title: ATG5-knockout mutants of Physcomitrella provide a platform for analyzing the involvement of autophagy in senescence processes in plant cells.
abstract: Autophagy is a pathway in which a cell degrades part of its cytoplasm in vacuoles or lysosomes. To identify the physiological functions of autophagy in plants, we disrupted ATG5, an autophagy-related gene, in Physcomitrella, and confirmed that atg5 mutants are deficient in the process of autophagy. On carbon or nitrogen starvation medium, atg5 colonies turned yellow earlier than the wild-type (WT) colonies, showing that Physcomitrella atg5 mutants, like yeast and Arabidopsis, are sensitive to nutrient starvation. In the dark, even under nutrient-sufficient conditions, colonies turned yellow and the net degradation of chlorophyll and Rubisco protein occurred together with the upregulation of several senescence-associated genes. Yellowing reactions were inhibited by the protein synthesis inhibitor cycloheximide, suggesting that protonemal colonies undergo dark-induced senescence like the green leaves of higher plants. Such senescence responses in the dark occurred earlier in atg5 colonies than WT colonies. The sugar content was almost the same between WT and atg5 colonies, indicating that the early-senescence phenotype of atg5 is not explained by sugar deficiency. However, the levels of 7 amino acids showed significantly different alteration between atg5 and WT in the dark: 6 amino acids, particularly arginine and alanine, were much more deficient in the atg5 mutants, irrespective of the early degradation of Rubisco protein. On nutrient-sufficient medium supplemented with casamino acids, the early-senescence phenotype was slightly moderated. We propose that the early-senescence phenotype in atg5 mutants is partly explained by amino acid imbalance because of the lack of cytoplasmic degradation by autophagy in Physcomitrella.
authors.affiliation: a Graduate School of Science and Engineering; Saitama University ; Saitama , Japan.
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Mukae
authors.affiliation: a Graduate School of Science and Engineering; Saitama University ; Saitama , Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Inoue
authors.affiliation: a Graduate School of Science and Engineering; Saitama University ; Saitama , Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Moriyasu
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects; D019068:Bryopsida / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D002241:Carbohydrates / Q000032:analysis; D016922:Cellular Senescence / Q000187:drug effects; D002734:Chlorophyll / Q000378:metabolism; D003470:Culture Media; D003513:Cycloheximide / Q000494:pharmacology; D003624:Darkness; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D055786:Gene Knockout Techniques; D017343:Genes, Plant; D009154:Mutation / Q000235:genetics*; D010641:Phenotype; D059828:Plant Cells / Q000187:drug effects / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D012273:Ribulose-Bisphosphate Carboxylase / Q000378:metabolism; D012995:Solubility
chemical_list: D000596:Amino Acids; D002241:Carbohydrates; D003470:Culture Media; D010940:Plant Proteins; C017721:casamino acids; D002734:Chlorophyll; D003513:Cycloheximide; D012273:Ribulose-Bisphosphate Carboxylase
keywords: Physcomitrella; amino acid; autophagy; bryophyte; senescence; sugar