A Δ9 desaturase (SlitDes11) is associated with the biosynthesis of ester sex pheromone components in Spodoptera litura

Ya Nan Zhang; Xiao Qing Zhang; Guan Heng Zhu; Mei Yan Zheng; Qi Yan; Xiu Yun Zhu; Ji Wei Xu; Yun Ying Zhang; Peng He; Liang Sun; Subba Reddy Palli; Long Wa Zhang; Shuang Lin Dong

doi:10.1016/j.pestbp.2019.02.018

A Δ9 desaturase (SlitDes11) is associated with the biosynthesis of ester sex pheromone components in Spodoptera litura

Ya Nan Zhang, Xiao Qing Zhang, Guan Heng Zhu, Mei Yan Zheng, Qi Yan, Xiu Yun Zhu, Ji Wei Xu, Yun Ying Zhang, Peng He, Liang Sun, Subba Reddy Palli, Long Wa Zhang, Shuang Lin Dong

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

Sex pheromone biosynthesis in moths relies on the activity of multiple enzymes, including Δ9 desaturase, which plays an important role in catalyzing desaturation at the Δ9 position of the carbon chain. However, the physiological function of moth Δ9 desaturase has not been elucidated in vivo. In this study, we used the CRISPR/Cas9 system to knockout the Δ9 desaturase gene (SlitDes11) of Spodoptera litura to analyze its role in sex pheromone biosynthesis. First, through the direct injection of SlitDes11-single guide RNA (sgRNA)/Cas9 messenger RNA into newly laid eggs, gene editing was induced in around 30% of eggs 24 h after injection and was induced in 20.8% of the resulting adult moths. Second, using a sibling-crossing strategy, insects with mutant SlitDes11 (bearing a premature stop codon) were selected, and homozygous mutants were obtained in the G5 generation. Third, pheromone gland extracts of adult female homozygous SlitDes11 mutants were analyzed using Gas chromatography (GC). The results showed that titers of all three ester sex pheromone components; Z9, E11-14:Ac, Z9,E12-14:Ac, and Z9-14:Ac; were reduced by 62.40%, 78.50%, and 72.50%, respectively. This study provides the first direct evidence for the role of SlitDes11 in sex pheromone biosynthesis in S. litura, and indicates the gene could be as potential target to disrupt sexual communication in S. litura for developing a new pollution-free insecticide.

Original language	English
Pages (from-to)	152-159
Number of pages	8
Journal	Pesticide Biochemistry and Physiology
Volume	156
DOIs	https://doi.org/10.1016/j.pestbp.2019.02.018
State	Published - May 2019

Bibliographical note

Funding Information:
We thank Dr. Zhao-Qun Li (Key Laboratory of Tea Biology and Resource Utilization, Ministry of Agriculture, Tea Research Institute, Chinese Academy of Agricultural Science) for help in GC analysis. This work was supported by the National Natural Science Foundation of China ( 31501647 and 31870640 ), Natural Science Fund of Education Department of Anhui Province, China ( KJ2017A387 and KJ2017B019 ).

Funding Information:
We thank Dr. Zhao-Qun Li (Key Laboratory of Tea Biology and Resource Utilization, Ministry of Agriculture, Tea Research Institute, Chinese Academy of Agricultural Science) for help in GC analysis. This work was supported by the National Natural Science Foundation of China (31501647 and 31870640), Natural Science Fund of Education Department of Anhui Province, China (KJ2017A387 and KJ2017B019).

Publisher Copyright:
© 2019 Elsevier Inc.

Keywords

CRISPR/Cas9
Gas chromatography
Sex pheromone biosynthesis
Tobacco cutworm

ASJC Scopus subject areas

Agronomy and Crop Science
Health, Toxicology and Mutagenesis

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10.1016/j.pestbp.2019.02.018

Cite this

Zhang, Y. N., Zhang, X. Q., Zhu, G. H., Zheng, M. Y., Yan, Q., Zhu, X. Y., Xu, J. W., Zhang, Y. Y., He, P., Sun, L., Palli, S. R., Zhang, L. W., & Dong, S. L. (2019). A Δ9 desaturase (SlitDes11) is associated with the biosynthesis of ester sex pheromone components in Spodoptera litura. Pesticide Biochemistry and Physiology, 156, 152-159. https://doi.org/10.1016/j.pestbp.2019.02.018

@article{8891a3ab479b42018cc48ce4be598d40,

title = "A Δ9 desaturase (SlitDes11) is associated with the biosynthesis of ester sex pheromone components in Spodoptera litura",

abstract = "Sex pheromone biosynthesis in moths relies on the activity of multiple enzymes, including Δ9 desaturase, which plays an important role in catalyzing desaturation at the Δ9 position of the carbon chain. However, the physiological function of moth Δ9 desaturase has not been elucidated in vivo. In this study, we used the CRISPR/Cas9 system to knockout the Δ9 desaturase gene (SlitDes11) of Spodoptera litura to analyze its role in sex pheromone biosynthesis. First, through the direct injection of SlitDes11-single guide RNA (sgRNA)/Cas9 messenger RNA into newly laid eggs, gene editing was induced in around 30% of eggs 24 h after injection and was induced in 20.8% of the resulting adult moths. Second, using a sibling-crossing strategy, insects with mutant SlitDes11 (bearing a premature stop codon) were selected, and homozygous mutants were obtained in the G5 generation. Third, pheromone gland extracts of adult female homozygous SlitDes11 mutants were analyzed using Gas chromatography (GC). The results showed that titers of all three ester sex pheromone components; Z9, E11-14:Ac, Z9,E12-14:Ac, and Z9-14:Ac; were reduced by 62.40%, 78.50%, and 72.50%, respectively. This study provides the first direct evidence for the role of SlitDes11 in sex pheromone biosynthesis in S. litura, and indicates the gene could be as potential target to disrupt sexual communication in S. litura for developing a new pollution-free insecticide.",

keywords = "CRISPR/Cas9, Gas chromatography, Sex pheromone biosynthesis, Tobacco cutworm",

author = "Zhang, {Ya Nan} and Zhang, {Xiao Qing} and Zhu, {Guan Heng} and Zheng, {Mei Yan} and Qi Yan and Zhu, {Xiu Yun} and Xu, {Ji Wei} and Zhang, {Yun Ying} and Peng He and Liang Sun and Palli, {Subba Reddy} and Zhang, {Long Wa} and Dong, {Shuang Lin}",

note = "Funding Information: We thank Dr. Zhao-Qun Li (Key Laboratory of Tea Biology and Resource Utilization, Ministry of Agriculture, Tea Research Institute, Chinese Academy of Agricultural Science) for help in GC analysis. This work was supported by the National Natural Science Foundation of China ( 31501647 and 31870640 ), Natural Science Fund of Education Department of Anhui Province, China ( KJ2017A387 and KJ2017B019 ). Funding Information: We thank Dr. Zhao-Qun Li (Key Laboratory of Tea Biology and Resource Utilization, Ministry of Agriculture, Tea Research Institute, Chinese Academy of Agricultural Science) for help in GC analysis. This work was supported by the National Natural Science Foundation of China (31501647 and 31870640), Natural Science Fund of Education Department of Anhui Province, China (KJ2017A387 and KJ2017B019). Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",

year = "2019",

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doi = "10.1016/j.pestbp.2019.02.018",

language = "English",

volume = "156",

pages = "152--159",

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T1 - A Δ9 desaturase (SlitDes11) is associated with the biosynthesis of ester sex pheromone components in Spodoptera litura

AU - Zhang, Ya Nan

AU - Zhang, Xiao Qing

AU - Zhu, Guan Heng

AU - Zheng, Mei Yan

AU - Yan, Qi

AU - Zhu, Xiu Yun

AU - Xu, Ji Wei

AU - Zhang, Yun Ying

AU - He, Peng

AU - Sun, Liang

AU - Palli, Subba Reddy

AU - Zhang, Long Wa

AU - Dong, Shuang Lin

N1 - Funding Information: We thank Dr. Zhao-Qun Li (Key Laboratory of Tea Biology and Resource Utilization, Ministry of Agriculture, Tea Research Institute, Chinese Academy of Agricultural Science) for help in GC analysis. This work was supported by the National Natural Science Foundation of China ( 31501647 and 31870640 ), Natural Science Fund of Education Department of Anhui Province, China ( KJ2017A387 and KJ2017B019 ). Funding Information: We thank Dr. Zhao-Qun Li (Key Laboratory of Tea Biology and Resource Utilization, Ministry of Agriculture, Tea Research Institute, Chinese Academy of Agricultural Science) for help in GC analysis. This work was supported by the National Natural Science Foundation of China (31501647 and 31870640), Natural Science Fund of Education Department of Anhui Province, China (KJ2017A387 and KJ2017B019). Publisher Copyright: © 2019 Elsevier Inc.

PY - 2019/5

Y1 - 2019/5

N2 - Sex pheromone biosynthesis in moths relies on the activity of multiple enzymes, including Δ9 desaturase, which plays an important role in catalyzing desaturation at the Δ9 position of the carbon chain. However, the physiological function of moth Δ9 desaturase has not been elucidated in vivo. In this study, we used the CRISPR/Cas9 system to knockout the Δ9 desaturase gene (SlitDes11) of Spodoptera litura to analyze its role in sex pheromone biosynthesis. First, through the direct injection of SlitDes11-single guide RNA (sgRNA)/Cas9 messenger RNA into newly laid eggs, gene editing was induced in around 30% of eggs 24 h after injection and was induced in 20.8% of the resulting adult moths. Second, using a sibling-crossing strategy, insects with mutant SlitDes11 (bearing a premature stop codon) were selected, and homozygous mutants were obtained in the G5 generation. Third, pheromone gland extracts of adult female homozygous SlitDes11 mutants were analyzed using Gas chromatography (GC). The results showed that titers of all three ester sex pheromone components; Z9, E11-14:Ac, Z9,E12-14:Ac, and Z9-14:Ac; were reduced by 62.40%, 78.50%, and 72.50%, respectively. This study provides the first direct evidence for the role of SlitDes11 in sex pheromone biosynthesis in S. litura, and indicates the gene could be as potential target to disrupt sexual communication in S. litura for developing a new pollution-free insecticide.

AB - Sex pheromone biosynthesis in moths relies on the activity of multiple enzymes, including Δ9 desaturase, which plays an important role in catalyzing desaturation at the Δ9 position of the carbon chain. However, the physiological function of moth Δ9 desaturase has not been elucidated in vivo. In this study, we used the CRISPR/Cas9 system to knockout the Δ9 desaturase gene (SlitDes11) of Spodoptera litura to analyze its role in sex pheromone biosynthesis. First, through the direct injection of SlitDes11-single guide RNA (sgRNA)/Cas9 messenger RNA into newly laid eggs, gene editing was induced in around 30% of eggs 24 h after injection and was induced in 20.8% of the resulting adult moths. Second, using a sibling-crossing strategy, insects with mutant SlitDes11 (bearing a premature stop codon) were selected, and homozygous mutants were obtained in the G5 generation. Third, pheromone gland extracts of adult female homozygous SlitDes11 mutants were analyzed using Gas chromatography (GC). The results showed that titers of all three ester sex pheromone components; Z9, E11-14:Ac, Z9,E12-14:Ac, and Z9-14:Ac; were reduced by 62.40%, 78.50%, and 72.50%, respectively. This study provides the first direct evidence for the role of SlitDes11 in sex pheromone biosynthesis in S. litura, and indicates the gene could be as potential target to disrupt sexual communication in S. litura for developing a new pollution-free insecticide.

KW - CRISPR/Cas9

KW - Gas chromatography

KW - Sex pheromone biosynthesis

KW - Tobacco cutworm

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JO - Pesticide Biochemistry and Physiology

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ER -

A Δ9 desaturase (SlitDes11) is associated with the biosynthesis of ester sex pheromone components in Spodoptera litura

Abstract

Bibliographical note

Keywords

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