【作者】 程焕欣；

【导师】 梁玉玲；

【作者基本信息】 河北大学 ， 植物学， 2010， 硕士

【摘要】 甘草为我国重要的传统药材，主要有三萜类化合物和甘草黄酮类活性成分，其中黄酮类物质具有较强的生物活性，现代药理学研究发现甘草黄酮类对肿瘤、胃溃疡和肝损伤等具有多方面的药效，所以提高甘草黄酮含量则变得尤为重要。甘草一般生长在干旱盐化的环境，而次生代谢产物的合成与积累是长期生存选择的结果，与环境密切相关，给予一定的环境胁迫可以提高次生代谢产物的含量。本实验采用非生物胁迫的方法对胀果甘草愈伤组织进行胁迫处理，研究胁迫条件对愈伤组织的生长量、生长曲线、生理指标和次生代谢产物含量的影响。结果表明随着胁迫程度的增加，甘草愈伤组织的生长量一般呈下降趋势，但在0.5%NaCl胁迫下子叶愈伤组织生长量出现了上升状态。随着胁迫程度的增加MDA呈上升趋势，POD活性呈现先上升后下降的趋势。经过NaCl胁迫处理愈伤组织中甘草黄酮含量和对照比都有所提高，其中0.5%时子叶愈伤组织甘草黄酮含量最高，为对照的2.31倍，为药材的1.1倍；0.8%和1.0%时分别是对照的2倍和1.2倍。下胚轴愈伤组织在0.5%时为对照的2倍，为药材的0.278倍；其它浓度黄酮含量都大约为对照的1倍多。干旱胁迫下子叶愈伤组织甘草黄酮含量只有在PEG浓度为25%时比对照提高了，约为对照1.4倍，但低于药材。ABA处理对甘草黄酮也有影响，子叶愈伤组织甘草黄酮含量和对照比有所下降，下胚轴愈伤组织甘草黄酮含量有所提高。查尔酮合成酶(CHS)是黄酮类化合物合成中的一个关键酶，利用半定量RT-PCR的方法，检测不同胁迫条件下胀果甘草愈伤组织中chs基因表达量，结果表明胀果甘草愈伤组织中总黄酮含量与chs基因的表达量成正相关。更多还原

【Abstract】 Liquorice is an important traditional Chinese medicinal，the mainly active ingredientsare triterpenoids and licorice flavonoid，Licorice flavonoid has stronger bio-activities. Theconclusions of pharmacological investigation are that they have anti-cancer，anti-ulcers，anti-liver damage and so on，it becomes particularly important to improve the concent oflicorice flavonoid. Secondary metabolites in plant are closely related to the arid environmentof salinization where Licorice usually grows，and that is the result of the choice of long-termsurvival. According to that，the content of secondary metabolites can be increased by givinga certain degree of environmental stress.It is necessary to study the effect of abiotic stress on the growth，growth curve，physiological indicators and the liquorice flavonoid of Glycyrrhiza inflata callus. With thestress level increased，the growth of the Licorice callus drops，and MDA concentration rises.The growth of cotyledon callus aslo arises only under 0.5%NaCl stress. At first，PODactivity shows up，then downward. Under 0.5% NaCl stress，the flavonoid content ofcotyledon callus is 2.31 fold and 1.1 fold to compare with the control and the herbsrespectively. However，to compare with the control by the 0.8%NaCl and the 1.0%NaCl，the flavonoid content of cotyledon callus is 2.0 and 1.2 fold respectively. Licorice flavonoidcontent of hypocotyl callus is 2.0 fold to compare with the control by the 0.5% NaCl，and isalso improved under the ABA treatment. Flavonoid content of cotyledon callus was 1.4 timesto compare with the control by 25% PEG-6000，but it was lower than that of herbs.Chalcone Synthase (CHS) is a key enzyme in the biosynthesis of flavonoids. Usingsemi-quantitative RT-PCR method，the expression of chs gene of Glycyrrhiza inflata calluswere detected under different treatment，the results show that the flavonoid content of callusis positively correlated with the expression of chs gene.更多还原