减少多年生麻栎茎段外植体褐化的材料采集与组织培养条件优化  

Material Collection of Stem Segment Explants from Perennial Quercus acutissima and Optimization of Tissue Culture Condition to Reduce Browning

作　　者：于艳[1] 夏兴宏[1] 石淑萍[1] 孙娟[1] 曲芳[1] 康丽娜[1] 郎庆龙[1] 

机构地区：[1]辽宁省蚕业科学研究所,辽宁凤城118100

出　　处：《蚕业科学》2015年第5期793-800,共8页ACTA SERICOLOGICA SINICA

基　　金：辽宁省重大科技攻关项目(No.2014209001-11)

摘　　要：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%。The stem segments with axillary buds from perennial shoots of 20 years-old Quercus acutissima were used as explants to culture saplings. The collection time,sampling position,sterilization method,pretreatment temperature and illumination condition of shoot explants,and the dosage of inorganic salt,hormone and antioxidant as well as the pH of culture medium were optimized to solve the technical difficulty of easy-browning during culture of shoot explants. The results indicated that the optimal conditions for collection and treatment of perennial Quercus acutissima shoots was as follows：collect young stem segment without growth points（ stem tip） of spring shoot as explant,and sterilize in 1 g /L HgCl2 solution for 7 min,shading treatment for 5 d,and primary culture of stem segments in dark environment for 5 to7 d. Under these conditions,necrosis rate of the explants was effectively reduced. The best medium formulation was1 /2MS0＋4. 0 mg /L 6-BA＋3. 0 mg /L KT＋0. 5 mg /L IBA＋0. 2 mg /L NAA＋1. 0 mg /L PVP,which contains low concentration of inorganic salt,appropriate hormone ratio and antioxidant addition amount. The optimal medium pH was 5. 4. In thismedium,necrosis of the explants was also obviously inhibited. Under the above optimal conditions,the survival rate of perennial shoot of Quercus acutissima was up to37. 5% and the necrosis rate declined from 34. 38%to 21. 88%.

关 键 词：麻栎 组织培养育苗 枝条外植体 褐化 取材 培养基 

分 类 号：S792.181[农业科学—林木遗传育种]