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文中分析了葡萄愈伤组织诱导和继代培养的最佳光照强度并探索葡萄愈伤组织褐变的机理。以金手指葡萄的幼嫩茎段为材料,设置0、500、1 000、1 500、2 000、2 500、3 000和4 000 Lx等8个光照强度,研究不同光照强度对葡萄愈伤组织的诱导率、褐化率及相关酶活性和基因表达的影响。结果表明,在0、500、1 000和1 500 Lx光照强度处理下,愈伤组织的诱导率均达到92%以上,显著高于其他处理(P<0.05),其中1 000和1 500 Lx光照强度处理愈伤组织及其继代培养的生长势良好且褐化程度较轻;绿原酸、咖啡酸、对羟基苯甲酸和香豆酸与葡萄愈伤组织褐变关系密切,其中绿原酸的含量与褐化程度呈显著正相关(P<0.05);POD和PPO酶活性与褐化率呈极显著负相关(P<0.01);POD、PPO和PAL酶基因的表达量与褐化率呈显著(P<0.05)或极显著(P<0.01)正相关。因此,葡萄愈伤组织诱导和继代培养适宜的光照强度为1 000-1 500 Lx,光照强度过高或过低对其正常的生理生长都有较大负面影响。
In this paper, we analyzed the best light intensity of callus induction and subculture in grape and explored the mechanism of browning of grape callus. The young shoots of golden finger grapes were used as materials to set up 8 light intensities of 0, 500, 1 000, 1 500, 2 000, 2 500, 3 000 and 4 000 Lx to study the effects of different light intensities on the quality of grape callus Induction rate, browning rate and related enzyme activity and gene expression. The results showed that the induction rate of callus reached more than 92% at 0, 500, 1 000 and 1 500 Lx light intensity, significantly higher than other treatments (P <0.05), of which 1 000 and 1 500 Lx light The growth vigor of callus and its subculture were good and the degree of browning was light. The chlorogenic acid, caffeic acid, p-hydroxybenzoic acid and coumaric acid were closely related to the browning of grape callus, (P <0.05). There was a significant negative correlation between POD and PPO activities and browning rate (P <0.01). The expression of POD, PPO and PAL genes and browning rate were significantly Significantly (P <0.05) or very significant (P <0.01) positive correlation. Therefore, the optimum light intensity for the induction and subculture of grape callus was 1000-1 500 Lx, and the light intensity was too high or too low to have a significant negative impact on its normal physiological growth.