Lailiang Cheng

The sun-exposed peel of apple fruit has higher xanthophyll cycle-dependent thermal dissipation and antioxidants of the ascorbate–glutathione pathway than the shaded peel

Abstract The objective of this study was to determine how xanthophyll cycle-dependent thermal dissipation and the antioxidant system in the peel of apple fruit respond to the natural light exposure within the tree canopy. Fruit from exterior and interior canopies of both mature ‘Gala’ and ‘Smoothee’ apple trees were sampled at noon and/or predawn to measure chlorophyll fluorescence, xanthophyll cycle pool size and composition, and enzymatic and non-enzymatic antioxidants. Compared with the shaded side, the sun-exposed peel of the fruit had more excess absorbed photon flux density (PFD) as a result of a lower photosystem II operating efficiency and a higher incident PFD at noon. The efficiency of excitation transfer was lower in the sun-exposed peel than in the shaded peel, indicative of higher thermal dissipation. The sun-exposed peel had a larger xanthophyll cycle pool size and a higher conversion state. It also had higher activities of ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase, and glutathione reductase, and a larger size and a higher reduction state of the ascorbate pool and the glutathione pool. However, catalase activity was lower in the sun-exposed peel than in the shaded peel. Superoxide dismutase did not show significant trend with regard to fruit peel type or position in the canopy. We conclude that both the xanthophyll cycle and the ascorbate–glutathione pathway in the apple fruit peel are acclimated to the prevailing light exposure within the tree canopy to meet the respective needs for dissipating excess absorbed PFD and detoxifying reactive oxygen species.

Expression Patterns of Genes Involved in Sugar Metabolism and Accumulation during Apple Fruit Development

Both sorbitol and sucrose are imported into apple fruit from leaves. The metabolism of sorbitol and sucrose fuels fruit growth and development, and accumulation of sugars in fruit is central to the edible quality of apple. However, our understanding of the mechanisms controlling sugar metabolism and accumulation in apple remains quite limited. We identified members of various gene families encoding key enzymes or transporters involved in sugar metabolism and accumulation in apple fruit using homology searches and comparison of their expression patterns in different tissues, and analyzed the relationship of their transcripts with enzyme activities and sugar accumulation during fruit development. At the early stage of fruit development, the transcript levels of sorbitol dehydrogenase, cell wall invertase, neutral invertase, sucrose synthase, fructokinase and hexokinase are high, and the resulting high enzyme activities are responsible for the rapid utilization of the imported sorbitol and sucrose for fruit growth, with low levels of sugar accumulation. As the fruit continues to grow due to cell expansion, the transcript levels and activities of these enzymes are down-regulated, with concomitant accumulation of fructose and elevated transcript levels of tonoplast monosaccharide transporters (TMTs), MdTMT1 and MdTMT2; the excess carbon is converted into starch. At the late stage of fruit development, sucrose accumulation is enhanced, consistent with the elevated expression of sucrose-phosphate synthase (SPS), MdSPS5 and MdSPS6, and an increase in its total activity. Our data indicate that sugar metabolism and accumulation in apple fruit is developmentally regulated. This represents a comprehensive analysis of the genes involved in sugar metabolism and accumulation in apple, which will serve as a platform for further studies on the functions of these genes and subsequent manipulation of sugar metabolism and fruit quality traits related to carbohydrates.

Delay in leaf senescence of Malus hupehensis by long‐term melatonin application is associated with its regulation of metabolic status and protein degradation

Melatonin has an important anti‐aging role in plant physiology. We tested the effects of long‐term melatonin exposure on metabolic status and protein degradation during natural leaf senescence in trees of Malus hupehensis Rehd. The 2‐month regular supplement of 100 μm melatonin to the soil once every 6 days altered the metabolic status and delayed protein degradation. For example, leaves from treated plants had significantly higher photosynthetic activity, chlorophyll concentrations, and levels of three photosynthetic end products (sorbitol, sucrose, and starch) when compared with the control. The significant inhibition of hexose (fructose and glucose) accumulation possibly regulated the signaling of MdHXK1, a gene for which expression was also repressed by melatonin during senescence. The plants also exhibited better preservation of their nitrogen, total soluble protein, and Rubisco protein concentrations than the control. The slower process of protein degradation might be a result of melatonin‐linked inhibition on the expression of apple autophagy‐related genes (ATGs). Our results are the first to provide evidence for this delay in senescence based on the metabolic alteration and protein degradation.

Polyphenolic profiles detected in the ripe berries of Vitis vinifera germplasm

Polyphenolic profiles in the berry samples of 344 European grape (Vitis vinifera) cultivars were evaluated for two consecutive years. These cultivars represent a diverse collection of V. vinifera germplasm maintained at the USDA-Agricultural Research Service Vitis Clonal Repository in Davis of California, USA. A total of 36 polyphenolic compounds, including 16 anthocyanins, 6 flavonols, 6 flavanols, 6 hydroxycinnamic acids and 2 hydroxybenzoic acids, were identified via HPLC-MS and quantified by HPLC-DAD. The mean contents for anthocyanins, flavanols, flavonols, hydroxycinnamic acids and hydroxybenzoic acids were 0.946 (coloured cultivars), 0.147, 0.043, 0.195 and 0.016mgg(-1) FW, respectively. On average, wine grapes had higher concentrations than had table grapes for all of these compounds except hydroxycinnamic acids. Berry colours affected the total contents of anthocyanins, but not others. Positive correlations (0.151-0.535) were found among these groups of compounds. As expected, these groups of compounds were all negatively correlated with berry weight.

Phloem Loading Strategies and Water Relations in Trees and Herbaceous Plants

Most herbaceous plants employ thermodynamically active mechanisms of phloem loading, whereas in many trees, the mechanism is passive, by diffusion. Considering the different water transport characteristics of herbs and trees, we hypothesized that water relations play a role in the adoption of phloem loading strategies. We measured whole-plant hydraulic conductance (Kp), osmolality, concentrations of polar metabolites, and key inorganic ions in recently mature leaves of 45 dicotyledonous species at midafternoon. Trees, and the few herbs that load passively, have low Kp, high osmolality, and high concentrations of transport sugars and total polar metabolites. In contrast, herbs that actively load sucrose alone have high Kp, low osmolality, and low concentrations of sugars and total polar metabolites. Solute levels are higher in sugar alcohol-transporting species, both herbs and trees, allowing them to operate at lower leaf water potentials. Polar metabolites are largely responsible for leaf osmolality above a baseline level (approximately 300 mm) contributed by ions. The results suggest that trees must offset low Kp with high concentrations of foliar transport sugars, providing the motivating force for sugar diffusion and rendering active phloem loading unnecessary. In contrast, the high Kp of most herbaceous plants allows them to lower sugar concentrations in leaves. This reduces inventory costs and significantly increases growth potential but necessitates active phloem loading. Viewed from this perspective, the elevation of hydraulic conductance marks a major milestone in the evolution of the herbaceous habit, not only by facilitating water transport but also by maximizing carbon use efficiency and growth.

Photosystem 2 is more tolerant to high temperature in apple (Malus domestica Borkh.) leaves than in fruit peel

Tolerance of photosystem 2 (PS2) to high temperature in apple (Malus domestica Borkh. cv. Cortland) leaves and peel was investigated by chlorophyll a fluorescence (OJIP) transient after exposure to 25 (control), 40, 42, 44, and 46 °C in the dark for 30 min. The positive L-step was more pronounced in a peel than in leaves when exposed to 44 °C. Heat-induced K-step became less pronounced in leaves than in peel when exposed to 42 °C or higher temperature. Leaves had negative L-and K-steps relative to the peel. The decrease of oxygen-evolving complex (OEC) by heat stress was higher in the peel than in the leaves. OJIP transient from the 46 °C treated peel could not reach the maximum fluorescence (Fm). The striking thermoeffect was the big decrease in the relative variable fluorescence at 30 ms (VI), especially in the leaves. Compared with the peel, the leaves had less decreased maximum PS2 quantum efficiency (Fv/Fm), photochemical rate constant (KP), Fm and performance index (PI) on absorption basis (PIabs) and less increased minimum fluorescence (F0) and non-photochemical rate constant (KN), but more increased reduction of end acceptors at PS1 electron acceptor side per cross section (RE0/CS0) and per reaction center (RE0/RC0), quantum yield of electron transport from QA− to the end acceptors (ϕ R0) and total PI (PIabs,total) when exposed to 44 °C. In conclusion, PS2 is more thermally labile than PS1. The reduction of PS2 activity by heat stress primarily results from an inactivation of OEC. PS2 was more tolerant to high temperature in the leaves than in the peel.

Heterogeneous behavior of PSII in soybean (Glycine max) leaves with identical PSII photochemistry efficiency under different high temperature treatments

The purpose of this study is to demonstrate the heterogeneous behavior of PSII in soybean (Glycine max) leaves and identical maximum PSII photochemistry efficiency (F(V)/F(M)) under different high temperature treatments. We observed that, with an identical decrease in F(V)/F(M) in soybean leaves caused by different high temperature treatments, chlorophyll a fluorescence differed significantly, indicating different behaviors in the photosynthetic apparatus. The quantitative analysis showed that, with an identical F(V)/F(M), leaves treated at 48 degrees C showed a higher W(K), an indicator of damage to the oxygen-evolving complex along with a lower O(2) evolution rate compared with leaves treated at 45 degrees C. This demonstrated that the donor side of PSII was damaged more severely at 48 degrees C than at 45 degrees C despite the same decrease in F(V)/F(M) in the two heat-treated leaves. The ratios of Q(A)- and Q(B)-reducing PSII reaction centers to total PSII reaction centers were both lower in leaves treated at 48 degrees C than in leaves treated at 45 degrees C with an identical F(V)/F(M), indicating that the acceptor side of PSII was also more damaged by heat treatment at 48 degrees C than at 45 degrees C. However, when damage to the donor side of PSII was similar in leaves treated at two different temperatures, the acceptor side of PSII was damaged less severely at 48 degrees C, which accounted for higher electron transport rate at the acceptor side of PSII in leaves treated at 48 degrees C than in leaves treated at 45 degrees C.

Phloem Loading Strategies in Three Plant Species That Transport Sugar Alcohols

Many plants translocate sugar alcohols in the phloem. However, the mechanism(s) of sugar alcohol loading in the minor veins of leaves are debated. We characterized the loading strategies of two species that transport sorbitol (Plantago major and apple [Malus domestica]), and one that transports mannitol (Asarina scandens). Plasmodesmata are abundant at all interfaces in the minor vein phloem of apple, and in one of two types of phloem in the minor veins of A. scandens. Few plasmodesmata are present in the minor veins of P. major. Apple differs from the other two species in that sugar alcohol and sucrose (Suc) are present in much higher concentrations in leaves. Apple leaf tissue exposed to exogenous [14C]sorbitol, [14C]Suc, or 14CO2 did not accumulate radiolabel in the minor veins, as determined by macroautoradiography. P. major minor veins accumulated radiolabel from [14C]Suc, [14C]sorbitol, and 14CO2. A. scandens minor veins accumulated 14C from [14C]Suc and 14CO2, but not from [14C]mannitol. We conclude that the movement of sugar alcohol from the mesophyll into the phloem in apple and A. scandens is symplastic and passive, but in P. major it involves an apoplastic step and is energized. We also suggest that apple leaves transport sorbitol in high concentrations to avoid the feedback limitation of photosynthesis that would result from driving passive movement of solute into the phloem with high levels of Suc alone. The loading pathways and the mechanisms by which hydrostatic pressure is maintained in the minor vein phloem of these species are discussed.

MdMYB1 Regulates Anthocyanin and Malate Accumulation by Directly Facilitating Their Transport into Vacuoles in Apples.

An MYB transcription factor influences organ coloration and acidity by activating the expression of the genes encoding vacuolar proton pump subunits as well as anthocyanin transporters and malate transporters. Tonoplast transporters, including proton pumps and secondary transporters, are essential for plant cell function and for quality formation of fleshy fruits and ornamentals. Vacuolar transport of anthocyanins, malate, and other metabolites is directly or indirectly dependent on the H+-pumping activities of vacuolar H+-ATPase (VHA) and/or vacuolar H+-pyrophosphatase, but how these proton pumps are regulated in modulating vacuolar transport is largely unknown. Here, we report a transcription factor, MdMYB1, in apples that binds to the promoters of two genes encoding the B subunits of VHA, MdVHA-B1 and MdVHA-B2, to transcriptionally activate its expression, thereby enhancing VHA activity. A series of transgenic analyses in apples demonstrates that MdMYB1/10 controls cell pH and anthocyanin accumulation partially by regulating MdVHA-B1 and MdVHA-B2. Furthermore, several other direct target genes of MdMYB10 are identified, including MdVHA-E2, MdVHP1, MdMATE-LIKE1, and MdtDT, which are involved in H+-pumping or in the transport of anthocyanins and malates into vacuoles. Finally, we show that the mechanism by which MYB controls malate and anthocyanin accumulation in apples also operates in Arabidopsis (Arabidopsis thaliana). These findings provide novel insights into how MYB transcription factors directly modulate the vacuolar transport system in addition to anthocyanin biosynthesis, consequently controlling organ coloration and cell pH in plants.

Purification and characterization of sorbitol-6-phosphate phosphatase from apple leaves

Abstract Sorbitol-6-phosphate phosphatase (SorPP; EC 3.1.3.50) catalyzes the final step in sorbitol biosynthesis in sorbitol-synthesizing plant species, but its kinetic and regulatory properties have not been characterized. In this study, the enzyme was purified 1727-fold to apparent homogeneity from apple leaves with a maximal specific activity of 89.8 μmol min−1 mg−1 protein measured at 2 mM sorbitol-6-phosphate (sorbitol-6-P). The enzyme is a monomer with a molecular mass of 61 kDa. The enzyme is highly specific for sorbitol-6-P with a Km of 0.85 mM and is unable to cleave other phosphate esters at a significant rate. The activity is absolutely dependent on Mg2+ with a Km of 0.29 mM at an optimal pH of 6.8. Fluoride, vanadate, molybdate, and inorganic phosphate inhibit SorPP activity. Sorbitol is a competitive inhibitor for SorPP with a Ki of 109 mM. The possible feedback mechanism for the regulation of sorbitol biosynthesis is also discussed.