首页 > 过刊浏览>2023年第42卷第2期 >131-138. DOI:10.13300/j.cnki.hnlkxb.2023.02.017
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金针菇中免疫调节蛋白编码基因FIP-fve的功能
CSTR:
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作者:
作者单位:

1.沈阳农业大学生物科学技术学院，沈阳110866;2.辽宁省林业科学研究院，沈阳110032

通讯作者:

林景卫，E-mail： 2009500003@syau.edu.cn

中图分类号:

S567.3+9

基金项目:

辽宁省教育厅科学研究基础项目（LSNQN202013）陈焕，E-mail：2507297357@qq.com


Native bio-function of FIP-fve towards Flammulina filiformis
Author:
Affiliation:

1.College of Biological Science and Technology， Shenyang Agricultural University， Shenyang 110866，China;2.Liaoning Academy of Forestry Sciences，Shenyang 110032，China

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    摘要:

    为探究FIP-fve基因在金针菇本体的生物学功能，构建了pCAMBIA1301-pGPD-FIP-fve超表达载体并将其转化至农杆菌LBA4404中，采用农杆菌介导的金针菇转化法，最终获得了FIP-fve基因超表达的金针菇；进行FIP-ve基因沉默（RNAi： FIP-fve）、FIP-fve超表达（pGPD： FIP-fve）和野生金针菇（CK）菌丝体培养和子实体栽培，并记录菌丝体和子实体时期的生物学性状。结果显示：pGPD： FIP-fve金针菇菌丝生长速度最快，CK金针菇次之，而RNAi： FIP-fve金针菇生长速度最为缓慢；子实体出菇栽培后，在子实体数量、菌柄长度、子实体产量和生物学效率方面，pGPD： FIP-fve金针菇都明显优于CK金针菇，而RNAi： FIP-fve金针菇的子实体生长状况最差。研究结果表明，FIP-fve基因能够促进金针菇菌丝体的生长，并对金针菇原基形成具有促进作用，最终影响金针菇的产量。

    Abstract:

    FIP-fve is a low molecular-mass protein identified from Flammulina filiformis， which has many biological activities including immunomodulation. Our previous studies found that FIP-fve had certain growth-promoting effect on F. filiformis mycelia. We constructed an over-expression vector pCAMBIA1301-pGPD-FIP-fve， and transformed it into Agrobacterium tumefaciens LBA4404 strain to study the native biological function of FIP-fve to F. filiformis. FIP-fve gene overexpressed F. filiformis was obtained via Agrobacterium transformation. By comparing the biological characteristics of FIP-fve gene-silent F. filiformis （RNAi：FIP-fve）， FIP-fve gene-overexpressed F. filiformis （pGPD：FIP-fve） and wild F. filiformis （CK） in mycelium and fruiting body stages， we found that the growth speed of pGPD：FIP-fve was faster than that of CK， whereas the growth speed of RNAi：FIP-fve was the lowest. After cultivating the fruiting body， yield-related traits of pGPD：FIP-fve were significantly better than that of CK in terms of fruiting body numbers， stalk length， fruiting body yield and biological efficiency， and those traits of RNAi： FIP fve were the worst. It is indicated that FIP-fve significantly promote the growth of F. filiformis mycelia and affect the final yields of F. filiformis.

    表 3 野生型（CK）、FIP- fve过表达和沉默菌株的金针菇产量Table 3 The mushroom yield of wild-type （CK），FIP- fve overexpression and silent strains
    表 2 实时定量PCR引物序列Table 2 Primer sequence for q-RT PCR
    图1 担子菌门不同真菌的FIP序列比对Fig.1 FIP sequence alignment of different fungi in Basidiomycetes
    图2 FIP的系统发育进化树Fig.2 The phylogenic tree of FIP
    图3 表达载体的酶切检测Fig.3 Restriction analysis of expression vector
    图4 表达载体pCAMBIA1303-pGPD-FIP-fve 构建Fig.4 pCAMBIA 1303-pGPD-FIP-fve vector construction
    图5 金针菇转化子潮霉素基因PCR检测结果Fig.5 PCR results of hygromycin gene transformed from Flammulina filiformis
    图6 野生菌株、过表达菌株和沉默菌株中的FIP-fve的转录水平Fig.6 Transcription levels of FIP-fve in the wild，overexpressing and silent strains
    图7 FIP-fve正向调控金针菇的生长发育Fig.7 FIP-fve positively regulates the growth and development of Flammulina velutipes
    图8 FIP-fve过表达和沉默对菌丝体生长的影响Fig.8 Effect of FIP-fve overexpression and silent on mycelium growth
    表 1 常规PCR所用引物及序列Table 1 Primer sequences used for PCR amplification
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陈焕,范天宁,白玉东,郭芡芡,王浩安,陈罡,高英旭,林景卫.金针菇中免疫调节蛋白编码基因FIP-fve的功能[J].华中农业大学学报,2023,42(2):131-138

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  • 收稿日期:2022-08-03
  • 在线发布日期: 2023-03-31
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