Identification of Genes in JA/SA Signaling Pathways in Flammulina velutipes and Their Response to Exogenous JA/SA
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摘要: 本研究鉴定了金针菇茉莉酸（JA）信号通路的4个关键基因：COI1、PDF1.2、MYC2-1、MYC2-2与水杨酸（SA）信号通路的4个关键基因：PR1-1、PR1-2、NPR1-1、NPR1-2。NBT染色法和荧光定量结果表明，金针菇JA/SA信号通路可应答外源JA/SA，50 μmol·L-1外源JA和500 μmol·L-1外源SA处理金针菇菌丝12 h可显著提高JA/SA信号通路基因的转录水平，基因PDF1.2响应JA诱导最明显，可作为JA信号通路标记基因；JA/SA信号转导途径间存在协同或拮抗作用：SA信号转导通路中NPR1蛋白对JA信号转导通路中PDF1.2基因表达有抑制作用，外源JA/SA的相对浓度决定其作用的强弱。Abstract: In this study, we identified 4 key genes of JA signaling pathway (COI1、PDF1.2、MYC2-1、MYC2-2) and SA signaling pathway (PR1-1、PR1-2、NPR1-1、NPR1-2) in Flammulina velutipes, respectively. The results of NBT staining and RT-qPCR demonstrated that JA/SA signaling pathways of F. velutipes responsed to exogenous JA/SA. In addition, the expression levels of these genes of JA/SA signaling pathway significantly increased under the treatment of exogenous 50 μmol · L-1 JA and 500 μmol · L-1 SA for 12 h, respectively. It was worth noting that PDF1.2 could be considered as the marker gene of JA signaling pathways for its response to exogenous JA obviously. The antagonistic effect between JA and SA signaling pathways may also exist in F.velutipes and the protein NPR1 of SA signaling pathways had inhibitory effect on the gene PDF1.2 expression level of JA signaling pathways which could be affected by the relative concentration of exogenous JA/SA.
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Key words:
- Flammulina velutipes /
- jasmonic acid /
- salicylic acid /
- NBT /
- gene expression
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图 1 A：不同浓度JA/SA处理菌丝后NBT染色；B：不同时间JA/SA处理菌丝后NBT染色
Figure 1. A: NBT staining after different concentrations of JA/SA treatment; B: NBT staining after different times of JA/SA treatment
图 2 A：不同浓度JA处理后基因相对表达量；B：不同浓度SA处理后基因相对表达量；C：不同时间JA处理后基因相对表达量；D：不同时间SA处理后基因相对表达量
Figure 2. A: Gene relative expression after different concentrations of JA treatment; B: Gene relative expression after different concentrations of SA treatment; C: Gene relative expression after different times of JA treatment; D: Gene relative expression after different times of SA treatment
图 3 JA/SA交叉处理相对定量结果
Figure 3. Relative gene expression level after JA/SA Co- treatment
表 1 金针菇JA/SA信号通路关键基因分析
Table 1. Main gene analysis of JA/SA signaling pathway in F. velutipes
信号途径 基因 名称 GenBank登录号 同源蛋白物种 同源蛋白GenBank登录号 结构域名称 E值 相似性/% 分值 荧光定量引物(5′-3′) JA gene 2615 COI1 KX129930 Rhodotorula toruloides EMS23488.1 AMN1 2.00E-134 44 438 F:CTATGGACGAACGAGAGACGAC R:GCCACCGTTTCTTTATGCC gene 6872 PDF1.2 KX129931 Arabidopsis thaliana AED95107.1 Knot1 4.00E-45 34 24 F: TGGAAGATACCCCGAGGACAC R:GGAAGAAGGCGAGCGAAGTC gene 6724 MYC2-1 KX129932 Laccaria bicolor XP_001879735.1 Helix-loop-helix domain 3.00E-38 43 147 F: TAACTCCTTCGCAAACCCTCG R:GCTCGTTTATGCCTTCGTTGA gene 2796 MYC2-2 KX129933 Laccaria bicolor XP_001879735.1 Helix-loop-helix domain 1.00E-77 50 252 F: ACAAGTGAGGCTCAGCAAGGTC R:GAAGACGAGTGATTTCGGCAGT SA gene1937 PR1-1 KX257996 Cylindrobasidium torrendii KIY69700.1 SCP_PRY1_like 1.00E-58 50 197 F: TGGAAAGGGACGACCGAAGT R:CGGACGGATGATTGTGGAGAA gene5143 PR1-2 KX129927 Moniliophthora perniciosa AEZ63361.1 SCP_PRY1_like 3.00E-57 54 200 F: GATACAACAGAGGTGGGATGCG R:TTGAAGTTTCCAGGTGGCAAGTAT gene4378 NPR1-1 KX129928 Saccharomyces cerevisiae CAA96076.1 NPR1_like_C 3.00E-81 56 272 F: GCGACAGTGTCACCACATTCC R:AACCAAAGAAGCGTCACATCC gene8890 NPR1-2 KX129929 Saccharomyces cerevisiae CAA96076.1 NPR1_like_C 4.00E-55 39 202 F: GGAGGAGTTGGTCGTCAGGC R:AAGGTCGGTTCGTCCCCAC 
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