以前期在超低温保存中鉴定出凋亡蛋白的春石斛类原球茎（PLBs）为实验材料，通过研究类原球茎玻璃化超低温保存中程序性细胞死亡（PCD）特征指标、PCD诱导物质（过氧化氢、超氧阴离子、一氧化氮、乙烯）内源含量变化及其抑制剂使用对PCD和存活率的影响，确定PCD内源信号分子；在此基础上，将PCD内源信号的抑制剂和线粒体通透性转换孔抑制剂分别导入玻璃化超低温保存程序，以未添加体系为对照，检测这些实验体系中PCD特征指标、线粒体PCD途径特征指标、信号分子内源含量和我们鉴定得到的定位在内质网的凋亡蛋白的表达变化，探讨信号分子和线粒体PCD途径及凋亡蛋白的关系，从而揭示超低温保存中PCD相关信号分子及其传导途径。

Protocorm-like bodies (PLBs) of Dendrobium nobile, from which 2 cryopreservation differential expression proteins related to apoptosis were identified, will be used as the material in our study. First, the programmed cell death (PCD) event, the contents of endogenous PCD inducers (including hydrogen peroxide, superoxide radical, nitric oxide and ethylene) and the effect of their inhibitors on PCD event and PLBs survival will be determined during the cryopreservation by vitrification to find out the PCD related signal molecules. Then, inhibitors of PCD related signal molecular and mitochondrial permeability transition pore will be used in protocal of PLBs cryopreservation by vitrification as additives, with the no added as control, while the characteristic index of PCD event and mitochondrial PCD pathway, the content of endogenous signals molecules and the expression of apoptosis-related protein will be measured. Last, the relationship between the signal molecules and mitochondrial PCD pathway and the apoptosis-related protein located in endoplasmic reticulum will be discussed. The aim is to explore the PCD related signal molecules and their regulation pathways during the cryopreservation.