Transformation and functional verification of Cry5Aa in cotton

Abstract Most of the cotton bollworm-resistant genes applied in cotton are more than 20 years and they all belong to Cry1Ab/c family, but the insect-resistant effects of Cry5Aa on cotton were rarely reported. The possible risk of resistance is increasing. The study synthesized a novel bollworm-resis...

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Autores principales:	Shihao Zhao, Feng Wang, Qiuping Zhang, Jiayi Zou, Zhangshu Xie, Kan Li, Jingyi Li, Bo Li, Wen He, Jinxiang Chen, Yunxin He, Zhonghua Zhou
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3d49aa06da58470e94ba908c2dfd2da6
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spelling	oai:doaj.org-article:3d49aa06da58470e94ba908c2dfd2da62021-12-02T14:06:11ZTransformation and functional verification of Cry5Aa in cotton10.1038/s41598-021-82495-82045-2322https://doaj.org/article/3d49aa06da58470e94ba908c2dfd2da62021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82495-8https://doaj.org/toc/2045-2322Abstract Most of the cotton bollworm-resistant genes applied in cotton are more than 20 years and they all belong to Cry1Ab/c family, but the insect-resistant effects of Cry5Aa on cotton were rarely reported. The possible risk of resistance is increasing. The study synthesized a novel bollworm-resistant gene Cry5Aa artificially based on preferences of cotton codon. The new gene was transferred to cotton through the method of pollen tube pathway. The transgenic strains were identified by kanamycin test in field and laboratory PCR analysis. Meanwhile, an insect resistance test was conducted by artificial bollworm feeding with transgenic leaves and GK19 was used as a control in this study. Results showed that rate of positive transgenic strains with kanamycin resistance in the first generation (T1), the second generation (T2) and the third generation (T3) respectively were 7.76%, 73.1% and 95.5%. However, PCR analysis showed that the positive strain rate in T1, T2 and T3 were 2.35%, 55.8% and 94.5%, respectively. The resistant assay of cotton bollworm showed that the mortality rate of the second, third and fourth instar larva feed by the transgenic cotton leaves, were 85.42%, 73.35% and 62.79%, respectively. There was a significant difference between transgenic plant of Cry5Aa and GK19 in insect resistance. Finally, we also conducted the further analysis of gene expression patterns, gene flow and the effect on non-target pest in the study. The results showed that Cry5Aa gene had less environmental impact, and Cry5Aa has been transferred successfully and expressed stably in cotton. Therefore, the novel bollworm resistance gene can partially replace the current insect-resistance gene of Lepidoptera insects.Shihao ZhaoFeng WangQiuping ZhangJiayi ZouZhangshu XieKan LiJingyi LiBo LiWen HeJinxiang ChenYunxin HeZhonghua ZhouNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Shihao Zhao Feng Wang Qiuping Zhang Jiayi Zou Zhangshu Xie Kan Li Jingyi Li Bo Li Wen He Jinxiang Chen Yunxin He Zhonghua Zhou Transformation and functional verification of Cry5Aa in cotton
description	Abstract Most of the cotton bollworm-resistant genes applied in cotton are more than 20 years and they all belong to Cry1Ab/c family, but the insect-resistant effects of Cry5Aa on cotton were rarely reported. The possible risk of resistance is increasing. The study synthesized a novel bollworm-resistant gene Cry5Aa artificially based on preferences of cotton codon. The new gene was transferred to cotton through the method of pollen tube pathway. The transgenic strains were identified by kanamycin test in field and laboratory PCR analysis. Meanwhile, an insect resistance test was conducted by artificial bollworm feeding with transgenic leaves and GK19 was used as a control in this study. Results showed that rate of positive transgenic strains with kanamycin resistance in the first generation (T1), the second generation (T2) and the third generation (T3) respectively were 7.76%, 73.1% and 95.5%. However, PCR analysis showed that the positive strain rate in T1, T2 and T3 were 2.35%, 55.8% and 94.5%, respectively. The resistant assay of cotton bollworm showed that the mortality rate of the second, third and fourth instar larva feed by the transgenic cotton leaves, were 85.42%, 73.35% and 62.79%, respectively. There was a significant difference between transgenic plant of Cry5Aa and GK19 in insect resistance. Finally, we also conducted the further analysis of gene expression patterns, gene flow and the effect on non-target pest in the study. The results showed that Cry5Aa gene had less environmental impact, and Cry5Aa has been transferred successfully and expressed stably in cotton. Therefore, the novel bollworm resistance gene can partially replace the current insect-resistance gene of Lepidoptera insects.
format	article
author	Shihao Zhao Feng Wang Qiuping Zhang Jiayi Zou Zhangshu Xie Kan Li Jingyi Li Bo Li Wen He Jinxiang Chen Yunxin He Zhonghua Zhou
author_facet	Shihao Zhao Feng Wang Qiuping Zhang Jiayi Zou Zhangshu Xie Kan Li Jingyi Li Bo Li Wen He Jinxiang Chen Yunxin He Zhonghua Zhou
author_sort	Shihao Zhao
title	Transformation and functional verification of Cry5Aa in cotton
title_short	Transformation and functional verification of Cry5Aa in cotton
title_full	Transformation and functional verification of Cry5Aa in cotton
title_fullStr	Transformation and functional verification of Cry5Aa in cotton
title_full_unstemmed	Transformation and functional verification of Cry5Aa in cotton
title_sort	transformation and functional verification of cry5aa in cotton
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/3d49aa06da58470e94ba908c2dfd2da6
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_version_	1718392014522810368

Transformation and functional verification of Cry5Aa in cotton

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