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青榨槭愈伤组织诱导分化及生根生理生化基础研究

标题: 青榨槭愈伤组织诱导分化及生根生理生化基础研究
英文标题: Studies on Induction and Differentiation of Callus and Physiological and Biochemical Basis of Roots of Acer Davidii Franch
作者: 郝爱丽
出版时间: 2010-01-01
所在大学: 河北农业大学
关键词: 青榨槭,愈伤组织,组织培养,扦插,代谢调节剂
英文关键词: Acer davidii Franch,the callus,culture organization,cutting,Metabolism regulator
论文级别: 硕士
学位: 学位论文
导师: 路丙社
专业: 园林植物与观赏园艺
提交时间: 2010
摘要: 青榨槭(Acer Davidii Franch)为槭树科槭树属,落叶乔木,高812m,其树形优美,树皮花纹奇特,秋叶变红、橙、紫色,是城市园林极具应用前景的绿化和观赏树种,近年来引起了人们的高度关注.本文分别以青榨槭无菌叶片和茎段为外植体,对外植体愈伤诱导、分化、不定芽增殖以及生根培养进行了系统研究,建立了再生体系,并对青榨槭的扦插繁殖进行了深入探讨,结果表明生长调节剂可显著提高插条生根率,生根数量和根长,并调节了插穗内部相关酶的活性.主要研究结果如下:1.研究了MS、1/2MS、WPM和B5四种基本培养基对愈伤组织的诱导,其中,以MS培养基对茎段和叶片愈伤组织诱导最佳,愈伤无褐变,表面多呈黄绿色松软颗粒状,愈伤发生率和产量均最好.2.生长调节剂对愈伤诱导的试验研究表明:TDZ和NAA有利于叶片及茎段愈伤组织的诱导,最佳愈伤诱导培养基为:MS + TDZ0.5mg/L + NAA 0.1 mg/L,愈伤组织出愈率分别达98.7%和93.2%,且叶片叶基部位较叶片中部和尖端出愈率高,褐化率低.3.在不定芽诱导时,叶片与茎段的诱导率存在着较大的差异,叶片的诱导率明显高于茎段.试验做了生长素对比试验,其中NAA效果要好于IBA.最佳诱导培养基为:MS+TDZ2.0mg/L+NAA0.1 mg/L,诱导率分别为78.85%和59.26%.4.在不定芽诱导中,pH值、蔗糖含量和暗培养时间对不定芽诱导有一定影响.pH值6.2、蔗糖含量30g/L和暗培养20d时,叶片和茎段不定芽再生率可分别提高到87.10%和76.67%.5.通过正交试验(L933)方法筛选出最佳增殖培养基配方为:MS培养基中添加6-BA1.5mg/L+NAA0.12mg/L+GA30.2mg/L,添加琼脂粉7g/L,蔗糖30g/L,增殖系数为4.34,增殖率最高为82.76%,茎芽生长正常.6.从生根数、根长和生根率等因素综合考虑,以MS附加IBA0.5mg/L时生根率最高66.8%,平均生根2.32条.7.用IBA400 mg/L+NAA400 mg/L+GA320 mg/L组合处理青榨槭插条比单独使用某一种生长调节剂插条生根效果好,生根率、生根数和根长分别达88.5%、6条和15.88 cm,分别比对照处理T1增加了108%、200%、285%.同时,组合处理提高了插穗POD酶活性,降低了IAAO酶活性,加速了PPO酶活性高峰的出现.8.研究了青榨槭嫩枝扦插过程中,喷施MHS代谢调节剂对插条成活率、生根率和保绿时间的影响,及其叶片中叶绿素、可溶性糖、MDA含量和SOD活性的变化.表明MHS对插穗生根生长有明显促进作用.经生根剂浸根并喷施MHS2后插条的成活率和生根率均最高,与对照相比分别提高了908.61%和104.76%,保绿时间延长了31.22d.插条的叶绿素和可溶性糖含量随处理天数的增加而呈增高趋势,均高于浸根对照;MDA呈下降趋势;均低于对照;SOD活性在处理初期活力有所下降,后期明显增强.可见,MHS是通过调控插条的生理生化代谢来提高青榨槭生根成活率的.
英文摘要: Acer davidii Franch which belongs to genus Acer, family Aceraceae, is 8-12meters high. The tree is beautiful and the bark is green with the stripes.The autumn leaves red, orange, purple. Because it's beautiful and suitable for sightseeing, the future implication is great. It is wildly applied in park-green engineering, therefore,it has an extensive prospect.At present, more and more people begin to pay attentionto it but simple introduction of its shape and the research of its resistant ability has been only made, however, the organization of its culture doesn't be reported. The paper has a comprehensive and thorough research from two sides of the inductive rate of adventitious buds, with explant Franch and Sterile leaf and stem of Acer davidii. External implant callus induction, adventitious bud induction, proliferation, development, and training to conduct a systematic study of root buds.Acer davidii and cutting propagation of in-depth study, results showed that the growth regulators can significantly increase the rooting rate, root number and root length, and regulates the activity of enzymes cutting inside.The result is as following:1. Among the MS、1/2MS、WPM and B5 four basic mediums, the MS is the best for the stem and leaf callus induction. The callus without browning, yellow-green soft granular surface often isolated, and the production rate of callus were the best.2. TDZ and NAA benefit of the leaf and stem callus induction. The best callus inducation is MS + TDZ0.5mg/L + NAA 0.1 mg/L, and the callus were 98.7%and 93.2%. The leaf blade better the central part of leaf base, and less for browning rate.3. There is a big difference in the induction of adventitious buds for leaves and stems,and the induction rate of leaves was higher than the induction rate of stem. Auxin contrast test test done, which NAA is better than IBA. The best medium for induction: MS + TDZ2.0mg / L + NAA0.1 mg / L, induction rates were 78.85%and 59.26%.4. In the induction of adventitious buds, pH, sucrose content and dark culture of adventitious buds to some extent. pH 6.2, sucrose 30g / L and dark culture 20d, the leaves and stems shoot regeneration rate was improved to 87.10%and 76.67%.5. Orthogonal test (L933) method of choosing the best medium for proliferation: MS medium containing 6-BA1.5mg/L + NAA0.12mg/L + GA30.2mg/L, adding the agar 7g/L, sucrose 30g/L, the multiplication coefficient is 4.34, growth rate is 82.76%, and growth of shoot is normal.6. From the root number, root length and rooting rate factors to consider, MS additional IBA0.5mg/L is the best. The highest rooting rate is 66.8%, and the average root is 2.32.7. With IBA 400 mg/L + NAA400 mg/L + GA320mg/L combined treatment davidii than cutting alone a particular growth regulator effective rooting, rooting rate, root number and root length respectively 88.5%, 6and 15.88 cm, respectively,compared with control treatment T1 increased by 108%, 200%, 285%. Meanwhile, the combination treatment increased the POD activity of cuttings, reducing the IAAO activity, accelerated the emergence of peak of PPO activity.8. The effect s of MHS metabolism regulator on cuttings survival rate, rooting percent and green time, were studied by spraying MHS in the cutting of Acer davidii Franch. The variation in the contents of chlorophyll, solubility carbohydrate and MDA and the activities of SOD were also discussed in the experiment. The results showed that MHS could improve growth of Acer davidii Franch. The highest survival rate and rooting rate were obtained after the cuttings of Acer davidii Franch were soaked with rooting reagent and sprayed by MHS. The survival rate and rooting rate with this treatment were increased by 908.61%and 104.76%separately. The green time was 31.22 d longer than the control .The contents of chlorophyll and solubility carbohydrate were increased gradually with the treated days. The contents of MDA was decreased with the treated days. The activities of SOD were descend firstly, then enhanced. This showed that MHS increased survival rate by regulating the metabolism of cutting sphysiology and biochemistry.