1.1.1),

comp7073 (aminopeptidase N) (EC 3.4.11.2), comp12788 (pancreatic triacylglycerol lipase) (EC 3.1.1.3), and comp13347 (vitellogenin-A1) (Tables 2 and S6) are shown in Fig. 1. All of the contigs, except for comp13347 (vitellogenin-A1), were specifically expressed in salivary glands; transcript ratios were 3.7 × 102 − 1.9 × 106 times higher in salivary glands than in stomach and Malpighian tubules. Of the 13 contigs examined, only comp13347 (vitellogenin-A1) was similarly expressed in salivary glands, stomach, and Malpighian tubules, with relative expression levels 1.54:1:1.72) (Fig. 1). The expression patterns were surveyed using PCR amplification for 63 of the 76 contigs (contig IDs from comp13378 to comp13413 check details and comp13407 to comp13545 in Tables S6 and 2) using cDNAs of salivary glands, stomach, and Malpighian tubules that were subjected to qRT-PCR. As a result (data not shown), 56 contigs showed amplification almost specific to salivary glands and 40 of these showed no similarity PFT�� mw to known proteins. Seven contigs showed amplification in all tissues (salivary, stomach, and Malpighian tubules): comp12773 (protein disulfide-isomerase), comp13517 (40S ribosomal protein S15), comp13506 (transferrin), comp11878 (proactivator polypeptide), comp13359 (heat shock 70 kDa protein cognate 3), comp13270 (allergen Cr-PI),

and comp13610 (peptidyl-prolyl cis–trans isomerase B). Of the 76 most highly expressed putative secretory contigs, 68 were salivary gland-specific or at least -predominant transcripts and 48 of the 66 were unknown proteins. Many highly

expressed transcripts were salivary gland-specific and unknown, which suggests that the proteins have specifically evolved in the relationship between GRH and various poaceous host plants including rice. In a previous study, NcSP84 (comp13102) was detected as the most abundant protein in both secreted saliva and salivary gland extracts of GRH Oxymatrine (Hattori et al., 2012). This protein was predicted to have three EF hand motifs and was shown to exhibit calcium-binding activities (Hattori et al., 2012). The function of salivary calcium-binding protein is expected to be the binding of calcium ions that trigger the plugging response of wounded sieve tubes on insect feeding (Knoblauch et al., 2001). In addition, calcium-binding proteins are contained in the saliva of the pea aphid (Carolan et al., 2011), although proteins with similarity to NcSP84 have not been reported. Carboxylesterases are detoxification enzymes, as are cytochrome P450 monooxygenases (P450s) and glutathione S-transferases (GSTs) in insects (Després et al., 2007), and are considered to play important roles in insecticide resistance (Silva et al., 2012 and Jackson et al., 2013). However, their functions in the salivary gland remain unknown.