【作者】 王雪芳；

【导师】 王锁民；

【作者基本信息】 兰州大学 ， 草坪生物学， 2012， 硕士

【摘要】 本文以小花碱茅(Puccinellia tenuiflora)为研究材料,以其成熟种子为外植体,通过对愈伤组织最适培养条件(种子不同处理和消毒方案、基本培养基、不同激素配比、蔗糖浓度、pH等),胚性愈伤组织分化及生根,再生苗的炼苗与移栽的研究,成功建立了小花碱茅组织培养再生体系。主要得到以下结果：(1)小花碱茅组织培养体系以成熟种子作为外植体,种子脱颖处理后,显著提高了出愈率,且其出愈率高于以幼穗、茎段、叶片为外植体的出愈率。(2)种子最佳消毒时间为75%酒精3min,50%次氯酸钠溶液10min。(3)与B5培养基相比,MS培养基适合作为小花碱茅组织培养的基本培养基,愈伤水化率明显降低。(4)诱导愈伤组织的最佳培养基为MS+5.0mg·L-12,4-D+3%蔗糖+0.7%琼脂,诱导率可达55.2%,2周后可见白色愈伤组织,较以茎段和叶片为外植体诱导愈伤组织所需时间缩短。(5)培养基pH值为7～8时,诱导的小花碱茅愈伤组织质量比较好,水化率较前人研究减少一半。(6)胚性愈伤组织分化芽点的最适培养基为MS+1.0mg·L-16-BA+0.5mg·L-1NAA+3%蔗糖+0.7%琼脂,分化率达31.7%,同时伴随大量根毛发生,生根率达81.7%。(7)分化后的愈伤组织在1/2MS+1%蔗糖+0.7%琼脂培养基上可100%生根成苗,植株生长健壮,而且移栽后全部成活。(8)小花碱茅再生周期为16～20周。更多还原

【Abstract】 In this paper, Puccinellia tenuiflora was selected as material. Tissue culture system was developed for plant regeneration from mature seed of Puccinellia tenuiflora. We have selected the most suitable conditions for the callus growth according to the effect of many factors, such as the different processing and disinfection of seed, basic medium, the compounding proportions of different hormones, sugar concentration and pH. And then the good embryo callus will be take differentiation and root.The main results are as follows:(1) The tissue culture system of Puccinellia tenuiflora was developed from mature seed, seed germination rate can improve greatly after peeling, so as to increase the callus induction rate obviously.And the callus induction rate from seed is higher than the one from young spikelet, stem and leaf.(2) The best seed disinfection time is3min of75%alcohol and10min of50%NaClO.(3) Compared with the B5, MS is more suitable for the basic medium of tissue culture of Puccinellia tenuiflora and the callus hydration rate reduced significantly.(4) The induction rates of primary callus from mature seed of Puccinellia tenuiflora were up to55.2%on callus induction medium contained MS basal salt and vitamins,5.0mgL-12,4-D and3%sucrose and solidified with0.7%agar, respectively. The little white callus can be seeing after2weeks,2weeks earlier than inducing by stem and leaf.(5) The optimal pH of the callus induction is7-8. Hydration rate decrease by half when Ph is from5.8to7-8.(6) The differentiation rates of embryo callus were31.7%on differentiation medium contained MS basal salts and vitamins,1.Omg·L-16-BA and0.5mg·L-1NAA and30%sucrose and solidified with0.7%agar. At the same time, most callus have a lot of root hair and the rooting rate were81.7%.(7) The rooting rate were100%on1/2MS basal salts and vitamins and1%sucrose and solidified with0.7%agar. The regenerants with roots can all survive after they were transplanted to soil.(8) The regeneration period of Puccinellia tenuiflora was about16-20weeks.更多还原