| 黄 海, 杜 娟, 李尚伟*, 龚 涛, 齐小浪.九香虫i型溶菌酶基因的克隆与细菌诱导表达分析[J].植物保护,2022,48(1):17-28. |
| 九香虫i型溶菌酶基因的克隆与细菌诱导表达分析 |
| Cloning and induced expression analysis of the i type lysozyme genes in Coridius chinensis |
| 投稿时间：2020-12-15 修订日期：2021-02-24 |
| DOI：10.16688/j.zwbh.2020669 |
| 中文关键词: 九香虫 i型溶菌酶 时空表达 先天免疫 细菌诱导 |
| 英文关键词:Coridius chinensis i type lysozyme spatiotemporal expression innate immunity bacterial induction |
| 基金项目:国家自然科学基金(31560610) |
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| 中文摘要: |
| 溶菌酶是一组进化上保守的酶, 在昆虫中主要分为c型和i型两类。为明确i型溶菌酶在九香虫 Coridius chinensis 体内的表达模式与功能, 我们对九香虫的两个i型溶菌酶基因 CcLys i1 和 CcLys i2 进行了克隆, 这两个基因的编码区分别为504 bp和432 bp, 分别编码167和143个氨基酸。序列分析显示, CcLys i1和CcLys i2缺乏与催化活性相关的谷氨酸(E)和丝氨酸(S)。同源性和聚类分析显示, CcLys i1和CcLys i2与斯氏珀蝽 Plautia stali 的i型溶菌酶相似性最高, 分别为80.36%和48.95%。采用实时荧光定量PCR(RT-qPCR)解析 CcLys i1 和 CcLys i2 基因的时空表达谱, 结果表明它们在九香虫不同发育阶段都有表达, CcLys i1 在成虫中表达水平最高, 而 CcLys i2 在5龄若虫中表达水平最高;它们在所检测的成虫不同组织中都有表达, 都在脂肪体中表达水平最高。九香虫被注射细菌后24 h, CcLys i1 和 CcLys i2 的表达显著上调;九香虫被饲喂细菌后, 这两种基因的表达水平没有显著变化。九香虫的这两种i型溶菌酶基因的表达受到自身免疫机制的调控, 可以被诱导而参与免疫应答, 但不具有参与消化的功能。该研究为进一步明确九香虫i型溶菌酶基因的功能奠定基础。 |
| 英文摘要: |
| Lysozymes are a group of evolutionarily conserved enzymes and can be divided into c type and i type in insects. In order to clarify the expression pattern and function of the i type lysozyme in Coridius chinensis, we cloned two i type lysozyme genes CcLys i 1 and CcLys i 2. The coding regions of these two genes were 504 bp and 432 bp, encoding 167 and 143 amino acids, respectively. Sequence analysis showed that CcLys i1 and CcLys i2 lacked glutamic acid (E) and serine (S) related to catalytic activity . Homology and cluster analyses showed that CcLys i1 and CcLys i2 shared the similarity of 80.36% and 48.95%, respectively, with the i type lysozyme from Plautia stali . The spatiotemporal expression profiles of CcLys i1 and CcLys i2 were analyzed by using real time quantitative PCR (RT-qPCR). The results indicated that these two genes were expressed at various developmental stages of C. chinensis, with the highest expression levels for CcLys i1 in the adults and for CcLys i2 in the fifth instar nymphs; they were expressed in different tissues of adults and had the highest expression level in the fat bodies. The expression levels of CcLys i1 and CcLys i2 were significantly upregulated within 24 h post bacterial injection, but there were no significant changes in the expression levels of these two genes post bacterial feeding. The expression of these two i type lysozyme genes in C. chinensis was regulated by its own immune mechanism and could be induced to participate in immune response, but they were not involved in digestion. This study laid a foundation for further elucidating the functions of the i type lysozyme genes in C. chinensis. |
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