Function analysis and characterisation of a novel chitinase, MdCht9, in Musca domestica.

Insect chitinases have been proposed as potential targets for pest control. In this work, a novel group IV chitinase gene, MdCht9, from Musca domestica was found to have multiple functions in the physiological activity, including chitin regulation, development and antifungal immunity. The MdCht9 gene was cloned and sequenced, its phylogeny was analysed and its expression was determined in normal and 20E treated larvae. Subsequently, RNA interference (RNAi)-mediated MdCht9 knockdown was performed, followed by biochemical assays, morphological observations and transcriptome analysis. Finally, the recombinant protein MdCht9 (rMdCht9) was purified and tested for anti-microbial activity and enzyme characteristics. The results showed that MdCht9 consists of three domains, highly expressed in a larval salivary gland. RNAi silencing of MdCht9 resulted in significant down-regulation of chitin content and expression of 15 chitin-binding protein (CBP) genes, implying a new insight that MdCht9 might regulate chitin content by influencing the expression of CBPs. In addition, more than half of the lethality and partial wing deformity appeared due to the dsMdCht9 treatment. In addition, the rMdCht9 exhibited anti-microbial activity towards Candida albicans (fungus) but not towards Escherichia coli (G-) or Staphylococcus aureus (G+). Our work expands on previous studies of chitinase while providing a potential target for pest management.

Proteomic Analysis of the Peritrophic Matrix from the Midgut of Third Instar Larvae, Musca domestica.

OBJECTIVE: To better comprehend the molecular structure and physiological function of the housefly larval peritrophic matrix (PM), a mass spectrometry approach was used to investigate the PM protein composition. METHODS: The PM was dissected from the midgut of the third instar larvae, and protein extracted from the PM was evaluated using SDS-PAGE. A 1D-PAGE lane containing all protein bands was cut from top to bottom, the proteins in-gel trypsinised and analysed via shotgun liquid chromatography- tandem mass spectrometry (LC-MS/MS). RESULTS: In total, 374 proteins, with molecular weights varying from 8.225 kD to 996.065 kD and isoelectric points ranging from 3.83 to 11.24 were successfully identified, most identified proteins were mainly related to immunity, digestion, nutrient metabolism and PM structure. Furthermore, many of these proteins were functionally associated with pattern binding, polysaccharide binding, structural constituent of peritrophic membrane and chitin binding, according to Gene Ontology annotation. CONCLUSION: The PM protein composition, which provides a basis for further functional investigations of the identified proteins, will be useful for understanding the housefly larval gut immune system and may help to identify potential targets and exploit new bioinsecticides.

Establishment of a Faba Bean Banker Plant System with Predator Orius strigicollis for the Control of Thrips Dendrothrips minowai on Tea Plants under Laboratory Conditions.

The stick tea thrip Dendrothrips minowai (Priesner) (Thysanoptera: Thripidae) is a destructive pest in tea plantations in south and southwest China. To control this pest, a non-crop banker plant system was developed using a polyphagous predator Orius strigicollis (Poppius) (Heteroptera: Anthocoridae) with the black bean aphid Aphis fabae (Scopoli) (Hemiptera: Aphididae) as an alternative prey and the faba bean Vicia faba as the banker plant to support the predator in targeting the pest. The fitness of A. fabae on tea plants and faba bean was evaluated to determine its host specificity. Moreover, the control efficacy of the banker plant system on D. minowai on tea plants was tested in the laboratory and compared with that of direct release of O. strigicollis. The experiments showed that faba bean was an excellent non-crop host for A. fabae because, while the aphid population increased quickly on faba bean, it could only survive for up to 9 days on tea plants. Compared with direct release of O. strigicollis, lower densities of pest were observed when introducing the banker plant system. Our results indicate that this banker plant system has the potential to be implemented in the field to improve the control of the pest thrips.

[Effects of temperature on the growth and development of Aglossa dimidiata parasitized on Litsea coreana].

In order to reveal the effects of temperature on the growth and development of Aglossa dimidiata parasitized on Litsea coreana, a laboratory experiment was conducted to study the mean development duration, development rate, and survival rate of A. dimidiata at its different growth stages at 31 degrees C, 28 degrees C, 25 degrees C, 22 degrees C, and 19 degrees C, with the development threshold temperature and effective accumulated temperature for different growth stages calculated. Temperature had significant effects on the developmental duration. Except that the development duration of egg was shortened with increasing temperature, the development durations of larva, pupa, and immature A. dimidiata were the shortest at 25 degrees C, being 249.53 +/- 23.83, 12.94 +/- 1.27, and 273.00 +/- 24.19 days, respectively. There existed significant relationships between the development rates of A. dimidiata at its different growth stages and temperature, with positive linear relationship at egg stage, and quadratic relationship at larva, pupa, and immature stages. Temperature also had significant effects on the survival rate of A. dimidiata. The survival rates of A. dimidiata at its different growth stages were all the highest at 25 degrees C, being 94.0%, 73.8%, 91.3%, and 63.4% for the egg, larva, pupa, and immature A. dimidiata, respectively, followed by at 22 degrees C and 19 degrees C, and the lowest at 31 degrees C. No larva and pupa could survive at 31 degrees C, suggesting that A. dimidiata was not resistant to high temperature. The development threshold temperature for egg, larva, pupa, and immature A. dimidiata was 13.21 degrees C, 17.12 degrees C, 14.76 degrees C, and 16.47 degrees C, and the effective accumulated temperature was 117.94, 870.88, 149.70, and 1442.75 day-degree, respectively. The results coincided with the fact that the A. dimidiata reproduced 2 or 3 generations a year in Xifeng area of Guizhou, Southwest China.