论文部分内容阅读
1987年1月17日,从植物园采取中华猕猴桃36号的枝条,进行表面消毒后,切成约1 cm长的茎段,并从中央交叉切开成4块,立即投入试管中,加入在0℃预冷的冰冻保护剂,在水浴中预处理45分钟,随后以1℃/min的降温速度从0℃降到-10℃;停留20分钟,继而以同样的降温速度从-10℃降到-40℃;停留3小时,然后投入液氮中超低温(-196℃)保存。经120天贮存后,在40℃水浴中化冻,在附加1 mg/L玉米素的MS培养基上培养25天左右即可看到从皮层和木质部之间产生愈伤组织,愈伤组织是来源于形成层细胞。猕猴桃茎段经120天的超低温保存后,其存活率几乎是100％,而且茎段产生的愈伤组织在生长状态上也与未经超低温保存的一样,致密、硬实、带绿色,分化出的新植株的形态、色泽完全正常。这一结果为猕猴桃体细胞种质的长期低温保存提供了可能性。(简
January 17, 1987, taken from the Botanical Garden branches of Chinese kiwifruit No. 36, surface disinfection, cut into about 1 cm long stems, and cut into four from the central cross, immediately put into test tubes, adding 0 Precooling cryoprotectant was pretreated for 45 minutes in a water bath and then decreased from 0 ° C to -10 ° C at a cooling rate of 1 ° C / min; held for 20 minutes and then decreased from -10 ° C to the same cooling rate -40 ℃; stay 3 hours, then put liquid nitrogen in ultra-low temperature (-196 ℃) preservation. After 120 days of storage, they were thawed in a 40 ° C water bath and cultured on MS medium supplemented with 1 mg / L zeatin for about 25 days to produce callus from cortex and xylem, which is the source of callus Forming layer cells. After 120 days of cryopreservation, the survival rate of kiwifruit stems was almost 100%, and the callus generated from the stems was in the same growth state as that of the non-cryopreserved, dense, hard, greenish, differentiated New plant morphology, color completely normal. This result offers the possibility of long-term cryopreservation of kiwifruit somatic germplasm. (simple