近年来的研究发现,葡萄对高锰胁迫具有极强的抗性,但是其耐锰生理仍不清楚。以2种对锰富集相反的葡萄品种(根系富集的金手指和地上部富集的康拜尔品种)为研究材料,采用沙培实验法,用不同浓度的MnSO4溶液进行锰毒胁迫处理,通过荧光定量PCR,分析锰胁迫相关基因在2种葡萄的叶片及根部表达的差异,挖掘耐锰相关基因,探讨基因表达的品种差异性及组织差异性。结果表明:在高锰(10 g·L-1)胁迫下,金手指葡萄根系中葡萄油菜素内酯-6-氧化酶基因表达量上调2倍,其余几个基因没有明显变化,在其叶片中葡萄铜伴超氧化物歧化酶基因、葡萄油菜素内酯-6-氧化酶基因都上调2倍,病程相关基因表达量上调5倍,而葡萄4-香豆酸:辅酶A连接酶基因下降为CK组的一半。康拜尔根系中油菜素内酯-6-氧化酶基因、3-脱氧-D-阿拉伯庚酮糖-7-磷酸合成酶基因和葡萄紫色酸性磷酸酶3类似酶基因都上调了2倍,叶片中的病程相关基因上调了2~3倍。实验表明,高锰胁迫下,2种葡萄都会通过抗逆基因的差异表达来应对锰毒,同时,磷酸水解酶类也协同作用。由于2种葡萄不同的锰富集方式,这些抗逆基因在根系和叶片也呈现出不同的调控方式。在这些差异表达中,病程相关蛋白1基因和油菜素内酯-6-氧化酶基因表现的最为明显,在抵抗锰胁迫中可能发挥了较大的作用。
Two grape varieties (Gold Finger and Campbell) were taken as the materials to carry out a sand culture experiment,study the different expressions of genes of roots and leaves under various MnSO4 stresses,and explore the differences in varieties and tissue in gene expression. It was found that,under the excess Mn stress,the up-expression folds of Vitis vinifera brassinosteroid-6-oxidase gene in the roots of Gold Finger were 2,and there was no significant change for other genes. In the leaves,the up-expression folds of V.vinifera copper chaperone for superoxide dismutase-like gene and V.vinifera brassinosteroid-6-oxidase gene were 2,and the up-expression fold of pathogenesis-related protein 1 gene were 5. For the expression of V.vinifera 4-coumarate:CoA ligase gene was a half of CK. At Campbell,the up-expression folds of V.vinifera brassinosteroid-6-oxidase gene,3-deoxy-D-arabino-heptulosonate 7-phosphate synthase gene and purple acid phosphatase 3-like gene in roots were 2,there was no obvious change for others,and those of pathogenesis-related protein 1 gene varied from 2 to 3. The experiment indicated that,under Mn stress,the tolerance of both the grape varieties to excess Mn stress responded through the different expressions of stress resistance gene,and phosphoric acid hydrolases also had a synergistic effect. There were the different regulatory approaches between roots and leaves of these genes due to the different ways of manganese enrichment in two grape varieties. From the different gene expressions,pathogenesis-related protein 1 gene and V.vinifera brassinosteroid-6-oxidase gene had the most significant manifestations,and they may be at an important position in the way of resistance to excess manganese.
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