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[目的]探索黄肉枇杷果实类胡萝卜积累的调控机制。[方法]该研究以枇杷突变体(黄肉枇杷一枝芽变结出白肉果实)为供试材料,利用高效液相色谱测定成熟果肉中β-胡萝卜素的积累,对野生型和突变型枇杷类胡萝卜素合成途径基因进行同源克隆及表达分析。[结果]野生型黄肉和突变体白肉枇杷成熟果肉中β-胡萝卜素含量分别为60.9和4.6μg/g;利用同源克隆获得的5个枇杷类胡萝卜素合成基因编码区序列在野生型和突变型没有发现核苷酸变异;实时荧光定量PCR分析显示类胡萝卜素β环羟化酶基因在突变体白肉果实中的表达显著降低,其它6个类胡萝卜素合成途径基因在两供试材料中的表达水平没有差异,与β-胡萝卜素的积累没有直接关联。[结论]突变体白肉枇杷缺少β-胡萝卜素的积累可能与类胡萝卜素β环羟化酶基因表达量下调有关。
[Objective] The research aimed to explore the regulatory mechanism of carrot accumulation in Loquat fruit. [Method] In this study, the loquat mutant (yellow meat loquat branch of a white bud fruit) as the test material, the determination of β-carotene accumulation in mature pulp by high performance liquid chromatography, the wild-type and mutant loquat Carotene synthesis pathway gene homologous cloning and expression analysis. [Result] The content of β-carotene in mature pulp of wild-type yellow meat and mutant white loquat were 60.9 and 4.6 μg / g, respectively. The coding sequence of five loquat carotenoid genes obtained by homologous cloning was wild type and Mutations showed no nucleotide variation. Real-time quantitative PCR analysis showed that the expression of carotenoid β-hydroxylase gene in mutant white meat was significantly decreased. The other six carotenoid pathway genes were found in the two tested materials There was no difference in the expression level of β-carotene, which was not directly related to the accumulation of β-carotene. [Conclusion] The lack of β-carotene accumulation in mutant white loquat may be related to the down-regulation of β-hydroxylase gene expression of carotenoids.