秋石斛胚性愈伤组织诱导及再生体系的建立  

Establishment of Embryogenic Callus Induction and Regeneration System for Phalaenopsis-hybrid Dendrobium

作　　者：李志晴 李亚梅 李崇晖[2,3] 尹俊梅 LI Zhiqing;LI Yamei;LI Chonghui;YIN Junmei(School of Tropical Agriculture and Forestry,Hainan University,Haikou,Hainan 570228,China;Tropical Crop Germplasm Research Institute,Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China,Ministry of Agriculture&Rural Affairs/Key Laboratory of Tropical Crops Germplasm Resources Genetic Improvement and Innovation of Hainan Province,Haikou,Hainan 571101,China;Engineering Technology Research Center of Tropical Ornamental Plant Germplasm Innovation and Utilization,Danzhou,Hainan 571737,China;Sanya Research Institute,Chinese Academy of Tropical Agricultural Sciences,Sanya,Hainan 572330,China;Haikou Experiment Station,Chinese Academy of Tropical Agricultural Sciences,Haikou,Hainan 571101,China)

机构地区：[1]海南大学热带农林学院,海南海口570228 [2]中国热带农业科学院热带作物品种资源研究所/农业农村部华南作物基因资源与种质创制重点实验室/海南省热带作物资源遗传改良与创新重点实验室,海南海口571101 [3]海南省热带观赏植物种质创新利用工程技术研究中心,海南儋州571737 [4]中国热带农业科学院三亚研究院,海南三亚572330 [5]中国热带农业科学院海口实验站,海南海口571101

出　　处：《热带作物学报》2024年第6期1175-1183,共9页Chinese Journal of Tropical Crops

基　　金：海南省自然科学基金青年基金项目(No.322QN359);中央级公益性科研院所基本科研业务费专项(No.1630032022002);海南省重点研发项目(No.ZDYF2024XDNY239)。

摘　　要：秋石斛(Phalaenopsis-hybrid Dendrobium)花姿优美、花色丰富,是热区重要观赏植物,具有极高的产业价值。本研究以秋石斛杂交种迷你(Den.Yaya Victoria)、水蜜桃(Den.Swirl)、三亚阳光(Den.Sonia Hiasakul)原球茎和无菌苗幼嫩茎段为外植体,探讨不同植物生长调节剂组合对不同品种秋石斛胚性愈伤诱导、不定芽分化及植株再生的影响。结果表明:在胚性愈伤组织诱导阶段,原球茎和生长2个月的幼苗茎段作为外植体诱导效果最佳,暗培养30 d即可获得胚性愈伤。3个品种的愈伤组织诱导最佳培养基是以MSD(MS+30 g/L葡萄糖+1 g/L花宝1号+8 g/L琼脂)为基本培养基,添加0.5 mg/L KT和0.2~0.5 mg/L 2,4-D。迷你、水蜜桃、三亚阳光的胚性愈伤诱导率最高分别为31.11%、22.78%和50%,其中迷你品种仅需15 d即可获得胚性愈伤。最佳胚性愈伤增殖培养基为MSD+0.5 mg/L IBA+0.5 mg/L KT,胚性愈伤状态保持良好且不发生分化,增殖倍数0.9。最佳分化培养基为1/2 MS+0.5 mg/L IBA+0.15 mg/L KT,在光照条件下,接种后的胚性愈伤组织能够快速诱导形成芽点,30 d内分化形成2~3片叶的幼苗,相同培养基中保持60 d能够自行生根获得完整再生苗。最佳生根培养基为1/2 MS+30 g/L蔗糖+0.5 mg/L NAA+8 g/L琼脂,生根率为100%,平均生根数5条,根长约1.3 cm。本研究所使用的胚性愈伤组织诱导、增殖和分化培养基具有一定的广谱性,可为后续秋石斛胚再生途径及分子育种研究奠定基础。Phalaenopsis-hybrid Dendrobium is an important ornamental plant in tropical area and has high market values,with beautiful flower appearance and abundant flower colors.In this study,the protocorms and young seedling’s stem segments of commercial varieties,Den.Yaya Victoria,Den.Swirl and Den.Sonia Hiasakul,were used as the explants.The effects of different phytohormone combination on embryogenic callus induction,adventitious bud differentiation and plantlet regeneration were investigated.During the embryogenic callus(EC)induction stage,the best induction explants were protocorms and 2-month-old seedling stem segments and EC could be obtained after 30 days culture in dark condition.The best medium for EC induction of was MSD(MS+30 g/L glucose+1 g/L Hyponex+8 g/L agar)as the basic medium with 0.5 mg/L KT and 0.2-0.5 mg/L 2,4-D.The highest average induction rate of EC was 31.11%,22.78%,and 50% for Den.Yaya Victoria,Den.Swirl and Den.Sonia Hiasakul,and the induction time for Den.Yaya Victoria was only 15 days from explants to EC.The best proliferation medium was MSD+0.5 mg/L IBA+0.5 mg/L KT,which maintaining EC in good condition without differentiation and the multiplication rate was 0.9.The best medium for differentiation was 1/2 MS+0.5 mg/L IBA+0.15 mg/L KT under light condition of 16 h one day,EC quickly differentiating and forming 2-3 leaf adventitious buds within 30 days,and finally regenerating into plantlet after 60 days in the same medium.The best rooting medium was 1/2 MS+30 g/L sugar+0.5 mg/L NAA+8 g/L agar,with rooting rate 100% and five roots in length 1.3 cm per plantlet.In this research,the EC induction,proliferation and differentiation media have a wide-spectrum and the EC regeneration pathway could lay a solid foundation for further molecular breeding of Phalaenopsis-hybrid Dendrobium.

关 键 词：秋石斛 茎段 胚性愈伤 植株再生 

分 类 号：S682.31[农业科学—观赏园艺]