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白姜花胚性愈伤组织的诱导与植株再生体系的建立 被引量:6

Embryogenic Callus Induction and Plant Regeneration from Hedychium coronarium via Somatic Embryogenesis
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摘要 将白姜花(Hedychium coronarium)未成熟花丝接种在MS+4 mg·L~(-1) 2,4-D+4 mg·L~(-1) NAA+1 mg·L~(-1) 6-BA的培养基上,经过180 d的培养,诱导出3种愈伤组织:Ⅰ,浅黄色松散易碎;Ⅱ,白色疏松半透明水渍状;Ⅲ,黄色致密颗粒状。对愈伤组织继代培养基中的2,4-D、NAA和6-BA浓度进行优化,结果表明培养基中含有1 mg·L~(-1) 2,4-D、0.25 mg·L~(-1) NAA、0.25 mg·L~(-1) 6-BA时可获得胚性状态良好的愈伤组织。胚性愈伤组织在MS无机盐+0.25 mg·L~(-1) NAA+0.5 mg·L~(-1) TDZ+维生素B5+100 mg·L~(-1)谷氨酰胺+230 mg·L~(-1)脯氨酸+100 mg·L~(-1)麦芽提取物+45 g·L~(-1)蔗糖的体胚诱导培养基中培养20d后,转移到MS基本培养基上培养30 d,1 g胚性愈伤组织可获得50~60个体胚。成熟体胚在MS+0.2mg·L~(-1) IAA+0.5 mg·L~(-1) 6-BA的萌发培养基上培养20 d时,体胚萌发率为85%。萌发的体胚转移到1/2MS+1 g·L~(-1)活性炭的成苗培养基中可发育成正常植株,植株室外栽培成活率达90%。对100株再生植株的染色体数进行分析,发现3株三倍体(2n=3x=51)、6株四倍体(2n=4x=68)和2株六倍体(2n=6x=102)。 A reproducible protocol for somatic embryogenesis was established for the ornamental ginger Hedychium coronarium.Immature filaments were cultured on Murashige and Skoog(MS) medium supplemented with 4 mg ? L~(-1) 2,4-D,4 mg ? L~(-1) NAA,1 mg ? L~(-1) 6-BA.After culture for 180 days,three types of callus were obtained,type I callus were light yellow and friable with tightly packed cell contained dense cytoplasm that appeared to be embryogenic;type Ⅱ callus were white with long,highly vacuolated cells that appeared to be non-embryogenic;type Ⅲ callus were yellow with large,less packed cells contained dense cytoplasm that appeared to between embryogenic and non-embryogenic.Medium containing 1 mg ? L~(-1)2,4-D,0.25 mg ? L~(-1) NAA,0.25 mg ? L~(-1) 6-BA was suitable for embryogneic callus proliferation.Embryogenic calli were cultured on somatic embryo induction medium containing MS basal salts,vitamin B_5,100 mg ? L~(-1) glutamine,230 mg ? L~(-1) proline,100 mg ? L~(-1) malt extract,0.25 mg ? L~(-1)NAA,0.5 mg ? L~(-1) TDZ,45 g ? L~(-1) sucrose and 7 g ? L~(-1) agar for 20 days,then transferred on MS medium free hormone for 30 days.About 50- 60 somatic embryos were induced from per gram callus.Germination percentage of 85%were observed in germination medium,which consisted of MS basal salts,0.2 mg ? L~(-1) NAA and 0.5 mg ? L~(-1) 6-BA.Regenerated plantlets with normal shoot and root were developed after 30 d on half strength MS medium with 1 g ? L~(-1) active charcoal.Well rooted plantlets were successfully acclimatized with a survival rate of 90%and grew vigorously.The chromosome number of100 regenerated plants showed a wide range of variation,89 of them were diploid(2n = 2x = 34),3 were triploid(2n = 3x = 51),6 were tetraploid(2n = 4x = 68) and 2 were hexaploid(2n = 6x=102).
作者 肖望 涂红艳 张爱玲 XIAO Wang, TU Hong-yan, and ZHANG Ai-ling (Biology and Food Engineering Institute, Guangdong University of Education, Guangdong Development Center of Applied Ecology and Ecological Engineering in Universities, Guangzhou 510303, China)
出处 《园艺学报》 CAS CSCD 北大核心 2016年第8期1605-1612,共8页 Acta Horticulturae Sinica
基金 广东省创新强校工程省级重大项目(2014KZDXM076) 广东省科技计划项目公益研究与能力建设专项(2015A030302097) 广东省高等院校学科与专业建设专项资金项目(2013KJCX0137) 广州市科技计划项目科学研究专项(2014J4100151) 国家级大学生创新训练计划项目(1427815060)
关键词 白姜花 胚性愈伤组织 植株再生 Hedychium coronarium embryogenic callus plant regeneration
作者简介 E-mail:xiaowang@gdei.edu.cn
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