论文总字数：18223字

摘 要

以金钗石斛原球茎为对象，研究信号分子茉莉酸甲酯对其生长、繁殖以及石斛碱积累的影响规律。基于茉莉酸甲酯的添加时间和浓度两个因素，采用植物生物反应器和固体瓶培养金钗石斛原球茎，分析金钗石斛原球茎中石斛碱含量规律。

结果表明：原球茎在培养20 d后，添加茉莉酸甲酯浓度为5μmol/L时，相较于其他添加浓度，石斛碱产量最高，为20.0597μg/mL；在考察添加时间时，添加10μmol/L茉莉酸甲酯，石斛碱含量在第20 d添加时最高，为18.5147μg/mL。综上表明，在植物生物反应器培养第20 d加入5μmol/L的茉莉酸甲酯为金钗石斛原球茎培养及其石斛碱积累的最佳方式。

在固体瓶培养时，添加茉莉酸甲酯浓度为2.5μmol/瓶时，金钗石斛原球茎的生长繁殖状态最好，增殖系数可达到86.38%；在添加时间上，第30 d加入茉莉酸甲酯，增殖系数可以达到68.67%。

金钗石斛原球茎的生长繁殖状态来看，植物生物反应器培养要优于固体瓶，增殖系数可以达到155.18%。

关键词：金钗石斛 原球茎 反应器 茉莉酸甲酯 石斛碱

Effect of Methyl Jasmonate on Dendrobiine Production from Protocorm of Dendrobium nobile

ABSTRACT

The protocorm of Dendrobium nobile was taken as an object to study the effect of methyl jasmonate, a signal molecule, on its growth, reproduction and accumulation of dendrobine. Based on the addition time and concentration of methyl jasmonate, the protocorm of Dendrobium nobile was cultured in plant bioreactor and solid bottle. The content rule of dendrobine in protocorm of Dendrobium nobile was analyzed.

The results showed that when the concentration of methyl jasmonate was 5μmol/L after the protocorm was cultured for 20 d, compared with other concentrations, the yield of dendrobine was the highest, which was 20.0597μg/mL. When examining the addition time, 10μmol/L methyl jasmonate was added, and the dendrobiine content was the highest at the 20th day, 18.5147μg/mL. To sum up, adding 5μmol/L methyl jasmonate at the 20th day of plant bioreactor culture is the best way to culture Dendrobium nobile protocorm and accumulate dendrobium alkaloids. When the concentration of methyl jasmonate was 2.5μmol/ bottle in solid bottle culture, the protocorm of Dendrobium nobile had the best growth and reproduction state, and the multiplication coefficient could reach 86.38%. When methyl jasmonate was added at the 30th day, the proliferation coefficient could reach 68.67%.

According to the growth and reproduction status of Dendrobium nobile protocorm, the bioreactor culture is better than the solid bottle, and the multiplication coefficient can reach 155.18%.

Key Words: Dendrobium nobile; Protocorm; Bioreactor; Methyl jasmonate; Dendrobium alkaloid

目 录

摘要…………………………………………………………………………………I

ABSTRACT………………………………………………………………………II

第一章 前言 ……………………………………………………………………1

1.1金钗石斛…………………………………………………………………… 1

1.2石斛碱……………………………………………………………………… 2

1.3反应器……………………………………………………………………… 3

1.4茉莉酸甲酯………………………………………………………………… 4

第二章 材料和研究方法………………………………………………………6

2.1培养基……………………………………………………………………… 6

2.2金钗石斛原球茎的接种培养……………………………………………… 6

2.3茉莉酸甲酯的配置和添加………………………………………………… 6

2.3.1反应器培养金钗石斛原球茎…………………………………… 6

2.3.2固体瓶培养金钗石斛原球茎…………………………………… 7

2.4金钗石斛原球茎生理生化指标测定 ………………………………………7

2.4.1增殖系数的测定………………………………………………… 7

2.4.2褐化率的测定 ……………………………………………………7

2.4.3石斛碱的提取与含量测定……………………………………… 8

第三章 结果与分析……………………………………………………………9

3.1反应器培养的金钗石斛原球茎…………………………………………… 9

3.1.1加入不同浓度茉莉酸甲酯的处理……………………………… 9

3.1.2不同时间加入茉莉酸甲酯的处理………………………………11

3.2固体瓶培养的金钗石斛原球茎……………………………………………15

3.2.1加入不同浓度茉莉酸甲酯的处理………………………………15

3.2.2不同时间加入茉莉酸甲酯的处理………………………………17

3.2.3反应器与固体瓶培养金钗石斛原球茎的比较…………………20

第四章 结论……………………………………………………………………22

第五章 展望……………………………………………………………………23

参考文献…………………………………………………………………………24

致谢 ………………………………………………………………………………27

第一章 前言

金钗石斛

石斛科植物被中医视为名贵药材，常常以茎入药，《本草纲目》等医书古籍对其功效用量等均有记载。石斛被认为对治疗目暗不明有疗效，有养胃润肺、强健筋骨甚至是延年益寿的功效。但因其对生长条件如湿度、温度、光照要求严苛，种子十分细小、不储存营养，不易繁殖，又因高价值被过度采集，导致野生资源稀少，又因石斛科植物种类繁多，品质良莠不齐，导致不能在市场大范围普遍应用。

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