Regulation of Mitochondrial Respiration by Hydrogen Sulfide.

Hydrogen sulfide (H2S), the third gasotransmitter, has positive roles in animals and plants. Mitochondria are the source and the target of H2S and the regulatory hub in metabolism, stress, and disease. Mitochondrial bioenergetics is a vital process that produces ATP and provides energy to support the physiological and biochemical processes. H2S regulates mitochondrial bioenergetic functions and mitochondrial oxidative phosphorylation. The article summarizes the recent knowledge of the chemical and biological characteristics, the mitochondrial biosynthesis of H2S, and the regulatory effects of H2S on the tricarboxylic acid cycle and the mitochondrial respiratory chain complexes. The roles of H2S on the tricarboxylic acid cycle and mitochondrial respiratory complexes in mammals have been widely studied. The biological function of H2S is now a hot topic in plants. Mitochondria are also vital organelles regulating plant processes. The regulation of H2S in plant mitochondrial functions is gaining more and more attention. This paper mainly summarizes the current knowledge on the regulatory effects of H2S on the tricarboxylic acid cycle (TCA) and the mitochondrial respiratory chain. A study of the roles of H2S in mitochondrial respiration in plants to elucidate the botanical function of H2S in plants would be highly desirable.

Nitric oxide-mediated DNA methylation enhances cold resistance in postharvest peach fruit.

Low temperature can affect DNA methylation. Since exogenous use of NO can reduce cold damage in peach fruit during cold storage, this study investigated the roles of NO on DNA methylation of peaches suffering cold stress. The results showed that exogenous NO effectively alleviated the decrease in total DNA methyltransferase (DNMT) activity and transcript levels induced by cold stress, whereas c-PTIO exacerbated the decrease in total DNMT activity and transcript levels. Further BSP analysis of the promoter regions of four cold resistance genes (PpCBF5-IS2, PpICE1-IS, PpMYC2-IS, PpCOR-IS1) in peaches showed that in peaches treated with exogenous NO, PpCBF5-IS2 and PpICE1-IS were modified by hypermethylation, PpMYC2-IS was modified by methylation, PpCOR-IS1 was modified by demethylation and insensitive to NO. It was suggested that NO could enhance the cold resistance of postharvest peaches by mediating DNA methylation.

The transcription factor OsMYBc and an E3 ligase regulate expression of a K+ transporter during salt stress.

Sodium (Na+) and potassium (K+) homeostasis is essential for plant survival in saline soils. A member of the High-Affinity K+ Transporter (HKT) family in rice (Oryza sativa), OsHKT1;1, is a vital regulator of Na+ exclusion from shoots and is bound by a MYB transcription factor (OsMYBc). Here, we generated transgenic rice lines in the oshkt1;1 mutant background for genetic complementation using genomic OsHKT1;1 containing a native (Com) or mutated (mCom) promoter that cannot be bound by OsMYBc. In contrast to wild-type (WT) or Com lines, the mCom lines were not able to recover the salt-sensitive phenotype of oshkt1;1. The OsMYBc-overexpressing plants were more tolerant to salt stress than WT plants. A yeast two-hybrid screen using the OsMYBc N-terminus as bait identified a rice MYBc stress-related RING finger protein (OsMSRFP). OsMSRFP is an active E3 ligase that ubiquitinated OsMYBc in vitro and mediated 26S proteasome-mediated degradation of OsMYBc under semi-in vitro and in vivo conditions. OsMSRFP attenuated OsMYBc-mediated OsHKT1;1 expression, and knockout of OsMSRFP led to rice salt tolerance. These findings uncover a regulatory mechanism of salt response that fine-tunes OsHKT1;1 transcription by ubiquitination of OsMYBc.

Interplay Among Hydrogen Sulfide, Nitric Oxide, Reactive Oxygen Species, and Mitochondrial DNA Oxidative Damage.

Hydrogen sulfide (H2S), nitric oxide (NO), and reactive oxygen species (ROS) play essential signaling roles in cells by oxidative post-translational modification within suitable ranges of concentration. All of them contribute to the balance of redox and are involved in the DNA damage and repair pathways. However, the damage and repair pathways of mitochondrial DNA (mtDNA) are complicated, and the interactions among NO, H2S, ROS, and mtDNA damage are also intricate. This article summarized the current knowledge about the metabolism of H2S, NO, and ROS and their roles in maintaining redox balance and regulating the repair pathway of mtDNA damage in plants. The three reactive species may likely influence each other in their generation, elimination, and signaling actions, indicating a crosstalk relationship between them. In addition, NO and H2S are reported to be involved in epigenetic variations by participating in various cell metabolisms, including (nuclear and mitochondrial) DNA damage and repair. Nevertheless, the research on the details of NO and H2S in regulating DNA damage repair of plants is in its infancy, especially in mtDNA.

Seed specifically over-expressing DGAT2A enhances oil and linoleic acid contents in soybean seeds.

Triacylglycerol (TAG), a main component of oil, is mainly biosynthesized by diacylglycerol acyltransferase (DGAT), which is critical for oil accumulation in plants. Intensive focus has been on DGAT2 functioning in unsaturated fatty acids biosynthesis. In this study, we analyzed the coding sequence (CDS) and amino acid sequence of GmDGAT2A and determined its key active sites through site-directed mutagenesis. As a consequence, H132, G201, and P152-X-I154-K155 were found to be essential active sites for GmDGAT2A. The spatial structure of the protein may bring the three active sites into close proximity, constituting an active domain. Additionally, N-terminus of GmDGAT2A was found to be an important regulator for the activity. Further, in vitro activity results uncovered GmDGAT2A was prone to utilize C18:2-CoA as the substrate. Consequently, overexpression of GmDGAT2A driven by a seed-specific promoter of Gmole1 in soybean significantly increased linoleic acid content specifically and total oil content, concomitant with accelerated elongation.

Overexpression of soybean GmPLDγ enhances seed oil content and modulates fatty acid composition in transgenic Arabidopsis.

Phospholipase D (PLD) hydrolyzes the phosphodiester bond of glycerophospholipids to yield phosphatidic acid (PA) and a free headgroup. PLDs are important for plant growth, development, and responses to external stresses. However, their roles in triacylglycerol (TAG) synthesis are still unclear. Here, we report that a soybean (Glycine max) PLDγ (GmPLDγ) is involved in glycerolipid turnover and seed oil production. GmPLDγ was targeted to mitochondria and exhibited PLD activity that was activated by oleate and phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2]. Overexpression of GmPLDγ (abbreviated GmPLDγ-OE) in Arabidopsis thaliana resulted in enhanced seed weight, elevated levels of TAGs with 18-, 20-, and 22-carbon fatty acids (FAs), and altered oil-body morphology. Furthermore, the levels of membrane lipids in vegetative tissues decreased significantly, whereas no overt changes were found in mature seeds except for a decrease in the digalactosyldiacylglycerol (DGDG) level in the GmPLDγ-OE lines. Additionally, the expression of genes involved in glycerolipid metabolism was significantly upregulated in developing siliques in GmPLDγ-OE lines. Together, our data indicate a regulatory role for GmPLDγ in TAG synthesis and fatty-acid remodeling, highlighting the importance of mitochondria-directed glycerophospholipid homeostasis in seed oil accumulation.

Genome-wide analysis and functional characterization of Acyl-CoA:diacylglycerol acyltransferase from soybean identify GmDGAT1A and 1B roles in oil synthesis in Arabidopsis seeds.

Acyl-CoA:diacylglycerol acyltransferase (DGAT) is a key enzyme in the Kennedy pathway of triacylglycerol (TAG) synthesis. It catalyzes the acyl-CoA-dependent acylation of sn-1, 2-diacylglycerol to form TAG. DGATs in soybean (Glycine max) have been reported, but their functions are largely unclear. Here we cloned three members of DGAT1 and four members of DGAT2 family from soybean, named GmDGAT1A to GmDGAT1C, and GmDGAT2A to GmDGAT2D, respectively. GmDGAT1A and GmDGAT1C were expressed at a high level in immature seeds, GmDGAT2B in mature seeds, and GmDGAT2C in older leaves. The seven genes were transformed into the H1246 quadruple mutant yeast strain, in which GmDGAT1A, GmDGAT1B, GmDGAT1C, GmDGAT2A, and GmDGAT2B had the ability to produce TAG. Six genes were transformed into Arabidopsis respectively, and constitutive expression of GmDGAT1A and GmDGAT1B resulted in an increase in oil content at the cost of reduced protein content in seeds. Overexpression of GmDGAT1A produced heavier weight of individual seed, but did not affect the weight of total seeds from a plant. Our results reveal the functions of soybean DGATs in seed oil synthesis using transgenic Arabidopsis. The implications for the biotechnological modification of the oil contents in soybeans by altering DGAT expression are discussed.

Effects of nitric oxide on mitochondrial oxidative defence in postharvest peach fruits.

BACKGROUND: It has been confirmed that the accumulation of reactive oxygen species (ROS) in fruit can cause oxidative damage and nitric oxide (NO) can regulate the accumulation of ROS and the antioxidative defence of fruit. However, little is known about the roles of NO on the antioxidant system in mitochondria of fruit. In this study, Feicheng peach fruits were dipped with 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO) and NO solutions to explore the effects of NO on the membrane permeability transition and antioxidant system in mitochondria of peach fruit. RESULTS: Treatment with 15 µmol L(-1) NO solution could delay the decrease of mitochondrial permeability transition and decrease the content of ROS in mitochondria. Besides, when the endogenous NO was scavenged by c-PTIO, the ROS in mitochondria increased greatly and superoxide dismutase activity decreased, while the content and activities of peroxidase and catalase changed slightly. CONCLUSION: By delaying the decrease of mitochondrial permeability transition, 15 µmol L(-1) NO treatment could promote a more stable internal medium in mitochondria of Feicheng peach fruit. The increases in the activities of antioxidant enzymes in mitochondria caused by the remove of endogenous NO suggested that NO also plays an important role in the mitochondrial antioxidant system of Feicheng peach fruit.