斜纹夜蛾Spodoptera litura是一种世界性分布的重要农业害虫,在生长发育过程中要经历幼虫一蛹的变态发育过程。由于变态发育前后昆虫的食性发生了明显的改变,作为食物消化吸收的中肠也发生了解体和重建。与此相适应,昆虫中肠的各种物质和能量代谢也可能会相应地发生改变。为研究斜纹夜蛾中肠变态发育过程中糖代谢途径的变化情况,我们从斜纹夜蛾中肠EST文库中鉴定出了12个糖代谢相关基因,克隆了其中3个基因的全长cDNA,并应用半定量PCR和定量PCR的办法检测了其在幼虫-蛹变态发育期中肠组织的转录表达以及对激素和饥饿等因素的响应情况。结果表明:这3个基因(α-L-岩藻糖苷酶、N-乙酰葡萄糖胺-6-磷酸去乙酰酶和烯醇化酶基因)的开放阅读框分别为1 461,1 200和1 299 bp,预测的分子量分别为56.3,43.3和46.7 kDa。这12个糖代谢相关的基因在变态发育期的中肠组织中具有5种不同的mRNA表达模式:(Ⅰ)只在幼虫期高表达(唾液麦芽糖酶前体蛋白、糖基水解酶31家族成员蛋白、线粒体乙醛脱氢酶、β-1,3-葡聚糖酶基因);(Ⅱ)只在预蛹期高表达(β-葡萄糖醛酸酶、β-N-酰基氨基葡萄糖苷酶3基因);(Ⅲ)只在蛹期高表达(葡萄糖胺-6-磷酸异构酶基因);(Ⅳ)在预蛹期和蛹期高表达(α-葡萄糖苷酶、α-淀粉酶、N-乙酰葡糖胺-6-磷酸脱乙酰酶和α-L-岩藻糖苷酶基因);(V)在变态发育期恒定表达(烯醇化酶基因)。这说明,为适应变态发育斜纹夜蛾中肠糖代谢途径发生了明显的改变。保幼激素对这些基因的表达没有明显的影响,但蜕皮激素对Ⅰ类基因(如糖基水解酶31家族成员蛋白基因)具有一定的抑制作用,对Ⅲ类基因(如葡萄糖胺-6-磷酸异构酶基因)有显著的上调作用。此外,我们还发现饥饿对几乎所有这些基因的表达都有显著的抑制作用。这些结果表明,昆虫中肠变态发育过程中糖代谢相关基因的动态变化可能受到蜕皮激素以及饥饿相关因素的共同调控。这一研究对从代谢角度揭示昆虫变态发育的分子机理具有重要意义。 Spodoptera litura is an important agricultural pest distributed worldwide. It undergoes the process of metamorphosis of a larval pupa during its growth and development. Because of the phenotypic changes of insects before and after metamorphosis, disintegration and reconstruction also occurred in the midgut as digestion and absorption of food. In line with this, the insect midgut various substances and energy metabolism may also be changed accordingly. In order to study the changes of glycometabolism pathway in the midgut midgut of Spodoptera litura, we identified 12 genes related to glucose metabolism from the midgut EST library of Spodoptera litura and cloned the full-length cDNAs of three of them Semi-quantitative PCR and quantitative PCR methods were used to detect the transcriptional expression of intestinal tissue during larval-pupal metamorphosis and the responses to hormones and starvation. The results showed that the open reading frames of these three genes (α-L-fucosidase, N-acetylglucosamine-6-phosphate deacetylase and enolase genes) were 1 461, 1 200 and 1 299 bp, respectively The predicted molecular weights were 56.3, 43.3 and 46.7 kDa, respectively. These 12 genes related to glucose metabolism have 5 different mRNA expression patterns in the midgut tissue of the metamorphosis: (I) Highly expressed only in larval stages (salivary maltase precursor protein, member of glycosyl hydrolase 31 family Protein, mitochondrial aldehyde dehydrogenase, β-1,3-glucanase gene); (Ⅱ) high expression of β-glucuronidase, β-N-acyl glucosaminidase 3 (¢ ó) was highly expressed only at pupal stage (glucosamine-6-phosphate isomerase gene); (¢ ó) was highly expressed at pre-pupal stage and pupal stage £ ¨α-glucosidase, α-amylase, -acetylglucosamine-6-phosphate deacetylase and alpha-L-fucosidase genes); (V) constitutive expression during the metamorphosis (enolase gene). This shows that in order to adapt to abnormal development of Spodoptera litura midgut glucose metabolism pathway has undergone significant changes. Juvenile hormones have no significant effect on the expression of these genes, but ecdysone has a certain inhibitory effect on class I genes (such as glycosyl hydrolase 31 family member protein genes), class III genes (such as glucosamine-6-phosphate Isomerase gene) has a significant up-regulation effect. In addition, we found that starvation significantly inhibited the expression of almost all of these genes. These results indicate that the dynamic changes of glucose metabolism-related genes in insect midgut midgut may be regulated by ecdysone and starvation-related factors. This study is of great significance to reveal the molecular mechanism of insect metamorphosis from the perspective of metabolism.