Agrobacterium-mediated genetic transformation of Chinese chestnut (Castanea mollissima Blume)

• Published in: Plant cell, tissue and organ culture Vol. 140; no. 1; pp. 95 - 103
• Main Authors: Sun, Zhi-lin, Li, Xiao, Zhou, Wan, Yan, Jun-di, Gao, Yue-rong, Li, Xiao-wei, Sun, Jia-chen, Fang, Ke-feng, Zhang, Qing, Xing, Yu, Qin, Ling, Cao, Qing-qin
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 2020; Springer Nature B.V
• Subjects: Biomedical and Life Sciences; Biotechnology; Breeding; Castanea mollissima; Chestnut; Chimeras; Genetic improvement; Genetic transformation; Germplasm; Life Sciences; Original Article; Plant Genetics and Genomics; Plant Pathology; Plant Physiology; Plant Sciences; Chinese chestnut; Regeneration; mediated transformation; Somatic embryo
• ISSN: 0167-6857; 1573-5044
• DOI: 10.1007/s11240-019-01713-4

Chinese chestnut ( Castanea mollissima ) is an important germplasm resource for the breeding of Castanea species worldwide with vital ecological and economic value. Biotechnology overcomes the limitations of traditional breeding and accelerates germplasm improvement. However, a genetic transformation system for Chinese chestnut has not yet been established. In this study, a stable and efficient Agrobacterium -mediated genetic transformation method for Chinese chestnut is described. Embryogenic calli of C. mollissima cv. ‘Yanshanhongli’ were used as the target material. The sensitivity of embryogenic calli to kanamycin was determined, whereby the proliferation of non-transformed calli was completely inhibited at 180 mg/L. Antibiotic inhibition results for Chinese chestnut embryogenic calli showed that 50 mg/L cefotaxime and 500 μM timentin completely inhibited the growth of Agrobacterium tumefaciens but did not affect the normal growth of Chinese chestnut embryogenic calli. When embryogenic calli were co-cultured for 2 days with Agrobacterium tumefaciens strain AGL1 harboring the PBI121-EGFP plasmid, an embryogenic callus transformation efficiency of 4.55% was obtained, and two transgenic chimera were acquired. This Agrobacterium -mediated transformation system for Chinese chestnut provides a fundamental platform for genetic improvement of core germplasm and for further verification of gene function. Key message A method of Agrobacterium -mediated transformation was established for Chinese chestnut, providing a basis for genetic improvement of core germplasm and an important platform for further verification of gene function in Chinese chestnut.