【摘 要】
:
本实验以当年生平邑甜茶(Malus hupehensis(Pamp) Rehd.)幼苗和三年生幼树为试材,克隆了平邑甜茶多胺合成的三个关键酶的基因(分别为精氨酸酶,精氨酸脱羧酶和鸟氨酸脱羧酶),
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本实验以当年生平邑甜茶(Malus hupehensis(Pamp) Rehd.)幼苗和三年生幼树为试材,克隆了平邑甜茶多胺合成的三个关键酶的基因(分别为精氨酸酶,精氨酸脱羧酶和鸟氨酸脱羧酶),探讨了这些基因在平邑甜茶幼树和幼苗的不同发育时期及不同器官的表达情况,研究了盐胁迫和重金属胁迫下平邑甜茶多胺的合成途径,探讨了外源多胺缓解重金属胁迫对平邑甜茶的伤害的机制。主要实验结果如下:1.根据已知蛋白基因序列设计简并引物,利用RT-PCR结合RACE技术获得了平邑甜茶的精氨酸酶、精氨酸脱羧酶基因和鸟氨酸脱羧酶基因,分别命名为MhArg、MhADC和MhODC,并在Genebank分别注册为EF427896、EU431331、EU431332。2.平邑甜茶幼树和幼苗的不同发育时期及不同器官的表达情况表明MhArg在贮藏营养转换期表达量高,而MhODC和MhADC多在幼嫩组织表达。3.在盐胁迫下,MhADC的表达量随处理时间而逐渐增大,叶片中处理6h达到高峰,而根中3h就达到表达高峰。MhODC在叶片中表达量受盐胁迫影响不明显,不过在根中24h时表达量明显升高。在盐胁迫下平邑甜茶体内腐胺含量明显上升,12h达到高峰,后下降,而精胺和亚精胺含量明显下降。在重金属胁迫明显促进了MhODC的表达,而对MhADC表达的诱导不明显。重金属胁迫下明显促进腐胺的积累,亚精胺含量下降,而精胺含变化较平稳。平邑甜茶在重金属和盐胁迫下多胺的合成主要是依靠ADC途径。相比盐胁迫在重金属胁迫下平邑甜茶腐胺的积累更多地依赖ODC途径。4.在氯化镉胁迫下,外源多胺可以明显增加平邑甜茶叶片中的内源Spd和Spm的含量,降低Put的含量,从而明显提高(Spd+Spm)/Put的值,通过提高抗氧化酶的活性,增强了直接清除活性氧的能力,从而减轻了氯化镉胁迫下的膜质过氧化,减轻了重金属胁迫对平邑甜茶的伤害。
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