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Characterization of carboxylesterase
PxαE8
and its role in multi-insecticide resistance in
Plutella xylostella
(L.)
LI Ran, SUN Xi, LIANG Pei, GAO Xi-wu
2022, 21 (
6
): 1713-1721. DOI:
10.1016/S2095-3119(21)63636-5
Abstract
(
212
)
PDF in ScienceDirect
Carboxylesterases (CarEs) were considered as important phase-I detoxifying enzymes which participated in detoxification of different types of insecticides. Up-regulation of CarE genes has been proved playing a major role in insecticide resistance in many pest insects, but its involvement in resistance to insecticides in
Plutella xylostella
has been rarely reported. In this study, a CarE cDNA named
PxαE8
was identified in
P. xylostella
, which has an open reading frame of 1599 nucleotides and putatively encodes 532 amino acids. The investigation of spatial and temporal expression profiles of
PxαE8
revealed that it was expressed in all developmental stages and body part/tissues, especially in larvae, adults as well as fat body, malpighian tubule and hemolymph of larvae. Further, the relative expression of
PxαE8
in two multi-resistant field populations, Hainan (HN) and Guangdong (GD) populations, was found 24.4- and 15.5-fold higher than that in susceptible population, respectively. Knockdown of
PxαE8
by RNAi dramatically increased the mortalities of larvae of HN population treated with LC
50
of beta-cypermethrin and phoxim by 25.3 and 18.3%, respectively. These results suggested that up-regulation of
PxαE8
was involved in resistance to both beta-cypermethrin and phoxim in
P. xylostella
, which shed light on further understanding of molecular mechanisms of multi-insecticide-resistance in
P. xylostella
and other pest insects.
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The impact of allelochemicals on the differential expression of symbiotic bacteria in cotton aphids
LIU Ying, LIANG Ping-zhuo, LI Fen, MA Kang-sheng, CHEN Xue-wei, CHEN An-qi, LIANG Pei, GAO Xi-wu
2018, 17 (
08
): 1815-1821. DOI:
10.1016/S2095-3119(17)61838-0
Abstract
(
328
)
PDF in ScienceDirect
Insects have developed a good adaptive mechanism in response to environmental stresses in the long-term evolution.
They have developed a helpful metabolism system to resist plant allelochemicals. Insects also harbor different kinds of
symbiotic bacteria, which provide them a competitive advantage. Here, using cotton aphid as an example, we investigated
the effects of four plant allelochemicals on the differential expression of symbiotic bacteria based on transcriptome data.
We also studied the composition of symbiotic bacteria and function on pathway level in three kinds of aphids. We found that
the bacteria have a significant role in resisting the plant allelochemicals stress and host plant selection by aphids. These
results should be useful to investigate the environmental adaption mechanism of aphids in the view of symbiotic bacteria.
These results would offer a new insight for improving strategy of aphids and developing new pest control systems.
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cDNA cloning and characterization of the carboxylesterase
pxCCE016b
from the diamondback moth,
Plutella xylostella
L.
HU Zhen-di, FENG Xia, LIN Qing-sheng, CHEN Huan-yu, LI Zhen-yu, YIN Fei, LIANG Pei, GAO Xi-wu
2016, 15 (
05
): 1059-1068. DOI:
10.1016/S2095-3119(15)61278-3
Abstract
(
1668
)
PDF in ScienceDirect
Carboxylesterase is a multifunctional superfamily and can be found in almost all living organisms. As the metabolic enzymes, carboxylesterases are involved in insecticides resistance in insects for long time. In our previous studies, the enhanced carboxylesterase activities were found in the chlorantraniliprole resistance strain of diamondback moth (DBM). However, the related enzyme gene of chlorantraniliprole resistance has not been clear in this strain. Here, a full-length cDNA of carboxylesterase
pxCCE016b
was cloned and exogenously expressed in Escherichia coli at the first time, which contained a 1 693 bp open reading frame (ORF) and encoded a protein of 542 amino acids. Sequence analysis showed that this cDNA has a predicted mass of 61.56 kDa and a theoretical isoelectric point value of 5.78. The sequence of deduced amino acid possessed the classical structural features: a type-B carboxylesterase signature 2 (EDCLYLNVYTK), a type-B carboxylesterase serine active site (FGGDPENITIFGESAG) and the catalytic triad (Ser186, Glu316, and His444). The real-time quantitative PCR (qPCR) analysis showed that the expression level of the
pxCCE016b
was significantly higher in the chlorantraniliprole resistant strain than in the susceptible strain. Furthermore,
pxCCE016b
was highly expressed in the midgut and epidermis of the DBM larvae. When the 3rd-instar larvae of resistant DBM were exposed to abamectin, alpha-cypermethrin, chlorantraniliprole, spinosad, chlorfenapyr and indoxacarb insecticides, the up-regulated expression of
pxCCE016b
was observed only in the group treated by chlorantraniliprole. In addition, recombinant vector pET-
pxCCE016b
was constructed with the most coding region (1 293 bp) and large number of soluble recombinant proteins (less than 48 kDa) were expressed successfully with prokaryotic cell. Western blot analysis showed that it was coded by
pxCCE016b
. All the above findings provide important information for further functional study, although we are uncertainty whether the
pxCCE016b
gene is actually involved in chlorantraniliprole resistance.
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Biochemical Mechanism of Chlorantraniliprole Resistance in the Diamondback Moth, Plutella xylostella Linnaeus
HU Zhen-di, FENG Xia, LIN Qing-sheng, CHEN Huan-yu, LI Zhen-yu, YIN Fei, LIANG Pei , GAO Xi-wu
2014, 13 (
11
): 2452-2459. DOI:
10.1016/S2095-3119(14)60748-6
Abstract
(
1337
)
PDF in ScienceDirect
The insecticide chlorantraniliprole exhibits good efficacy and plays an important role in controlling the diamondback moth, Plutella xylostella Linnaeus. However, resistance to chlorantraniliprole has been observed recently in some field populations. At present study, diamondback moths with resistance to chlorantraniliprole (resistant ratio (RR) was 82.18) for biochemical assays were selected. The assays were performed to determine potential resistance mechanisms. The results showed that the selected resistant moths (GDLZ-R) and susceptible moth could be synergized by known metabolic inhibitors such as piperonyl butoxide (PBO), triphenyl phosphate (TPP) and diethyl-maleate (DEM) at different levels (1.68-5.50-fold and 2.20-2.89-fold, respectively), and DEM showed the maximum synergism in both strains. In enzymes assays, a high level of glutathione-S-transferase (GST) was observed in the resistant moth, in contrast, moths that are susceptible to the insecticide had only 1/3 the GST activity of the resistant moths. The analysis of short-term exposure of chlorantraniliprole on biochemical response in the resistant strain also showed that GST activity was significantly elevated after exposure to a sub-lethal concentration of chlorantraniliprole (about 1/3 LC50, 12 mg L-1) 12 and 24 h, respectively. The results show that there is a strong correlation between the enzyme activity and resistance, and GST is likely the main detoxification mechanism responsible for resistance to chlorantraniliprole in P. xylostella L., cytochrome P450 monooxygenase (P450) and carboxy-lesterase (CarE) are involved in to some extent.
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Omethoate-Induced Changes of (+)-d-Cadinene Synthase Activity and Gossypol Content in Cotton Seedlings
YANG Wen-ling, SHI Xue-yan, ZHONG Li, LIANG Pei, SONG Dun-lun, GAO Xi-wu
2012, 12 (
10
): 1682-1690. DOI:
10.1016/S1671-2927(00)8701
Abstract
(
1457
)
PDF in ScienceDirect
The gene expression and activity of (+)-d-cadinene synthase during cotton development and in response to stress, as well as the spatial and temporal pattern of sesquiterpene biosynthesis, constitute one of chemical defense mechanisms in cotton plants. In order to explore the effects of omethoate on the cotton defense in relation to (+)-d-cadinene synthase and gossypol, effects of omethoate treatments on activity of (+)-d-cadinene synthase and gossypol content in cotton seedlings were investigated. Cotton seedlings treated with 400 mg L-1 omethoate exhibited a significant decrease in the specific activity of (+)-d-cadinene synthase from 12 to 120 h after treating when compared to the untreated control; significantly lower (+)-d-cadinene synthase activity was also observed in cotton seedlings treated with 200 mg L-1 omethoate from 72 to 120 h after treating; but for cotton seedlings treated with 100 mg L-1 omethoate, from 12 to 120 h after treating, no significant changes were observed for activity of (+)-d-cadinene synthase. The gossypol content in cotton seedlings treated with 100, 200 or 400 mg L-1 omethoate for different time periods showed no significant changes compared to that of the control. These results indicated that the activity of (+)-d-cadinene synthase in cotton seedlings in responses to exposure of omethoate at three concentrations for different time periods followed dose- and time-dependent responses to omethoate exposure. With omethoate as a chemical stress factor for cotton seedlings, the cotton defense in relation to the activity of (+)-d-cadinene synthase in cotton seedlings may be affected by omethoate application.
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