Effect of light and activated charcoal on tracheary element differentiation in callus cultures of Pinus radiata D. Don
Effect of light and activated charcoal on tracheary element differentiation in callus cultures of Pinus radiata D. Don
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DOI：
10.1007/s11240-005-9065-z
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发表时间：
2006-05-01
影响因子：
3
通讯作者：
Find, J
中科院分区：
文献类型：
--
作者：
Möller, R;Ball, RD;Find, J
已发现，在源自辐射松木质部薄壁组织的愈伤组织培养物中，在含有活性炭但不含植物激素的诱导培养基上生长的愈伤组织培养物中，光可增加气管成分分化的比例。分化率从在黑暗中生长愈伤组织时的20％增加到在16小时或24小时光周期下生长愈伤组织时的45％。当愈伤组织在 16 小时的光周期下生长时，将愈伤组织转移到诱导培养基后 2 天即可观察到气管元件，而在黑暗中培养愈伤组织则需要 5 天。分化率还受诱导培养基中活性炭添加浓度的影响，最佳浓度为5 g·l(-1)。从诱导培养基中排除活性炭使分化率降低至 2%。与在黑暗中生长相比，在 16 小时光周期下生长的细胞培养物中，木质素相关酶 L-苯丙氨酸解氨酶和肉桂醇脱氢酶的活性显着更高。结果表明，光对辐射松愈伤组织培养物中气管元件的分化和木质素相关酶的活性具有刺激作用。新的生长条件显着改善了该细胞培养系统，使其特别适用于针叶裸子植物体外生长的气管元件的功能基因测试和细胞壁分析。
Light has been found to increase the proportion of tracheary elements differentiating in callus cultures derived from xylem-parenchyma of Pinus radiata D. Don grown on an induction medium containing activated charcoal but no phytohormones. The differentiation rate increased from 20% when callus was grown in darkness to 45% when callus was grown with a 16 h or 24 h photoperiod. When callus was grown with a 16 h photoperiod, tracheary elements were observed 2 days after transfer of callus to the induction medium, as compared to 5 days when callus was cultured in darkness. The differentiation rate was also influenced by the concentration of activated charcoal added to the induction medium, the optimum concentration being 5 g l(-1). Exclusion of activated charcoal from the induction medium decreased the differentiation rate to 2%. The activities of the lignin-related enzymes L-phenylalanine ammonia lyase and cinnamyl alcohol dehydrogenase were significantly higher in cell cultures grown with a 16 h photoperiod as compared to when grown in darkness. The results show that light had a stimulating effect on tracheary element differentiation and the activities of lignin-related enzymes in P. radiata callus cultures. The new growth conditions markedly improve this cell culture system and make it particularly useful for functional gene testing and cell-wall analysis of in vitro grown tracheary elements of coniferous gymnosperms.