Arabidopsis possesses nine ATG8 isoforms, and the dominant isoform influences the overall substrate specificity and characteristics of autophagosomes. Plants frequently experience heat stress and subsequently recover to their original temperature. During these processes, autophagosomes undergo continuous changes, and the targeted proteins are dynamically altered. Our study reveals that the disappearance of ATG8a during recovery from heat stress, accompanied by the predominance of other ATG8 isoforms, plays a critical role. To investigate how these changes impact the protein profile, we conducted LC-MS/MS analysis.