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Identification and characterization of a prophenoloxidase‐1 ( PPO 1) c DNA in the cabbage butterfly P ieris rapae L .
Author(s) -
Lu YanLi,
Li Bin,
Gong Wei,
Gao Li,
Zhang Xin,
Xue ChaoBin
Publication year - 2015
Publication title -
entomological science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 28
eISSN - 1479-8298
pISSN - 1343-8786
DOI - 10.1111/ens.12094
Subject(s) - prophenoloxidase , biology , complementary dna , biochemistry , microbiology and biotechnology , pieris rapae , open reading frame , peptide sequence , botany , larva , gene , receptor , innate immune system
Phenoloxidase ( PO ) is a melanin‐synthesizing enzyme widely distributed in animals, plants and microorganisms. It plays an important role in insect immune response. Here we report the cloning of a prophenoloxidase ( PPO ) cDNA from P ieris rapae , prophenoloxidase‐1 ( P r PPO 1 ). The full‐length c DNA consisted of 3338 bp, containing an open reading frame of 2049 bp encoding 682 amino acid (aa) residues. Two putative copper‐binding sites, a proteolytic activation site, three conserved hemocyanin domains and a thiolester motif, but no signal sequence, were found in the deduced aa sequence. A BLAST p search and neighbor‐joining analysis showed the deduced aa sequence had high identity to the published sequence of PPO 1 from other lepidopteran insects, ranging from 71.31 to 73.15%. The results of real‐time polymerase chain reaction ( PCR ) showed that the highest expression of P r PPO 1 was in the 4th instar larvae and the highest density of oenocytoids was observed in the 4th instar larvae also. P r PPO 1 transcripts in the 4th instar larvae were significantly decreased at 6 h and 12 h with B eauveria bassiana infection; however, they were sharply increased at 72 h. P r PPO 1 transcriptional level in the 5th instar larvae showed no significant changes compared with the control at 4 h with quercetin injection; however it was significantly increased at 12 h. The expression level of P r PPO 1 was mainly related to the treatment time of the quercetin, but not the dose.