Chlorophyll synthesis and the photoprotective mechanism in leaves of mulberry (Morus alba L.) seedlings under NaCl and NaHCO3 stress revealed by TMT-based proteomics analyses

Abstract

Chlorophyll (Chl) and effective photoprotective mechanism are important prerequisites to ensure the photosynthetic function of plants under stress. In this study, the effects of 100 mmol L^-1 NaCl and NaHCO₃ stress on chlorophyll synthesis and photosynthetic function of mulberry seedlings were studied by physiological combined with proteomics technology. The results show that: NaCl stress had little effect on the expression of Chl synthesis related proteins, and there were no significant changes in Chl content and Chl a:b ratio. However, 13 of the 15 key proteins in the process of Chl synthesis were significantly decreased under NaHCO₃ stress, and the contents of Chl a and Chl b were significantly decreased (especially Chl a). Although stomatal conductance (G_s) decreased significantly under NaCl stress, net photosynthetic rate (P_n), PSII maximum photochemical efficiency (F_v/F_m) and electron transfer rate (ETR) did not change significantly, but under NaHCO₃ stress, not only G_s decreased significantly, PSII activity and photosynthetic carbon were the same. In the photoprotective mechanism under NaCl stress, NAD(P)H dehydrogenase (NDH)-dependent cyclic electron flow (CEF) enhanced, the expression of related proteins subunit, ndhH, ndhI, ndhK, and ndhM, the key enzyme of the xanthophyll cycle, violaxanthin de-epoxidase (VDE) were up-regulated, the ratio of (A + Z)/(V + A + Z) and non-photochemical quenching (NPQ) was increased. The expressions of proteins FTR and Fd-NiR were also significant up-regulated under NaCl stress, Fd-dependent ROS metabolism and nitrogen metabolism can effectively reduce the electronic pressure on Fd. Under NaHCO₃ stress, the expressions of NDH-dependent CEF related proteins subunit (ndhH, ndhI, ndhK, ndhM and ndhN), VDE, ZE, FTR, Fd-NiR and Fd-GOGAT, were significant down-regulated, and ZE, CP26, ndhK, ndhM, Fd-NiR, Fd-GOGAT and FTR genes expression also significantly decreased, the photoprotective mechanism, like the xanthophyll cycle，CEF and Fd-dependent ROS metabolism and nitrogen metabolism might be damaged, resulting in the inhibition of PSII electron transfer and carbon assimilation in mulberry leaves under NaHCO₃ stress.

Keywords: Morus alba L.; NaCl and NaHCO(3) stress; Photoprotective mechanism; Photosynthesis; Proteomics.