甘蔗蔗糖合酶基因Susy半定量表达体系的建立及其应用  

Establishment of Semi-quantitative RT-PCR System for Susy Gene in Sugarcane and Its Application

作　　者：黄文静[1] 檀小辉[2] 黄静丽[1] 何龙飞[1] 杨丽涛[1] 李杨瑞 王爱勤[1] 

机构地区：[1]广西大学农学院,广西南宁530004 [2]广西壮族自治区亚热带作物研究所,广西南宁530001 [3]广西作物遗传改良与生物技术重点实验室,广西南宁530007

出　　处：《热带作物学报》2013年第7期1288-1293,共6页Chinese Journal of Tropical Crops

基　　金：国家科技支撑计划项目(No.2007BAD30B00);广西自然科学基金项目(No.2011GXNSFA018069)

摘　　要：为进一步研究高等植物体内控制蔗糖合成的关键酶基因蔗糖合酶基因Susy在甘蔗中的表达情况,本研究以甘油醛-3-磷酸脱氢酶(GAPDH)基因为内参基因,通过对退火温度、循环数的优化建立了一个稳定、方便的半定量RT-PCR体系。通过该体系检测干旱处理下甘蔗叶的Susy基因表达,并结合叶片酶活性和蔗糖含量进行分析。结果表明:甘蔗Susy基因的半定量RT-PCR扩增,以循环数30个、退火温度为54℃、内参基因与目的基因同批异管进行扩增为宜。甘蔗Susy基因在不同叶位中的表达有很大差异,总体上表现为:+1叶、+2叶>+5叶、+6叶>心叶,干旱胁迫可诱导幼嫩叶中Susy基因的表达量上升,基因表达的变化与其酶活性和蔗糖含量变化的情况相一致。To further study the expression of Susy gene,which is the key gene in sucrose synthesis,a stable and convenient semi-quantitative RT-PCR system was established by using GAPDH gene as the inner control,optimizing the cycle number and annealing temperature for PCR system.The expression of Susy gene in sugarcane was analyzed by using this system under drought treatment combined with analysis of enzyme activity and the sucrose content.The result showed that the most suitable semi-quantitative RT-PCR cycle was 30,annealing temperature was 54 ℃,and the PCR amplification of inner control gene GAPDH and the target gene Susy should be done with the same cycle numbers in different tubes.The expression of sucrose synthas gene was quite different in sugarcane leaves with different positions in the plant,which showed leaves ＋1,＋2 leaves ＋5,＋6 heartleaf.Drought treatment induced the expression of Susy gene in the immature leaves.The changes in gene expression,enzyme activity of Susy and sugar content were basically identical.

关 键 词：甘蔗 蔗糖合酶 干旱 基因表达 

分 类 号：S59[农业科学—作物学] Q943.2[生物学—植物学]