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林业科学 ›› 2002, Vol. 38 ›› Issue (3): 1-6.doi: 10.11707/j.1001-7488.20020301

• 论文及研究报告 •    下一篇

胡杨愈伤组织继代增殖和器官发生中蛋白分子标记的研究

谷瑞升1 蒋湘宁3 裴东2   

  1. 中国科学院植物研究所,北京100093;北京林业大学森林生物学中心,北京100083;中国林业科学研究院林业研究所,北京100091
  • 收稿日期:2000-12-27 修回日期:1900-01-01 出版日期:2002-05-25 发布日期:2002-05-25

PROTEINS ASSOCIATED WITH CALLUS PROLIFERATION AND ADVENTITIOUS BUD DIFFERENTIATION OF POPULUS EUPHRATICA

Gu Ruisheng,Jiang Xiangning,Pei Dong   

  1. Institute of Botany, The Chinese Academy of Sciences Beijing100093;Experimental Center of Forest Biology, Beijing Forestry University Beijing100083;Institute of Forestry, Chinese Academy of Forestry Beijing100091
  • Received:2000-12-27 Revised:1900-01-01 Online:2002-05-25 Published:2002-05-25

摘要:

利用十二烷基磺酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)和等电聚焦双向电泳(IEF/2D)对胡杨不同类型愈伤组织和愈伤组织分化不定芽过程中表达的蛋白进行了研究。发现:不同类型愈伤组织中表达的蛋白存在着明显的差异。在光照和培养基中BA/NAA值为1时诱导产生的有极强器官发生能力的茎基愈伤组织,其蛋白组分明显地少于经过继代培养、器官发生能力明显下降的愈伤组织,表明了茎基愈伤组织本身分化程度低。实验获得了不同类型愈伤组织和愈伤组织分化不定芽过程中不同阶段所表达的特异蛋白,表现为经过黑暗和培养基中BA/NAA值为0.5的继代增殖培养,愈伤组织产生了特异的24.5kDa和58.6kDa的标记蛋白带,并且也表达了其器官发生时表达的19kDa和31kDa蛋白带。茎基愈伤组织在光照和BA NAA值为0.5的条件下进行器官发生诱导并且随着愈伤组织形成分生细胞团块和不定芽原基,明显地表达了20kDa和5 5kDa蛋白带,在20kDa蛋白带中含有pI为5.5~6.5的特异蛋白。pI为6.5~7.5的43kDa为不定芽发生前期表达的特异蛋白。文中就愈伤组织器官发生能力与其表达蛋白的关系进行了讨论。

关键词: 胡杨, 标记蛋白, 愈伤组织增殖, 不定芽分化

Abstract:

The changes in protein patterns were monitored with SDS PAGE and IEF/2D during callus proliferation and adventitious bud differentiation of Populus euphratica. The results showed callus differentiation potential was closely related to its expressed proteins. Callus induced from shoot bases on the medium with the ratio of BA to NAA 1 under light, had a high differentiation capability, and expressed less sorts of proteins than those being subcultured, indicating its differentation degree was lower. Marker proteins for different kinds of calli and different phases of callus differentiation were identified. Two subculture specific markers protein bands, 24.5 kDa and 58.6 kDa, were observed in callus proliferated in dark on the medium with the rate of BA to NAA 0.5. Meanwhile, the 19 kDa and 31 kDa protein bands that expressed during callus differentation were also found in the subcultured callus, suggesting that gradual loss of organogenesis ability of the subcultured callus may be contributed to the increasing of its differentiation. When the callus induced from shoot bases was cultured on the regeneration medium with the ratio of BA to NAA 0.5 under light, it firstly differentiated into a few smaller green meristematic islands in its surface layer, then adventitious shoot primordial from the islands. Two protein bands of 20 kDa and 55?kDa, as distinctive differentiation markers, expressed obviously in the differentiated callus. The 20 kDa protein with pI of 5.5~6.5 was specific to callus differentiation. A 43 kDa protein with pI of 6.5~7.5 generated during the early stage of differentiation. The relation between callus organogenesis potential and its expressed proteins was discussed.

Key words: Populus euphratica, Marker protein, Callus proliferation, Adventitious bud differentation