Abstract

Plantation forestry of Eucalyptus urophylla × Eucalyptus grandis supplies high-quality raw material for pulp, paper, wood, and energy and thereby reduces the pressures on native forests and their associated biodiversity. Nevertheless, owing to the heterozygosity of the E. urophylla × E. grandis genetic background, germplasm improvement by crossbreeding tends to be inefficient. As an alternative approach, genetic engineering of Eucalyptus can be used to effectively improve germplasm resources. From a strategic standpoint, increasing the plantation productivity and wood quality by transgenic technology has become increasingly important for forest industry. In this study, we established a fluorescence labelling method using CRISPR/Cas9 technology to obtain positive transformed progenies. The positive transformed progenies were easily obtained from the genetically modified population via fluorescence screening. This system can be used as a plant genome site-specific editing tool and may be useful for improving Eucalyptus genetic resources.

Highlights

  • Eucalyptus is a fast growing genus, with high economic value, that is frequently planted over large areas worldwide [1]

  • Eucalyptus urophylla × E. grandis, a hybrid species characterized by rapid growth and strong stress resistance, covers the largest cultivated area in China

  • Agrobacterium tumefaciens GV3102 and competent cells of Escherichia coli DH5α were purchased from Guangzhou Dingguo Biology. pHDE-mCherry and pHDE-Cas9 plasmids were a kind gift from Professor Zhao, University of California, San Diego

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Summary

Introduction

Eucalyptus is a fast growing genus, with high economic value, that is frequently planted over large areas worldwide (over 20 million hm2) [1]. Eucalyptus was introduced in China over 120 years ago. It remains a frequently-planted tree in 17 provinces (regions) of China, with a yearly timber yield of over 40 million m3. Eucalyptus provides more than 50% of the raw materials for pulpwood in China, making it an important afforestation tree species for maintaining timber safety in this country [2,3]. Eucalyptus urophylla × E. grandis, a hybrid species characterized by rapid growth and strong stress resistance, covers the largest cultivated area in China.

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