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BmSUC1 对家蚕糖代谢的调控是必需的。

BmSUC1 is essential for glycometabolism modulation in the silkworm, Bombyx mori.

机构信息

School of Life Sciences, Anhui Agricultural University, Hefei 230036, Anhui, China; Anhui International Joint Research and Development Center of Sericulture Resources Utilization, Hefei 230036, Anhui, China.

School of Life Sciences, Anhui Agricultural University, Hefei 230036, Anhui, China.

出版信息

Biochim Biophys Acta Gene Regul Mech. 2018 Jun;1861(6):543-553. doi: 10.1016/j.bbagrm.2018.04.002. Epub 2018 Apr 14.

Abstract

Sucrose is the most commonly transported sugar in plants and is easily assimilated by insects to fulfill the requirement of physiological metabolism. BmSuc1 is a novel animal β-fructofuranosidase (β-FFase, EC 3.2.1.26)-encoding gene that was firstly cloned and identified in silkworm, Bombyx mori. BmSUC1 was presumed to play an important role in the silkworm-mulberry enzymatic adaptation system by effectively hydrolyzing sucrose absorbed from mulberry leaves. However, this has not been proved with direct evidence thus far. In this study, we investigated sucrose hydrolysis activity in the larval midgut of B. mori by inhibition tests and found that sucrase activity mainly stemmed from β-FFase, not α-glucosidase. Next, we performed shRNA-mediated transgenic RNAi to analyze the growth characteristics of silkworm larvae and variations in glycometabolism in vivo in transgenic silkworms. The results showed that in the RNAi-BmSuc1 transgenic line, larval development was delayed, and their body size was markedly reduced. Finally, the activity of several disaccharidases alone in the midgut and the sugar distribution, total sugar and glycogen in the midgut, hemolymph and fat body were then determined and compared. Our results demonstrated that silencing BmSuc1 significantly reduced glucose and apparently activated maltase and trehalase in the midgut. Together with a clear decrease in both glycogen and trehalose in the fat body, we conclude that BmSUC1 acts as an essential sucrase by directly modulating the degree of sucrose hydrolysis in the silkworm larval midgut, and insufficient sugar storage in the fat body may be responsible for larval malnutrition and abnormal petite phenotypes.

摘要

蔗糖是植物中最常见的运输糖,很容易被昆虫吸收,以满足生理代谢的需要。BmSuc1 是一种新型动物β-呋喃果糖苷酶(β-FFase,EC 3.2.1.26)编码基因,最初在蚕,家蚕中克隆和鉴定。BmSUC1 被认为在蚕-桑酶适应系统中发挥重要作用,通过有效水解从桑叶中吸收的蔗糖。然而,到目前为止,还没有直接证据证明这一点。在这项研究中,我们通过抑制试验研究了家蚕幼虫中肠的蔗糖水解活性,发现蔗糖酶活性主要来自β-FFase,而不是α-葡萄糖苷酶。接下来,我们进行了 shRNA 介导的转基因 RNAi 分析,以研究转基因家蚕幼虫的生长特性和体内糖代谢的变化。结果表明,在 RNAi-BmSuc1 转基因系中,幼虫发育延迟,体型明显减小。最后,单独测定了中肠中几种二糖酶的活性以及中肠、血液和脂肪体中的糖分布、总糖和糖原。我们的结果表明,沉默 BmSuc1 显著降低了葡萄糖的活性,明显激活了中肠的麦芽糖酶和海藻糖酶。伴随着脂肪体中糖原和海藻糖的明显减少,我们得出结论,BmSUC1 作为一种必需的蔗糖酶,通过直接调节家蚕幼虫中肠蔗糖水解的程度而起作用,而脂肪体中糖储存不足可能是幼虫营养不良和异常小体表型的原因。

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