中华芦荟（Aloe vera L. var. chinensis （Haw.） Berger）无性系的离体快速繁殖技术及对水分胁迫的适应性研究

【作者】 廖志华；

【导师】 谈锋；

【作者基本信息】 西南师范大学 ， 植物学， 2001， 硕士

【摘要】 本试验以中华芦荟（Aloe vera L. var. chinensis（Haw.）Berger）为材料，研究了中华芦荟无性系的离体快速繁殖；并在此基础上成功地快速繁殖了库拉索芦荟（Aloe Vera L.）、木立芦荟（Aloe arborescens Mill.）、皂质芦荟（Aloe saponria（Ait.）Haw.）；同时通过人工水分胁迫处理，研究了中华芦荟的抗旱生理生化指标、生物量和药用成分的变化，对中华芦荟在水分胁迫条件下的适应机理进行了探讨。 根据前期的工作经验，以MS培养基为基本培养基，选取6-BA、NAA和蔗糖这3个因子，每个因子取3个水平。选用L₉（3⁴）正交表，考察上面三个因素对中华芦荟丛生芽诱导的影响。筛选到中华芦荟丛生芽诱导的最优培养基为：MS＋2.0mg／L 6-BA＋0.3mg／L NAA＋30g／L蔗糖。用筛选出的最优培养基进行验证实验，中华芦荟丛生芽诱导既快又多，年繁殖系数可达15¹²以上，为目前国内中华芦荟无性系快速克隆繁殖的最高水平；而且在这种培养基中，丛生芽的生长也很正常。 以1／2 MS培养基为基本培养基，选取NAA、IBA、蔗糖和抗氧化剂PVP这4个因子，每个因子取3个水平，选用L₉（3⁴）正交表，考察上面四个因素对中华芦荟丛生芽生根的影响。筛选到中华芦荟丛生芽生根的最优培养基是：1／2 MS＋0.5％ PVP＋0.2mg／L NAA＋0.2mg／L IBA＋30g／L蔗糖。用筛选出的最优培养基进行验证实验，中华芦荟丛生芽生根既快又多。10天左右即可生根，且根系生长发育较自然条件下好，而且在这种培养基中，丛生芽的生长也很正常。 中华芦蔓无性系的离体快速拄殖技术及对水分胁迫的适应性研究 得到的快繁幼苗十分健壮，移栽后很容易，存活达97％。 在中华芦苔快速繁殖的基础上，成功繁殖了库拉索芦苔、木立芦苔、皂质 芦苔的试管茵；最后，用筛选到的两个培养基进行了中华芦苔试管苗的大规模 繁殖取得成功。 在干旱逆境下，中华芦苔的生物量降低；而药用成分芦苔多糖和芦苔贰的 变化截然不同。芦苔多糖表现为单位鲜重叶片中的含量随土壤含水量的减少而 下降，单位干重叶片中芦苔贰含量却增加。这说明在逆境条件下，中华芦苔初 生代谢受到抑制，从而导致生物量和芦苔多糖含量的下降；次生代谢增强，芦 苔贰的合成加强使芦苔戌含量增加；但是由于生物量的减少速率大于芦苔贰的 增加速率使芦苔贰的产率降低。土壤干旱使中华芦苔叶绿素含量下降，Hill 反 应活力下降，气孔阻力变大，蒸腾速率降低，CO。固定速率降低，同化产物积累 减少，这是导致中华芦姿的生物量和经济产量减少的主要原因。 人工模拟土壤干旱条件下，中华芦苔对水分胁迫的适匝能力及影响生物量 指标的研究结果表明：在干旱条件下，中华芦苔叶片的长、宽、厚都减小，减 小了蒸腾面积；气孔频度降低，水分的散失减少，利于保水；根冠比变大，利 于叶片维持较高的自由水含量，以保证细胞内生理生化过程的顺利进行。这说 明中华芦苔在干旱条件下，能产生形态学的变化以适应干旱维持生存。在水分 胁迫下，中华芦苔叶片的含水量、叶片相对含水量和自由水／束缚水的值降低。 这主要是由于自由水含量的减少而致。中华芦苔具有一定的渗透调节能力。叶 片中作为细胞内渗透调节物质的游离脯氨酸、可溶性糖。K”等细胞内溶质含量 在水分胁迫下都有明显增加，均与土壤含水量呈负相关。但叶片相对含水量和 质膜相对透性下降表明渗透调节作用很有限。 在轻度水分胁迫下，中华芦苔叶片中的保护酶类500、POD、CAT的活性升 高，表明中华芦著在清除体内的活性氧，降低膜脂过氧化，稳定膜结构和功能 等方面，表现出对水分胁迫的适应性；但在中度水分胁迫下非酶促保护物质ASA 含量降低，SOD、POD、CAT的活性降低，导致活性氧产生和清除的平衡失调，0。·－ 的积累迅速增加，膜脂过氧化加剧，MDA含量提高，从而降低了膜的稳定性， 最终导致膜伤害，这是水分胁迫下造成中华芦苔伤害的主要原因。 内源激素是植物体内的正负化学信号，调控着植物的多种生理生化过程。 随土壤含水量的减少，水分胁迫的加剧，植物内源激素ZRs和GA含量降低，ABA 2l且 H．H更多还原

【Abstract】 The study is about rapid propagation in vitro of doing line Aloe vera L. var. chinesis (Haw.) Berger and its adaptability to water stress. By one orthogonal design(L934) of three factors with three levels each, such as 6- BA,NAA and sucrose, a best medium for tufted-bud induction of Aloe vera L. var. chinesis (Haw.) Berger is attained, which is MS+2.Omg/L 6-BA+ 0.3mgIL NAA+30g/L sucrose.The further proving experiment indicates that the propagation- coefficient is as high as 1 ~慽2 per year which is the highest one in the same research field, and tufted-buds grow very well on it; the other orthogonal design(L934) of four factors, including NAA,IBA,sucrose and PVP, the best medium for rooting of tufted buds of Aloe vera L. var. chinesis(I-law. )Berger is got, which is 1/2 MS+0.05% PVP+ 0.2mg/L NAA?.2mg/L IBA+30g/L sucrose. Tufted-buds root more and better on the selectd medium. Tube seedlings are transplanted successfully. On the base of rapid propagation of Aloe vera L. var. chinesis (Haw.) Berger, large-scale culture in vitro of Aloe vera L. var. chinesis( I-law. )Berger is carried on successfully. Finally, Aloe Vera L., Aloe arborescens Mill., and Aloe saponria (Ait.) Haw. are cultured in vitro successfully in Wakes. Under the condition of drought, bio-mass of Aloe vera L. var. chinesis (Haw.) Berger is on the decease, as can be said that its financial production is on the decrease. Medicinal components, aloe polysaccharide and aloin, change quite differently. The content of former in unit FW decreases, while the later increases. These show that inhabitation of primary metabolism leads to lessening of bio-mass and aloe polysaccharide; and enhancement of secondary metabolism causes more accumulation of alo in. Due to the rate of bio-mass decrease is faster than that of aloin accumulation, 54 zJ~Jf~ productivity of aloin is on the decrease. Because of soil drought, the contents of TChI., Chl a and Chl. b go down, as well as activity of Hill reaction, rate of transpiration and CO2 fixation, and assimilated products, companied by increase of stomatal resistance By artificial soil drought treatment, the result of researching adaptability of Aloe vera L. var. chinesis (Haw.) Berger to water stress show decrease of relative water content, and WflWb under water stress.To some extend, Aloe vera L. var. chinesis (Haw.) Berger has capacity of osmotic regulation because some relating materials in cells, such as free Pro, soluble sugar and K~, become more under water stress. However, decrease of RWC and RMP indicate that osmotic regulation is not very effective. The length, width and height of leaves of Aloe vera L. var. chinesis (Haw.) Berger decrease which means that leaf square decrease. Density of stoma becomes smaller, and the value of weight of roots ratio weight of up-ground part becomes lesser which helps maintain high free water content n cells. All of these are helpful to make use of water. These prove that Aloe vera L. var. chinesis (Haw.) Berger can change its formation to maintain its existence. Under light water stress, activities of protection enzymes, SOD, POD and CAT, become higher. Aloe vera L. var. chinesis (Haw.) Berger can eliminate active oxygen, release membrance lipid peroxidation, and make the structure and function of membrance stable. However, under middle water stress, concent of AsA and activit更多还原