摘要
为建立大花三色堇(Viola×wittrockiana)植株再生体系,以其8个品种的叶片和叶柄为实验材料,筛选并确定了再生率高的品种和部位。结果表明,PXP是供试品种中用于大花三色堇再生体系建立的最佳品种,其叶柄是最佳外植体。以品种PXP的叶柄为外植体,进行再生体系优化。结果表明,愈伤组织诱导最适培养基为1/2MS(不含糖)+30 g·L^(-1)蔗糖+1.5 mg·L^(-1)2,4-D+1.5 mg·L^(-1) KT,不定芽诱导最优培养基为1/2MS(不含糖)+30 g·L^(-1)海藻糖+0.05 mg·L^(-1)2,4-D+3 mg·L^(-1)6-BA,再生苗经多次诱导分化,再生率可达(67.33±3.06)%。不定芽增殖最优培养基为MS(不含糖)+30 g·L^(-1)海藻糖+0.5 mg·L^(-1)2,4-D+1 mg·L^(-1)6-BA,增殖系数为3.29±0.22。再生苗在配方为1/2MS(不含糖)+30 g·L^(-1)海藻糖+0.1 mg·L^(-1) NAA的生根培养基上培养2周后开始生根,生根率为(84.44±6.93)%。该研究建立了分化率较高的大花三色堇再生体系,解决了大花三色堇不定芽分化困难的难题,为其育种改良和优良品种快繁提供了技术支持。
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 prop-agation 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 systems for this species have been reported,but issues including inconsistent callus differentiation,low adventitious bud forma-tion 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 rege-neration 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 carbohydrate sources(sucrose,maltose,trehalose),were systematically evaluated.Repeated induction cycles were employed 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^(-1)6-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.
作者
张汝鑫
李晨荣
王童欣
黎洁
李霆格
许慧娴
李梅儿
赵莹
彭婷
王健
Ruxin Zhang;Chenrong Li;Tongxin Wang;Jie Li;Tingge Li;Huixian Xu;Meier Li;Ying Zhao;Ting Peng;Jian Wang(Key Laboratory of Genetics and Germplasm Innovation of Tropical Forest Trees and Ornamental Plants,Ministry of Educa-tion/Key Laboratory of Germplasm Resources Biology of Tropical Special Ornamental Plants of Hainan/School of Tropical Agriculture and Forestry,Hainan University,Haikou 570228,China;College of Agriculture,Guizhou University,Guiyang 550025,China;College of Life Science and Technology,Huazhong Agricultural University,Wuhan 430070,China)
出处
《植物学报》
北大核心
2025年第6期968-977,共10页
Chinese Bulletin of Botany
基金
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)
海南省研究生创新科研课题(No.Qhys20230-215)。
关键词
大花三色堇
叶柄
再生体系
组织培养
多次诱导
Viola×wittrockiana
petiole
regeneration system
tissue culture
repeated induction
