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The occurrence of Bemisia tabaci poses an increasingly serious threat to cotton and vegetable crops in Xinjiang, China. Currently, neonicotinoid insecticides are commonly used to control the insect, to which resistance is inevitable due to intensive use. However, the resistance status and mechanism of B. tabaci to neonicotinoid insecticides in Xinjiang are poorly understood. Cytochrome P450 monooxygenases represent a key detoxification mechanism in the neonicotinoid resistance of B. tabaci. In this study, the resistance level to imidacloprid and thiamethoxam was investigated using the leaf dipping method in five field populations of B. tabaci from Turpan (TP, two sampling sites), Shache (SC), Hotan (HT) and Yining (YN) in northern and southern Xinjiang. The expression changes of eighteen cytochrome P450 genes from the select B. tabaci populations were determined by real‐time fluorescence quantitative PCR (qPCR). The bioassay revealed that the five populations tested had developed moderate to high levels of resistance to imidacloprid (12.26–46.07‐fold), while the populations remained sensitive to thiamethoxam except for HT, which had a low level of resistance. The qPCR results showed that the expression levels of five P450 genes, CYP4G68, CYP6CM1, CYP303A1‐like, CYP6DZ7 and CYP6DZ4, were significantly higher in some resistant field populations than in the susceptible strain. Resistance to imidacloprid in field populations of B. tabaci might be associated with the increased expression of these five cytochrome P450 genes. The results are useful for further understanding the mechanism of neonicotinoid resistance and will contribute to the management of insecticide‐resistant B. tabaci in Xinjiang.

A water stable one-dimensional (1D) ladder-shaped coordination polymer (CP) has been synthesized and exhibits a strong affinity to two fluorescein-tagged single-stranded probe DNAs (P-DNAs), giving a sensing platform of P-DNAs@1. Such a hybrid sensing platform is capable of simultaneous detection of breast cancer related microRNA-221 (miRNA-221) and miRNA-222 in a specific and synchronous manner, without observable cross-reactions, as supported by experimental evidences. The interaction mode and the electronic energy between CP 1 with nucleic acid were confirmed by molecular simulation and the universal force field (UFF).

CRISPR/Cas9 has recently been developed as an efficient genome engineering tool. The rabbit is a suitable animal model for studies of metabolic diseases. In this study, we generated ATP7B site-directed point mutation rabbits to simulate a major mutation type in Asians (p. Arg778Leu) with Wilson disease (WD) by using the CRISPR/Cas9 system combined with single-strand DNA oligonucleotides (ssODNs). The efficiency of the precision point mutation was 52.94% when zygotes were injected 14 hours after HCG treatment and was significantly higher than that of zygotes injected 19 hours after HCG treatment (14.29%). The rabbits carrying the allele with mutant ATP7B died at approximately three months of age. Additionally, the copper content in the livers of rabbits at the onset of WD increased nine-fold, a level similar to the five-fold increase observed in humans with WD. Thus, the efficiency of precision point mutations increases when RNAs are injected into zygotes at earlier stages, and the ATP7B mutant rabbits are a potential model for human WD disease with applications in pathological analysis, clinical treatment and gene therapy research.

The Colorado potato beetle (Leptinotarsa decemlineata [Say]) in the north Xinjiang Uygur autonomous region in China has evolved resistance to pyrethroids and carbamates. Mutations resulting in target site insensitivity, namely L1014F in LdVssc1 voltage-sensitive sodium channel and S291G in acetylcholine esterase, confer, at least partially, the resistance. Insecticides with different modes of action may serve as likely replacements. Endosulfan and fipronil are GABA-gated chloride channel-blocking insecticides. In this article, we found that the contact toxicities of the two compounds were among the highest ever estimated to the 4th-instar larvae and the adults and affirmed that they were useful for L. decemlineata control in north Xinjiang. Regarding stomach toxicities of the two compounds, adult beetles were less sensitive than 2nd-, 3rd-, and 4th-instar larvae, suggesting that the appropriate timing for spraying is the early larval stage. Mixtures of endosulfan and α-cypermethrin at 1:160, 1:80, and 1:40 ratios, of endosulfan and phoxim at 1:24 ratio, and of endosulfan and isocarbophos at 1:72 and 1:288 ratios, significantly increased toxicity in a field population. The combination indices were significantly below 1 at both LD50 and LD90 levels, revealing synergistic effects. Our results demonstrated that endosulfan and fipronil could be applied alone and endosulfan may also be used in binary mixtures to restore pyrethroid susceptibility. These findings may have considerable practical implications for L. decemlineata resistance management.

The Colorado potato beetle (Leptinotarsa decemlineata (Say)) in the north Xinjiang Uygur autonomous region has evolved resistance to various types of insecticides. Chlorantraniliprole is a novel anthranilic diamide insecticide that binds and activates ryanodine receptors. It exhibited excellent efficacy against L. decemlineata in several field trails in Europe. In the present paper, the susceptibility of L. decemlineata fourth-instar larvae derived from six field populations and L. decemlineata adults derived from three field populations to chlorantraniliprole was determined by a topical application. The fourth-instar larvae were substantially more susceptible to chlorantraniliprole than adults, although the range of susceptibility was far greater among the fourth-instar larvae. Regarding stomach toxicities, adult beetles were less susceptible to chlorantraniliprole than larvae. Chlorantraniliprole was most toxic to second-instar larvae, followed by third- and fourth-instar larvae. These data suggested that the appropriate timing for chlorantraniliprole spraying is the early larval stage. Moreover, the synergistic activities of chlorantraniliprole in combination with triphenyl phosphate, diethyl maleate, or piperonyl butoxide against fourth-instar larvae from two field populations and adults from one field population were tested. Piperonyl butoxide had synergistic effects with chlorantraniliprole against fourth-instar larvae but not against adult beetles. Conversely, triphenyl phosphate and diethyl maleate exerted little synergistic effects. It appears that there is a potential risk of resistance against chlorantraniliprole resulting from cytochrome P450 monooxygenase activity.

A survey of resistance to five conventional insecticides was conducted in 2009 and 2010 for the first generation 4th-instar larvae of Leptinotarsa decemlineata from Urumqi, Changji, Qitai and Qapqal. Compared with the Tekes population, a reference susceptible population, the Changji and Qapqal populations exhibited very high to moderate levels of resistance to cyhalothrin and deltamethrin, moderate to high levels of resistance to carbosulfan and carbofuran, and low levels of resistance to azinphosmethyl. Moreover, the Urumqi and the Qitai populations reached a high and a moderate level of resistance to carbosulfan, respectively. Synergistic effects of triphenyl phosphate, diethylmeleate, and piperonyl butoxide on cyhalothrin and carbosulfan in Changji population revealed that cytochrome P450s were involved in the resistance to cyhalothrin but not carbosulfan. A modified bi-PASA was developed to simultaneously detect point mutations of S291G in the AChE and L1014F in the LdVssc1 genes. The former mutation resulted in the resistance to carbamates and the latter in the resistance to pyrethroids. The rates of homozygous and heterozygous resistant individuals to carbamates (S291G mutation) were 17.6% and 14.7%, 50.6% and 42.2%, 49.9% and 41.7%, 51.3% and 41.4%, and 44.8% and 47.4%; to pyrethroids (L1014F mutation) were 5.8% and 8.7%, 36.1% and 27.0%, 41.8% and 24.8%, 12.2% and 9.7%, and 7.9% and 10.6%, respectively, in samples from Tekes, Changji, Qapqal, Urumqi and Qitai. I392T point mutation in the AChE was detected by RT-PCR among 18 individuals from Changji, Qapqal, Urumqi and Qitai. These results demonstrated that point mutations of S291G in the AChE and L1014F in the LdVscc1 are responsible for, at least partially, the resistance to carbamates and pyrethroids in L. decemlineata in some field populations in northern Xinjiang Uygur autonomous region.

J. Agric. Urban Entomol. 27: 61-73 (2010/2011) ABSTRACT The Colorado potato beetle Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae) in the northern part of the Xinjiang Uygur Autonomous Region has evolved resistance to several pyrethroid and carbamate insecticides. Biological control methods should be a major component of integrated pest management for L. decemlineata. Spinosad and abamectin are two biopesticides that have unique mechanisms of action. In this study, the contact toxicities of spinosad and abamectin to L. decemlineata fourth instars and adults were determined by topical applica- tions to several field populations. The average LD50 values of spinosad and abamectin for adults were 0.1275 and 0.0101 mg (a.i.) per individual, and for fourth instars they were 0.0181 and 0.0016 mg (a.i.) per individual, respectively. These data were among the lowest LD50s ever estimated, which affirmed that the two biopesticides are useful for L. decemlineata control in north Xinjiang. Susceptibilities to the two biopesticides varied slightly but significantly among tested field populations and the variations did not result from cross-resistance to conventional insecticides. Regarding stomach toxici- ties, the LC50 values of spinosad applied to excised potato leaves for second instars, third instars, fourth instars, and adults were 0.2840, 0.4093, 1.2413, and 2.3783 mg/L (a.i.), respectively. The LC50 values of abamectin for second instars, third instars, fourth instars, and adults were 0.0036, 0.0088, 0.0177, and 0.2591 mg/L (a.i.), respectively. The two biopesticides were most toxic to second instars, followed by third, then fourth instars, and they were least toxic to adults. These data suggested that the appropriate timing for spinosad or abamectin spraying is to early larval stages.

Nicotinic acetylcholine receptors (nAChRs) are pentameric ACh-gated ion channels. It is believed that nAChRs composed of different subunits may vary in their function and toxicological characteristics. Neonicotinoids are activators of nAChRs and important insecticides that are extensively used for crop protection and resistance has been developed by some pests. They are also major insecticides for the control of Leptinotarsa decemlineata, which is a destructive defoliator pest that invaded the Xinjiang region of China in the 1990s. However, little is known about the constitution or subunits of the target in this pest. In this study, the full-length cDNAs encoding four new nAChR subunits (named Ldα3, Ldα6, Ldα10, and Ldβ1) were cloned from L. decemlineata. These genes encode 822-, 753-, 672-, and 759-amino acid proteins, respectively, which share typical features of insect nAChRs subunits and closely resemble the corresponding subunits of the nAChRs from Tribolium castaneum. Temporal and spatial expression analyses showed that these genes, as well as the previously identified Ldα1, Ldα2, and Ldα8 genes, are widely expressed in all developmental stages, including eggs, larvae of various instars, pupae, and adults. All genes monitored were expressed at higher levels in the head than in the thorax and abdomen, except for Ldα10. Dietary ingestion of double-stranded RNA bacterially expressed for Ldα1 (dsLdα1) significantly reduced the mRNA level of Ldα1 in treated larvae and adults by 48.0% and 78.6%, respectively. Among the non-target genes, Ldα3, Ldα9, and Ldβ1 were significantly up-regulated in larvae. A toxicity bioassay showed that dsLdα1 treatment greatly decreased the sensitivity to imidacloprid and thiamethoxam in adults. The larval susceptibility to thiamethoxam but not to imidacloprid was also reduced because of the lower down-regulation of Ldα1. Thus, our results suggest that Ldα1 encodes a subunit of a functional nAChR that mediates the toxicity of imidacloprid and thiamethoxam against L. decemlineata and that the down-regulation of Ldα1 might be an important mechanism for resistance and/or tolerance of L. decemlineata to neonicotinoids.

Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), has become the economically most important insect defoliator of potatoes, Solanum tuberosum L., in northern Xinjiang Uygur autonomous region in China. Currently, control of Colorado potato beetle relies mainly on chemical insecticides. And this may result in insecticide resistance. In this study, LD50 values were measured by a topical bioassay for 14 conventional insecticides in seven local populations from Urumqi, Changji, Tacheng, Nilka, Gongliu, Qapqal, and Tekes counties (cities). The Tekes field population was the most susceptible population and was selected as a reference strain. Compared with the Tekes strain, the Changji, Qapqal, Nilka, Tacheng, and Gongliu populations exhibited moderate to very high levels of resistance to cyhalothrin. The Qapqal and Changji populations showed a moderate and a very high resistance to deltamethrin, respectively. And the Changji population developed a high resistance against alpha-cypermethrin. Moreover, the Qapqal population had a moderate resistance to carbofuran, and the Urumqi population reached high level of resistance to endosulfan. Possible resistance mechanisms of the Changji and Qapqal populations were determined using three enzyme inhibitors. Triphenyl phosphate (TPP), diethylmeleate, and piperonyl butoxide (PBO) had little synergism to cyhalothrin in the two populations. In contrast, PBO and TPP exhibited some synergistic effects to carbofuran in the Qapqal population, indicating the involvement of monooxygenases and esterases in conferring carbofuran resistance. It seems that additional mechanisms, such as target site insensitivity, should play an important role in Colorado potato beetle resistances to cyhalothrin and carbofuran in northern Xinjiang local populations.