当前位置: 首页 > 文章 > 湿加松胚性愈伤组织的程序降温技术研究 南京林业大学学报(自然科学版) 2013,37 (6) 4-8
Position: Home > Articles > Cryopreservation of embryogenic callus of Pinus elliottii × Pinus caribaea using control freezing Journal of Nanjing Forestry University(Natural Sciences Edition) 2013,37 (6) 4-8

湿加松胚性愈伤组织的程序降温技术研究

作  者:
刘伟东;陈金慧;周艳威;赵亚琦;施季森
单  位:
林木遗传与生物技术省部共建教育部重点试验室;南京林业大学
关键词:
湿加松;超低温冷冻保存;胚性愈伤组织
摘  要:
对湿加松胚性愈伤组织进行了超低温冷冻保存的研究。结果表明:继代培养9~12 d的湿加松胚性愈伤组织经0.5 mol/L山梨醇预培养4 d,在0.6 mol/L山梨醇+10%DMSO的冷冻保护液下0℃预处理20 min,然后以-1℃/min的降温速率降至-40℃,停留10 min后再以-5℃/min的降温速率降至-90℃,投放入液氮中保存;复苏时于37℃水浴2 min,1 mol/L山梨醇的液体培养基清洗3次,以滤纸作支持物转到固体继代培养基再培养。在此程序处理下1个月内可获得生长状态良好的再生愈伤组织。
译  名:
Cryopreservation of embryogenic callus of Pinus elliottii × Pinus caribaea using control freezing
作  者:
LIU Weidong;CHEN Jinhui;ZHOU Yanwei;ZHAO Yaqi;SHI Jisen;Key Laboratory of Forest Genetics and Biotechnology Ministry of Education,Nanjing Forestry University;
关键词:
Pinus elliottii × Pinus caribaea;;embryogenic callus;;cryopreservation
摘  要:
Cryopreservation of callus of Pinus elliottii × Pinus caribaea were conducted in this research. The results showed that the best process of cryopreservation for the embryogenic callus of P. elliottii × P. caribaea was as follows:the callus were sub-cultured for 9-12 days in callus proliferation medium firstly and were pre-cultured in 0. 5 mol/L sorbol for 4 days,and then pretreated 20 minutes in buffer of 0. 6 mol/L sorbol + 10% DMSO,cooled to -40 ℃ at rate of -1 ℃/min,maintained at -40 ℃ for 10 min,preserved in liquid nitrogen after cooled to -90 ℃ at rate of -5 ℃/min. The resuscitate callus were kept in 37 ℃ water bath,clean three times in 1 mol/L sorbol liquid medium,passage in solid medium support by filter paper. New embryogenic callus will be produced within one month.

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