尾巨桉良种无性系DH32-28高效植株再生体系的建立  

Establishment of an Efficient Plant Regeneration System for the Elite Clone DH32-28 of Eucalyptus urophylla×E.grandis

作　　者：韦利达 刘正兴 梁康明 吴永富 林凯 蒋兴成 吴雯霏 黎明慧 胥佳静 徐增富 WEI Lida;LIU Zhengxing;LIANG Kangming;WU Yongfu;LIN Kai;JIANG Xingcheng;WU Wenfei;LI Minghui;XU Jiajing;XU Zengfu(State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources,Key Laboratory of National Forestry and Grassland Administration on Cultivation of Fast-Growing Timber in Central South China,College of Forestry,Guangxi University,Nanning,530004;Guangxi Dongmen Forest Farm,Chongzuo,532108)

机构地区：[1]广西大学林学院,亚热带农业生物资源保护与利用国家重点实验室,中南速生材繁育国家林业和草原局重点实验室,南宁530004 [2]广西壮族自治区国有东门林场,崇左532108

出　　处：《基因组学与应用生物学》2024年第8期1353-1363,共11页Genomics and Applied Biology

基　　金：广西重点研发计划揭榜制科技项目(桂科JB22035001);广西科技基地和人才专项(桂科AD23026337)共同资助。

摘　　要：本研究以尾巨桉(Eucalyptus urophylla×E.grandis)三种良种无性系DH32-26、DH32-28和DH32-29的无菌增殖苗为实验材料,旨在探讨植物生长调节剂对增殖苗节间伸长和植株再生的影响,并进一步研究了无性系、植株类型和外植体类型对植株再生能力的影响。此外,还以易再生无性系DH32-28为材料,进一步研究植物生长调节剂和愈伤组织诱导时间对再生的影响。结果表明,无性系是影响尾巨桉离体再生的主要因素,DH32-28的再生效率显著高于DH32-26和DH32-29的;GA_(3)或IBA处理后增殖苗的节间均能显著伸长,其中GA_(3)处理获得的DH32-28伸长苗的芽再生率最高,并且再生芽个数最多;不同外植体类型的芽再生率差异显著,其中DH32-28伸长苗茎段芽再生率最高,其次是叶柄和叶片。最适宜DH32-28离体植株再生的条件为:增殖苗经过GA_(3)处理20 d后的伸长苗节间茎段作为外植体,最适宜的愈伤组织培养基为:1/2 MS+0.22 mg/L TDZ+0.019 mg/L NAA+30 g/L蔗糖+7 g/L琼脂,诱导时间为20 d;最适宜的再生芽诱导培养基为:1/2 MS+0.25 mg/L 6-BA+0.125 mg/L NAA+30 g/L蔗糖+7 g/L琼脂,诱导30 d的芽再生率达95.00%;在根诱导培养基(1/2 MS+0.5 mg/L IBA+30 g/L蔗糖+7 g/L琼脂)上再生芽均可生根。本研究成功建立了尾巨桉良种无性系DH32-28茎段高效植株再生体系,为进一步的遗传转化体系建立及后续基因功能和基因工程等研究奠定了基础。This study investigated the effects of plant growth regulators on the elongation of internodes in proliferated seedlings and plant regeneration using aseptic seedlings from three elite clones of Eucalyptus urophylla×E.grandis(DH32-26,DH32-28 and DH32-29).The effects of clones,plant types and explant types on plant regeneration capability were further studied.The easily regenerated clone DH32-28 was used as the research material to further investigate the effects of plant growth regulators and callus induction time on shoot regeneration.The results showed that the clone was the primary factor influencing the in vitro regeneration in E.urophylla×E.grandis.The regeneration efficiency of DH32-28 was significantly higher than those of DH32-26 and DH32-29.The internodes of all proliferated seedlings exhibited significant elongation after treatment with GA_(3) or IBA,and the GA_(3)-treated DH32-28 seedlings had the highest shoot regeneration rate and the highest number of regenerated shoots.Significant differences were observed in the shoot regeneration rates among different types of explants,with the highest regeneration rate found in the stem segments of elongated seedlings of DH32-28,followed by petioles and leaves.The optimal conditions for DH32-28 in vitro shoot regeneration were as follows.The internode stem segments of elongated seedlings after 20 days of GA_(3) treatment were used as explants.The callus-inducing medium is 1/2 MS supplemented with 0.22 mg/L TDZ,0.019 mg/L NAA,30 g/L sucrose,and 7 g/L agar with a 20 days incubation.The optimal shoot-inducing medium is 1/2 MS supplemented with 0.25 mg/L 6-BA,0.125 mg/L NAA,30 g/L sucrose,and 7 g/L agar,and the shoot regeneration rate reaching 95.00%after 30 days of induction.The regenerated shoots were placed on root-inducing medium 1/2 MS supplemented with 0.5 mg/L IBA,30 g/L sucrose,and 7 g/L agar for root induction,where all regenerated shoots established roots.This study successfully established an efficient plant regeneration system from stem segments of the

关 键 词：DH32-28 GA3处理 再生体系 

分 类 号：S792.39[农业科学—林木遗传育种]