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为了建立苎麻外源DNA导入和遗传转化的实验体系,本试验以“湘苎3号”为材料,就激素、培养方法、培养条件对叶片外植体脱分化与再分化的影响作了较系统的研究。试验结果表明:1.较低浓度(0.1mg/l)的NAA能加快叶片启动。较高浓度(2.0~3.0mg/1)的BA有利于叶组织脱分化和愈伤组织形成,并能促进芽的再生,但BA浓度过高则抑制愈伤组织和芽的生长发育。2.NAA与BA配合使用能改善愈伤组织的质量和生长速度,促进芽的形成,高浓度BA与适当浓度NAA配合能分化丛生芽。配合使用时NAA与BA表现了互作效应,其互作效应受光温条件的影响。3.低温短光照预处理能提早叶组织启动;低温短光照预处理和昼夜温差预处理均能显著提高叶外植体分化成芽率;暗培养能促进愈伤组织分化和芽的发生。
In order to establish an experimental system of ramie exogenous DNA introduction and genetic transformation, this experiment made a systematic study on the effects of hormones, culture methods and culture conditions on dedifferentiation and re-differentiation of leaf explants with “Xiangzhu 3” Research. The test results show that: 1. Lower NAA concentration (0.1 mg / l) accelerated leaf start-up. Higher concentrations (2.0-3.0 mg / l) of BA were beneficial to dedifferentiation of leaf tissue and callus formation, and promoted regeneration of buds, but higher concentrations of BA inhibited callus and bud growth and development . 2. NAA and BA can improve the quality and growth rate of callus and promote bud formation. BA with high concentration can cooperate with NAA to differentiate the buds. With the use of NAA and BA showed an interaction effect, the interaction effect by light temperature conditions. 3. Low-temperature short-light pre-treatment can pre-activate leaf tissue. Low-temperature short-light pre-treatment and diurnal temperature difference pre-treatment can significantly improve the differentiation rate of leaf explants into shoots; dark culture can promote callus differentiation and bud formation.