滇牡丹花粉粒发育细胞学观察与雄蕊愈伤组织诱导及增殖培养  

Pollen Cytology,Stamen Callus Induction,and Seedling Propagation of Paeonia delavayi

作　　者：牟亚萍 张平芳 杜春 李进 刘子榕 王娟 MOU Yaping;ZHANG Pingfang;DU Chun;LI Jin;LIU Zirong;WANG Juan(College of Landscape Architecture,Southwest Forestry University,Kunming,Yunnan 650224,China;Key Laboratory of Biodiversity Conservation in Southwest China,National Forestry and Grassland Bureau,Kunming,Yunnan 650224,China;Forestry College,Southwest Forestry University,Kunming,Yunnan 650224,China;Green Development Research Institute,Southwest Forestry University,Kunming,Yunnan 650224,China)

机构地区：[1]西南林业大学园林园艺学院,云南昆明650224 [2]西南地区生物多样性保育国家林业和草原局重点实验室,云南昆明650224 [3]西南林业大学林学院,云南昆明650224 [4]西南林业大学绿色发展研究院,云南昆明650224

出　　处：《福建农业学报》2023年第9期1064-1072,共9页Fujian Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(32060089);云南省重大基础专项生物资源数字化开发应用项目(202002AA100007);云南省千人计划“高层次人才”专项(2019);云南省万人计划“云岭产业技术领军人才”专项(2018-212)。

摘　　要：【目的】研究滇牡丹花蕾与花粉粒发育间的相关性,建立外观形态与花粉粒主导发育时期间的关系,为后期取材提供参考;筛选出滇牡丹雄蕊愈伤组织、增殖和分化诱导的最适培养基,以期建立滇牡丹雄蕊的再生体系。【方法】以红色滇牡丹硬蕾期和透色期的花蕾为研究材料,通过石蜡切片加光学显微观察的途径确定各阶段花粉粒的主导发育时期;结合MS固体培养基,研究不同植物生长调节剂及其质量浓度对滇牡丹雄蕊愈伤组织诱导、增殖和分化的影响。【结果】(1)红色滇牡丹花蕾直径10~17 mm且尚未透色的Y2−Y4阶段,其花粉粒主导发育时期是单核中期;花蕾直径12~15 mm且透色面积小于30%的T1−T2阶段为单核靠边期,花蕊直径15~19 mm且透色面积大于30%的T3−T4阶段为双核期。(2)滇牡丹雄蕊的最佳消毒方式为花蕾经流水冲洗30 min后,使用1%次氯酸钠消毒8 min。(3)滇牡丹雄蕊的最佳诱导时期为单核靠边期。(4)雄蕊愈伤组织诱导适宜的培养基为MS+0.2 mg·L^(−1)NAA+0.5 g·L^(−1)PVP;最适增殖培养基为MS+0.5 mg·L^(−1)NAA+0.25 mg·L^(−1)6-BA+0.5 mg·L^(−1)GA3+1 g·L^(−1)PVP。【结论】明确了花蕾大小和外观透色形态与各阶段花粉粒的主导发育时期的关系,筛选出红花滇牡丹雄蕊愈伤组织诱导与增殖适宜的培养基和培养条件,为滇牡丹组织培养技术的深入研究提供基础。【Objective】Developments of flower buds and pollen grains,optimum medium for stamen callus induction,and proper conditions for seedling propagation and differentiation of Paeonia delavayi were investigated.【Method】Dominant pollen grain development stages were examined under an optical microscope on the paraffin sections of red P.delavayi hard buds with visible color.On an MS solid medium,various plant growth regulators in different concentrations were added to determine the optimal formulation for the induction,proliferation,and differentiation of stamen callus.【Result】(1)The prevailing stage of pollen grain development for the red P.delavayi,Y2-Y4,occurred in the mid-mononuclear period when the bud diameter was 10-17 mm but the color had yet to show.At T1-T2,the bud grew to a diameter ranging from 12 mm to 15 mm with less than 30%of the bud showing its color.That was in the single-core sideline period.When the diameter of a stamen reached 15-19 mm with a colored area greater than 30%,the T3-T4 stage was in the binuclear period.(2)The stamens were most effectively disinfected with running water for 30min followed by a rinse with 1%sodium hypochlorite for 8min.(3)The stamen could be optimally induced during the single-core sideline period.(4)The medium for optimal stamen callus induction was formulated with MS+0.2 mg NAA·L^(−1)+0.5 g PVP·L^(−1),and that for the proliferation MS+0.5 mg NAA·L^(−1)+0.25 mg 6-BA·L^(−1)+0.5 mg GA3·L^(−1)+1 g PVP·L^(−1).【Conclusion】This study clearly revealed the floral bud size,appearance,and dominant pollen grain development stages of the red P.delavayi and determined the optimal medium formulations and culture conditions for inducing,proliferating,and differentiating the stamen callus.

关 键 词：滇牡丹 雄蕊 愈伤组织 增殖 

分 类 号：S685.11[农业科学—观赏园艺]