植物学报2025，Vol.60Issue(6)：968-977,10.DOI:10.11983/CBB24164
大花三色堇再生体系建立
Establishment of a Regeneration System for Viola × wittrockiana
摘要
Abstract
INTRODUCTION:Viola × wittrockiana,a member of the Violaceae family,is recognized as a commercially valuable ornamental species due to its diverse flower colors and potential medicinal applications.However,its vegetative propagation via tissue culture has been limited by challenges such as low regeneration efficiency,procedural complexity in existing protocols,and genotype-dependent regeneration capacity.Previous attempts to establish regeneration sys-tems for this species have been reported,but issues including inconsistent callus differentiation,low adventitious bud formation rates,and high dependency on explant quality remain unresolved.Consequently,the development of a stable and efficient regeneration system is considered critical for enabling biotechnological advancements,including genetic transformation and large-scale propagation of elite cultivars.A systematic approach focusing on genotype screening,explant selection,and optimization of plant growth regulator combinations is required to address these limitations and facilitate the species' genetic improvement. RATIONALE:Regeneration capacity in plants is highly influenced by genotype.This study aimed to optimize the re-generation system for V.× wittrockiana by screening eight cultivars and selecting petioles as superior explants due to their higher callus differentiation potential.Key factors,including plant growth regulators(2,4-D,KT,6-BA)and carbo-hydrate sources(sucrose,maltose,trehalose),were systematically evaluated.Repeated induction cycles were em-ployed to enrich high-regeneration genotypes,enhancing overall efficiency and reproducibility. RESULTS:Petioles were identified as superior explants,exhibiting a significantly higher callus induction rate(91.90％)compared to leaves(49.65％)across eight V.× wittrockiana cultivars,with the highest differentiation efficiency(15.83％)observed in the PXP cultivar.The optimal callus induction medium was determined to be 1/2MS(sugar-free)supple-mented with 30 g·L-1 sucrose,1.5 mg·L-1 2,4-D,and 1.5 mg·L-1 KT,achieving a differentiation rate of 18.52％.For ad-ventitious bud induction,the highest regeneration efficiency(67.33±3.06)％was obtained through repeated induction cycles using 1/2MS(sugar-free)containing 30 g·L-1 trehalose,0.05 mg·L-1 2,4-D,and 3 mg·L-1 6-BA.Proliferation of adventitious buds was maximized on MS(sugar-free)medium with 30 g·L-1 trehalose,0.5 mg·L-1 2,4-D,and 1 mg·L-16-BA,yielding a proliferation coefficient of 3.29±0.22.Rooting of regenerated shoots was successfully achieved(84.44±6.93)％on 1/2MS medium containing 0.1 mg·L-1 NAA,followed by acclimatization with a survival rate exceeding 85％,validating the efficacy of the established regeneration protocol. CONCLUSION:This study established a petiole-based high-efficiency regeneration system for V.× wittrockiana through genotype screening,media optimization,and repeated induction of high-regeneration genotypes.The protocol significan-tly improved adventitious bud differentiation rates(67.33％),addressing long-standing challenges in regeneration of this species.The system provides a robust technical foundation for genetic transformation,gene function studies,and large-scale propagation,thereby facilitating the commercial development and biotechnological advancement of this economically valuable species.关键词
大花三色堇/叶柄/再生体系/组织培养/多次诱导Key words
Viola × wittrockiana/petiole/regeneration system/tissue culture/repeated induction引用本文复制引用
张汝鑫,王健,李晨荣,王童欣,黎洁,李霆格,许慧娴,李梅儿,赵莹,彭婷..大花三色堇再生体系建立[J].植物学报,2025,60(6):968-977,10.基金项目
贵州省自然科学基金(No.ZK[2022]095)、国家自然科学基金(No.32160719,No.32060365)、海口市重点科技创新项目(No.2022-013)和海南省研究生创新科研课题(No.Qhys20230-215) （No.ZK[2022]095）
