水曲柳的无菌发芽与胚轴的离体再生
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摘要
本论文首次系统研究了水曲柳无菌发芽和胚轴外植体的离体再生。通过切
开处理克服了水曲柳成熟种子的休眠；从直接不定芽增生途径，确立了一套
水曲柳组织培养再生系统。
将未经层积处理的水曲柳成熟种子切去子叶端的1／3，在添加了BA的MS
或1／2MS培养基中培养38天，切开种子的发芽率达65.3％，未切开种子的发
芽率为0。BA显著促进了切开种子的萌发，包含BA的培养基中切开种子的
发芽率为75.0％，是未添加处理发芽率的2倍以上。蔗糖和基本培养基对切
开种子的发芽率没有显著影响，但蔗糖可以促进发芽后幼苗子叶的伸长生长。
以水曲柳成熟种子的胚轴为外植体，实现了植株再生。WPM培养基对不
定芽的诱导是非常有效的。只有添加了TDZ或BA后才能诱导产生不定芽，
1.0mg／L是TDZ与BA诱导效果变化的转折点。低于此浓度，TDZ处理的诱
导率高，反之，低于BA的诱导效果。生长素对不定芽的产生没有任何好的
影响。在不定芽的伸长上，1／2MS优于MS和WPM培养基、TDZ优于BA
的诱导伸长效果。在整个培养过程中，培养物的死亡率是一个重要的影响因
子。1／2MS不适合在生根培养时使用，WPM培养基中的生根率为19.1％。再
生植株很容易驯化成活。
Aseptic germination and plant regeneration of Fraxinus mandshurica Rupr. was studied. Overcoming of dormancy and germination of seeds were obtained by cut seeds. And a culture system via adventitious buds regeneration, was established.
Non-stratified mature seeds of F. mandshurica, were cut and removed 1/3 of the seed coat near to the cotyledons. Then they were cultured aceptically onto MS or half strength MS medium (1/2 MS) supplemented with BA. Through 38 days of culture, 65% of the seeds germinated. While none of the intact seeds germinated. BA was strikingly effective for germination of cut-seeds. On medium containing BA, 75% of the seeds germinated, which was 2 times over non-treated seeds. Sucrose and basal media had no effects on germination in this experiment. However, sucrose could stimulate elongation of germinated buds.
Plant regeneration of F. mandshurica Rupr. from hypocotyls explants, was obtained. Woody plant medium (WPM), was effective for induction of adventitious buds. Multiplication of buds can only be induced with the supplemented of TDZ or BA. TDZ was more effective than BA when the concentration was lower than 1.0 mg/L, but less effective than BA when over 1.0 mg/L. Auxins seemed to have no effects on multiplication of adventitious buds. For the elongation of buds, 1/2 MS was more effective than WPM, and TDZ was better than BA. Compared to 1/2 MS, WPM was better for rooting, and the percentage of the rooted shoots was 19. 1%. Rooted plantlets were easy to survive after acclimation.
开处理克服了水曲柳成熟种子的休眠；从直接不定芽增生途径，确立了一套
水曲柳组织培养再生系统。
将未经层积处理的水曲柳成熟种子切去子叶端的1／3，在添加了BA的MS
或1／2MS培养基中培养38天，切开种子的发芽率达65.3％，未切开种子的发
芽率为0。BA显著促进了切开种子的萌发，包含BA的培养基中切开种子的
发芽率为75.0％，是未添加处理发芽率的2倍以上。蔗糖和基本培养基对切
开种子的发芽率没有显著影响，但蔗糖可以促进发芽后幼苗子叶的伸长生长。
以水曲柳成熟种子的胚轴为外植体，实现了植株再生。WPM培养基对不
定芽的诱导是非常有效的。只有添加了TDZ或BA后才能诱导产生不定芽，
1.0mg／L是TDZ与BA诱导效果变化的转折点。低于此浓度，TDZ处理的诱
导率高，反之，低于BA的诱导效果。生长素对不定芽的产生没有任何好的
影响。在不定芽的伸长上，1／2MS优于MS和WPM培养基、TDZ优于BA
的诱导伸长效果。在整个培养过程中，培养物的死亡率是一个重要的影响因
子。1／2MS不适合在生根培养时使用，WPM培养基中的生根率为19.1％。再
生植株很容易驯化成活。
Aseptic germination and plant regeneration of Fraxinus mandshurica Rupr. was studied. Overcoming of dormancy and germination of seeds were obtained by cut seeds. And a culture system via adventitious buds regeneration, was established.
Non-stratified mature seeds of F. mandshurica, were cut and removed 1/3 of the seed coat near to the cotyledons. Then they were cultured aceptically onto MS or half strength MS medium (1/2 MS) supplemented with BA. Through 38 days of culture, 65% of the seeds germinated. While none of the intact seeds germinated. BA was strikingly effective for germination of cut-seeds. On medium containing BA, 75% of the seeds germinated, which was 2 times over non-treated seeds. Sucrose and basal media had no effects on germination in this experiment. However, sucrose could stimulate elongation of germinated buds.
Plant regeneration of F. mandshurica Rupr. from hypocotyls explants, was obtained. Woody plant medium (WPM), was effective for induction of adventitious buds. Multiplication of buds can only be induced with the supplemented of TDZ or BA. TDZ was more effective than BA when the concentration was lower than 1.0 mg/L, but less effective than BA when over 1.0 mg/L. Auxins seemed to have no effects on multiplication of adventitious buds. For the elongation of buds, 1/2 MS was more effective than WPM, and TDZ was better than BA. Compared to 1/2 MS, WPM was better for rooting, and the percentage of the rooted shoots was 19. 1%. Rooted plantlets were easy to survive after acclimation.
引文
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