CAJ | 학술논문

　　以继代培养60 d的大苞鞘石斛原球茎（矱＝2～3 mm）为材料，对比研究低温高渗（HT＋CA）和室温高渗（HT-CA）2种预处理方法对原球茎玻璃化法超低温保存冷冻前含水量以及预处理、脱水和解冻3个关键步骤中相对电导率、丙二醛质量摩尔浓度、可溶性糖和可溶性蛋白质量分数的影响，并结合显微结构观测和组织化学分析揭示高渗预处理影响超低温保存后原球茎相对存活率的生理生化基础。结果表明：在冷冻前，干样含水量显著降低，当干样含水量下降为1．06 g? g-1时，冻后相对存活率达最大值，为46．6％；预处理和脱水显著地提高了相对电导率；脱水和解冻使丙二醛质量摩尔浓度显著升高；可溶性糖和可溶性蛋白质量分数呈显著上升后明显下降的单峰变化曲线。多元相关分析表明，丙二醛质量摩尔浓度与相对存活率之间呈极显著负相关（相关系数为-0．992），膜脂过氧化作用是影响大苞鞘石斛原球茎冻后成活的关键因素之一。显微结构观察到细胞膜在预处理和脱水过程中受到明显破坏；组织化学分析显示淀粉粒和蛋白质的分布多集中在未受损伤的顶端分生组织中，分布变化呈先增多后减少的趋势，这与生理生化分析的结果是一致的。
　　이계대배양60 d적대포초석곡원구경（확＝2～3 mm）위재료，대비연구저온고삼（HT＋CA）화실온고삼（HT-CA）2충예처리방법대원구경파리화법초저온보존냉동전함수량이급예처리、탈수화해동3개관건보취중상대전도솔、병이철질량마이농도、가용성당화가용성단백질량분수적영향，병결합현미결구관측화조직화학분석게시고삼예처리영향초저온보존후원구경상대존활솔적생리생화기출。결과표명：재냉동전，간양함수량현저강저，당간양함수량하강위1．06 g? g-1시，동후상대존활솔체최대치，위46．6％；예처리화탈수현저지제고료상대전도솔；탈수화해동사병이철질량마이농도현저승고；가용성당화가용성단백질량분수정현저상승후명현하강적단봉변화곡선。다원상관분석표명，병이철질량마이농도여상대존활솔지간정겁현저부상관（상관계수위-0．992），막지과양화작용시영향대포초석곡원구경동후성활적관건인소지일。현미결구관찰도세포막재예처리화탈수과정중수도명현파배；조직화학분석현시정분립화단백질적분포다집중재미수손상적정단분생조직중，분포변화정선증다후감소적추세，저여생리생화분석적결과시일치적。
We used protocorms as material in subculture for 60 days and compared the physiological changes of two pretreatment methods, namely hypertonic treatment under cold acclimation(HT+CA)and room temperature (HT-CA), respectively, in vitrification cryopreservation, such as the water content before freezing and relative conductiv-ity, malondialdehyde concentration, soluble sugar, soluble protein levels during pretreatment, dehydration and thaw.We analyzed the physiological basis affected relative survival rate of protocorm after cryopreservation by microscopic structure observation and histochemical analysis.The water content of protocorms reduces significantly.The relative survival rate reaches a maximum of 46.6%, when the water content is 1.06 g? g-1 DW before freezing.Pretreatment and dehydration can improve the relative conductivity with a drastic increase in malondialdehyde content during dehydration and thaw.Sin-gle peak curve is manifested for the contents of soluble sugar and soluble protein, the trend ascendes markedly, and then decreases signally.Moreover, by the multiple correlation analysis the survival rate and the content of malondialdehyde are significantly negative correlation with the correlation coefficient of -0.992.The lipid peroxidation is a key factor influen-cing the survival rate in cryopreserved protocorms significantly.Microscope structural observation shows that the plasma membranes are damaged markedly during the pretreatment anddehydration .By the histochemical analysis of starch grain and protein, the distribution accumulates mostly in intact apical meristem with the variation trend of ascending and then de-scending in coincident with the physiological and biochemical characteristics above.