Toxic effects of heavy metal Cd and Zn on chlorophyll, carotenoid metabolism and photosynthetic function in tobacco leaves revealed by physiological and proteomics analysis.

To explore the mechanisms underlying the action of the heavy metals Cd and Zn on the photosynthetic function of plant leaves, the effects of 100 µmol L-1 Cd and 200 µmol L-1 Zn stress (the exposure concentrations of Cd and Zn in the culture medium were 2.24 mg kg-1 and 5.36 mg kg-1) on the chlorophyll and carotenoid contents as well as the photosynthetic function of tobacco leaves (Long Jiang 911) were studied. The key proteins in these physiological processes were quantitatively analyzed using a TMT-based proteomics approach. Cd stress was found to inhibit the expression of key enzymes during chlorophyll synthesis in leaves, resulting in a decrease of the Chl content. However, Zn stress did not significantly influence the chlorophyll content. Leaves adapted to Zn stress by upregulating CAO expression and increase the Chl b content. Although the Car content in leaves did not significantly change under either Cd or Zn stress, the expressions of ZE and VDE during Car metabolism decreased significantly under Cd stress. This was accompanied by damages to the xanthophyll cycle and the NPQ-dependent energy dissipation mechanism. In contrast, under Zn stress, leaves adapted to Zn stress by increasing the expression of VDE, thus improving NPQ. Under Cd stress, the expressions of three sets of proteins were significantly down-regulated, including PSII donor-side proteins (PPD3, PPD6, OEE1, OEE2-1, OEE2-2, OEE2-3, and OEE3-2), receptor-side proteins (D1, D2, CP43, CP47, Cyt b559α, Cyt b559ß, PsbL, PsbQ, PsbR, Psb27-H1, and Psb28), and core proteins of the PSI reaction center (psaA, psaB, psaC, psaD, psaE-A, PsaE-B, psaF, psaG, psaH-1, psaK, psaL, psaN, and psaOL). In comparison, only eight of the above proteins (PPD6, OEE3-2, PsbL, PsbQ, Psb27-H1, psaL, and psaOL) were significantly down-regulated by Zn stress. Under Cd stress, both the donor side and the receptor side of PSII were damaged, and PSII and PSI experienced severe photoinhibition. However, Zn stress did not decrease either PSII or PSI activities in tobacco leaves. In addition, the expression of electron transport-related proteins (cytb6/f complex, PC, Fd, and FNR), ATPase subunits, Rubisco subunits, and RCA decreased significantly in leaves under Cd stress. However, no significant changes were observed in any of these proteins under Zn stress. Although Cd stress was found to up-regulate the expressions of PGRL1A and PGRL1B and induce an increase of PGR5/PGRL1-CEF in tobacco leaves, NDH-CEF was significantly inhibited. Under Zn stress, the expressions of ndhH and PGRL1A in leaves were significantly up-regulated, but there were no significant changes in either NDH-CEF or PGR5/PGRL-CEF. Under Cd stress, the expressions of proteins related to Fd-dependent nitrogen metabolism and reactive oxygen species (ROS) scavenging processes (e.g., FTR, Fd-NiR, and Fd-GOGAT) were significantly down-regulated in leaves. However, no significant changes of any of the above proteins were identified under Zn stress. In summary, Cd stress could inhibit the synthesis of chlorophyll in tobacco leaves, significantly down-regulate the expressions of photosynthesis-related proteins or subunits, and suppress both the xanthophyll cycle and NDH-CEF process. The expressions of proteins related to the Fd-dependent nitrogen metabolism and ROS scavenging were also significantly down-regulated, which blocked the photosynthetic electron transport, thus resulting in severe photoinhibition of both PSII and PSI. However, Zn stress had little effect on the photosynthetic function of tobacco leaves.

Retinal microvascular changes in patients with pancreatitis and their clinical significance.

Acute pancreatitis, a common exocrine inflammatory disease affecting the pancreas, is characterized by intense abdominal pain and multiple organ dysfunction. However, the alterations in retinal blood vessels among individuals with acute pancreatitis remain poorly understood. This study employed optical coherence tomography angiography (OCTA) to examine the superficial and deep retinal blood vessels in patients with pancreatitis. Sixteen patients diagnosed with pancreatitis (32 eyes) and 16 healthy controls (32 eyes) were recruited from the First Affiliated Hospital of Nanchang University for participation in the study. Various ophthalmic parameters, such as visual acuity, intraocular pressure, and OCTA image for retina consisting of the superficial retinal layer (SRL) and the deep retinal layer (DRL), were recorded for each eye. The study observed the superficial and deep retinal microvascular ring (MIR), macrovascular ring (MAR), and total microvessels (TMI) were observed. Changes in retinal vascular density in the macula through annular partitioning (C1-C6), hemispheric quadrant partitioning (SR, SL, IL, and IR), and early diabetic retinopathy treatment studies (ETDRS) partitioning methods (R, S, L, and I). Correlation analysis was employed to investigate the relationship between retinal capillary density and clinical indicators. Our study revealed that in the superficial retinal layer, the vascular density of TMI, MIR, MAR, SR, IR, S, C2, C3 regions were significantly decreased in patients group compared with the normal group. For the deep retinal layer, the vascular density of MIR, SR, S, C1, C2 regions also reduced in patient group. The ROC analysis demonstrated that OCTA possesses significant diagnostic performance for pancreatitis. In conclusion, patients with pancreatitis may have retinal microvascular dysfunction, and OCTA can be a valuable tool for detecting alterations in ocular microcirculation in pancreatitis patients in clinical practice.

TMT-based quantitative proteomic analysis of the effects of Pseudomonas syringae pv. tabaci (Pst) infection on photosynthetic function and the response of the MAPK signaling pathway in tobacco leaves.

To reveal the mechanism of photosynthesis inhibition by infection and the response of the MAPK signaling pathway to pathogen infection, tobacco leaves were inoculated with Pseudomonas syringae pv. tabaci (Pst), and the effects of Pst infection on photosynthesis of tobacco leaves were studied by physiological and proteomic techniques, with a focus on MAPK signaling pathway related proteins. Pst infection was observed to lead to the degradation of chlorophyll (especially Chl b) in tobacco leaves and the down-regulation of light harvesting antenna proteins expression, thus limiting the light harvesting ability. The photosystem II and I (PSII and PSI) activities were also decreased, and Pst infection inhibited the utilization of light and CO2. Proteomic analyses showed that the number of differentially expressed proteins (DEPs) under Pst infection at 3 d were significantly higher than at 1 d, especially the number of down-regulated proteins. The KEGG enrichment of DEPs was mainly enriched in the energy metabolism processes such as photosynthesis antenna proteins and photosynthesis. The down-regulation of chlorophyll a-b binding protein, photosynthetic electron transport related proteins (e.g., PSII and PSI core proteins, the Cytb6/f complex, PC, Fd, FNR), ATP synthase subunits, and key enzymes in the Calvin cycle were the key changes associated with Pst infection that may inhibit tobacco photosynthesis. The effect of Pst infection on the PSII electron acceptor side was significantly greater than that on the PSII donor side. The main factor that decreased the photosynthetic ability of tobacco leaves with Pst infection at 1 d may be the inhibition of photochemical reactions leading to an insufficient supply of ATP, rather than decreased expression of enzymes involved in the Calvin cycle. At 1 d into Pst infection, the PSII regulated energy dissipation yield Y(NPQ) may play a role in preventing photosynthetic inhibition in tobacco leaves, but the long-term Pst infection significantly inhibited Y(NPQ) and the expression of PsbS proteins. Proteins involved in the MAPK signaling pathway were up-regulated, suggesting the MAPK signaling pathway was activated to respond to Pst infection. However, at the late stage of Pst infection (at 3 d), MAPK signaling pathway proteins were degraded, and the defense function of the MAPK signaling pathway in tobacco leaves was damaged.

Physiological and proteomics responses of nitrogen assimilation and glutamine/glutamine family of amino acids metabolism in mulberry (Morus alba L.) leaves to NaCl and NaHCO3 stress.

In order to find out the response mechanism of nitrogen assimilation and glutamine/glutamine family of amino acids metabolism in mulberry (Morus alba L.) leaves under NaCl and NaHCO3 stress, and to reveal its role in salt alkali adaptation. The effects of the nitrogen metabolism of mulberry leaves were studied under 100 mmol L-1 NaCl and NaHCO3 stress.The results showed that the activity of NR and the content of TN and SP did not change significantly, the expression of NiR, Fd-NiR, Fd-NiR gene and theactivity of NiR increased significantly under NaCl stress, but nitrogen assimilation was inhibited under NaHCO3 stress. NaCl stress had no significant effect on the expression and activity of GS and GOGAT in mulberry leaves. Under NaHCO3 stress, the expression of Fd-GOGAT, Fd-GOGAT2, Fd-GOGAT gene, and the activity of GS and GOGAT were significantly decreased. NaCl stress can promote the accumulation of Pro, Put and Spd in mulberry leaves. The accumulation of Pro under NaHCO3 stress is greater than that under NaCl stress. NaCl stress also induced the up-regulation of GAD, GAD1 and GAD1 gene expression, so promoting the synthesis of GABA may be an adaptive mechanism for mulberry to cope with NaCl stress, but the expression of GAD did not change significantly and GAD gene expression lower than CK under NaHCO3 stress. Although both NaCl and NaHCO3 stress could promote the synthesis of GSH by up-regulation of GCLM expression, GSH under NaHCO3 stress was significantly higher than that under NaCl stress, the content of H2O2 was still significantly higher than that of NaCl stress, that means GSH may not play a key role in alleviating the oxidative damage in mulberry leaves caused by salt and alkali.

The degradation of BPA on enhanced heterogeneous photo-Fenton system using EDDS and different nanosized hematite.

Photo-Fenton processes have been widely studied in wastewater treatment. In this research, the degradation of bisphenol A (BPA) was carried out in a new heterogeneous photo-Fenton process. The ethylenediamine-N,N'-disuccinic acid (EDDS) was used as chelating agent in this system with two different kinds of commercially available nanosized hematite (30 nm and 80 nm) addition. The results showed that the present of EDDS could enhance the degradation efficiency. And can be concluded that the degradation efficiency is better in the system with 30 nm hematite. The TEM, XRD, and specific surface area were conducted to understand the different characteristics of the two size hematite. The adsorption experiments of BPA and EDDS on hematite proved that there was little adsorption of BPA while the EDDS was adsorbed much more on hematite, which has confirmed Fe(III) and EDDS can form Fe(III)-EDDS complex. The effects of different parameters including hematite loading, H2O2, and EDDS concentrations on the degradation process were investigated. According to the results, the optimum condition for BPA degradation using 30 nm (0.8 g L-1 hematite, 0.1 mmol L-1 H2O2, and 1.2 mmol L-1 EDDS) and 80 nm (0.6 g L-1 hematite, 0.05 mmol L-1 H2O2, and 1.2 mmol L-1 EDDS) hematite were selected. It was confirmed that the ·OH plays an important role in the oxidation process through attacking the BPA molecule and produce hydroxyl addition derivative. In addition, O2 can react with electron (e-) and holes (h+) produced by iron oxide under UV irradiation to create 1O2, which could work as potential reactive species to oxidize BPA.

Overexpression of Trx CDSP32 gene promotes chlorophyll synthesis and photosynthetic electron transfer and alleviates cadmium-induced photoinhibition of PSII and PSI in tobacco leaves.

Cadmium stress causes a decrease in chlorophyll content and inhibits photosynthesis in tobacco leaves. The role of thioredoxin-like protein CDSP32 expressed in plant chloroplasts is to alleviates the reduced enzymes expression involved in chlorophyll synthesis of tobacco leaves due to Cd exposure, effectively preventing chlorophyll degradation and promoting increased tobacco biomass. Overexpression of Trx CDSP32 can protect the oxygen-evolving complex on the PSII donor side and promote electron transfer on the PSII acceptor side of tobacco leaves under Cd stress. Trx CDSP32 not only significantly increase the PSI activity of tobacco leaves, but also alleviate cadmium-induced PSI photoinhibition. Although Trx CDSP32 has no significant effect on the expression of PC and FNR proteins in tobacco leaves under Cd stress, it can alleviate the decreased expression of protein subunits involved in photosynthetic electron transfer such as Cyt b6/f complex subunits, Fd, and ATP synthase subunits. Trx CDSP32 can promote the synthesis of chlorophyll, stabilize the electron transfer chain, and promote ATP synthase activity to alleviate cadmium-induced photoinhibition of PSII and PSI in tobacco leaves.