文献类型：期刊文献

英文题名：Establishment and application of efficient protoplast isolation and transformation system from leaves of multi-genotype poplars

作者：Tang, Fang[1,2] Zhong, Shanchen[1] Qu, Huishan[1,3] Zhao, Shutang[1,2] Hu, Jianjun[1,2]

第一作者：Tang, Fang;唐芳

通信作者：Tang, F[1]

机构：[1]Chinese Acad Forestry, Res Inst Forestry, State Key Lab Tree Genet & Breeding, Beijing 100091, Peoples R China;[2]Nanjing Forestry Univ, Coinnovat Ctr Sustainable Forestry Southern China, Nanjing 210037, Peoples R China;[3]Dalian Univ, Coll Life & Hlth, Dalian 116622, Liaoning, Peoples R China

年份：2026

卷号：362

外文期刊名：PLANT SCIENCE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001604843600002)】；

基金：This work was supported by The Fundamental Research Funds of CAF (CAFYBB2022SY004) and The Science and Technology Innovation 2030 (2023ZD04057) .

语种：英文

外文关键词：Poplar; Genotype; Protoplast; Enzyme Solution; Purification method; Transformation efficiency

摘要：Seven widely cultivated, representative populus varieties across four taxonomic sections were employed to establish a multi-genotype protoplast isolation and transformation system, with systematic evaluation of differences in protoplast dissociation, purification, and transformation efficiency. As representatives, Populus alba (P. davidiana x P. simonii) x P. tomentosa '741' from the Sect. Leuce, Populus euramericana from the Sect. Aigeiros, Populus deltoides x P. cathayana 'Senhai 4' from the Sect. Tacamahaca, and Populus euphratica from the Sect. Turanga were selected to compare two enzyme solutions. Enzyme Solution I (1.50 % cellulase R-10 + 0.50 % macerozyme R-10) consistently yielded protoplasts with significantly higher viability than Enzyme Solution II (1.50 % cellulase R-10 + 0.50 % pectinase Y-23). Taking Populus x '741' as an example, direct washing with W5 solution outperformed sucrose density gradient centrifugation in preserving protoplast viability, demonstrating broader applicability across genotypes. Under optimized conditions (Enzyme Solution I and W5 purification), all seven genotypes produced >= 7 x 106 cells/g, though protoplast viability and GFP transformation efficiency exhibited striking variation. Sect. Leuce varieties (e.g., Populus x '741' and Populus alba var. pyramidalis 'xinjiang') showed both high viability (93.87 %, 92.16 %) and transformation efficiency (49.55 %, 46.23 %), whereas Sect. Aigeiros, Tacamahaca, and Turanga genotypes displayed lower performance. Notably, Populus x 'Senhai 4' achieved a relatively high transformation efficiency (37.14 %) despite low viability (11.28 %), highlighting the complex interplay of factors governing transformation beyond simple viability. This study establishes a robust foundation for gene editing and molecular breeding in poplars, with implications for improving transgenic efficiency across woody plant species.