Dissecting the role of Krüppel homolog 1 in the metamorphosis and female reproduction of the cotton bollworm, Helicoverpa armigera.

In insects, metamorphosis and reproduction are controlled by juvenile hormone (JH) and 20-hydroxyecdysone (20E). Krüppel homolog 1 (Kr-h1), a transcription factor, is regarded as a JH-early inducible gene responsible for the repression of metamorphosis. However, the role of Kr-h1 in reproduction of holometabolic insects is relatively less understood. In this study, we studied the role of Kr-h1 in larvae-pupae transition and female reproduction in the major agricultural pest Helicoverpa armigera. Two HaKr-h1 isoforms (HaKr-h1α and HaKr-h1ß) were identified, with HaKr-h1α predominant in the cotton bollworm. In larvae, HaKr-h1 was predominately expressed in the epidermis and markedly up-regulated during the moult stage, whereas in adults HaKr-h1 was mainly expressed in females and the highest transcription was detected in the ovaries. Considering the function of hormones in larval metamorphosis, we examined the modulation of gene expression in response to hormones, which showed that HaKr-h1 was significantly induced by both JH analogue (JHA) and 20E. Knockdown of HaKr-h1 in fifth-instar larvae resulted in precocious metamorphosis from larvae to pupae. Moreover, a fluorescence immunoassay coupled with heterologous expression revealed that HaKr-h1 was localized in the nucleus of oocyte membrane. In female adults, depletion of HaKr-h1 severely repressed the transcription of vitellogenin, disrupted oocyte maturation and reduced the number of eggs laid, suggesting that HaKr-h1 is required for vitellogenesis and egg production in H. armigera. The present study provides insight into the roles of HaKr-h1 in JH-mediated reproduction and highlights HaKr-h1 as a target for suppression of lepidopteran pests.

Analysis of symptom clusters in Chinese cervical cancer patients undergoing radiotherapy, chemoradiotherapy, or postoperative chemoradiotherapy.

OBJECTIVE: To assess the common symptom clusters in cervical cancer patients treated with radiotherapy alone, chemorachothierapy, or postoperative chemoradiotherapy, and evaluate differences among multi-modality treatment in these symptom clusters. MATERIALS AND METHODS: Ninety-three patients diagnosed with cervical cancer were recruited to the survey, in which destination time was the fourth friday of radiotherapy. Symptoms of patients were evaluated by the Chinese version of the 13-item M.D. Anderson Symptom Inventory (MDASI). RESULTS: Among the symptoms, lack of appetite was the most common and the most severe symptome, treatment modalities may have important effects on symptom clusters in cervical cancer patients undergoing radiotherapy. CONCLUSION: The results offer valuable information for the future development treatment-based symptom cluster intervention.

Reproductive Cost Associated With Juvenile Hormone in Bt-Resistant Strains of Helicoverpa armigera (Lepidoptera: Noctuidae).

Transgenic Bacillus thuringiensis (Bt) crops are increasingly significant in pest control, but resistance development of target pests is a major issue in the sustainable deployment of Bt crops. The fitness cost of resistance in target pests is regarded as one of the main factors delaying resistance when adopting the refuge strategy. In this study, we compared the life-history traits of three independent sets of Helicoverpa armigera (Hübner, 1809) adults, of each there were a susceptible population and a Cry1Ac-resistant population derived by selection from it. Confirming to the previous studies, resistant individuals exhibited fewer progeny, less fecundity, lower egg hatching rate, and longer adult longevity. And poor fecundity in resistant strains was associated with the decline of the mature follicular amount, the ovarian weight ratio, and the length of the longest ovarian tubule. Interestingly, the juvenile hormone (JH) level appeared higher in resistant strains relative to susceptible strains. Application of methoprene (JH analogue) in vivo was effective in reducing fecundity and hatchability with the up-regulation of detected JH titer. These results suggested that resistance against Bt toxin reduced the reproductive capacity of H. armigera, and JH level is affected in the tradeoff between reproductive capacity and Bt resistance.

The effect of silencing arginine kinase by RNAi on the larval development of Helicoverpa armigera.

Arginine kinase (AK) is an important regulation factor of energy metabolism in invertebrate. An arginine kinase gene, named HaAK, was identified to be differentially expressed between Cry1Ac-susceptible (96S) and Cry1Ac-resistant (Bt-R) Helicoverpa armigera larvae using cDNA-amplification fragment length polymorphism analysis. The full-length open reading frame sequence of HaAK gene with 1068 bp was isolated from H. armigera. Quantitative reverse transcription polymerase chain reaction assay revealed that HaAK gene is specifically expressed in multiple tissues and at larval developmental stages. The peak expression level of HaAK was detected in the midgut of the fifth-instar larvae. Moreover, the expression of HaAK was obviously down-regulated in Bt-R larvae. We further constructed a dsRNA vector directly targeting HaAK and employed RNAi technology to control the larvae. The feeding bioassays showed that minute quantities of dsRNA could greatly increase the larval mortality and delay the larval pupation. Silencing of HaAK significantly retarded the larval development, indicating that HaAK is a potential target for RNA interference-based pest management.

Tradeoff between reproduction and resistance evolution to Bt-toxin in Helicoverpa armigera: regulated by vitellogenin gene expression.

Evolution of resistance to insecticides usually has fitness tradeoffs associated with adaptation to the stress. The basic regulation mechanism of tradeoff between reproduction and resistance evolution to Bacillus thuringiensis (Bt) toxin in the cotton bollworm, Helicoverpa armigera (Ha), based on the vitellogenin (Vg) gene expression was analyzed here. The full-length cDNA of the Vg gene HaVg (JX504706) was cloned and identified. HaVg has 5704 base pairs (bp) with an open reading frame (ORF) of 5265 bp, which encoded 1756 amino acid protein with a predicted molecular mass of 197.28 kDa and a proposed isoelectric point of 8.74. Sequence alignment analysis indicated that the amino acid sequence of HaVg contained all of the conserved domains detected in the Vgs of the other insects and had a high similarity with the Vgs of the Lepidoptera insects, especially Noctuidae. The resistance level to Cry1Ac Bt toxin and relative HaVg mRNA expression levels among the following four groups: Cry1Ac-susceptible strain (96S), Cry1Ac-resistant strain fed on artificial diet with Bt toxin for 135 generations (BtR stands for the Cry1Ac Bt resistance), progeny of the Cry1Ac-resistant strain with a non-Bt-toxin artificial diet for 38 generations (CK1) and the direct descendants of the 135th-generation resistant larvae which were fed on an artificial diet without the Cry1Ac protein (CK2) were analyzed. Compared with the 96S strain, the resistance ratios of the BtR strain, the CK1 strain and the CK2 strain were 2917.15-, 2.15- and 2037.67-fold, respectively. The maximum relative HaVg mRNA expression levels of the BtR strain were approximately 50% less than that of the 96S strain, and the coming of maximum expression was delayed for approximately 4 days. The overall trend of the HaVg mRNA expression levels in the CK1 strain was similar to that in the 96S strain, and the overall trend of the HaVg mRNA expression levels in the CK2 strain was similar to that in the BtR strain. Our results suggest that the changes in reproduction due to the Bt-toxin resistance evolution in the BtR strain may be regulated by the Vg gene expression. The down-regulation of HaVg at the early stages resulted in a period of delayed reproduction and decreased fecundity in the BtR strain. This performance disappeared when the Bt-toxin selection pressure was lost.