Differentially expressed protein and gene analysis revealed the effects of temperature on changes in ascorbic acid metabolism in harvested tea leaves

Tea is an important non-alcoholic beverage worldwide.Tea quality is determined by numerous secondary metabolites in harvested tea leaves,including tea polyphenols,theanine,caffeine,and ascorbic acid(AsA).AsA metabolism in harvested tea leaves is affected by storage and transportation temperature.However,the molecular mechanisms underlying AsA metabolism in harvested tea leaves exposed to different storage and transportation temperature conditions remain unclear.Here we performed RP-HPLC to detect dynamic changes in AsA content in tea leaves subjected to high-(38°C),low-(4°C),or room-temperature(25°C)treatments.The AsA distribution and levels in the treated tea leaves were analyzed using cytological–anatomical characterization methods.The differentially expressed CsAPX1 and CsDHAR2 proteins,which are involved in the AsA recycling pathway,were identified from the corresponding proteomic data using iTRAQ.We also analyzed the expression profiles of 18 genes involved in AsA metabolism,including CsAPX1 and CsDHAR2.AsA was mainly distributed in tea leaf mesophyll cells.High-and low-temperature treatments upregulated the CsAPX1 and CsDHAR2 proteins and induced CsAPX and CsDHAR2 gene expression.These results indicated that the CsAPX1 and CsDHAR2 proteins might have critical roles in AsA recycling in tea leaves.Our results provide a foundation for the in-depth investigation of AsA metabolism in tea leaves during storage and transportation,and they will promote better tea flavor in tea production.