidine Fungicide Pyrimethanil

When mycelium of Botrytis cinerea was treated with low concentrations of the anilinopyrimidine fungicide pyrimethanil the total amount of free amino acids increased. Qualitative variations were also induced: alanine, glutamine, lysine, glycine, histidine, asparagine, arginine, threonine and moreover, α-aminobutyrate and β-alanine were accumulated; cyst(e)ine, valine, leucine and citrulline were reduced. When mycelium of B. cinerea was incubated with Na₂[³⁵S]O₄, pyrimethanil at 1·5 μM induced a decrease of [³⁵S]methionine and simultaneously an increase of [³⁵S]cystathionine. These data indicate that the anilinopyrimidine fungicide pyrimethanil inhibits the biosynthesis of methionine and suggest that the primary target could be the cystathionine β-lyase. © 1997 SCI.     

新型杀菌剂辛唑嘧菌胺的合成研究

为了探索杀菌剂辛唑嘧菌胺的工艺路线和合成方法,并寻找一条适用于工业化生产的路线。以丙酰乙酸甲酯和溴代正辛烷为原料,经过取代、环合、氯化、胺化4步反应合成了目标化合物辛唑嘧菌胺。反应总收率为31.2%,含量为96.8%,产物经1H NMR进行表征。该方法操作简单,条件温和,适用于工业化生产。     

杀菌剂戊唑醇的合成研究

本文介绍了杀菌剂戊唑醇的合成原理、合成方法及达到的技术指标。     

Action of pesticides on earthworms. Part II: Elimination of parathion by the earthworm Eisenia foetida (Savigny)

The earthworm, Eisenia foetida, eliminated parathion and carbofuran at first order rates when continually rinsed in water after treatment with the pesticides. This experiment was also carried out on Lumbricus rubellus for comparison. Carbofuran which is more soluble in water, was eliminated quicker than parathion. The later rate of elimination was very similar for the two species, but immediately after injection the rate was much higher in E. foetida. The metabolism of 1-ethyl¹⁴C labelled parathion and paraoxon (diethyl 4-nitrophenyl phosphate) was studied in E. foetida. The worm was able to convert parathion to paraoxon by a rather slow process although this metabolite could not be detected in the worms due to its rapid transformation to diethyl hydrogen phosphate. Indirectly, paraoxon can be postulated as a parathion metabolite because of a progressive depression of cholinesterase level observed after treatment with parathion. Small amounts of diethyl hydrogen phosphate were detected as a metabolite of parathion; this is also an indication of paraoxon formation. During the 30 h following injection of parathion, only 4.4% of the applied dose was recovered as water-soluble metabolites (2.8% in the worms and 1.6% in the sand surrounding them), while 52% was recovered as unmetabolised parathion. Because of inefficient injection, only 70-59% of the dose thought to be injected was recovered. Therefore the part of the actual applied dose that remained unmetabolised was probably even greater (88%). Five days after injection of parathion, 15 and 9.3 % of the recovered radioactivity in the surrounding sand and in the worm extracts, respectively, was identified as O,O-diethyl O-hydrogen phosphorothioate, 3.7 and 7.0% as diethyl hydrogen phosphate, 8.8 and 3.3% as O-ethyl O-4-nitrophenyl O-hydrogen phosphorothioate (desethylparathion) and/or O-4-aminophenyl O,O-diethyl phosphorothioate, while 70.3 and 80.4% was unmetabolised parathion. Paraoxon was very quickly hydrolysed to diethyl hydrogen phosphate in vivo and in vitro. The in-vitro hydrolysis was associated with a microsomal fraction and was not inhibited by ethylenediaminetetra-acetic acid or 4-(chloromercuri)benzoic acid, and incompletely by aldicarb. Cholinesterase and arylesterase were therefore excluded as enzymes responsible for the activity.     

Field Resistance to Strobilurin (Q(o)I) Fungicides in Pyricularia grisea Caused by Mutations in the Mitochondrial Cytochrome b Gene

ABSTRACT Gray leaf spot caused by Pyricularia grisea is a highly destructive disease of perennial ryegrass turf. Control of gray leaf spot is dependent on the use of preventative fungicide treatments. Strobilurin-based (Q(o)I) fungicides, which inhibit the cytochrome bc(1) respiratory complex, have proven to be very effective against gray leaf spot. However, in August 2000, disease was diagnosed in Q(o)I-treated perennial ryegrass turf on golf courses in Lexington, KY, Champaign, IL, and Bloomington, IL. To determine if resistance was due to a mutation in the fungicide target, the cytochrome b gene (CYTB) was amplified from baseline and resistant isolates. Nucleotide sequence analysis revealed an intronless coding region of 1,179 bp. Isolates that were resistant to Q(o)I fungicides possessed one of two different mutant alleles, each of which carried a single point mutation. The first mutant allele had a guanine-to-cytosine transition at nucleotide position +428, resulting in a replacement of glycine 143 by alanine (G143A). Mutant allele two exhibited a cytosine-to-adenine transversion at position +387, causing a phenylalanine-to-leucine change (F129L). Cleavable amplified polymorphic sequence analysis revealed that neither mutation was present in a collection of baseline isolates collected before Q(o)I fungicide use and indicated that suspected Q(o)I- resistant isolates found in 2001 in Indiana and Maryland possessed the F129L mutation. The Pyricularia grisea isolates possessing the G143A substitution were significantly more resistant to azoxystrobin and trifloxystrobin, in vitro, than those having F129L. DNA fingerprinting of resistant isolates revealed that the mutations occurred in just five genetic backgrounds, suggesting that field resistance to the Q(o)I fungicides in Pyricularia grisea is due to a small number of ancestral mutations.     

7种季铵盐阳离子表面活性剂对啶虫脒在防治烟粉虱的增效作用评价

采用温室盆栽法研究了7种季铵盐阳离子表面活性剂对啶虫脒在防治烟粉虱的增效作用,为阳离子助剂在杀虫剂领域的开发利用提供一定的依据。结果表明：季铵盐阳离子1227、12143、31506与啶虫脒混用,在高浓度（200mg/L）处理下增效作用明显;1427、31551、31527与啶虫脒混用可延长其持效期;31503无增效作用。     

Resistance of rose rootstocks to crown gall (Agrobacterium tumefaciens)

Various types of rose rootstocks were tested for their resistance to crown gall. The rootstock Iowa State University (ISU) 60–5 was the most resistant, followed by Brooks 48, Clarke 1957 and Welch. Rosa multiflora, R. noisettiana (Manetti) and Basye No. 3 were very susceptible. The inoculations were made with four isolates ofAgrobacterium tumefaciens (Smith et Townsend) Conn, respectively from aDahlia sp.,Rosa spp. andPrunus persica. It was found that the isolate fromDahlia was a different race to the isolates fromRosa andPrunus spp.Samenvatting Bij onderstammen van rozen, kunstmatig geïnoculeerd metAgrobacterium tumefaciens (Smith et Townsend) Conn., werden verschillen in resistentie tegen wortelknobbel gevonden. Het meest resistent was Iowa State University (ISU) 60–5, gevolgd door Brooks 48, Clarke 1957 en Welch, Zeer vatbaar warenRosa multiflora, R. noisettiana (Manetti) en Basye No. 3. De vier isolaten vanA. tumefaciens, gebruikt voor de inoculaties, waren respectievelijk afkomstig van eenDahlia sp.,Rosa spp. enPrunus persica. Het isolaat vanDahlia en de isolaten vanRosa enPrunus spp. behoorden tot twee verschillende fysiologische rassen. De vorming van tumoren was in sommige gevallen afhankelijk van de methode van inoculatie; inoculaties bij de stambasis waren meer succesvol dan in het midden van de stam.     

Resistance to bacterial wilt in tomato as discerned by spread of Pseudomonas (Burholderia) solanacearum in the stem tissues

Different criteria were compared for assessing bacterial wilt resistance in 13 tomato genotypes varying in disease susceptibility. Wilt severity and bacterial invasiveness at collar and midstem were compared in the field under cooler (March to May, 20–28°C) and warmer months (June to August, 23–29°C), which were unfavourable and favourable to wilt symptom expression, respectively. A model was proposed for determining resistance regardless of climatic conditions prevalent during field experimentation. This model was based on an estimate of bacterial invasiveness termed the colonization index. Using a qualitative imprint method we confirmed that the more resistant the genotype, the lower the bacterial colonization of the stem. The colonization index accounted both for wilted plants and for infected asymptomatic plants in which Pseudomonas solanacearum populations failed to produce wilt. The colonization index at midstem was the more useful indicator of resistance under favourable conditions. When environmental conditions were unfavourable to wilt, colonization index at collar level discerned resistant genotypes more clearly. The results formed the basis for a model for predicting the degree and stability of resistance in tomato.