Low temperature inhibits pectin degradation by PpCBFs to prolong peach storage time.

Low-temperature storage is a widely used method for peach fruit storage. However, the impact of PpCBFs on pectin degradation during low-temperature storage is unclear. As such, in this study, we stored the melting-flesh peach cultivar "Fuli" at low temperature (LT, 6°C) and room temperature (RT, 25°C) to determine the effect of different temperatures on its physiological and biochemical changes. Low-temperature storage can inhibit the softening of "Fuli" peaches by maintaining the stability of the cell wall. It was found that the contents of water-soluble pectin and ionic-soluble pectin in peach fruit stored at RT were higher than those stored at LT. The enzyme activities of polygalacturonase (PG), pectate lyase (PL), and pectin methylesterase (PME) were all inhibited by LT. The expressions of PpPME3, PpPL2, and PpPG were closely related to fruit firmness, but PpCBF2 and PpCBF3 showed higher expression levels at LT than RT. The promoters of PpPL2 and PpPG contain the DER motif, which suggested that PpCBF2 and PpCBF3 might negatively regulate their expression by directly binding to their promoters. These results indicated that LT may maintain firmness by activating PpCBFs to repress pectin-degradation-related enzyme genes during storage.

The Malus domestica metal tolerance protein MdMTP11.1 was involved in the detoxification of excess manganese in Arabidopsis thaliana.

Ion homeostasis is maintained in plant cells by specialized transporters. However, functional studies on Mn transporters in apple trees have not been reported. MdMTP11.1, which encodes a putative Mn-MTP transporter in Malus domestica, was expressed highly in leaves and induced by Mn stress. Subcellular localization analysis of the MdMTP11.1-GFP fusion protein indicated that MdMTP11.1 was targeted to the Golgi. Meanwhile, overexpression of MdMTP11.1 in Arabidopsis thaliana conferred increased resistance to plants under toxic Mn levels, as evidenced by increased biomass of whole plant and length of primary root. Analysis of Mn bioaccumulation indicated that overexpression of MdMTP11.1 effectively reduced the content of Mn in every subcellular component and chemical forms when the plants were subjected with Mn stress. The majority of Mn of action were bound to cell wall and combined with un-dissolved phosphate. Besides, contents of malondialdehyde (MDA), proline and hydrogen peroxide (H2O2) were significantly lower, while content of chlorophyll and activities of CAT, SOD, POD and APX were significantly higher in MdMTP11.1-over-expressing plants compared with that in wild type plants under Mn stress. Taken together, these results suggest that MdMTP11.1 is a Mn specific transporter localized to the Golgi can maintain the phenotype, reduce the Mn accumulation and alleviate damage of oxidative stress, conferring the positive role of Mn tolerance.

Brassinosteroids enhance resistance to manganese toxicity in Malus robusta Rehd. via modulating polyamines profile.

Manganese (Mn) toxicity in soil is a widely observed phenomenon, which seriously restricts growth, quality, and yield of various crops and fruits including apples. However, mechanisms underlying the regulation of polyamines (PAs) by brassinosteroids (BRs) to improve tolerance to Mn stress are still unclear. In this study, we investigated the effects of 2,4-epibrassinolide (EBL; a BR) on the expression of genes involved in BR signaling pathway, Mn accumulation, PAs-mediated responses (PA precursor levels, metabolic enzymes, and genes), and growth parameters in Mn-stressed Malus robusta Rehd. EBL application significantly modulated the expressions of genes related to BR signaling (MdBRI, MdBSK, etc.) and reduced Mn accumulation, along with improving the rate of increase in root length and plant height, relative water content, chlorophyll content, maximum photochemical efficiency of PSII (Fv/Fm), and actual photochemical efficiency (ΦPSII) and decreasing electrical conductivity. Furthermore, EBL application significantly reduced putrescine (Put) accumulation and increased spermine (Spm) content and (Spd + Spm)/Put ratio. EBL weakened ornithine (Orn) pathway, decreased ornithine decarboxylase (ODC) activity, and increased biosynthesis of Spm from Put via elevating the PA oxidase (PAO) activity and expression of MdSPDS, MdSPMS, and MdPAO. The trends for free, PS-conjugated, and PIS-bound PAs were similar to that of total PAs, except that no significant change was observed in free Spm, PS-conjugated Spd, and Spm, as well as PIS-bound Spd. This study revealed that BR-regulated PAs help in mitigating Mn toxicity and clarified the mechanisms of regulation of PAs by BRs in apple trees.

The Chloroplastic Small Heat Shock Protein Gene KvHSP26 Is Induced by Various Abiotic Stresses in Kosteletzkya virginica.

Small heat shock proteins (sHSPs) are a group of chaperone proteins existed in all organisms. The functions of sHSPs in heat and abiotic stress responses in many glycophyte plants have been studied. However, their possible roles in halophyte plants are still largely known. In this work, a putative sHSP gene KvHSP26 was cloned from K. virginica. Bioinformatics analyses revealed that KvHSP26 encoded a chloroplastic protein with the typical features of sHSPs. Amino acid sequence alignment and phylogenetic analysis demonstrated that KvHSP26 shared 30%-77% homology with other sHSPs from Arabidopsis, cotton, durian, salvia, and soybean. Quantitative real-time PCR (qPCR) assays exhibited that KvHSP26 was constitutively expressed in different tissues such as leaves, stems, and roots, with a relatively higher expression in leaves. Furthermore, expression of KvHSP26 was strongly induced by salt, heat, osmotic stress, and ABA in K. virginica. All these results suggest that KvHSP26 encodes a new sHSP, which is involved in multiple abiotic stress responses in K. virginica, and it has a great potential to be used as a candidate gene for the breeding of plants with improved tolerances to various abiotic stresses.

Lead tolerance mechanism in sterilized seedlings of Potamogeton crispus L.: subcellular distribution, polyamines and proline.

The effects of increasing concentrations of lead (Pb) on malondialdehyde (MDA) content, soluble protein, Pb accumulation, nutrients, polyamines (PAs) and proline metabolism were investigated in sterilized seedlings of Potamogeton crispus L. after 5d exposure. Significant oxidative stress was not caused, indicated by a little induction of MDA content and soluble proteins. Pb accumulation increased in a concentration-dependent manner and most of Pb was stored in the cell wall. Total P, Mg, Na and Zn rose and total Fe fell; total Ca increased at 25 µM Pb but then declined. The nutrients in cell wall fraction changed in the same pattern as total nutrients, whereas those in soluble and organelle fraction declined. Total putrescine (Put) decreased markedly, while total spermidine (Spd), spermine (Spm) and (Spd+Spm)/Put ratio increased progressively but then declined. The trends for free, perchloric acid soluble conjugated (PS-conjugated) and perchloric acid insoluble bound (PIS-bound) PAs were similar to those on total PAs, except that PIS-bound Spm increased significantly. Arginine decarboxylase (ADC) and ornithine decarboxylase (ODC) activities rose gradually, while diamine oxidase (DAO) initially increased but then declined. Proline content increased initially only to decline later, due to the increase of r-glutamyl kinase (GK) activity. Meanwhile, ornithine-d-aminotransferase (OAT) activity gradually reduced, while no significant change was observed in proline dehydrogenase (PDH) activity. Our results indicated that the tolerance of P. crispus to Pb stress was based on cell wall compartmentalization combined with increase of nutrients, alterations of PAs, and induction of proline.

Photochemical performance of thylakoid membrane in lead-treated Nymphoides peltatum.

Photochemical responses in the thylakoid membrane of Nymphoides peltatum to increasing lead (Pb) concentrations were investigated after 5 days of exposure. Pb accumulation increased in a concentration-dependent manner, with a maximum of 118.44 µg g(-1) at 100 µM. Nutrients (Zn, Mg, Mo, Ca, Fe and Mn), ATPase activity and pigment generally increased progressively at Pb concentrations of 12.5 and 25 µM, but then declined at concentrations of 50 and 100 µM. Moreover, Pb stress induced an increase in chlorophyll (Chl) a/b ratio in a different extent. No outstanding changes were observed in several Chl a fluorescence parameters at low Pb concentrations (12.5 and 25 µM), while significant changes (p < 0.05) were observed in these parameters at higher concentrations. The alterations of nutrients, ATPase activity and pigment content were associated with disturbances in the thylakoid membrane, indicated by the quenching of Chl a fluorescence. These results were indicative of a disarray in photochemical activities exerted by Pb phytotoxicity.

Biochemical defense strategies in sterilized seedlings of Nymphoides peltatum adapted to lead stress.

In order to study potential antioxidant defense mechanisms, the effects of increasing concentrations of lead (Pb) on polyamines (PAs), various thiols, vitamins C and E, and proline contents in sterilized seedlings of Nymphoides peltata (S.G. mel.) Kuntze were investigated after 5 days of exposure. The levels of total putrescine (Put), spermidine (Spd), and spermine (Spm) decreased significantly, while the ratio of (Spd + Spm)/Put first increased but then declined as the concentration of Pb increased. The trends for free, perchloric acid soluble-conjugated (PS-conjugated), and perchloric acid insoluble-bound (PIS-bound) PAs were similar to the trend seen for total PAs. Moreover, reduced glutathione (GSH), nonprotein thiols (NP-SH), phytochelatins (PCs), and vitamin C were induced at high Pb concentrations. No significant change was observed in vitamin E. An initial decline in proline content was followed by an increase as the Pb concentration rose. The reduced level of Put and elevated contents of GSH, NP-SH, PCs, vitamin C, and proline were found to be associated with antioxidant efficiency, which supports the hypothesis that they could play a significant role in the adaptation mechanisms of N. peltatum under Pb stress.

Lead-induced oxidative damage in steriled seedlings of Nymphoides peltatum.

The effects of increasing concentrations of lead (Pb) on Pb accumulation and its influence on nutrient elements, malondialdehyde (MDA) content, generation of superoxide anion (O2(-·)), hydrogen peroxide (H2O2) content, antioxidant enzymes activities, soluble protein, and photosynthetic pigment, as well as chloroplast ultrastructure in steriled seedlings of Nymphoides peltata (S. G. Gmel.) Kuntze were investigated in order to understand Pb-induced toxicity. The accumulation of Pb was found to increase in a concentration-dependent manner. Nutrient elements (Ca, K, Fe, Mn, and Mo) were also affected. MDA content and O2(-·) generation rate increased progressively, while H2O2 content first boosted up at a low Pb concentration of 12.5 µM but then declined. Guaiacol peroxidase and catalase activities increased alternately, while superoxide dismutase activity gradually fell. Negative correlations were found between Pb and soluble protein and photosynthetic pigment. Moreover, Pb exposure resulted in a significant damage of chloroplasts. Taken together, these findings supported the hypothesis that Nymphoides peltatum underwent oxidative stress induced by Pb. In addition, both the disorder of nutrient elements and the damage to the ultrastructure of chloroplasts were indicative of general disarray in the cellular functions exerted by Pb.

[Subcellular distribution and phytotoxicity of cadmium in Alternanthera philoxeroides leaves].

A hydroponic experiment was conducted to study the subcellular distribution of Cd and mineral elements in Alternanthera philoxeroides leaves and the leaves anti-oxidative capacity and chlorophyll, soluble protein, and phytochelatins (PCs) contents under the stress of different concentration (0, 0.05, 0.1, 0.2, 0.4 mmol x L(-1)) Cd. With the increasing concentration of Cd in culture medium, the Cd content in all subcellular components of A. philoxeroides leaves increased significantly, and mainly distributed in cell wall, followed by in soluble fractions, and in chloroplast and mitochondria. When the Cd concentration in the medium exceeded 0.2 mmol x L(-1), the Cd was more allocated in soluble fractions than in cell wall. Cd stress resulted in an obvious imbalance of mineral elements uptake. With the increasing concentration of Cd, the Ca content in all subcellular components, especially in cell wall, increased significantly, whereas the P and K contents in cell wall and soluble fractions as well as the Mg and Fe contents in chloroplast decreased. In the meantime, the soluble protein and chlorophyll contents showed a decreasing trend, the glutathione and ascorbic acid contents decreased after an initial increase, the total anti-oxidative capacity (T-AOC) increased progressively, and the PCs accumulated in large quantity. These results suggested that A. philoxeroides had definite resistance to the water body Cd, and there was a dosage-effect relationship between the Cd enrichment in the subcellular components of A. philoxeroides leaves and the phytotoxicity of Cd. The imbalance of the mineral elements in subcellular components and the decrease of soluble protein and chlorophyll contents in chloroplast indicated the obvious phytotoxicity of Cd, while the massive accumulation of Ca in cell wall and the increased levels of PCs and T-AOC suggested the stronger resistance of A. philoxeroides to Cd stress. There was a definite correlation between the PCs production by A. philoxeroides and the toxicity of Cd, suggesting that the PCs could be considered as a sensitive biomarker for estimating the Cd phytotoxicity.