柞蚕己糖激酶基因的组织表达特征及在解除滞育和蛹发育期的表达与酶活性变化
发布时间：2018-11-13 12:19
【摘要】：昆虫能够将消化吸收的葡萄糖在脂肪体中转化为海藻糖或作为糖原储存,当机体需要能量时,海藻糖再转化为葡萄糖进入糖酵解途径,己糖激酶(hexokinase,HK)是在此转化过程合成中间代谢物葡萄糖-6-磷酸的限速酶。为研究柞蚕蛹解除滞育及发育过程中的糖类代谢规律,克隆了柞蚕2个己糖激酶同工酶基因,分别命名为Ap HK1和Ap HK2(Gen Bank登录号:KY624592,KY624593)。2个基因的ORF全长分别为1 455 bp、1 362 bp,编码484、453个氨基酸。系统进化分析柞蚕HK1和HK2蛋白分布于2个不同的进化分支,不同种类昆虫的同一类型HK具有更高的同源性。柞蚕5龄幼虫各组织中,2个HK基因在脂肪体中的表达水平都较高,Ap HK2在各组织中均有表达,分布比Ap HK1更广,且在中肠、肌肉内也有较高表达。以一化性柞蚕滞育蛹的脂肪体组织为材料,采用qRT-PCR分析长光照(17 h/d)处理解除柞蚕蛹滞育及蛹发育过程中HK基因的表达规律,结果表明:2个HK基因的表达量在处理后21 d明显升高,其中Ap HK2的表达量升高了3.1倍;化蛹后42 d,Ap HK1的表达量升高了2.8倍,而Ap HK2的表达量无显著变化。同时测定蛹脂肪体中己糖激酶的活性在长光照处理下高于对照组,前期呈现上升趋势,处理后28 d达到最高值,此后逐渐下降并于处理后35 d趋于稳定;测定蛹血淋巴中葡萄糖含量呈先下降后上升的趋势,这与脂肪体中己糖激酶的活性具有相关性。研究结果显示柞蚕滞育蛹解除滞育及发育过程中己糖激酶基因表达上调,在蛹发育中期己糖激酶酶活力及葡萄糖含量之间呈现出互补关系,即己糖激酶的活性增强,血淋巴中葡萄糖的含量减少。
[Abstract]:Insects can convert digested and absorbed glucose into trehalose in the fat body or store it as a glycogen. When the body needs energy, trehalose is converted into glucose to enter the glycolytic pathway, hexokinase, HK is a rate-limiting enzyme for the synthesis of glucose-6 phosphoric acid, an intermediate metabolite. In order to study the rules of carbohydrate metabolism during deferment and development of tussah silkworm pupae, two isozyme genes of hexokinase were cloned and named Ap HK1 and Ap HK2 (Gen Bank accession number: KY624592,. KY624593). The ORF length of the two genes was 1 455 bp,1 362 bp, encoding 484453 amino acids, respectively. Phylogenetic analysis showed that HK1 and HK2 proteins were distributed in two different branches of evolution, and the homology of the same type of HK in different species of insects was higher. In every tissue of 5th instar larvae of Antheraea pernyi, the expression of two HK genes was higher in fat body, and the expression of Ap HK2 was more extensive than that of Ap HK1 in all tissues, and it was also expressed in midgut and muscle. The fat body tissue of diapause pupae of Antheraea pernyi was used as the material, and the expression of HK gene during diapause and development of tussah silkworm pupae was relieved by qRT-PCR analysis with long illumination (17 h / d). The results showed that the expression of two HK genes increased significantly 21 days after treatment, and the expression of Ap HK2 increased 3.1-fold. At 42 d after pupation, the expression of AP HK1 increased 2.8 times, but the expression of Ap HK2 did not change significantly. At the same time, the activity of hexokinase in fat body of pupae was higher than that of control group under long light treatment. The activity of hexokinase in fat body of pupae increased in early stage, reached the highest value at 28 days after treatment, then decreased gradually and became stable at 35 days after treatment. The content of glucose in hemolymph of pupae decreased first and then increased, which was related to the activity of hexokinase in fat body. The results showed that the expression of hexokinase gene was up-regulated during diapause and development of Antheraea pernyi pupae, and there was a complementary relationship between the activity of hexokinase and the content of glucose, that is, the activity of hexokinase was enhanced. The content of glucose in hemolymph was decreased.
【作者单位】： 沈阳农业大学生物科学技术学院 辽宁省昆虫资源工程技术研究中心;河南省蚕业科学研究院;
【基金】：现代农业产业技术体系建设专项(No.CARS-22) 辽宁省农业领域青年科技创新人才培养计划项目(No.2014010) 辽宁省教育厅一般项目(No.L2015488)
【分类号】：S885.1
，
本文编号：2329085
[Abstract]:Insects can convert digested and absorbed glucose into trehalose in the fat body or store it as a glycogen. When the body needs energy, trehalose is converted into glucose to enter the glycolytic pathway, hexokinase, HK is a rate-limiting enzyme for the synthesis of glucose-6 phosphoric acid, an intermediate metabolite. In order to study the rules of carbohydrate metabolism during deferment and development of tussah silkworm pupae, two isozyme genes of hexokinase were cloned and named Ap HK1 and Ap HK2 (Gen Bank accession number: KY624592,. KY624593). The ORF length of the two genes was 1 455 bp,1 362 bp, encoding 484453 amino acids, respectively. Phylogenetic analysis showed that HK1 and HK2 proteins were distributed in two different branches of evolution, and the homology of the same type of HK in different species of insects was higher. In every tissue of 5th instar larvae of Antheraea pernyi, the expression of two HK genes was higher in fat body, and the expression of Ap HK2 was more extensive than that of Ap HK1 in all tissues, and it was also expressed in midgut and muscle. The fat body tissue of diapause pupae of Antheraea pernyi was used as the material, and the expression of HK gene during diapause and development of tussah silkworm pupae was relieved by qRT-PCR analysis with long illumination (17 h / d). The results showed that the expression of two HK genes increased significantly 21 days after treatment, and the expression of Ap HK2 increased 3.1-fold. At 42 d after pupation, the expression of AP HK1 increased 2.8 times, but the expression of Ap HK2 did not change significantly. At the same time, the activity of hexokinase in fat body of pupae was higher than that of control group under long light treatment. The activity of hexokinase in fat body of pupae increased in early stage, reached the highest value at 28 days after treatment, then decreased gradually and became stable at 35 days after treatment. The content of glucose in hemolymph of pupae decreased first and then increased, which was related to the activity of hexokinase in fat body. The results showed that the expression of hexokinase gene was up-regulated during diapause and development of Antheraea pernyi pupae, and there was a complementary relationship between the activity of hexokinase and the content of glucose, that is, the activity of hexokinase was enhanced. The content of glucose in hemolymph was decreased.
【作者单位】： 沈阳农业大学生物科学技术学院 辽宁省昆虫资源工程技术研究中心;河南省蚕业科学研究院;
【基金】：现代农业产业技术体系建设专项(No.CARS-22) 辽宁省农业领域青年科技创新人才培养计划项目(No.2014010) 辽宁省教育厅一般项目(No.L2015488)
【分类号】：S885.1
，
本文编号：2329085
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