抑制谷胱甘肽合成对大豆胞囊线虫发育的影响

摘要

为明确谷胱甘肽对大豆胞囊线虫(Heterodera glycines,SCN)发育的影响,利用谷胱甘肽合成抑制剂L-丁硫氨酸-亚砜亚胺(L-Buthionine-sulfoximine,BSO)处理大豆感病品种辽豆15,观察SCN的发育;同时,利用高效液相色谱法(HPLC)和实时荧光定量PCR(qRT-PCR)检测谷胱甘肽含量和合成关键酶基因的表达.结果表明,在接种线虫14d,SCN在辽豆15根系内的发育受到了抑制,二龄幼虫(J2)比例显著高于对照53.86％,四龄幼虫(J4)比例显著低于对照10.54％;但在接种线虫28d时,J4数量与对照比无显著差异.在接种线虫14d,辽豆15根系内谷胱甘肽含量及合成关键酶基因γ-ECS、GSHS和hGSHS的表达量均显著低于对照.BSO有效地抑制了谷胱甘肽合成途径关键酶基因的表达,降低了谷胱甘肽的生物合成产物的含量.在谷胱甘肽缺乏的条件下,大豆胞囊线虫的发育受到了一定程度的抑制.说明谷胱甘肽是参与调控SCN发育的重要因子.%For better understanding the development of soybean cyst nematode (SCN,Heterodera glycines) af fected by glutathione,L-Buthionine-sulfoximine (BSO),an inhibitor of glutathione biosynthesis,was used for treating on Liaodoul5,a SCN susceptible cultivar,and the development of SCN was examined.The contents and levels of gene expression were detected using qRT-PCR and HPLC,respectively.The results indicated that the development of SCN was inhibited significantly at 14 days post inoculation.Compared with control,the number of juvenile at second stage and juvenile at fourth stage increased to 53.86％ and decreased to 10.54％ respectively;but,the number of J4 showed no difference at 28 days post SCN infection.The expression level of y-ECS,GSHS and hGSHS were inhibited significantly during BSO treatment at 14 days post SCN infection.The contents of glutathione decreased significantly comparing with control.In summary,the development of SCN was inhibited under glutathione deficiency.And glutathione was a significant factor in development of SCN.These results provided a theoretic support for SCN resistance breeding in future.