麻疯树精细胞分离
Isolation of Sperm Cells of Jatropha curcas L.

作者: 朱云 , 陈美灵 , 田惠桥 :厦门大学生命科学学院,厦门; 何恩铭 :福建省亚热带植物研究所,厦门;

关键词: 麻疯树精细胞分离Jatropha curcas L. Sperm Isolation

摘要: 将麻疯树成熟花粉置于一定渗透剂溶液中,可使其破裂,释放出其中的一对精细胞。释放的精细胞在20~30 min后用FDA染色仍可显示具有活性的荧光。用显微操作仪将释放的精细胞收集,得到一定数量具有活性的精细胞群体。研究了不同渗透剂浓度下释放的精细胞状态,比较了不同渗透剂的精细胞释放率及保持精细胞活性的最适浓度,以7.5%的葡萄糖溶液分离麻疯树精细胞的最佳浓度。麻疯树精细胞的成功分离为其离体受精实验奠定了一定基础,也为研究麻疯树精细胞发育的分子生物学实验提供了材料。

Abstract: Mature pollen of Jatropha curcas L. was broken in solutions with different osmolalities, and re-leased its cytoplasmic content, including two sperms. The isolated sperms could keep viability fluorescence for 20 - 30 min and were collected to concentration using a micromanipulator. The state of isolated sperms in different osmolalities was analyzed. Optimal concentration of isolation solution was compared according to the isolated sperm number and its duration of keeping viability and 7.5% glucose was best for isolating the sperm cells. The sperm isolation of Jatropha curcas L. makes a basis for its reproduction engineering study and provides study material to research its sperm development.

文章引用: 朱云 , 陈美灵 , 何恩铭 , 田惠桥 (2012) 麻疯树精细胞分离。 植物学研究, 1, 36-40. doi: 10.12677/BR.2012.12006

参考文献

[1] 田惠桥. 高等植物离体受精研究进展[J]. 物生理与分子生物学学报, 2003, 29(1): 3-10.

[2] E. Kranz, H. Lörz. In vitro fertilization with isolated, single gametes results in zygotic embryogenesis and fertile maize plants. Plant Cell, 1993, 5: 739-746.

[3] T. Uchiumi, I. Uemura, T. Okamoto, et al. Establishment of an in vitro fertilization system in rice (Oryza sativa L.). Planta, 2007, 226(3): 581-586.

[4] Y. Y. Wang, A. Kuang, S. D. Russell, et al. In vitro fertilization as a tool for investigating sexual reproduction of angiosperms. Sexual Plant Reproduction, 2006, 19(3): 103-115.

[5] D. Southworth, S. Kwiatkowski. Arabinogalactan proteins at the cell surface of Brassica sperm and Lilium sperm and generative cells. Sex Plant Reproduction, 1996, 9(5): 269-272.

[6] I. Tanaka. Differentiation of generative and vegetative cells in angiosperm pollen. Sexual Plant Reproduction, 1997, 10(1): 1-7

[7] C. Miao, X. P. Gou, L. Q. Lan, et al. Molecular cloning and expression of RSSG58 gene in rice sperm cells. Acta Bot Sin, 2003, 45(2): 234-241.

[8] L. L. Ge, X. P. Gou, T. Yuan, et al. Migration of sperm cells during pollen tube elongation in Arabidopsis thaliana: Behavior during transport, maturation and upon dissociation of male unit associations. Planta, 2011, 233(2): 325-332.

[9] P. Mukherjee, A. Varshney, T. S. Johnson, et al. Jatropha curcas: A review on biotechnological status and challenges. Plant Bio- tech Reports, 2011, 5(3): 197-215.

[10] S. Sato, H. Hirakawa, S. Isobe, et al. Sequence analysis of the genome of an oil-bearing tree, Jatropha curcas L. DNA Re- search, 2011, 18(1): 65-76.

[11] 王秀荣, 丁贵杰, 李平等. 麻疯树花的形态和解剖结构[J]. 林业科学, 2011, 9: 57-61.

[12] 莫永胜, 杨弘远. 紫菜苔精细胞的大量分离和生活力保存[J]. 植物学报, 1991, 33: 649-657.

[13] 刘焕芳, 廖景平, 吴国江. 麻疯树小孢子发育的超微结构研究[J]. 亚热带植物学报, 2007, 2: 107-114.

[14] J. E. Faure. Double fertilization in flowering plants: Origin, mechanisms and new information from in vitro fertilization. In: M. Cresti, et al., Eds., Fertilization in Higher Plants, Molecular and Cytological Aspects. Berlin: Springer-Verlag, 1999: 79-89.

[15] H. P. Xu, T. H. Tsao. Detection and immunolocalization of gly- coproteins of the plasma membrane of maize sperm cells. Proto- plasma, 1997, 198(3-4): 125-129.

[16] 胡适宜, 田惠桥. 雄性生殖单位的结构与功能[J]. 被子植物受精生物学, 北京: 科学出版社, 2002: 59-77.

分享
Top