湿加松胚性愈伤组织的程序降温技术研究

刘伟东; 陈金慧; 周艳威; 赵亚琦; 施季森

doi:10.3969/j.issn.1000-2006.2013.06.001

湿加松胚性愈伤组织的程序降温技术研究

刘伟东,陈金慧,周艳威,赵亚琦,施季森

南京林业大学学报（自然科学版） ›› 2013, Vol. 37 ›› Issue (06) : 1-5.

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南京林业大学学报（自然科学版） ›› 2013, Vol. 37 ›› Issue (06) : 1-5. DOI: 10.3969/j.issn.1000-2006.2013.06.001

研究论文

湿加松胚性愈伤组织的程序降温技术研究

刘伟东,陈金慧,周艳威,赵亚琦,施季森^*

作者信息 +

Cryopreservation of embryogenic callus of Pinus elliottii × Pinus caribaea using control freezing

LIU Weidong, CHEN Jinhui, ZHOU Yanwei, ZHAO Yaqi, SHI Jisen^*

Author information +

文章历史 +

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。

Abstract

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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刘伟东,陈金慧,周艳威,赵亚琦,施季森. 湿加松胚性愈伤组织的程序降温技术研究[J]. 南京林业大学学报（自然科学版）. 2013, 37(06): 1-5 https://doi.org/10.3969/j.issn.1000-2006.2013.06.001

LIU Weidong, CHEN Jinhui, ZHOU Yanwei, ZHAO Yaqi, SHI Jisen. Cryopreservation of embryogenic callus of Pinus elliottii × Pinus caribaea using control freezing[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2013, 37(06): 1-5 https://doi.org/10.3969/j.issn.1000-2006.2013.06.001

中图分类号： S723.1+34

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基金

收稿日期:2013-03-07 修回日期:2013-08-27
基金项目:国家林业公益性行业科研专项项目(201004049); 国家自然科学基金重点项目(30930077); 江苏高校优势学科建设工程资助项目(PAPD)
第一作者:刘伟东,博士生。*通信作者:施季森,教授。 E-mail:jshi@njfu.edu.cn。
引文格式:刘伟东,陈金慧,周艳威,等. 湿加松胚性愈伤组织的程序降温技术研究[J]. 南京林业大学学报:自然科学版,2013,37(6):1-5.