An Efficient Transformation System for Fast Production of VcCHS Transgenic Blueberry Callus and Its Expressional Analysis

Qin, Xuejing; Hu, Jing; Xu, Guohui; Song, Huifang; Zhang, Lingyun; Cao, Yibo

doi:10.3390/plants12162905

Open AccessArticle

An Efficient Transformation System for Fast Production of VcCHS Transgenic Blueberry Callus and Its Expressional Analysis

by

Xuejing Qin

^1,†,

Jing Hu

^1,†,

Guohui Xu

²,

Huifang Song

¹,

Lingyun Zhang

^1,*

and

Yibo Cao

^1,*

¹

State Key Laboratory of Efficient Production of Forest Resources, Key Laboratory of Forest Silviculture and Conservation of the Ministry of Education, The College of Forestry, Beijing Forestry University, Beijing 100083, China

²

College of Life and Health, Dalian University, Dalian 116000, China

^*

Authors to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Plants 2023, 12(16), 2905; https://doi.org/10.3390/plants12162905

Submission received: 17 July 2023 / Revised: 6 August 2023 / Accepted: 7 August 2023 / Published: 9 August 2023

(This article belongs to the Special Issue Advances in Blueberry Research)

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Abstract

The Agrobacterium tumefaciens-mediated transformation for blueberries remains less efficient than is desirable. A new leaf callus regeneration and genetic transformation system was investigated in blueberries in this study. The leaf explants of cv. ‘Legacy’ and ‘Northland’ were used to establish the stable callus induction system when placed on the woody plant medium (WPM) supplemented with 1.0 mg·L⁻¹ 2, 4-D, 0.4 mg·L⁻¹ 6-BA for 30 d; then, the callus was sub-cultured in the proliferation medium supplemented with 1.5 mg·L⁻¹ 2, 4-D, 0.4 mg·L⁻¹ 6-BA in the darkness at 25 °C every 30 days. The co-cultivation of callus with A. tumefaciens was operated on WPM plus 100 μM acetosyringone for 4 days; then, the transferred callus was grown in WPM supplemented with 1.5 mg·L⁻¹ 2,4-D, 0.4 mg·L⁻¹ 6-BA, 50 mg·L⁻¹ hygromycin, and 200 mg·L⁻¹ cefotaxime. The VcCHS transgenic blueberry callus with both GFP signal and Hyg resistance was obtained from the transformed callus of cv. ‘Northland’. The rate of GFP signal detected in the transformed callus was as high as 49.02%, which was consistent with the PCR assay. Collectively, this study provides a highly efficient genetic transformation system in blueberry callus and a powerful approach for the molecular breeding of blueberries.

Keywords: callus induction; callus proliferation; Agrobacterium tumefaciens-mediated transformation; highbush blueberry

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MDPI and ACS Style

Qin, X.; Hu, J.; Xu, G.; Song, H.; Zhang, L.; Cao, Y. An Efficient Transformation System for Fast Production of VcCHS Transgenic Blueberry Callus and Its Expressional Analysis. Plants 2023, 12, 2905. https://doi.org/10.3390/plants12162905

AMA Style

Qin X, Hu J, Xu G, Song H, Zhang L, Cao Y. An Efficient Transformation System for Fast Production of VcCHS Transgenic Blueberry Callus and Its Expressional Analysis. Plants. 2023; 12(16):2905. https://doi.org/10.3390/plants12162905

Chicago/Turabian Style

Qin, Xuejing, Jing Hu, Guohui Xu, Huifang Song, Lingyun Zhang, and Yibo Cao. 2023. "An Efficient Transformation System for Fast Production of VcCHS Transgenic Blueberry Callus and Its Expressional Analysis" Plants 12, no. 16: 2905. https://doi.org/10.3390/plants12162905

APA Style

Qin, X., Hu, J., Xu, G., Song, H., Zhang, L., & Cao, Y. (2023). An Efficient Transformation System for Fast Production of VcCHS Transgenic Blueberry Callus and Its Expressional Analysis. Plants, 12(16), 2905. https://doi.org/10.3390/plants12162905

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An Efficient Transformation System for Fast Production of VcCHS Transgenic Blueberry Callus and Its Expressional Analysis

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