Involvement of esterases in diazinon resistance and biphasic effects of piperonyl butoxide on diazinon toxicity to Haematobia irritans irritans (Diptera: Muscidae)

Resistance to insecticides remains a major problem for the successful control of the horn fly, Haematobia irritans irritans (L.), one of the most important pests of cattle in many countries including the United States. The organophosphate (OP) insecticide diazinon has been used to control pyrethroid-resistant populations of the horn fly. There are only a few reported cases of horn fly resistance to diazinon in the United States and Mexico. Piperonyl butoxide (PBO) has been used successfully as a synergist of pyrethroid insecticides to control horn flies. PBO-synergized diazinon products are also available for horn fly control in the United States, although PBO is known to inhibit the bio-activation of certain OP insecticides including diazinon. A study was conducted to evaluate the effect of PBO on diazinon toxicity to horn flies using a filter paper bioassay technique. These bioassays in both the susceptible and diazinon-resistant horn fly strains revealed a biphasic effect of PBO on diazinon toxicity to horn flies. PBO inhibited diazinon toxicity when the PBO concentration used was high (5%), and no effect was observed when PBO concentration was intermediate (2%). However, at low concentrations (1% and lower), PBO significantly synergized diazinon toxicity. We demonstrated that enhanced esterase activity was associated with survivability of horn flies exposed to diazinon alone. PBO has been shown to inhibit esterase activity in other insect species. However, results of biochemical assays with esterases from this study suggest that PBO did not have significant effect on the overall esterase activity in the horn fly. The observed synergistic effect of PBO at lower concentrations on diazinon toxicity to horn flies could not be explained by reduced esterase activity due to PBO inhibition. It is likely that PBO synergized diazinon toxicity at lower concentrations by facilitating penetration of diazinon through the cuticle and/or inhibiting the oxidative detoxification of diazinon, and reduced diazinon toxicity at high PBO concentration by inhibiting the bio-activation of diazinon.     

陶赛特羊和小尾寒羊在青海湟源地区的生态适应性分析

为评定引入青海省湟源县良种肉羊繁育基地的陶赛特羊和小尾寒羊两个绵羊品种的生态适应性,对其18个生理生化指标进行了系统测定分析,结果表明：①引入的陶赛特羊和小尾寒羊两个绵羊品种大多数生理生化指标在品种内或不同品种间的不同性别、不同年龄个体间以及同性别、同年龄个体间差异不显著（P〉0.05）。②两绵羊品种所测血浆CO2结合力远超出本地藏绵羊水平;体温（T）、呼吸频率（R）、脉搏（P）和瘤胃蠕动频率以及白细胞数（WBC）、红细胞数（RBC）、血红蛋白含量（HGB）的测定结果均大致在本地藏绵羊的各生理指标测定范围之内。③与已引入青海牧区的小尾寒羊大多数生理生化指标相比,本次引进的小尾寒羊在经过一段时间的适应性饲养后,其大多数生理生化指标已与其基本接近或一致;同时,引入的陶赛特羊和小尾寒羊与原产地相应品种在基础生理指标上相比较接近。     

Molecular characterization of Miraflores peach variety and relatives using SSRs

Some traditional peach varieties, originated from the region of Aragón (Spain), were analysed by SSRs (simple sequence repeats). The aim of this research was to characterize 19 clones related to Miraflores variety, with unknown pedigrees, to assess their genetic diversity and to elucidate their possible relationships with 10 traditional peach varieties. Twenty SSR primer pairs with high levels of polymorphism, which have been previously developed for peach, were used in this study. A total of 46 alleles were obtained for all the microsatellites studied, ranging from one to six alleles per locus, with a mean value of 2.3 alleles per locus. Fourteen SSRs were polymorphic in the set of varieties studied and permitted to distinguish 16 different genotypes out of the 30 initially studied, although fourteen ‘Miraflores’ clones showed identical gel profiles. The genetic distance matrix was used to construct neighbor joining cluster and to perform principal coordinate analysis which allowed the arrangement of all the genotypes according to their genetic relationships. The genetic relationships among these traditional peach varieties, and in particular among ‘Miraflores’ clones are discussed. The obtained results confirm that microsatellite markers are very useful for these purpose.     

棉苜间作棉田天敌群落结构与动态及其对棉蚜的控制效应

采取不同的棉苜间作方式,研究了棉苜间作棉田天敌群落结构及种群动态规律、棉蚜与天敌的消长动态,以及棉苜间作对棉田蚜虫的控制效应。与常规单种棉田相比,间作苜蓿棉田内的瓢虫、蜘蛛、草蛉种群数量大幅度增长,尤以每隔1膜间作75cm苜蓿带处理区为甚,分别增长了318.0%、120.9%和79.6%。间作或邻作苜蓿的棉田天敌群落的物种丰富度、丰富度指数及多样性指数均高于常规单种棉田,表明间作和邻作苜蓿带并适时刈割,可以提高棉田内天敌群落的丰富度和多样性。刈割苜蓿带对棉田棉蚜及瓢虫类、蜘蛛类和草蛉类天敌的数量动态影响较大。在棉蚜上升初期刈割苜蓿带,使棉田内瓢虫类、蜘蛛类和草蛉类天敌数量急剧上升,棉蚜数量大幅下降,从而有效地控制了棉田棉蚜的暴发。     

Some compatible Rhizobium leguminosarum strains in peas decrease infections when parasitised by Orobanche crenata

Orobanche crenata (crenate broomrape) produces serious damage to many legume crops and particularly becomes a limiting factor for pea production in the Mediterranean basin. Nodulation effects on pea–broomrape relationships were studied using the commercial pea cultivar Douce de Provence and different Rhizobium strains using pot and Petri dish experiments. First, the benefit of bacterial inoculation on plant growth and efficiency of N incorporation were demonstrated for two isolates, P.SOM and P.1236. These isolates did not influence parasite germination induced by the artificial stimulant, GR24. In contrast, pea root inoculation with P.SOM and P.1236 isolates led to a reduced root infection by O. crenata , resulting from a lower Orobanche germination rate close to pea roots and a limited capacity of the parasitic seedlings to develop tubercles. Broomrape necrosis was observed both before and after parasite attachment to inoculated pea roots. Concomitantly, reduction in infection was accompanied by enhanced peroxidase activity and constantly high phenylalanine ammonia lyase activity in pea roots. These data suggest the involvement of these enzymes in pea resistance to crenate broomrape induced by the compatible rhizobia. Management of Orobanche via crop selection based on these enzyme systems is a viable option.     

Status of black grass (Alopecurus myosuroides) resistance to acetyl-coenzyme A carboxylase inhibitors in France

We assessed the contributions of target site‐ and non‐target site‐based resistance to herbicides inhibiting acetyl‐coenzyme A carboxylase (ACC) in Alopecurus myosuroides (black grass). A total of 243 A. myosuroides populations collected across France were analysed using herbicide sensitivity bioassay (24 300 seedlings analysed) and ACC genotyping (13 188 seedlings analysed). Seedlings resistant to at least one ACC‐inhibiting herbicide were detected in 99.2% of the populations. Mutant, resistant ACC allele(s) were detected in 56.8% of the populations. Among the five resistant ACC alleles known in A. myosuroides, alleles containing an isoleucine‐to‐leucine substitution at codon 1781 were predominant (59.5% of the plants containing resistant ACC alleles). Comparison of the results from herbicide sensitivity bioassays with genotyping indicated that more than 75% of the plants resistant to ACC‐inhibiting herbicides in France would be resistant via increased herbicide metabolism. Analysis of herbicide application records suggested that in 15.9% of the populations studied, metabolism‐based resistance to ACC‐inhibiting herbicides was mostly selected for by herbicides with other modes of action. Our study revealed the importance of non‐target site‐based resistance in A. myosuroides. Using herbicides with alternative modes of action to control populations resistant to ACC‐inhibiting herbicides, the recommended management approach, may thus be jeopardised by the widespread occurrence of metabolism‐based resistance mechanisms conferring broad‐spectrum cross‐resistance.     

Quantification of airborne spores of <Emphasis Type="Italic">Monilinia fructicola</Emphasis> in stone fruit orchards of California using real-time PCR

The fungal pathogen Monilinia fructicola causes blossom blight and fruit brown rot of stone fruits in California. In this study, spore densities in the air were monitored in six orchard/year combinations with Burkard spore traps. A real-time PCR assay was developed to efficiently quantify the dynamics of spore density in these orchards during the growing season. Different patterns of dynamics of spore density were observed in these orchards. A linear relationship between numbers of spores counted with a compound microscope and those determined with the real-time PCR assay was obtained, using the same samples of spore traps. Spore density in five of six orchard/year combinations ranged from 0.0 to 0.05 spores l⁻¹, except for that in orchard 4, which showed much higher values of spore density in the air, as well as higher values and wider range of incidences of blossom infection and fruit rot than those in the other orchards. The results demonstrated a potential method to quantitatively determine spore inoculum potential in orchards by using a real-time PCR assay.     

Expression of engineered antibodies in plants: a possible tool for spiroplasma and phytoplasma disease control

ABSTRACT In an attempt to develop maize plants with resistance to corn stunt spi-roplasma (CSS), a single-chain Fv fragment (scFv) gene that was constructed from antibodies with strong inhibitory activity against CSS, was expressed in a cell line of maize. However, plants regenerated from this transgenic cell line showed no distinct resistance to CSS infection under the greenhouse conditions. The affinity and functionality of scFv in vivo and the locations of CSS and expressed scFv in maize could be a part of the contributing factors affecting this result. Progress of expressing antibodies in plants for plant pathogen resistance is also discussed.     

Inoculation of cucumber roots with zoospores of mycoparasitic and plant pathogenic Pythium species: Differential zoospore accumulation, colonization ability and plant growth response

Hydroponically grown cucumber (Cucumis sativus) seedlings were inoculated with zoospores of 1 mycoparasitic (Pythium oligandrum) and 2 pathogenic (Pythium aphanidermatum and Pythium group F) Pythium spp. During the first 2 days after inoculation, all the Pythium spp. caused reduction in the root length. However, roots treated with Pythium oligandrum quickly reached the length of the control and on the 8th day, and for the rest of the experimental period, stimulation of root elongation was noted. Pythium oligandrum was not pathogenic on cucumber and no differences in the fresh weights of control and Pythium oligandrum inoculated plants were observed in the course of the experiment. Pythium group F and Pythium aphanidermatum were pathogenic on cucumber seedlings, but their pathogenicities differed. Thus, while Pythium group F had a constant, negative influence on root length and plant growth, measured as fresh weight, Pythium aphanidermatum caused generalized necroses of the root system, inhibiting consistently root elongation and plant growth and finally causing plant death. Moreover, the zoospores of 2 mycoparasitic species, Pythium oligandrum and Pythium periplocum, were not attracted to roots of cucumber and accumulated on the roots in very low numbers compared to those of the pathogenic species, Pythium aphanidermatum, which were strongly attracted and accumulated in large numbers. Finally, it was also found that Pythium oligandrum colonized the roots very poorly, while Pythium group F and Pythium aphanidermatum were significantly better root colonizers. The significance of these findings is discussed in relation to the ecology of Pythium species and biocontrol.     

Yolk utilisation in the newly hatched poult

1. Routes of yolk utilisation and some aspects of intestinal digestion and absorption were determined in the hatching poult and compared to the chick. 2. Transfer from yolk to blood was observed pre-hatch and up to 72 h post-hatch. From hatch the transport pattern was similar to the chick. 3. Transport from the yolk sac into the intestine via the yolk stalk was observed up to 120 h after hatch. Secretion was initially to the proximal ileum and reflux occurred, transporting contents to the proximal small intestine and gizzard. Antiperistaltic movements increased after hatch and secretion continued for longer post-hatch than in the chick. 4. In situ uptake of glucose per unit of duodenum did not change with age, whereas uptake of methionine and oleic acid decreased with age. In contrast, in the chick glucose and methionine uptake capacity increased slightly between hatch and 7 d. 5. The lipid class distribution of intestinal contents resembled that of yolk close to hatch, however, some lipolysis was observed in the duodenum. With age the proportion of free fatty acids increased rapidly, first in the duodenum then in the ileum and finally in the caecum. Yolk in the distal small intestine close to hatch did not appear to be utilised.