Annotated ESTs from various tissues of the brown planthopper Nilaparvata lugens: a genomic resource for studying agricultural pests

Abstract

Background: The brown planthopper (BPH), Nilaparvata lugens (Hemiptera, Delphacidae), is a serious insect pests of rice plants. Major means of BPH control are application of agricultural chemicals and cultivation of BPH resistant rice varieties. Nevertheless, BPH strains that are resistant to agricultural chemicals have developed, and BPH strains have appeared that are virulent against the resistant rice varieties. Expressed sequence tag (EST) analysis and related applications are useful to elucidate the mechanisms of resistance and virulence and to reveal physiological aspects of this non-model insect, with its poorly understood genetic background.

Gene knockdown by intro-thoracic injection of double-stranded RNA in the brown planthopper, Nilaparvata lugens

“…Actually, for every target gene, the optimal concentration should be different and has to be determined respectively (Huvenne and Smagghe, 2010). The exceeding concentration might not result in more silencing and cause further side-effect, such as high mortality (Meyering-Vos and Muller, 2007;Shakesby et al, 2009). Third, the nucleotide sequence and length of dsRNA fragment should be chosen based on each target gene itself.…”

“…Many progresses have been achieved in basic and applied aspects of this pest and the recent release of BPH EST Database http://bphest.dna. affrc.go.jp/ by Noda et al (2008) provides the powerful information for gene function studies in this pest. Functional analyses tools such as inactivation of gene expression will be essential for further investigations in this insect species.…”

“…In order to evaluate the universality of RNAi technology, three target genes (calreticulin, cathepsin B-like protease and Nlb2) with different tissue expression were selected and analysed. Calreticulin is a ubiquitously expressed gene (Jaubert-Possamai et al, 2007), Cathepsin B-like protease is a gut specific gene (Noda et al, 2008) and Nlb2 is the central nervous system (CNS) specific gene (Liu et al, 2005).…”

Proteomic analysis of the salivary glands of the rice brown planthopper, Nilaparvata lugens (Stål) (Homoptera: Delphacidae)

“…Since the N. lugens genome has not been sequenced, the use of tandem mass spectroscopy (MS/MS) for identifying pro-teins in this species is unreliable; therefore, the Nterminal and internal amino acid sequences obtained by Edman degradation were subjected to a search for the corresponding gene in our EST database for N. lugens (http://bphest.dna.affrc.go.jp/). To construct the EST database, mRNA was extracted from various tissues and organs of N. lugens, such as salivary glands, midgut, ovary, and the head-thorax complex (Noda et al, 2008). BlastX analysis was performed against the EST database.…”

Characterization of soluble and membrane‐bound alkaline phosphatase in Nilaparvata lugens and their potential relation to development and insecticide resistance

“…The first strand cDNA (1 ml) was used as a template for PCR. Searching the BPH EST Database (http://bphest.dna.affrc.go.jp/; Noda et al, 2008) with the amino acid sequences of two B. mori ALPs (sALP and mALP) gave two homolog fragments (Clone ID: NLMA5211 and C_NLMA1028) with 40-50% identity at the amino acid level to B. mori sALP and mALP. Based on these two fragments, gene-specific primers (GSPs) were designed and the rapid amplification of cDNA ends (RACE) technique was used to amplify the full-length cDNAs for two fragments according to Generacer TM Kit (Invitrogen).…”