Characterization of an Intradiol Ring-Cleavage Dioxygenase Gene Associated With Abamectin Resistance in Tetranychus cinnabarinus (Acari: Tetranychidae)

2019 ◽  
Vol 112 (4) ◽  
pp. 1858-1865
Author(s):  
Zhifeng Xu ◽  
Peilin Liu ◽  
Yuan Hu ◽  
Jia Hu ◽  
Cuicui Qi ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Spider Mite ◽  
Gene Sequence ◽  
Ring Cleavage ◽  
Tetranychus Cinnabarinus ◽  
Limited Data ◽  
Dioxygenase Gene ◽  
Carmine Spider Mite ◽  
Rnai Technique

AbstractTetranychus cinnabarinus (Boisduval), i.e., carmine spider mite, is a worldwide pest that can cause serious damage to plants. Problems of resistance have arisen since abamectin usage in the control of T. cinnabarinus. Unfortunately, there are only limited data on the extent of this problem. To understand the development of abamectin resistance in the carmine spider mite, we prokaryotically expressed an intradiol ring-cleavage dioxygenase (ID-RCD) gene sequence, TcID-RCD1, which had a significant upregulated expression of over 7.7 times in an abamectin-resistant strain (AbR) when compared with that of a susceptible strain (SS). The crude enzyme activity also indicated that the AbR had a higher activity than that exhibited in SS. When susceptible individuals were treated with abamectin, TcID-RCD1 was also overexpressed. Furthermore, using the RNA interference (RNAi) technique, TcID-RCD1 was successfully knocked down, with the expression level decreasing significantly to approximately 39% in the SS strain compared with the control. And the mortality of mites feeding on dsTcID-RCD1 increased significantly when treated with abamectin. These results strongly suggest that TcID-RCD1 is involved in abamectin resistance in T. cinnabarinus.

scholarly journals Characterization of the Gallate Dioxygenase Gene: Three Distinct Ring Cleavage Dioxygenases Are Involved in Syringate Degradation by Sphingomonas paucimobilis SYK-6

2005 ◽  
Vol 187 (15) ◽  
pp. 5067-5074 ◽  
Author(s):  
Daisuke Kasai ◽  
Eiji Masai ◽  
Keisuke Miyauchi ◽  
Yoshihiro Katayama ◽  
Masao Fukuda
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequences ◽  
Terminal Region ◽  
Ring Cleavage ◽  
Sphingomonas Paucimobilis ◽  
Acid Protein ◽  
Demethylation Pathway ◽  
Dioxygenase Gene ◽  
Amino Acid Protein

ABSTRACT Sphingomonas paucimobilis SYK-6 converts vanillate and syringate to protocatechuate (PCA) and 3-O-methylgallate (3MGA) in reactions with the tetrahydrofolate-dependent O-demethylases LigM and DesA, respectively. PCA is further degraded via the PCA 4,5-cleavage pathway, whereas 3MGA is metabolized via three distinct pathways in which PCA 4,5-dioxygenase (LigAB), 3MGA 3,4-dioxygenase (DesZ), and 3MGA O-demethylase (LigM) are involved. In the 3MGA O-demethylation pathway, LigM converts 3MGA to gallate, and the resulting gallate appears to be degraded by a dioxygenase other than LigAB or DesZ. Here, we isolated the gallate dioxygenase gene, desB, which encodes a 418-amino-acid protein with a molecular mass of 46,843 Da. The amino acid sequences of the N-terminal region (residues 1 to 285) and the C-terminal region (residues 286 to 418) of DesB exhibited ca. 40% and 27% identity with the sequences of the PCA 4,5-dioxygenase β and α subunits, respectively. DesB produced in Escherichia coli was purified and was estimated to be a homodimer (86 kDa). DesB specifically attacked gallate to generate 4-oxalomesaconate as the reaction product. The Km for gallate and the V max were determined to be 66.9 ± 9.3 μM and 42.7 ± 2.4 U/mg, respectively. On the basis of the analysis of various SYK-6 mutants lacking the genes involved in syringate degradation, we concluded that (i) all of the three-ring cleavage dioxygenases are involved in syringate catabolism, (ii) the pathway involving LigM and DesB plays an especially important role in the growth of SYK-6 on syringate, and (iii) DesB and LigAB are involved in gallate degradation.

Sign in / Sign up

Export Citation Format

Share Document