农杆菌介导的建兰品种市长红遗传转化体系的建立

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中图分类号：S682.310.1 文献标志码：A 文章编号：1004-390X（2025）04-0111-09

Establishment of Agrobacterium-mediated Genetic Transformation System for Cymbidium ensifolium ‘Shizhanghong'

ZHOU Bin1²，CAO Hua²，LU Lin²，ZHANG Yuying³，LI Han²，LI Donghui1

[1. College of Landscape and Horticulture， Yunnan Agricultural University， Kunming 65o201， China; 2.National Engineering Research Center for Ormamental Horticulture，Flower Research Institute， Yunnan Academy of Agricultural Sciences， Kunming 650205， China; 3.Forestry （Flower） Industry Development Center， Zhouning County，Ningde City， Fujian Province， Zhouning 355400， China]

Abstract： [Purpose] To explore the genetic transformation system of Cymbidium ensifolium ‘Shizhanghong' mediated by Agrobacterium. [Methods] The rhizomes of C . ensifolium‘Shizhanghong’ obtained from single pseudobulb propagation were used as experimental materials. Agrobacterium strain， concentration of acetosyringone， and mass concentration of antibiotics were screened. The main factors affecting the genetic transformation efficiency and regeneration efficiency of C ensifolium‘Shizhanghong' mediated by Agrobacterium were studied. [Results] The optimal strain for genetic transformation of C. ensifolium‘Shizhanghong’ was EHA1o5， the optimal concentration of acetosyringone was ， and the optimal mass concentration of hygromycin was 40mg/L . To ensure a high transformation rate and high regeneration rate of C . ensifolium ‘Shizhanghong'，the optimal infection system was： after 1O days of pre-culture， the rhizomes were shaken in Agrobacterium infection solution （OD600=0.4-0.6） ）for 20 minutes，and then co-cultured in the dark for three days. Through the screening of the optimal genetic transformation conditions， the transgenic positive plants of C. ensifolium ‘Shizhanghong’ were identified by green fluorescent protein， antibiotic screening， PCR and qRT-PCR detection. Finally， 17 transgenic positive plants of C .ensifolium‘Shizhanghong' were obtained from 18O rhizomes， with a transformation rate of 9.44% .[Conclusion] This study establishes a genetic transformation system mediated by Agrobacterium for C. ensifolium ‘Shizhanghong'，laying the foundation for transgenic breeding of this orchid cultivar.

Keywords： Cymbidium ensifolium ‘Shizhanghong'; rhizome; Agrobacterium infection; genetic transformation; green fluorescent protein （GFP）

建兰品种市长红（Cymbidiumensifolium‘Shizh-anghong'）是兰科（Orchidaceae）兰属（Cymbidium）地生草本植物，为纯种建兰，因其奇特的花形、独特的花姿、叶艺以及浓郁的花香深受广大消费者喜爱[]。（剩余12171字）