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家蚕血淋巴体外黑化观察及相关黑色素合成催化酶基因
引用本文:李田,张亮,申琦,赵威,李黎,吕银,蒋贵兵,闫登峰,肖俊杰,陈萍.家蚕血淋巴体外黑化观察及相关黑色素合成催化酶基因[J].生物工程学报,2016,32(8):1093-1103.
作者姓名:李田  张亮  申琦  赵威  李黎  吕银  蒋贵兵  闫登峰  肖俊杰  陈萍
作者单位:1 西南大学 生物技术学院,重庆 400716,1 西南大学 生物技术学院,重庆 400716,1 西南大学 生物技术学院,重庆 400716,1 西南大学 生物技术学院,重庆 400716,1 西南大学 生物技术学院,重庆 400716,1 西南大学 生物技术学院,重庆 400716,2 重庆市蚕业管理总站,重庆 400716,1 西南大学 生物技术学院,重庆 400716,1 西南大学 生物技术学院,重庆 400716,1 西南大学 生物技术学院,重庆 400716
基金项目:重庆市自然科学基金重点项目 (No. 2013jjB80004),重庆市研究生科研创新项目 (No. CYS14048),重庆市2015国家茧丝绸发展专项资金资助。
摘    要:观察不同生长期家蚕幼虫血淋巴在体外的黑化速度和对大肠杆菌生长的影响结果显示,随食桑生长幼虫血淋巴的黑化速度逐渐变快,对大肠杆菌生长的抑制作用逐渐增强。RT-PCR实验显示,黑色素合成催化酶Bm Tan、Bm Po-1、Bm Yellow-f和Bm Ddc等的基因在家蚕5 L 3 d血淋巴中表达量高,Bm Black、Bm Yellow和Bm Pah等的基因也有明显表达。q PCR分析显示,黑化病蚕中Bmtan、Bmddc、Bmyellow、Bmebony和Bmblack,尤其Bmddc表达发生了显著上调。与对照相比,Ddc酶的抑制剂能显著抑制脂多糖对血淋巴的诱导黑化作用。用大肠杆菌注射家蚕幼虫,血淋巴中多巴和多巴胺的含量明显上升。这些表明家蚕幼虫血淋巴黑化与防御免疫有关,Bmddc很可能在幼虫血淋巴的免疫黑化中发挥作用。

关 键 词:家蚕血淋巴,黑化反应,黑色素合成催化酶,免疫应答
收稿时间:2015/11/25 0:00:00

In vitro observation of haemolymph melanization and melanin-related biosynthesis enzyme genes in silkworm, Bombyx mori
Tian Li,Liang Zhang,Qi Shen,Wei Zhao,Li Li,Yin Lv,Guibing Jiang,Dengfeng Yan,Junjie Xiao and Ping Chen.In vitro observation of haemolymph melanization and melanin-related biosynthesis enzyme genes in silkworm, Bombyx mori[J].Chinese Journal of Biotechnology,2016,32(8):1093-1103.
Authors:Tian Li  Liang Zhang  Qi Shen  Wei Zhao  Li Li  Yin Lv  Guibing Jiang  Dengfeng Yan  Junjie Xiao and Ping Chen
Institution:1 College of Biotechnology, Southwest University, Chongqing 400716, China,1 College of Biotechnology, Southwest University, Chongqing 400716, China,1 College of Biotechnology, Southwest University, Chongqing 400716, China,1 College of Biotechnology, Southwest University, Chongqing 400716, China,1 College of Biotechnology, Southwest University, Chongqing 400716, China,1 College of Biotechnology, Southwest University, Chongqing 400716, China,2 sericulture Management Station of Chongqing, Chongqing 400716, China,1 College of Biotechnology, Southwest University, Chongqing 400716, China,1 College of Biotechnology, Southwest University, Chongqing 400716, China and 1 College of Biotechnology, Southwest University, Chongqing 400716, China
Abstract:The observation statistics suggested that the haemolymph melanization speed of larvae became fast and the growth inhibition of Escherichia coli was strong as the quantities of feeding on mulberry leaves increased. The RT-PCR result showed that the mRNA expressions of melanin biosynthesis enzyme BmTan, BmPo-1, BmYellow-f and BmDdc were high in the haemolyph of 5 L 3 d larvae. The qPCR analysis showed Bmtan, Bmddc, Bmyellow, Bmebony and Bmblack, especially Bmddc expression were significantly higher in black disease larvae than in normal larvae. Compared with control, Ddc inhibitors drastically inhibited the lipopolysaccharide-induced haemolymph melanization. In addition, the content of Dopa and Dopamine markedly rose after E. coli injection. These indicated that haemolymph melanization was linked to immune defenses and Bmddc may play a role in melanization response of haemolymph immune in silkworm.
Keywords:haemolymph of Bombyx mori  melanization  melanin biosynthesis enzyme  immune response
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