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作 者:Xiaohong YANG Wen LIU Lin TANG Xiaojuan XIE 杨晓红;刘雯;唐琳;解小娟(北京农学院园林学院,北京102206)
机构地区:College of Landscape Architecture, Beijing University of Agriculture, Beijing 102206, China
出 处:《Agricultural Science & Technology》2017年第1期56-59,66,共5页农业科学与技术(英文版)
基 金:Supported by The Agricultural Science and Technology Project of Beijing(PXM2016-014207-000053)~~
摘 要:In order to find the optimal bacteriostatic agents and selectors and their concentrations in genetic transformation of Lespedeza bicolor with BADH as a marker gene, an resistance experiment was carried out with four chemicals, NaCl, betaine aldehyde, PEG and cefotaxime (Cef) added into the media, Different concentrations and combinations of different chemicals were added into the media in the stage of adventitious bud regeneration of cotyledonary nodes, Shoot segment multiplication, and rooting. The results showed that NaCI and betaine aldehyde could inhibit tissue differentiation of L. bicolor, and could thus be used as selectors. Furthermore, 0.8 g/L NaCI could completely inhibit the cotytedonary node differentiation and shoot segment multiplication, and 0.5 g/L NaCI was optimal for inhibition of rooting. It was also found that with the concentration increase of betaine aldehyde in media, adventitious buds regenerated from cotyledonary nodes decreased, and 1.5 g/L was confirmed to be the optimum concentration. Moreover, the mixture of 0.7 g/L NaCI and 0.3 g/L betaine aldehyde significantly inhibited shoot segment multiplication and rooting, so this mixture could be used asa selector. The Cef ex- periment showed that 200-300 mg/L C, ef in media did not inhibit significantly the regeneration of adventitious buds from cotyledonary nodes, and 100 mg/L Cef had little effect on the shoot segment multiplication and rooting.以BADH基因为选择标记基因进行遗传转化最适宜的选择剂、抑菌剂种类和浓度,通过不同浓度NaCl、甜菜碱醛、PEG及甜菜碱醛和NaCl的不同组合等进行胡枝子子叶节分化,茎段增殖以及生根培养,发现NaCl和甜菜碱醛都能抑制二色胡枝子的子叶节分化、茎段增殖和生根,可作为遗传转化中的选择剂。培养基中添加0.8 g/L NaCl就能完全抑制二色胡枝子子叶节和茎段增殖,可作为遗传转化过程中子叶节和茎段增殖阶段的选择剂浓度。培养基中添加0.5 g/L NaCl能抑制生根,适合作为离体培养生根阶段的选择剂浓度。随着培养基中甜菜碱醛浓度的增加,子叶节分化能力下降,当甜菜碱醛达到1.5 g/L时子叶节不再分化,这可作为以甜菜碱醛为选择剂时子叶节分化阶段的最佳筛选。培养基中同时添加NaCl和甜菜碱醛时,NaCl 0.7 g/L+甜菜碱醛0.3 g/L就抑制了离体培养过程中茎段增殖和生根,比较适合二色胡枝子离体培养时茎段增殖和生根的筛选浓度。对头孢霉素的研究发现:培养基中添加200~300 mg/L头孢霉素对离体培养的二色胡枝子子叶节分化影响不大,培养基中添加100mg/L的头孢霉素对离体培养的二色胡枝子茎段增殖和生根无显著影响。
关 键 词:Lespedeza bicolor NaCl Betaine aldehyde CEFOTAXIME POLYETHYLENEGLYCOL In-vitro culture
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