群众杨39无性系耐盐悬浮细胞系的建立和体细胞变异体完整植株的诱导

Establishment of Saline-tolerant Cell Lines of Populus × xiaozhuanica cv.‘Popularis’-39 and the Induction of Somaclonal Variant Plant

本试验以群众杨３９号为试材，通过耐盐胁迫悬浮培养建立耐盐悬浮细胞系，经愈伤组织成功培育出耐ＮａＣｌ３．０‰～３．５‰的群众杨体细胞变异体的完整植株。实验表明，ＭＳ培养基附加０．４５ｍｇ／Ｌ２，４－Ｄ、０．３ｍｇ／ＬＮＡＡ和０１ｍｇ／ＬＫｉｎｅｔｉｎｅ的Ｍ４培养基能较好地获得松脆、易分散的愈伤组织。液体以ＭＳ培养基只附加０．５ｍｇ／Ｌ２，４－Ｄ的ＬＭ３为最好，附加氨基酸有益于悬浮细胞的正常生长。还对悬浮培养细胞的有关参数进行了测定。耐盐悬浮细胞培养实验结果表明ＮａＣｌ对细胞生长有抑制作用，并随着ＮａＣｌ升高而加强。对获得的耐３‰～６‰各水平ＮａＣｌ悬浮细胞经高密度植板，都能形成愈伤组织。耐盐愈伤组织诱导只获得耐３‰～４‰ＮａＣｌ的不定芽，高于４‰ＮａＣｌ未能诱导出不定芽，说明高浓度ＮａＣｌ对芽组织分化有明显的抑制作用。不定根分化对ＮａＣｌ反应非常敏感，只在耐３‰和３．５‰ＮａＣｌ浓度培养基诱导出不定根。

Somaclonal variants of Populus × xiaozhuanica cv.‘popularis’-39,which istolerant to 30‰～3.5‰NaCl,is successfully obtained through the establishment of NaCl-tolerant pressure cell culture to produce calli and control the conditions of adventitious budand root induction. The friable calli are well produced on the medium M4 MS+0.45 mg/L2,4-D+0.3mg/LNAA+ 0.1mg/L Kinetine.The best suspension liquid medium is LM3 :MS + 0. 5mg/L2,4-D+ 30g/L sucrose.The addition of amino acids was beneficial for thegrowth of the suspension cells.The relative parameters about cell growth are also measured.The culture test of NaCl-tolerant cells indicates that NaCl restrains cell growth and the high-er the concentration of NaCl,the stronger the restraint.Calli can be produced from all thecell lines tolerant to the every NaCl level from 3.0‰～6‰;The test,in which the shoots canbe induced only from the calli tolerant to 3.0‰～4.0‰,indicates that the high concentrationof NaCl exerts an obvious restraining impact on the shoot induction.The root differentiationis also highly sensible to NaCl and the roots can only be induced on the medium with 3.0‰and 3.5‰NaCl.