湿加松胚性愈伤组织的程序降温技术研究

doi:10.3969/j.issn.1000-2006.2013.06.001

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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 • 下一篇

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,林木遗传与生物技术省部共建教育部重点试验室,江苏南京 210037

出版日期:2013-12-18 发布日期:2013-12-18
基金资助:
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.

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

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

Key Laboratory of Forest Genetics and Biotechnology Ministry of Education,Nanjing Forestry University, Nanjing 210037, China

Online:2013-12-18 Published:2013-12-18

摘要/Abstract

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.

中图分类号:

S723.1⁺34

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(06): 1-5.

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 (Natural Science Edition), 2013, 37(06): 1-5.DOI: 10.3969/j.issn.1000-2006.2013.06.001.

参考文献

[1] 陈代喜,莫钊志,李贵玉,等.杂交松育苗对比试验效果分析[J].广西林业科学,2002,31(2):76-77. Chen D X, Mo Z Z, Li G Y, et al. The controlled experiment on Pinus elliottii × Pinus oaribaea seedling-raising[J]. Guangxi Forestry Science,2002,31(2):76-77.
[2] 庞正轰,陈代喜,莫钊志,等.澳大利亚杂交松产脂量及松脂成分研究[J].林业科技开发,2009,23(6):23-26. Pang Z H, Chen D X, Mo Z Z, et al. Study on the output of resin and its chemical composition from Australia pine hybrid [J]. China Forestry Science and Technology, 2009, 23(6): 23-26.
[3] 凌绍明.澳大利亚杂交松在广西引种试验研究[J].广西林业科学,2007,36(4):192-195. Ling S M. A study on the introduction of Australia hybrid pine in Guangxi[J]. Guangxi Forestry Science,2007, 36(4): 192-195.
[4] 方利娟.杂交松组织培养技术研究[D].广西: 广西大学,2006. Fang L J. Study on technology about the tissue culture of Pinus elliottii×P.caribaea[D]. Guangxi:Guangxi University, 2006.
[5] 施翔.杂交松组织培养研究[D].南京: 南京林业大学,2005. Shi X. Study on tissue culture of Pinus elliottii×P.caribaea[D]. Nanjing: Nanjing Forestry University, 2005.
[6] 项东云,王以红,吴幼媚, 等.杂交松组培技术研究初探[J].广西林业科学,2009, 38(3):134-136. Xiang D Y, Wang Y H, Wu Y M, et al. Preliminary study on tissue culture of Pinus elliottii×P.caribaea[J].Guangxi Forestry Science,2009, 38(3):134-136.
[7] 施季森,于敏,边黎明,等.湿地松和加勒比松杂种体细胞胚胎发生和植株再生方法. CN200710132707.8[P]. 北京:知识产权出版社,2008. Shi J S, Yu M, Bian L M, et al. Method for somatic embryogenesis and plant regeneration of hybrid between Pinus elliotti and Pinus caribaea. CN200710132707.8[P].Beijing: Intellectual Property Publishing House,2008.
[8] 李峰卿.湿加杂种松体细胞胚胎发生同步化控制和植株再生[D].南京: 南京林业大学, 2009. Li F Q. Synchronization controlling system for the embryogenesis proliferationand plantlets regeneration of the new hybrid between Pinus elliottii and Pinus caribaea[D]. Nanjing: Nanjing Forestry University, 2009.
[9] 叶芳,夏胜,梅兴国.红豆杉愈伤组织超低温保存有关因素的研究[J].武汉植物学研究,2001,19(4):327-331. Ye F, Xia S, Mei X G. Studies on factors in cryopreservation of Taxus callus [J]. Journal of Wuhan Botanical Research,2001,19(4):327-331.
[10] Nieves V, Conchi S, Lorena J, et al. Cryopreservation of chestnut by vitrification of in vitro-grown shoot tips[J]. In Vitro Cellular & Developmental Biology-Plant,2005,41(1):63-68.
[11] 史峰厚, 喻方圆, 沈永宝, 等. 超低温贮存对油松种子的影响[J].南京林业大学学报:自然科学版, 2005, 29(6): 119-122. Shi F H, Yu F Y, Shen Y B, et al. Cryopreservation of chinesepine seeds[J]. Journal of Nanjing Forestry University:Natural Sciences Edition, 2005, 29(6): 119-122.
[12] 任淑娟, 王航, 刘浩, 等. 七叶树种子离体胚的超低温冷冻保存[J].南京林业大学学报:自然科学版, 2010, 34(2): 19-23. Ren S J, Wang H, Liu H, et al. Cryopreservation of excised embryos of Aesculus chinensis seeds [J]. Journal of Nanjing Forestry University:Natural Sciences Edition, 2010, 34(2): 19-23.
[13] 施季森. 迎接21世纪现代林木生物技术育种的挑战[J].南京林业大学学报, 2000, 24(1): 1-6. Shi J S. The challenges to forestry biotechnology and tree breeding in the 21st century[J]. Journal of Nanjing Forestry University, 2000, 24(1): 1-6.
[14] 邵崇斌.概率论与数理统计[M].北京:中国林业出版社,2005.
[15] 徐刚标,易文,李美娥, 等.银杏愈伤组织超低温保存的研究[J].林业科学,2001, 37(3):30-34. Xu G B, Yi W, Li M E, et al. The studies of cryopreservation of callus for maidenhair tree(Gingko bilobal L.)[J]. Scientia Silvae Sinicae,2001, 37(3):30-34.
[16] 苗琦,谷运红,王卫东, 等.植物组织培养物的超低温保存[J].植物生理学通讯,2005,41(3): 350-354. Miao Q, Gu H Y, Wang W D, et al.The cryopreservation of plant tissue culture materials [J].Plant Physiology Journal, 2005, 41(3): 350-354.
[17] Karlsson J O, Toner M. Long-term storage of tissues by cryopreservation: critical issues [J]. Biomaterials, 1996, 17(3):243-256.
[18] 关静,龚承元,崔占峰.玻璃化冷冻保存细胞组织研究进展[J].国外医学:生物医学工程分册,2004, 27(4):252-256. Guan J, Gong C Y, Cui Z F.Recent advances in cryopreservation of cells and tissues by vitrification [J]. International Journal of Biomedical Engineering,2004, 27(4):252-256.
[19] 翟晓巧,程斐,朱延林.二乔刺槐愈伤组织超低温保存及适宜降温方法[J].林业科学,2009, 45(10):49-54. Zhao X Q, Cheng F, Zhu Y L. Methods of lowering temperature for cryopreservation of calli of Robinia bella-rosea[J]. Scientia Silvae Sinicae, 2009, 45(10):49-54.
[20] 刘剑锋,阎秀峰,程云清,等.高山红景天愈伤组织的超低温保存[J].林业科学,2007, 43(6):57-60. Liu J F, Yan X F, Cheng Y Q, et al. Cryopreservation of callus of Rhodiolasa chalinensis[J].Scientia Silvae Sinicae,2007, 43(6):57-60.
[21] 李广武,郑从义,唐兵.低温生物学[M].长沙:湖南科学技术出版社,1998.
[22] Barbara M Reed. Plant Cryopreservation: A Practical Guide[M]. New York:Springer, 2007.
[23] Malabadi R B, Nataraja K. Cryopreservation and plant regeneration via somatic embryogenesis using shoot apical domes of mature Pinus roxburghii sarg. trees[J]. In Vitro Cellular & Developmental Biology-Plant, 2006, 42(2):152-159.
[24] Mathur G, Alkuter V A, Nadgauda R S. Cryopreservation of embryogenic culture of Pinus roxburghii[J]. Biologia Plantarum, 2003, 46(2):205-210.
[25] Salaj T, Panis B, Swennen R, et al. Cryopreservation of embryogenic tissues of Pinus nigra Arn. by a slow freezing method[J]. CryoLetters, 2007, 28(2):69-76.
[26] Gale S, John A, Benson E E. Cryopreservation of Picea sitchensis(Sitka spruce)embryogenic suspensor masses[J]. CryoLetters, 2007,28(4):225-239.
[27] krlep K, Bergant M, De Winter G M. Cryopreservation of cell suspension cultures of Taxus media and Taxus floridana[J]. Biologia Plantarum, 2008, 52(2):329-333.
[28] 叶芳,夏胜,梅兴国.红豆杉愈伤组织超低温保存有关因素的研究[J].武汉植物学研究,2001,19(4):327-331. Ye F, Xia S, Mei X G. Studies on factors in cryopreservation of Taxus callus [J]. Journal of Wuhan Botanical Research,2001,19(4):327-331.
[29] 王高,范现丽,申晓辉,等.红松胚性愈伤组织的超低温保存技术[J].上海交通大学学报:农业科学版,2009,27(3):223-230. Wang G, Fan X L, Shen X H, et al. Cryopreservation of Korean pine somatic calli [J].Journal of Shanghai Jiaotong University:Agricultural Science,2009,27(3):223-230.

湿加松胚性愈伤组织的程序降温技术研究

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

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