【作者】 刘坤；

【导师】 汤庚国；

【作者基本信息】 南京林业大学 ， 园林植物与观赏园艺， 2008， 硕士

【摘要】 我国北方大部分地区水和土壤偏碱性导致了百合切花质量下降,制约了百合生产的发展。因此,培育耐盐碱百合品种就成为解决这一问题的有效方法。岷江百合(Lilium regale Wils.)是百合属中耐盐碱的优良种类。本文选用岷江百合为试材,以鳞片诱导愈伤组织为起始材料,培育出稳定的悬浮细胞系,得到再生植株;以岷江百合生长良好的胚性愈伤组织为试验材料,羟脯氨酸(HYP)和NaCl两种作为筛选剂,采用一步筛选法得到岷江百合耐盐细胞系。主要研究结果如下:1.以试管苗鳞片作为外植体,获得诱导愈伤组织的适宜培养条件为:光照条件下,MS+6-BA 1.0~2.0mg/L+NAA 0.1mg/L+3%蔗糖。在黑暗条件下,MS+6-BA 2.0 mg/L+NAA 0.1mg/L+3%蔗糖有利于愈伤组织的继代培养,易形成淡黄色疏松的胚性愈伤组织。光照培养下,愈伤组织的颜色出现绿色、褐色不同程度的变化,不利于愈伤组织的继代培养。2.通过对不同的接种量、蔗糖浓度、激素配比的研究得出了悬浮细胞适宜的生长条件:接种量2.0g/40mL,蔗糖浓度以30g~60g/L为宜,激素组合为MS+1.0 mg/L6-BA。根据悬浮细胞生长曲线得出,7天继代一次较适宜。3.采用一步筛选法将生长良好的胚性愈伤组织接种到不同NaCl和HYP浓度下进行筛选,得到不同盐浓度下的岷江百合耐盐细胞系。本试验只对在最高盐浓度下(70mmol/LNaCl与0.15mmol/LHYP)得到的耐盐变异体进行分析。4.通过测定耐盐变异体的生理生化指标发现,0.15mmol/LHYP耐盐变异体的脯氨酸、可溶性糖的含量显著高于对照,分别是对照的1.88和1.54倍,丙二醛的含量低于对照,并且各项生理指标两者之间都达到极显著性差异。而70mmol/LNaCl耐盐变异体的各项指标与对照之间没有达到显著性差异,表明0.15mmol/LHYP耐盐变异体为真正的耐盐变异体,而70mmol/LNaCl耐盐变异体为盐生理适应性愈伤组织。5.在盐胁迫下,耐盐变异体的生长量都高于对照,K~+、Na~+含量也高于对照,并能保持与对照相近的K~+/Na~+比。表明筛选得到的0.15mmol/LHYP耐盐变异体具有耐盐性和稳定性。更多还原

【Abstract】 On the condition of alkalescence in soils and water in North China,the cut flower quality of lily decreased seriously and the development of lily was restricted.Therefore,breeding of salt tolerant lily variety was a effective way to solve this question.Lilium regale Wils. was the excellent material for salt-toletant selection in Lilium species.In this paper,the bulbscales of L regale were used as explants to induce callus.Suspension cell lines were established and plants regenerated from the cell lines.By posotive selection methods,the callus were selected to obtain the salt-tolerant lines with the selective pressure of NaCl and HYP.The main results described as follows:1. The bulbscales of L regale were used as explants to induce callus in diffrent hormones.Finally,the results showed that the eligible medium to callus induction was MS+6-BA 2.0mg/L+NAA 0.1mg/L+3%sugar.Light was suitable for growth of callus;It was showed that the optimal medium to callus subculture was MS+6-BA 2.0mg/L+NAA 0.1mg/L+3%sugar in the dark.On the condition of dark, callus growed quickly and the color of callus was yellow.On the condition of light,the color of callus changed to green and brown differently.2. By researching the inoculum densities,sucrose concentrations and hormones,it showed that the optimal conditions for the growth of L regale cell lines were as follows:2.0g/40mL,30-60g/L sucrose,optimal medium was MS+1.0mg/L 6-BA. According to the growth curve of cell,it was better to subculture in seven-day cycles.3. The callus were planted onto the media containing maintained different NaCl and HYP concentrations.By posotive selection methods,the salt-tolerant callus of different salt concentrations were obtained.However,the salt-tolerant callus of highest salt concentations (70mmol/LNaCl and 0.15mmol/LHYP) were only researched.4. Through physiological and biochemical analyse,the contents of proline and soluble sugar of the salt-tolerant callus of 0.15mmol/LHYP were 1.88,1.54 times higher than control respectively,the content of MDA of the salt-tolerant callus was lower than control.It was included that the salt-tolerant callus of 0.15mmol/LHYP had significant salt tolerance,but the salt-tolerant callus of 70mmol/LNaCl did not have.So the salt-tolerant callus of 0.15mmol/LHYP was the true salt-tolerant variants,but the salt-tolerant callus of 70mmol/LNaCl was only the cells adaptability to salt. 5. The salt-tolerant callus of 0.15mmol/LHYP and the control were planted onto the media containing maintained different NaCl concentrations.It showed, in 20 days, that the growth rate of the salt-tolerant callus and the content of K~+、Na~+ were higher than the control and the ratio of K~+/Na~+ was the similar to the control.更多还原