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以20年生麻栎当年的枝条截取带腋芽的茎段为外植体进行组织培养育苗,为解决培养过程中枝条外植体易褐化的技术难题,对枝条的采集时间、取材部位、灭菌方法、预处理温度和光照条件,培养基中无机盐、激素、抗氧化剂等组分的用量及培养基pH进行优化试验。能有效降低外植体褐死率的多年生麻栎枝条采集与处理条件是:采集春枝不带生长点(茎尖)的幼嫩茎段作为外植体,用1 g/L HgCl2溶液灭菌7 min,将枝条遮光处理5 d和在茎段外植体初代培养阶段暗培养5~7 d。设计的低浓度无机盐及适当激素配比和抗氧化剂添加量的较佳培养基配方是1/2MS0+4.0 mg/L 6-BA+3.0 mg/L KT+0.5 mg/L IBA+0.5 mg/L NAA+1.0 mg/L PVP,培养基最优pH为5.4,使用该配方的培养基也可有效降低外植体的褐死率。在上述优化条件下,多年生麻栎枝条外植体的成活率达到37.5%,其中褐死率由对照的34.38%下降至21.88%。
In order to solve the technical problem of browning of shoot explants in the process of cultivation, the stems of axillary buds of 20-year-old branches of Quercus acutissima were intercalated with explants. The collection time, Methods, pretreatment temperature and light conditions, the amount of inorganic salts, hormones, antioxidants and other components in the medium and the pH of the medium were optimized. The conditions for collecting and treating perennial Quercus fabri branches which can effectively reduce the browning rate of the explants are as follows: the young shoots without the growing point (shoot tip) are taken as explants and sterilized with 1 g / L HgCl2 solution min. The branches were shaded for 5 days and darkly cultured for 5 to 7 days in the initial culture stage of stem explants. The optimal medium for the design of low concentration inorganic salts and appropriate hormones and antioxidants is 1 / 2MS0 + 4.0 mg / L 6-BA + 3.0 mg / L KT + 0.5 mg / L IBA + 0.5 mg / L NAA + 1.0 mg / L PVP, the optimal medium pH was 5.4, and the medium with this formula was also effective in reducing the browning rate of explants. Under the above optimized conditions, the survival rate of perennial Q. lua explants reached 37.5%, among which the brown mortality rate decreased from 34.38% to 21.88% of the control.