毛竹实生苗对铝毒反应的研究
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摘要
铝毒害是酸性土壤中限制植物生长的重要逆境因子。铝毒害主要通过抑制植物根伸长进而抑制植物对水分、养分的吸收最终抑制植物的生长和发育。本研究以毛竹种子和实生苗为材料研究不同铝浓度对毛竹种子发芽率、根长生长以及植物胁迫保护酶SOD、POD和MDA的影响,并通过同为禾本科且已研究较透彻的铝耐性不同的2个水稻品种(敏感型IR1552和耐受型Azucena)对比,确定毛竹的耐铝程度;采用切片染色技术,观察铝在毛竹实生苗根尖分布情况和对细胞的影响。研究结果如下:
毛竹种子发芽率在铝浓度为200μM时并没有受到显著性抑制,当达到500μM时,开始出现抑制现象,2000μM达到半致死浓度。毛竹种子的耐铝程度低于铝敏感型水稻IR1552。毛竹实生苗在铝胁迫(500μM) 1 d时出现根伸长显著被抑制现象,且3 d内抑制逐步加深,未见恢复。而两种水稻均能在3 d内不同程度的恢复一定生长能力。毛竹实生苗经铝胁迫3 d后SOD活性变化不显著(p=0.1531),两种水稻均为极显著(p均为0)。毛竹的POD活性变化显著(p=0.018),两种水稻均为极显著(IR1552为0.0002,Azucena为0.0084)。相对于两种水稻,0μM浓度下毛竹苗根系中的POD活性和MDA的含量均较高。总体而言,毛竹耐铝程度低于水稻敏感型IR1552。
而切片染色观察结果表明,毛竹实生苗根尖上的铝主要分布在1-2 mm部位,而且铝在根尖上主要分布在细胞壁等非原生质体中。但是在分生区和伸长区,有部分铝进入到原生质体中,核膜和核内尤其明显。试验结果也显示,铝能诱导毛竹实生苗根尖细胞凋亡,在500μM浓度下毛竹实生苗处理1d即能出现凋亡小体。而且只有当细胞核内出现铝,其后才会出现凋亡小体。
Based on the effects of different aluminum(Al) concentrations on the seed germination, seedling root growth, SOD(superoxide dismutase), POD(peroxidase) and MDA(malondialdehyde) in Moso bamboo, this study was to determine the level of Al tolerance for Moso bamboo through the comparison with two rice varieties IR1552 (Al-sensitive)and Azucena (Al-tolerant). With cytological techniques we observed the distribution of Al in the Moso bamboo seedlings, and the influence of Al on the Moso bamboo. The results are as follows:
Al concentration of 200μM did not significantly inhibit the seed germination of the Moso bamboo, while 500μM inhibited the germination rate. And the concentration of 2000μM resulted in 50% survival. With 500μM Al, the root growth was inhibited in 1d. The growth did not recover under Al stress within 3 d. While the root growth in IR1552 and Azucena would be more or less recovered within 3d. Under the Al stress for 3 d, the increase of SOD activity in root tips for Moso bamboo was insignificant (p=0.1531), while for IR1552 and Azucena were significant (p=0). The increase of POD activity was significant (p=0.018) for Moso bamboo, highly significant for IR1552 and Azucena. Moso bamboo’s POD activity and MDA content in root tips were higher than those in IR1552. It concluded that Moso bamboo had a lower Al-tolerant level than IR1552 .
With lumogallion staining and confocal microscopy, the localization of Al in seedling root tips of Phyllostachys pubescens was observed. The results showed that Al was mainly distributed in non-protoplasts, and most of it located in 1~2mm of the root tips. But in the meristematic zone and elongation zone, Al was found in protoplasts, especially in the nucleus. Within a certain range of Al concentrations (0~2000μM), the filtering of Al into root tips would be accelerated as the treated Al concentration increased and/or the treatment prolonged. In addition, Al could induce apoptosis in the root tips of Moso bamboo’s seedlings. When the root tips were under 500μM Al for 24 h, there always appeared apoptotic bodies. Only after the Al appeared in nuclei and the apoptotic bodies would appear.
毛竹种子发芽率在铝浓度为200μM时并没有受到显著性抑制,当达到500μM时,开始出现抑制现象,2000μM达到半致死浓度。毛竹种子的耐铝程度低于铝敏感型水稻IR1552。毛竹实生苗在铝胁迫(500μM) 1 d时出现根伸长显著被抑制现象,且3 d内抑制逐步加深,未见恢复。而两种水稻均能在3 d内不同程度的恢复一定生长能力。毛竹实生苗经铝胁迫3 d后SOD活性变化不显著(p=0.1531),两种水稻均为极显著(p均为0)。毛竹的POD活性变化显著(p=0.018),两种水稻均为极显著(IR1552为0.0002,Azucena为0.0084)。相对于两种水稻,0μM浓度下毛竹苗根系中的POD活性和MDA的含量均较高。总体而言,毛竹耐铝程度低于水稻敏感型IR1552。
而切片染色观察结果表明,毛竹实生苗根尖上的铝主要分布在1-2 mm部位,而且铝在根尖上主要分布在细胞壁等非原生质体中。但是在分生区和伸长区,有部分铝进入到原生质体中,核膜和核内尤其明显。试验结果也显示,铝能诱导毛竹实生苗根尖细胞凋亡,在500μM浓度下毛竹实生苗处理1d即能出现凋亡小体。而且只有当细胞核内出现铝,其后才会出现凋亡小体。
Based on the effects of different aluminum(Al) concentrations on the seed germination, seedling root growth, SOD(superoxide dismutase), POD(peroxidase) and MDA(malondialdehyde) in Moso bamboo, this study was to determine the level of Al tolerance for Moso bamboo through the comparison with two rice varieties IR1552 (Al-sensitive)and Azucena (Al-tolerant). With cytological techniques we observed the distribution of Al in the Moso bamboo seedlings, and the influence of Al on the Moso bamboo. The results are as follows:
Al concentration of 200μM did not significantly inhibit the seed germination of the Moso bamboo, while 500μM inhibited the germination rate. And the concentration of 2000μM resulted in 50% survival. With 500μM Al, the root growth was inhibited in 1d. The growth did not recover under Al stress within 3 d. While the root growth in IR1552 and Azucena would be more or less recovered within 3d. Under the Al stress for 3 d, the increase of SOD activity in root tips for Moso bamboo was insignificant (p=0.1531), while for IR1552 and Azucena were significant (p=0). The increase of POD activity was significant (p=0.018) for Moso bamboo, highly significant for IR1552 and Azucena. Moso bamboo’s POD activity and MDA content in root tips were higher than those in IR1552. It concluded that Moso bamboo had a lower Al-tolerant level than IR1552 .
With lumogallion staining and confocal microscopy, the localization of Al in seedling root tips of Phyllostachys pubescens was observed. The results showed that Al was mainly distributed in non-protoplasts, and most of it located in 1~2mm of the root tips. But in the meristematic zone and elongation zone, Al was found in protoplasts, especially in the nucleus. Within a certain range of Al concentrations (0~2000μM), the filtering of Al into root tips would be accelerated as the treated Al concentration increased and/or the treatment prolonged. In addition, Al could induce apoptosis in the root tips of Moso bamboo’s seedlings. When the root tips were under 500μM Al for 24 h, there always appeared apoptotic bodies. Only after the Al appeared in nuclei and the apoptotic bodies would appear.
引文
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