【作者】 吴丹丹；

【导师】 林义章；

【作者基本信息】 福建农林大学 ， 蔬菜学， 2008， 硕士

【摘要】 黄秋葵学名Abelmoschus esculentus L．（Hibiscus esculentus L．），属锦葵科（Malvaceae）秋葵属（belmoschus Medicus）一年生草本植物。它既是营养丰富的保健型蔬菜，又是疗效显著的常用中药，还是观赏价值颇高的绿化、美化材料。随着对黄秋葵的营养价值及其药用价值认识的不断深入，应用前景日益广阔，需求量与日俱增，市场效益显著。因此对黄秋葵的组织培养技术的研究亦愈发得到重视，通过组织培养，获得优质材料，降低提取成本，为黄秋葵的利用提供参考。本研究以黄秋葵为试验材料，采用黄秋葵当年生茎段为外植体，诱导胚性愈伤组织，建立植株再生体系和胚性悬浮体系。获得试验结果如下：1、以黄秋葵茎段为材料，诱导产生了胚性愈伤组织，建立了高频再生体系。筛选出了黄秋葵外植体消毒灭菌的最佳处理组合为75％乙醇消毒30 S后，转入0．1％HgCl₂溶液中消毒8 min。适于胚性愈伤诱导及扩增最佳培养基MS（NH₄NO₃/2）+2．0mg·L^-12，4-D+3．0 mg·L^-1ZT+3％蔗糖+0．7％琼脂。2，4-D在诱导胚性愈伤组织中是必需的。植物激素、氮源、糖源及光照条件对黄秋葵胚性愈伤组织形成有较大影响。2、以胚性愈伤组织为材料，建立了稳定的胚性细胞悬浮体系。试验结果表明培养时期悬浮细胞生长规律符合典型的“S”形生长曲线，其最佳继代周期为4～8 d。培养期悬浮细胞干重和pH之间、振荡培养转速和悬浮细胞形态之间都有一定的相关性。接种量和AgN0₃含量对悬浮细胞的生长有影响：最佳接种量为1．5％，最适AgN0₃浓度为5．0mg·L^-1。3、通过对胚性愈伤组织的分化培养，获得黄秋葵再生植株。筛选出黄秋葵不定芽的诱导培养基为MS+3．0 mg·L^-16-BA+0．5 mg·L^-12，4-D+3％蔗糖+0．7％琼脂；壮苗培养基：MS+1．5 mg·L^-16-BA+0．2 mg·L^-12，4-D+1．0 mg·L^-1GA₃+3％蔗糖+0．7％琼脂；生根培养基：1／2MS+0．1 mg·L^-1NAA。炼苗和移栽是提高黄秋葵移栽成活率的关键。自然光室温条件下炼苗7 d，然后打开瓶盖炼苗3 d。用腐质土：河沙：蛭石=1：1：1的混合物作为培养组培苗的基质利于组培苗成功移栽。更多还原

【Abstract】 Abelmoschus esculentus L. is an annual herbage plant in the Mal Vaceae family. It is used as vegetable,Chinese traditional medicine and ornamental plant,which has very high nutrient, medicine and ornamental value. As people come to know more about Okra,the applied prospect is getting more and more vast, which increase demand and perform good in market. And tissue culture technique of Okra has been given even more attention. Obtain good material and reduce the cost by tissue culture, which used as references for the usuage of Okra. A regeneration system of tissue culture in vitro including embryogenic callus induction and embryogenic susppension using stem as explant was established on Abelmoschus esculentus L. The main results were as follows:1 Somatic embryogenic calli of Abelmoschus esculentus L. was produced. For the stem, 75% ethanol treating for 30 seconds first and 0.1% mercuric cholride treating for 8 minutes could be the best design of sterilization combination. Somatic embryogenic callus was induced from stem on MS medium supplemented with 2.0 mg·L^-12,4-D and 3.0 mg·L^-1 ZT. 2,4-D was the necessity phytohormones. The effect of phytohormones,nitrogen and light on embryogenic callus formation was discussed.2 Somatic embryogenic suspension culture of Abelmoschus esculentus L. was established. In suspension culture stage, the rule of cell growth was shown S accorded with Sigmoid Curve, the best days of sub-cultured was 4 to 8. The relationship between the dry weight of suspension cultures and pH changes in medium,the rotational speed and suspension cellulat morphology werw discussed. The effects of inoculation and AgN0₃ on the growth of embryogenic suspensions were alsodiscuss ed.The optium concentration of inoculation and level of AgN0₃ were 1.5% and 5.0 mg·L^-1,respectively.3 Regeneratated plantlets were obtained from callus differentiation. The medium of shoot induction was MS + 3.0 mg·L^-1 6-BA + 0.5 mg·L^-12,4-D+3% sucrose + 0.7% agar. The exercising seedling medium was MS+ 1.5 mg·L^-16-BA + 0.2 mg·L^-12,4-D + 1.0 mg·L^-1GA3 +3% sucrose + 0.7% agar. The medium for tube seedling root-generation was 1/2MS + 0.1 mg·L^-1 NAA. The key steps of transplanting rate is exercising seedling. The transplanting medium in rotten quality soil, vermiculite and river sand by 1 to 1 to 1 is the best treament on smelt seedling.更多还原