Effects of sublethal doses of dinotefuran on genes expression and enzymes activities related to immune and detoxification in foragers of Apis mellifera ligustica

doi:10.3969/j.issn.1004-1524.2023.03.10

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (3): 575-581.DOI: 10.3969/j.issn.1004-1524.2023.03.10

• Animal Science • Previous Articles Next Articles

Effects of sublethal doses of dinotefuran on genes expression and enzymes activities related to immune and detoxification in foragers of Apis mellifera ligustica

ZHANG Yuhao¹(), MA Weihua², LIU Jinjia¹, MA Xiumei¹, JIANG Yusuo¹^,^*()

1. College of Animal Sciences, Shanxi Agricultural University, Taigu 030801, Shanxi, China
2. College of Horticulture, Shanxi Agricultural University, Taigu 030801, Shanxi, China

Received:2021-09-06 Online:2023-03-25 Published:2023-04-07

Abstract

Abstract:

To determine sublethal doses of dinotefuran to Apis mellifera ligustica and explore the effects of sublethal doses of dinotefuran on the expression of immune detoxification genes and enzyme activities on Apis mellifera ligustica, in this study, the feeding trials was used to determine the virulence curve of dinotefuran to Apis mellifera ligustica. The effects of LC₂₀ and LC₅₀ dinotefuran on the relative expressions of Abaecin, Apidaecins, Defensin-1, Hymenoptaecin, AChE-2, PPO, GST and Cyp450 mRNA were investigated by qRT-PCR. Determination of changes in the activities of PPO, AChE, SOD, GST, CAT in Apis mellifera ligustica. The LC₂₀ and LC₅₀ of dinotefuran on Apis mellifera ligustica determined by feeding trials were 0.458 mg·L^-1 and 0.988 mg·L^-1. Compared with the control group, the expression levels of PPO, Apidaecins, Defensin-1, Hymenoptaecin genes were significantly (P<0.05) up-regulated in the LC₂₀ group. Apidaecins, In LC₅₀ group, the expression of AChE-2, PPO, GST, Cyp450, Apidaecins, Defensin-1 showed a significant up-regulation trend, and the expression of Hymenoptaecin and Abaecin showed a significant down-regulation trend. In additon, the expression levels of Abaecin and Hymenoptaecin in the LC₅₀ group were significantly lower than those in the LC₂₀ group. The expression levels of AChE-2, PPO, GST, Cyp450, Apidaecins in the LC₅₀ group were significantly higher than those in the LC₂₀ group. Compared with the control group, the activities of AChE, CAT and SOD of LC₂₀ group decreased significantly. Compared with the control group, the activities of AChE, PPO, CAT and SOD of LC₅₀ group decreased significantly. In conclusion, dinotefuran inhibits gene expression of Abaecin and Hymenoptaecin, and has an inducing effect on Apidaecins, Defensin-1, AChE-2, PPO, GST, Cyp450 affects the normal immune system of Apis mellifera ligustica. The decline of AChE, PPO, CAT and SOD enzyme activities indicates the damage of dinotefuran to the detoxification system of Apis mellifera ligustica. Our stduy provides a further basis for further research on the effects of sublethal dose dinotefuran on the metabolism and physiology of Apis mellifera ligustica.

Key words: Apis mellifera ligustica, dinotefuran, sublethal effect, immune detoxification, enzyme activity

CLC Number:

S893.3

ZHANG Yuhao, MA Weihua, LIU Jinjia, MA Xiumei, JIANG Yusuo. Effects of sublethal doses of dinotefuran on genes expression and enzymes activities related to immune and detoxification in foragers of Apis mellifera ligustica[J]. Acta Agriculturae Zhejiangensis, 2023, 35(3): 575-581.

Figures/Tables 4

Table 1 Sequence information of primer pairs used in this experiment

基因名称 Gene name	上游引物 Forward primer (5'→3')	下游引物 Reverse primer (5'→3')
Abaecin	ATCTTCGCACTACTCGCCAC	CTGACCAGGAAACGTTGGAA
Apidaecin	AGATTGTCGATCGGTCCACG	TCTGGTTCGAGCATGCTACC
Defensin-1	GTTGAGGATGAATTCGAGCC	TTAACCGAAACGTTTGTCCC
Hymenoptaecin	ATATCCCGACTCGTTTCCGA	TCCCAAACTCGAATCCTGCA
AChE-2	GACGCGAAGACCATATCCGT	TCTGTGTCCTTGAAGTCCGC
PPO	ATGTGGATGGCCGCAACATA	CGCCATATTTCCGGTGAGGA
GST	TGCATATGCTGGCATTGATT	TCCTCGCCAAGTATCTTGCT
Cyp450	CAAAATGGTGTTCTCCTTACCG	ATGGCAACCCATCACTGC
β-actin	ATGCCAACACTGTCCTTTCTGG	GACCCACCAATCCATACGGA

Fig.1 Toxicity regression curve of dinotefuran acute toxicity to Apis mellifera ligustica y, Common logarithm value of dinotefuran concentration; x, Probit value to mortality.

Fig.2 Expression patterns of Hymenoptaecin, Abaecin, Apidaecin, Defensin-1, AChE-2,PPO,GST and Cyp450 mRNA in different concentrations of dinotefuran on Apis mellifera ligustica CK, 50% sucrose solution; LC20, 0.458 mg·L-1 dinotefuran sucrose solution; LC50, 0.988 mg·L-1 dinotefuran sucrose solution. The same as below. Bars marked without the same letters indicate significant (P<0.05) difference within treatments under the same gene.

Fig.3 Immune and detoxification enzyme activity in foragers of Apis mellifera ligustica after LC20 and LC50 dinotefuran stress Bars marked without the same letters indicate significant (P<0.05) difference within treatments.

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Effects of sublethal doses of dinotefuran on genes expression and enzymes activities related to immune and detoxification in foragers of Apis mellifera ligustica

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