Physiological and comparative transcriptome analysis of the response and adaptation mechanism of the photosynthetic function of mulberry (Morus alba L.) leaves to flooding stress

Flooding has become one of the major abiotic stresses that seriously affects plant growth and development owing to changes in the global precipitation pattern. Mulberry (Morus alba L.) is a desirable tree spePhysocarpus amurensis Maxim andcies with high ecological and economic benefits. To reveal the response and adaptive mechanisms of the photosynthetic functions of mulberry leaves to flooding stress, chlorophyll synthesis, photosynthetic electron transfer and the Calvin cycle were investigated by physiological studies combined with an analysis of the transcriptome. Flooding stress inhibited the synthesis of chlorophyll (Chl) and decreased its content in mulberry leaves. The sensitivity of Chl a to flooding stress was higher than that of Chl b owing to the changes of CHLG (LOC21385082) and CAO (LOC21408165) that encode genes during chlorophyll synthesis. The levels of expression of Chl b reductase NYC (LOC112094996) and NYC (LOC21385774), which are involved in Chl b degradation, were upregulated on the fifteenth day of flooding, which accelerated the transformation of Chl b to Chl a, and upregulated the expression of PPH (LOC21385040) and PAO (LOC21395013). This accelerated the degradation of chlorophyll. Flooding stress significantly inhibited the photosynthetic function of mulberry leaves. A Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of differentially expressed genes under different days of flooding stress indicated significant enrichment in Photosynthesis-antenna proteins (map00196), Photosynthesis (map00195) and Carbon fixation in photosynthetic organisms (map00710). On the fifth day of flooding, 7 and 5 genes that encode antenna proteins were identified on LHCII and LHCI, respectively. They were significantly downregulated, and the degree of downregulation increased as the trees were flooded longer. Therefore, the power of the leaves to capture solar energy and transfer this energy to the reaction center was reduced. The chlorophyll fluorescence parameters and related changes in the expression of genes in the transcriptome indicated that the PSII and PSI of mulberry leaves were damaged, and their activities decreased under flooding stress. On the fifth day of flooding, electron transfer on the PSII acceptor side of mulberry leaves was blocked, and the oxygen-evolving complex (OEC) on the donor side was damaged. On the tenth day of flooding, the thylakoid membranes of mulberry leaves were damaged. Five of the six coding genes that mapped to the OEC were significantly downregulated. Simultaneously, other coding genes located at the PSII reaction center and those located at the PSI reaction center, including Cytb6/f, PC, Fd, FNR and ATP, were also significantly downregulated. In addition, the gas exchange parameters (P(n), G(s), T(r), and C(i)) of the leaves decreased after 10 days of flooding stress primarily owing to the stomatal factor. However, on the fifteenth day of flooding, the value for the intracellular concentration of CO(2) was significantly higher than that on the tenth day of flooding. In addition, the differentially expressed genes identified in the Calvin cycle were significantly downregulated, suggesting that in addition to stomatal factors, non-stomatal factors were also important factors that mediated the decrease in the photosynthetic capacity of mulberry leaves. In conclusion, the inhibition of growth of mulberry plants caused by flooding stress was primarily related to the inhibition of chlorophyll synthesis, antenna proteins, photosynthetic electron transfer and the Calvin cycle. The results of this study provide a theoretical basis for the response and mechanism of adaptation of the photosynthetic function of mulberry to flooding stress.