Edible and Medicinal Progress of Cryptotympana atrata (Fabricius) in China.

As an important resource insect, the Cryptotympana atrata is widely distributed in the eastern and central parts of China. The cicada slough is one of the traditional crude drugs in East Asia, and the main component is polysaccharide, which has the functions of anti-convulsion, relieving asthma and improving lipid metabolism. The parasitoid fungus Cordyceps cicadae, which grows inside the cicada nymphs and forms the fruiting bodies on the surface of the host's carcass, is also known as the "cicada flower" in China. The Cordyceps cicadae is another old, traditional Chinese medicine, which has been used as a tonic and medicine to nourish and regulate human immunity for centuries. For the further development and utilization of the golden cicada, this paper summarized the C. atrata from the aspects of their biological characteristics, distribution area, life cycle, history of edible and medicinal use, edible methods and nutritional compositions; emphatically introduced the edible and potential medicinal value of the C. atrata; and specifically expounded the research progress of its application. As one popular insect food, the prospects for the development of C. atrata have also been put forward, especially in artificial breeding technology, food safety risk assessment and medicinal value utilization.

Genome-wide analysis of G-quadruplex in Spodoptera frugiperda.

Spodoptera frugiperda (Lepidoptera: Noctuidae) is a globally distributed lepidopteran crop pest that has developed resistance to most insecticides. The G-quadruplex (G4) is a secondary structure in the genome enriched in the promoters for regulating gene expression. However, little is known about G4 in S. frugiperda, especially whether G4 is involved in insecticide resistance and pest control. In this study, 387,875 G4 motifs in the whole genome of S. frugiperda were identified by bioinformatics prediction. We found that 66.90 % of theseG4 structures were located in genic regions and highly enriched in the upstream regions of start codons. Functional and pathway analyses showed that the genes with G4 enriched in promoter regions participate in several metabolic processes. Further analyses showed that G4 structures occurred more frequently in the promoters of P450 and CarE gene families. It was also investigated that G4 ligand N-methyl mesoporphyrin IX (NMM) decreased P450 protein activity in larval midgut tissue. Cytotoxicity and bioassay results revealed that NMM and pesticides had synergistic effects on toxicity. In conclusion, our findings suggest that G4 motif could be a new potential target for pest control.

Effects of Juvenile Hormone Analog and Days after Emergence on the Reproduction of Oriental Armyworm, Mythimna separata (Lepidoptera: Noctuidae) Populations.

Mythimna separata (Walker) is a main cereal crop pest that causes extensive damage to the world grain production. The effects of juvenile hormone on M. separata populations remain poorly understood. Here, we explored how JH analog (JHA) affected reproductive traits of both migrant and resident populations in this pest. Our results showed that the influence of JHA treatment on reproduction depended on adult age (days after emergence) of exposure to JHA and populations. Exposure of M. separata residents to JHA (methoprene) on day 1 and day 2 after adult emergence significantly shortened the pre-oviposition period, but increased the lifetime fecundity, mating frequency and grade of ovarian development compared to the controls. However, M. separata migrants exposed to JHA only on day 1 facilitated their reproduction, resulting in a reduction in the pre-oviposition period but an increase in lifetime fecundity, mating frequency and grade of ovarian development. In addition, exposure to JHA from day 2 to day 4 did not significantly affect the ovarian and reproductive development in both migrant and resident populations. These results indicated that the first two days after adult emergence were the sensitive period for residents. In contrast, only the first day after adult emergence was the sensitive stage for migrants. Our findings will contribute to a better understanding of JHA function on M. separata populations.

A dual-luciferase reporter system for characterization of small RNA target genes in both mammalian and insect cells.

MicroRNAs (miRNAs) are regulatory RNA molecules that bind to target messenger RNAs (mRNAs) and affect the stability or translational efficiency of the bound mRNAs. Single or dual-luciferase reporter systems have been successfully used to identify miRNA target genes in mammalian cells. These reporter systems, however, are not sensitive enough to verify miRNA-target gene relationships in insect cell lines because the promoters of the target luciferase (usually Renilla) used in these reporter systems are too weak to drive sufficient expression of the target luciferase in insect cells. In this study, we replaced the SV40 promoter in the psiCHECK-2 reporter vector, which is widely used with mammalian cell lines, with the HSV-TK or AC5.1 promoter to yield two new dual-luciferase reporter vectors, designated psiCHECK-2-TK and psiCHECK-2-AC5.1, respectively. Only psiCHECK-2 and psiCHECK-2-AC5.1 had suitable target (Renilla)/reference (firefly) luciferase activity ratios in mammalian (HeLa and HEK293) and insect (Sf9, S2, Helicoverpa zea fat body and ovary) cell lines, while psiCHECK-2-TK had suitable Renilla/firefly luciferase activity ratios regardless of the cell line. Moreover, psiCHECK-2-TK successfully detected the interaction between Helicoverpa armigera miRNA9a and its target, the 3'-untranslated region of heat shock protein 90, in both mammalian and H. zea cell lines, but psiCHECK-2 failed to do so in H. zea cell lines. Furthermore, psiCHECK-2-TK with the target sequence, HzMasc (H. zea Masculinizer), accurately differentiated between H. zea cell lines with or without the negative regulation factor (miRNA or piRNA) of HzMasc. These data demonstrate that psiCHECK-2-TK can be used to functionally characterize small RNA target genes in both mammalian and insect cells.

Identification of Wild-Type CYP321A2 and Comparison of Allelochemical-Induced Expression Profiles of CYP321A2 with Its Paralog CYP321A1 in Helicoverpa zea.

One possible way to overcome the diversity of toxic plant allelochemicals idiosyncratically distributed among potential host plants is to have more counterdefense genes via gene duplication or fewer gene losses. Cytochrome P450 is the most important gene family responsible for detoxification of the diversity of plant allelochemicals. We have recently reported the identification and cloning of the transposon (HzSINE1)-disrupted non-functional CYP321A2, a duplicated paralog of the xenobiotic-metabolizing P450 CYP321A1 from a laboratory colony of Helicoverpa zea. Here we report the identification of the wild-type intact allele of CYP321A2 from another H. zea colony. This CYP321A2 allele encodes a deduced protein of 498 amino acids and has the P450 signature motifs. Quantitative RT-PCR experiments showed that this CYP321A2 allele was highly expressed in midgut and fat body and achieved the highest expression level in the developmental stage of 5th and 3rd instar larvae. CYP321A2 and CYP321A1 were constitutively expressed in low levels but can be differentially and significantly induced by a range of the plant allelochemicals and plant signal molecules, among which xanthotoxin, flavone, and coumarin were the most prominent inducers of CYP321A2 both in midgut and fat body, whereas flavone, coumarin, and indole-3-carbinol were the prominent inducers of CYP321A1 in midgut and fat body. Moreover, xanthotoxin- and flavone-responsive regulatory elements of CYP321A1 were also detected in the promoter region of CYP321A2. Our results enrich the P450 inventory by identifying an allelochemical broadly induced CYP321A2, a paralog of CYP321A1 in H. zea. Our data also suggest that the CYP321A2/CYP321A1 paralogs are a pair of duplicated genes of multigene families and CYP321A2 could potentially be involved in the detoxification of plant allelochemicals and adaptation of H. zea to its chemical environment.