黄色金针菇SOD基因家族成员鉴定及表达分析

Identification and Expression Analysis of SOD Gene Family Members in Yellow Flammulina filiformis

对黄色金针菇(Flammulina filiformis)中超氧化物歧化酶(superoxide dismutase,SOD)编码基因进行鉴定和生物信息学分析,并采用荧光定量PCR研究光照(200 lx)条件下菌丝向原基转变过程中以及不同发育阶段(菌丝、原基、幼菇)和子实体不同部位(菌盖,菌柄自上而下0.5~1.5 cm、3~4.5 cm、5~7 cm)SOD表达情况,结果表明:鉴定到1个Fe-SOD基因FfSOD2、2个Mn-SOD基因FfSOD1和FfSOD6、3个Cu/Zn-SOD基因FfSOD3、FfSOD4和FfSOD5,在4条染色体上不均匀分布,内含子数量为2~6,且启动子区域均含有生长发育和胁迫响应元件;Cu/Zn-SOD以无规则卷曲结构为主,而Mn-SOD和Fe-SOD中α螺旋占比较大。与对照组相比,光照组FfSOD4和FfSOD5相对表达量上调,FfSOD1相对表达量下调,FfSOD2、FfSOD3和FfSOD6相对表达量无显著差异。在黄色金针菇不同发育时期和子实体不同部位中,FfSOD1和FfSOD5在菌柄下部,FfSOD2在原基、幼菇、菌盖和菌柄上部,FfSOD3在菌盖、菌柄上部,FfSOD4和FfSOD6在幼菇期相对表达量较高。研究结果为黄色金针菇发育过程中SOD基因功能的深入研究提供参考。

Superoxide dismutase(SOD)genes in yellow Flammulina filiformis were identified and subjected to bioinformatics analysis.Then the expression of the SOD genes under light illumination(200 lx)were analyzed by qRT-PCR during different growth and developmental stages,i.e.,mycelium,primordium,young fruiting body,and different parts of fruiting body,i.e.pileus,stipe from top to bottom 0.5–1.5 cm,3–4.5 cm and 5–7 cm.The results showed that a total of six SOD genes were identified in the yellow F.filiformis genome,including one Fe-SOD gene FfSOD2,two Mn-SOD genes FfSOD1 and Ff SOD6,and three Cu/Zn-SOD genes FfSOD3,FfSOD4 and FfSOD5.They are distributed on chromosomes No.1,2,5 and 11,and contain 2-6 introns and elements related to growth and development,and stress response at their promoter regions.For secondary protein structure,Cu/Zn-SOD is dominantly composed of random coils,whileα-helix accounts for a large proportion in Mn-SOD and Fe-SOD.Compared with the control in darkness,the relative expression levels of FfSOD4 and FfSOD5 were up regulated under light illumination,whereas the relative expression level of FfSOD1 was down regulated.The relative expression levels of FfSOD2,FfSOD3,and FfSOD6 remained the same under light illumination.In terms of expression at different developmental stages and fruiting body parts,FfSOD1 and FfSOD5 showed relatively high expression levels at 5-7 cm of stipe;FfSOD2 was highly expressed in primordium,young fruiting body,pileus,and 0.5-1.5 cm of stipe;FfSOD3 was also highly expressed in pileus and 0.5-1.5 cm of stipe;FfSOD4 and FfSOD6 were highly expressed in young fruiting bodies.These results provided a reference for further study on the function of the SOD genes during the growth and development of yellow F.filiformis.