细胞自噬是一种进化上高度保守的溶酶体/液泡降解途径,对机体适应各种生物或非生物胁迫,以及维持自身正常生长发育具有重要的意义。自噬相关蛋白8(autophagy-related protein 8,ATG8)是检测自噬的金标准。目前由于缺乏特异性好的小麦ATG...细胞自噬是一种进化上高度保守的溶酶体/液泡降解途径,对机体适应各种生物或非生物胁迫,以及维持自身正常生长发育具有重要的意义。自噬相关蛋白8(autophagy-related protein 8,ATG8)是检测自噬的金标准。目前由于缺乏特异性好的小麦ATG8抗体,导致小麦细胞自噬研究进展缓慢。本研究通过人工合成小麦ATG8蛋白序列上的一段18个氨基酸的多肽作为抗原去免疫兔子,成功获得了高度特异性的小麦ATG8多克隆抗体。该抗体不仅能够识别小麦根、叶及种子中的ATG8蛋白条带,而且可以在根、叶中检测到细胞自噬结构,并首次在小麦籽粒中实现了细胞自噬结构的检测。本研究研制的小麦ATG8抗体,除了能够检测到小麦中常见的ATG8a-h等8种典型类型外,还能检测到一种非典型的新的ATG8蛋白,为深入开展植物细胞自噬调控机制研究以及优异新基因挖掘提供了最佳方法学基础。展开更多
The basidiomycetous fungus Sporisorium scitamineum causes sugarcane smut that leads to severe economic losses in the major sugarcane growing areas in China,India and Brazil,etc.Autophagy is a conserved pathway in euka...The basidiomycetous fungus Sporisorium scitamineum causes sugarcane smut that leads to severe economic losses in the major sugarcane growing areas in China,India and Brazil,etc.Autophagy is a conserved pathway in eukaryotes for bulk degradation and cellular recycling,and was shown to be important for fungal cell growth,development,and pathogenicity.However,physiological function of autophagy has not been studied in S.scitamineum.In this study,we identified a conserved Atg8 protein,named as SsAtg8 and characterized its function.Our results showed that autophagy was blocked in the ssatg8Δ mutant,in nitrogen starvation.The ssatg8Δ mutant formed pseudohypha frequently and was hypersensitive to oxidative stress.However,mating or filamenation was unaffected in the ssatg8Δ mutant in vitro.Overall we demonstrate that autophagy is dispensable for S.scitamineum mating/filamentation,while critical for oxidative stress tolerance and proper morphology in sporidial stage.展开更多
文摘细胞自噬是一种进化上高度保守的溶酶体/液泡降解途径,对机体适应各种生物或非生物胁迫,以及维持自身正常生长发育具有重要的意义。自噬相关蛋白8(autophagy-related protein 8,ATG8)是检测自噬的金标准。目前由于缺乏特异性好的小麦ATG8抗体,导致小麦细胞自噬研究进展缓慢。本研究通过人工合成小麦ATG8蛋白序列上的一段18个氨基酸的多肽作为抗原去免疫兔子,成功获得了高度特异性的小麦ATG8多克隆抗体。该抗体不仅能够识别小麦根、叶及种子中的ATG8蛋白条带,而且可以在根、叶中检测到细胞自噬结构,并首次在小麦籽粒中实现了细胞自噬结构的检测。本研究研制的小麦ATG8抗体,除了能够检测到小麦中常见的ATG8a-h等8种典型类型外,还能检测到一种非典型的新的ATG8蛋白,为深入开展植物细胞自噬调控机制研究以及优异新基因挖掘提供了最佳方法学基础。
基金supported by the National 973 Program of China(2015CB150600)the Natural Science Foundation of Guangdong Province,China(2017A030310144)
文摘The basidiomycetous fungus Sporisorium scitamineum causes sugarcane smut that leads to severe economic losses in the major sugarcane growing areas in China,India and Brazil,etc.Autophagy is a conserved pathway in eukaryotes for bulk degradation and cellular recycling,and was shown to be important for fungal cell growth,development,and pathogenicity.However,physiological function of autophagy has not been studied in S.scitamineum.In this study,we identified a conserved Atg8 protein,named as SsAtg8 and characterized its function.Our results showed that autophagy was blocked in the ssatg8Δ mutant,in nitrogen starvation.The ssatg8Δ mutant formed pseudohypha frequently and was hypersensitive to oxidative stress.However,mating or filamenation was unaffected in the ssatg8Δ mutant in vitro.Overall we demonstrate that autophagy is dispensable for S.scitamineum mating/filamentation,while critical for oxidative stress tolerance and proper morphology in sporidial stage.
