包埋脱水法冷冻保存裙带菜配子体克隆的研究
Cryopreservation of clonal gametophytes of Undaria pinnatifida by encapsulation-dehydration
-
摘要: 将裙带菜配子体克隆包埋在含3%褐藻酸钙的胶球中,研究了预培养中的蔗糖浓度和培养时间,脱水过程中的脱水速率和胶球含水量以及化冻速度对配子体克隆的存活率和发育能力的影响,结果表明含裙带菜配子体克隆的胶球在0.3 mol/dm3的蔗糖培养基中培养9 h可以获得较高的存活率;胶球最适平均脱水速率为含水量的5%每小时,最适含水量为25%;较快的化冻速度可以显著提高配子体克隆的存活率,在优化条件下存活率可高达66%;经过冷冻保存的裙带菜配子体克隆在适合发育的条件下能够正常形成孢子体.Abstract: Clonal gametophytes of Undaria pinnatifida were encapsulated in 3% calcium alginate beads. The main factors influencing the viability and the developm entability of clonal gametophy tes,such as sucrose concentration,preculture time,dehydration rate,water content of beads and thawing rate were studied.The results show that the high viability can be obtained when alginate beads are precultured for 9 h in the medium containing 0.3 mol/dm3 sucrose.The optimum average dehydration rate of beads is reduction of 5% water content per hour and the optimum water content of beads is 25%.Fast rate of thawing can significantly improve the viability of clonal gametophytes.Under optimum conditions,the cry opreservation viability of clonal gametophytes can reach 66% at the most.After cry opreservation,the clonal game to phytes can produce the normal sporophytes in the conditions suitable for development.
-
Key words:
- clonal Undaria pinnatifida /
- cryo preservation /
- encapsulation dehydration /
- viability
-
王素娟.海藻生物技术[M上海:上海科学技术出版社,1994.112-115. 方宗熙,戴继勋,陈登勤.裙带菜的孤雌生殖及其后代的遗传特点[J].海洋学报,1979,1(1):112-116. 李大鹏,吴超元,刘晚昌,等.海带单倍体无性繁殖系育苗技术的研究[J].海洋学报,2003,25(5):141-146. TAYLOR R, FLETCHER R. Cryopreservation of eukaryotic alga——a review of methodologies[J]. Journal of Applied Phycology,1999,10:481-501. 王起华,石若夫,程爱华.3种饵料金藻的超低温保存研究[J].中国水产科学,1999,6(2):89-92. 王起华,张恩栋,李大鹏,等.盐生杜氏藻和青岛大扁藻的超低温保存[J] 植物学报,2000,42(4):399-402. 王起华,张恩栋,周春影.藻类种质超低温保存研究概况[J].植物学通报,2002,19(1):21-29. ARBAULT S, RENARD P, PÉREZ R,et al. Essai de cryopreservation des Gamétophytes de l'algue alimentaire Undaria pinnatifida(Laminariales)[J]. Aquatic Living Resources, 1990,3: 207-215. GINSBURGER-VOGEL T, ARBAULT S, PÉREZ R. Ultrastructural study of the effect of freezing-thawing on the gametophyte of the brown alga Undaria pinnatifida[J]. Aquaculture, 1992, 106:171-181. 王君晖,边红武,黄纯农.植物样品包埋脱水法超低温保存的研究进展[J].植物学通报,1999,16(5):582-586. 仵小南、谭桂英、周百成,等.几种海洋微藻的固定化培养[J].海洋学报,1992,14(1):129-133. HIRATA K, PHUNCHINDAWAN M, TAKAMOTO J,et al. Cryopreservation of microalgae using encapsulation-dehydration[J].Cryo-Letters, 1996, 17:321-328. VIGNERON T, ARBAULT S, KAAS R. Cryopreservation of gametophytes of Laminaria digitata (L) Lamouroux by encapsulation dehydration[J]. Cryo-Letters, 1997, 18:93-98. 王起华,刘明,程爱华.坛紫菜(Porphyra haitanensis)自由丝状体的胶囊化冰冻保存[J].辽宁师范大学学报(自然科学版),2000,23(4):387-390. 李莹,王起华,李贺,等.用包埋脱水法冰冻保存牟氏角刺藻[J].海洋科学,2003,7:48-51. HORNUNG R, DOMAS R, LYNCH P T. Cryopreservation of plumular explants of coconut (Cocos nucifera L.) to support programmes for mass clonal propagation through somatic embryogenesis[J]. Cyro-Letters, 2001, 22:211-220. MALAURIE B, TROUSLOT M F, ENGELANN F, et al. Effect of pretreatment conditions on the cryopreservation of in vitro-cultured yam (Dioscorea alata "Brazo Fuerte" and D. Bulbifera "Nouméa Imboro") shoot apices by encapsulation-dehydration[J]. Cryo-Letter,1998,19:15-26. SHIBLI R A, AL-JUBOORY K H. Cryopreservation of ‘nabali' Olive (Olea europea L.) somatic embryos by encapsulation-dehydration and encapsulation-vitrification[J]. Cryo-Letters, 2000, 21: 357-366. BLAKESLEY D, KIERNAN R J. Cryopreservation of axillary buds of a Eucalyptus grandis X Eucalyptus camaldulensis hybrid[J].Cryo-Letters, 2001, 22:13-18. LI C, LOH C S, SUN W Q. An improved dehydration protocol for cryopreservation of Brassica napus somatic embryos[J]. Cryo-Letters, 1999, 20:263-268. CAÑAVATE J P, LUBIAN L M. Effects of slow and rapid warming on the cryopreservation of marine microalgae[J]. Cryobiology,1997, 35:143-149.
点击查看大图
计量
- 文章访问数: 756
- HTML全文浏览量: 10
- PDF下载量: 1076
- 被引次数: 0