摘要
山茶是世界上三大饮料之一，又有重要的食用及医疗保健价值，具有重要的经济意义。本文以山茶初春新生枝条的幼叶为外植体，采用不同的培养基，探讨了不同的植物生长调节物质对愈伤组织的诱导和不同的理化因子对愈伤组织的增殖效应。研究了在愈伤组织增殖过程中的生长周期以及生长周期中的生理生化指标的变化。分析了在愈伤组织生长周期中山茶总黄酮、总皂甙和茶多酚的含量变化，为山茶的进一步开发，细胞培养和次生物质的生产提供理论和实验依据。
试验结果如下：
1．愈伤组织的诱导与增殖：
ER培养基是山茶愈伤组织诱导与增值的最佳培养基。MS、N6、B5、三种培养基也可以诱导出愈伤组织。
不同浓度、不同种类的植物生长调节物质对山茶的愈伤组织具有不同的诱导效应。单因子以2，4-D2.0mg／L的效果最好。组合因子NAA1.0mg／L+6-BA1.0mg／L最好。
愈伤组织增殖的培养基也是ER培养基+NAA1.0mg／L+6-BA1.0mg／L为最好。在愈伤组织增殖过程中，其生长周期为30天，生长曲线“S”形，基本上可以分为三个时期即延迟期(0～12天)、指数增长期(12～24天)、稳定期(24～30天)。碳源以葡萄糖40mg／L为最佳。光照对愈伤组织的增殖有积极的作用。
用以防褐化时，活性炭不起作用，以0.5％的水解酪蛋白为好。
3．愈伤组织生长周期中生理生化指标的变化
可溶性蛋白的变化、过氧化物酶(POD)活性的变化、超氧化物歧化酶(SOD)活性的变化均与生长周期密切相关。同时受光照条件的影响。
4．次生代谢物含量的变化
山茶总黄酮、总皂甙以及茶多酚的含量变化也与愈伤组织的生长状态有密切的联系。次生代谢物的合成与积累与光照条件有一定的联系。愈伤组织次生代谢物的积累主要在细胞增长周期的稳定期。光照培养下，三种次生代谢物的含量均高于暗培养。
Camellia is one of three main drinks in the world, it also has great edible and medical value. The paper reported something about it. The callus was induced and proliferated using young leave as explant, which comes out at early spring. In this paper, the author discussed the induction effect and the proliferation effect by using different kinds of hormone and different physical-chemical factors. The growth circle and some physiology-biochemical indexes were illustrated. As the same time, the tendencies of flavonids, saponins and tea-polyphenols during growth circle are analysed. The paper provides scientific and experimental basis for cell culture and secondary metabolism matters yield.
The results are following:
1.The effects of induction and the proliferation which mediums are applied different kinds of hormone or different concentration hormone are different.
Medium ER is optimum for callus inducing and proliferation, but medium MS B5, N6 also can be used to induce callus. In callus inducing, 2,4-D2.0mg/L is the best of all single hormone, NAA1.0mg/L+6-BA1.0mg/L is the best of all combined hormone. In callus proliferation, NAA1.0mg/L+6-BA1.0mg/L is the best.
The effects of different physical-chemical factors are different. The callus cultured in white light grows better than that in dark. When the concentration of glucose is 40mg/L, the effect of callus proliferation is the best of all.
Activated carbon has no use in preventing callus browning, but protein tyrosine hydrolysate 0.5%does good.
2.The change of physiology-biochemical norm:
The growth circle of callus is 30 days and the growth-curve looks like "S". The growth circle could be divided three parts: Delay phase(0-12d), Index phase(12-24d)and Steady phase (24-30d).
During the growth circle, the tendency soluble protein contents, SOD, POD activities were different during the three parts. They show some physiology -biochemical indexes character during proliferation and cell activities. It is different between growth-circle of callus cultured brightly and that of callus cultured darkly.
3.The change of secondary metabolism matters content:
During growth circle, flavonids, saponins and tea-polyphenols were connected
with callus growing condition. Secondary metabolism matters synthetic and accumulated are also connect with growing condition. Secondary metabolism matters accumulated mainly during steady phage (24-30d). And when the callus cultured brightly, the three secondary metabolism matters content are higher than that of callus cultured darkly.
引文
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