Biochemical study of resistance to imidacloprid in B biotype Bemisia tabaci from Guatemala

Systemic uptake bioassays using excised cotton leaves confirmed resistance to imidacloprid in a Guatemalan population of the tobacco whitefly Bemisia tabaci Gennadius. Polyacrylamide gel electrophoresis of naphthyl esterases identified the insects as B-types. Upon collection from the field, resistance was determined to be 58-fold relative to a susceptible strain originating in the Imperial Valley of California. Resistance levels increased to 126-fold in this population during its continuous exposure to systemically treated cotton. In biochemical investigations, there was no detectable NADPH-dependent mixed function oxidase metabolism of 14C-imidacloprid at any time during the selection process. In contrast, microsomal preparations from housefly abdomens readily produced significant amounts of the mono-hydroxy and olefin derivatives of the parent compound. Detoxification of imidacloprid by housefly MFOs may account for reports of lower toxicity of the insecticide towards this insect compared with whiteflies, despite similar binding properties between imidacloprid and the nicotinic acetylcholine receptors in both species.     

Effects of a juvenile hormone analog,pyriproxyfen, on Thrips tabaci (Thysanoptera: Thripidae)

Effects of a juvenile hormone analog, pyriproxyfen, on various developmental stadia of the onion thrips, Thrips tabaci Lindeman, were determined on cabbage leaves in the laboratory. Pyriproxyfen was applied at 0.064 or 0.128 g AI liter(-1) on leaves (residual contact-ingestion), thrips (direct contact) and leaf-thrips (residue contact-ingestion-direct contact). Pyriproxyfen did not have any significant lethal effects on thrips pupae in any treatment. Lethal effects on thrips larvae varied depending on application method and dosage. In the leaf and the leaf-thrips treatments, few larvae and pre-pupae molted to the next stage, and none developed to adults. In contrast, in the thrips-only treatment, pyriproxyfen did not show any significant lethal effects. The developmental times of larvae and pre-pupae were prolonged when larvae were treated with pyriproxyfen, and those of pre-pupae and pupae were shortened when pre-pupae and pupae were treated. The longevity and survival rates of thrips adults were generally shorter when they contacted and ingested pyriproxyfen-treated leaves than those in water control. Significantly fewer progeny (0.22-1.15 larvae per female) were produced by females that had fed on and been in contact with the pyriproxyfen-treated leaves than by those in the water control (11.94 larvae per female). However, the number of progeny produced by the thrips females increased significantly (3.32-7.28 larvae per female) when the females were transferred to untreated leaves after feeding on treated leaves for 5 days; the daily larval hatching pattern was similar to those in water control, indicating that female adults were able to produce viable eggs when untreated food was offered.     

Effects of time and concentration on mortality of phosphine-resistant Sitophilus oryzae (L) fumigated with phosphine

The effects of exposure period and phosphine concentration on mortality of susceptible and resistant Sitophilus oryzae (L) were investigated. Although S oryzae is one of the world's most serious pests of stored grain there are few data on the practical significance of phosphine resistance in this species. The strains investigated were an Australian susceptible strain, a homozygous resistant strain exhibiting a level of resistance common in Australia and an unselected field strain from China with a much stronger resistance. Fumigations were carried out at 25 degrees C on adults and mixed-age cultures. For adults of all three strains and mixed-age cultures of the susceptible and resistant Australian strains, the relationship between concentration and time could be described by equations of the form Cnt = k. In all cases n < 1, indicating that time was a more important variable than concentration. In all fumigations of adults the resistant strains were harder to kill than the susceptible strain. However, in fumigations of mixed-age cultures, which contained the tolerant pupal stage, the difference between susceptible and resistant strains was more pronounced at lower concentrations than higher concentrations. For example, at 0.02 mg litre-1 the estimated LT99.9 for mixed-age cultures of the Australian resistant strain (27 days) is 3.4 times that of the susceptible strain (8 days), but at 1 mg litre-1 there is no difference between the two strains (4 days). Limited data on the Chinese resistant strain supported this finding. Twenty-three days exposure at 0.02 mg litre-1 had no effect on mixed-age cultures of this strain, but there were no survivors after 5 days exposure to 1 mg litre-1.     

New developments in insecticide resistance in the glasshouse whitefly (Trialeurodes vaporariorum) and the two-spotted spider mite (Tetranychus urticae) in the UK

A recent survey of insecticide resistance in two of the most problematic pests in UK glasshouses revealed some new developments. At least some individuals in all UK samples of Trialeurodes vaporariorum that were tested resisted the insect growth regulator (IGR) buprofezin. The most strongly resistant strains were unaffected by the field application rate of this compound, and even samples from populations that had never been exposed to buprofezin contained individuals that survived the highest concentration applied (10,000 mg litre-1). The field rate of buprofezin was shown to select for resistance through vapour action alone. The benzophenylurea teflubenzuron, an unrelated IGR, was cross-resisted by buprofezin-resistant individuals. There was no evidence of resistance to imidacloprid, but all T vaporariorum strains tested, regardless of origin, exhibited a high innate tolerance to nicotine, when compared with another whitefly species, Bemisia tabaci. Marked resistance to fenbutatin oxide and tebufenpyrad was found in single glasshouse populations of Tetranychus urticae, but these compounds and abamectin appeared to remain highly effective against all other strains collected.     

鲁西黄牛体细胞克隆保种技术的研究

以体外培养的鲁西黄牛耳皮肤成纤维细胞作核供体，研究利用体细胞克隆技术保存我国地方黄牛优良品种的技术方法。通过组织块培养法建立的鲁西黄牛（1♂，4♀）成纤维细胞系，作为核移植供体细胞，利用屠宰场母牛卵巢卵母细胞经体外培养成熟后作为核受体进行核移植，试验结果表明：重构胚的融合率为62．5％（242／387），分裂率为63．6％（154／242），体外培养第7天囊胚发育率为42．9％（66／154）。体外培养第7天的囊胚的内细胞团细胞（ICM）与滋养层细胞数平均为37和47，ICM占44．2％。体外发育到7d的囊胚新鲜胚胎的移植妊娠率（新鲜胚胎）移植受体10头，60d妊娠率为20％（2／10）。     

Histological investigation of stripe rust (Puccinia striiformis f.sp. tritici) development in resistant and susceptible wheat cultivars

The wheat cultivar Kariega expresses complete adult plant resistance against stripe rust, whereas cv. Avocet S is susceptible. Using confocal laser scanning microscopy, initial fungal penetration into flag leaves was identical in both cultivars, with directional germ-tube growth towards stomata that were penetrated without the formation of an appressorium, followed by differentiation of a substomatal vesicle, infection hyphae, haustorial mother cells and haustoria. During the following 4 days, further fungal development occurred more quickly in the resistant than in the susceptible cultivar. However, by 7 days postinoculation (dpi) the situation changed, with exponential growth of the pathogen occurring only in the susceptible line. Induced cellular lignification, a typical defence reaction of cereals, was observed at 4 dpi in the resistant cultivar, and 2 days later lignified tissue completely surrounded the fungal colonies. In the susceptible cultivar, isolated lignified host cells occurred at 6 dpi, and long, unbranched fungal hyphae outgrowing the resistance reaction were observed.     

Mutants in wheat showing multipathogen resistance to biotrophic fungal pathogens

Five fast-neutron-derived mutants were isolated from the wheat line Hobbit 'sib' that show enhanced field resistance towards Puccinia striiformis f.sp. tritici , the causal agent of yellow rust. Subsequent testing showed the yellow rust resistance phenotypes to differ between mutants, to be expressed at different growth stages and, in some cases, to show an isolate interaction. Three mutants, I3-48, I3-49 and I3-54, exhibited an enhanced yellow rust resistance phenotype from the third seedling leaf onwards, while mutants I3-27 and I3-30 did not show an altered yellow rust phenotype until later growth stages. Additional resistance for brown rust (causal agent Puccinia triticina ) was identified in mutants I3-27, I3-30, I3-48 and I3-49, and for powdery mildew caused by Blumeria graminis f.sp. tritici in mutants I3-27, I3-30, I3-48 and I3-54, although in some cases the resistance was isolate-specific.     

Characterisation of a Quantitative Resistance to Vector Transmission of Tomato yellow leaf curl virus in Lycopersicon pimpinellifolium

Two wild genotypes from the same species Lycopersicon pimpinellifolium, WVA106 (susceptible) and INRA-Hirsute (so-called ‘resistant’), were compared with respect to their reaction to Tomato yellow leaf curl virus isolate Réunion (TYLCV-Mld[RE]), using both whitefly-mediated inoculation and graft inoculation. Disease incidence and symptom severity were scored. Presence and quantification of viral DNA were assessed by dot blot hybridisation. Upon insect inoculation, accession INRA-Hirsute showed a moderate resistance against TYLCV that was overcome by a high inoculation pressure obtained by increasing the cumulative number of inoculative whiteflies. Temporal analyses of the disease progress in relation to this criterion exhibited that the protection was quantitative, mainly reducing the TYLCV-Mld[RE] incidence by at maximum 50% at low inoculation pressure. When graft inoculated, the final TYLCV-Mld[RE] disease incidence was 100% in both susceptible and resistant genotypes with severe symptoms, suggesting a reduction of virus transmission by a vector resistance as a possible mechanism. Implications of using such type of resistance in breeding programmes are discussed.     

Behavioral hormoligosis in oviposition preference of Bemisia tabaci on cotton

The study on behavioral hormoligosis in oviposition preference in Bemisia tabaci (Genn.) on cotton was conducted, at Entomological Research Farm, Punjab Agricultural University (PAU), Punjab, India, during 2001 crop season and repeated in the same season. Multiple-choice test was followed for conducting the experiment. Quinalphos (250, 375, and 500), carbaryl (625, 938, and 1250), acephate (750, 1125, and 1500), endosulfan (438, 656, and 875), and fenvalerate (25, 38, and 50 g ai/ha) were repeatedly sprayed on potted plants of American cotton (var. LH-1556). The impact of these insecticides was evaluated in term of oviposition preference by B. tabaci to treated plants. Also, it investigated changes in biochemical components of treated cotton leaves and the correlation with oviposition preference. The results revealed that, fenvalerate treated plants were more preferred by whitefly for oviposition. Maximum number of eggs was observed on fenvalerate treated plants, 38, 50, and 25 g/ha (39.3, 37.3, and 36.1 eggs/leaf, respectively) followed by acephate 1500 g/ha (26.9 eggs/ leaf) compared with untreated control (14.1 eggs/leaf). Almost similar trend of results was observed in the repeated experiment. The results obtained from biochemical studies revealed that all the insecticidal treatments caused reduction in total sugars compared with untreated control except fenvalerate and low dose of quinalphos. All insecticides caused increase in total free amino acids and brought significant changes in total phenols and pH value of treated plants. These results have confirmed the behavioral hormoligosis in oviposition preference that induced by fenvalerate and acephate in B. tabaci, which may be one of the causes behind its resurgence on plants repeatedly treated with these insecticides.     

Identification of mutations in the para sodium channel of Bemisia tabaci from Crete, associated with resistance to pyrethroids

We investigated the mechanisms of resistance to α-cypermethrin in a Q biotype, highly resistant Bemisia tabaci strain (GRMAL-RP) isolated from Crete. Cytochrome P450-dependent monoxygenase activity with the substrate ethoxycoumarin, and carboxylesterase activity with the substrates α-naphthyl-acetate, β-naphthyl-acetate, and para-nitrophenol acetate were substantially elevated in the GRMAL-RP, compared to the susceptible SUD-S strain, while glutathione-S-transferase activity with the substrate 1-chloro-2,4-dinitrobenzene was not different. The metabolic inhibitors piperonyl butoxide and S,S,S-tributyl phosphorotrithioate synergised cypermethrin toxicity in the GRMAL-RP strain, however, mortality was still lower than that of the susceptible strain, indicating the presence of an additional resistance mechanism. Analysis of the sequence of the IIS4-IIS6 region of the para sodium channel gene of the GRMAL-RP strain revealed two amino acid replacements compared to that of the SUD-S susceptible strain. One is the leucine to isoleucine substitution at position 925 (L925I) previously implicated in B. tabaci pyrethroid resistance and the other is a novel kdr resistant mutation for B. tabaci, a threonine to valine substitution at position 929 (T929V). Genotype analysis showed that the L925I and T929V were present in all GRMAL-RP males tested, at an approximately 1:1 frequency, but never in combination in the same haplotype.