Proteomics and SSH Analyses of ALA-Promoted Fruit Coloration and Evidence for the Involvement of a MADS-Box Gene, MdMADS1

Skin color is a key quality attribute of fruits and how to improve fruit coloration has long been a major concern. 5-Aminolevulinic acid (ALA), a natural plant growth regulator, can significantly increase anthocyanin accumulation in fruit skin and therefore effectively improve coloration of many fruits, including apple. However, the molecular mechanism how ALA stimulates anthocyanin accumulation in fruit skin remains unknown. Here, we investigated the impact of ALA on apple skin at the protein and mRNA levels. A total of 85 differentially expressed proteins in apple skins between ALA and water treatment (control) were identified by complementary gel-based and gel-freeseparation techniques. Most of these differentially expressed proteins were up-regulated by ALA. Function analysis suggested that 87.06% of the ALA-responsive proteins were associated with fruit ripening. To further screen ALA-responsive regulators, we constructed a subtracted cDNA library (tester: ALA treatment; driver: control) and obtained 104 differentially expressed unigenes, of which 38 unigenes were indicators for the fruit ripening-related gene. The differentially changed proteins and transcripts did not correspond well at an individual level, but showed similar regulated direction in function at the pathway level. Among the identified fruit ripening-related genes, the expression of MdMADS1, a developmental transcription regulator of fruit ripening, was positively correlated with expression of anthocyanin biosynthetic genes (MdCHS, MdDFR, MdLDOX and MdUFGT) in apple skin under ALA treatment. Moreover, overexpression of MdMADS1 enhanced anthocyanin content in transformed apple calli, which was further enhanced by ALA. The anthocyanin content in MdMADS1-silenced calli was less than that in the control with ALA treatment, but higher than that without ALA treatment. These results indicated that MdMADS1 is involved in ALA-induced anthocyanin accumulation. In addition, anthocyanin-related verification in apple calli suggested that the regulation of MdMADS1 on anthocyanin biosynthesis was partially independent of fruit ripening process. Taken together, our findings provide insight into the mechanism how ALA regulates anthocyanin accumulation and add new information on transcriptase regulators of fruit coloration

The effects of short-term rainfall variability on leaf isotopic traits of desert plants in sand-binding ecosystems

Author's manuscript made available in accordance with the publisher's policy.Sand-binding vegetation is effective in stabilizing sand dunes and reducing soil erosion, thus helps minimize the detrimental effects of desertification. The aim of this study is to better understand the relationships between water and nutrient usage of sand-binding species, and the effects of succession and rainfall variability on plants’ water–nutrient interactions. We examined the effects of long-term succession (50 years), inter-annual rainfall variability (from 65% of the mean annual precipitation in 2004 to 42% in 2005) and seasonality on water–nutrient interactions of three major sand-binding species (Artemisia ordosica, Hedysarum scoparium and Caragana korshinskii) by measuring foliar δ13C, δ15N and [N]. Long-term succession in general did not significantly alter δ13C, δ15N and [N] of the three species. Short-term rainfall variability, however, significantly increased foliar δ13C levels of all three species by 1.0–1.8‰ during the severely dry year. No significant seasonal patterns were found in foliar δ13C and δ15N values of the three species, whereas foliar [N] varied by season. For the two leguminous shrubs, the correlations between δ13C and δ15N were positive in both sampling years, and the positive correlation between [N] and δ13C was only found in the severely dry year. The results indicate that these sand-binding plants have developed into a relatively stable stage and they are able to regulate their nitrogen and water use in responding to environmental conditions, which reinforces the effectiveness of plantation of native shrubs without irrigation in degraded areas. However, the results also indicate that short-term climate variability could have severe impact on the vegetation functions

Contribution of recycled moisture to local precipitation in the inland Heihe River Basin

Recycled moisture contributed by continental evaporation and transpiration plays an important role in regulating the hydrological processes and atmospheric humidity budget in arid inland river basins. However, knowledge of moisture recycling within many large inland basins and the factors that control moisture recycling is generally lacking. Based on a three-component isotopic mixing model, we assessed the characteristics of moisture recycling in China’s semi-arid Heihe River Basin. During the active growing season, almost half of the precipitation in the upper reaches was provided by local moisture recycling, and the main contribution came from transpiration. In the middle reaches, almost half of the precipitation in the artificial oasis and the desert-oasis ecotone was also provided by local moisture recycling, and the transpiration fraction (fTr) and evaporation fraction (fEv) of the artificial oasis differed from those of the desert-oasis ecotone. In the lower reaches, less than 25% of the precipitation was provided by local moisture recycling. Mean fTr values were relatively low in the Gobi (15.0%) in the middle reaches and in the riparian forest at Ejina (25.6%) in the lower reaches. The positive correlations between fTr and both precipitation and relative humidity suggest that higher precipitation and relative humidity promote transpiration fraction, whereas higher vapor pressure deficit reduces transpiration fraction. The positive correlation between fEv and temperature and vapor pressure deficit, and the negative correlation between fEv and relative humidity indicate that higher temperature and vapor pressure deficit promotes evaporation fraction, whereas higher relative humidity reduces the evaporation fraction. Our results show that contributions of recycled moisture (especially transpiration) to local precipitation play an important role in regional water resource redistribution in the arid and semi-arid region of northwestern China

Tree ring δ18O reveals no long-term change of atmospheric water demand since 1800 in the northern Great Hinggan Mountains, China

Global warming will significantly increase transpirational water demand, which could dramatically affect plant physiology and carbon and water budgets. Tree ring δ18O is a potential index of the leaf-to-air vapor-pressure deficit (VPD) and therefore has great potential for long-term climatic reconstruction. Here we developed δ18O chronologies of two dominant native trees, Dahurian larch (Larix gmelinii Rupr.) and Mongolian pine (Pinus sylvestris var. mongolica), from a permafrost region in the Great Hinggan Mountains of northeastern China. We found that the July–August VPD and relative humidity were the dominant factors that controlled tree ring δ18O in the study region, indicating strong regulation of stomatal conductance. Based on the larch and pine tree ring δ18O chronologies, we developed a reliable summer (July–August) VPD reconstruction since 1800. Warming growing season temperatures increase transpiration and enrich cellulose 18O, but precipitation seemed to be the most important influence on VPD changes in this cold region. Periods with stronger transpirational demand occurred around the 1850s, from 1914 to 1925, and from 2005 to 2010. However, we found no overall long-term increasing or decreasing trends for VPD since 1800, suggesting that despite the increasing temperatures and thawing permafrost throughout the region, forest transpirational demand has not increased significantly during the past two centuries. Under current climatic conditions, VPD did not limit growth of larch and pine, even during extremely drought years. Our findings will support more realistic evaluations and reliable predictions of the potential influences of ongoing climatic change on carbon and water cycles and on forest dynamics in permafrost regions

Near-net forming complex shaped Zr-based bulk metallic glasses by high pressure die casting

Forming complex geometries using the casting process is a big challenge for bulk metallic glasses (BMGs), because of a lack of time of the window for shaping under the required high cooling rate. In this work, we open an approach named the “entire process vacuum high pressure die casting” (EPV-HPDC), which delivers the ability to fill die with molten metal in milliseconds, and create solidification under high pressure. Based on this process, various Zr-based BMGs were prepared by using industrial grade raw material. The results indicate that the EPV-HPDC process is feasible to produce a glassy structure for most Zr-based BMGs, with a size of 3 mm × 10 mm and with a high strength. In addition, it has been found that EPV-HPDC process allows complex industrial BMG parts, some of which are hard to be formed by any other metal processes, to be net shaped precisely. The BMG components prepared by the EVP-HPDC process possess the advantages of dimensional accuracy, efficiency, and cost compared with the ones formed by other methods. The EVP-HPDC process paves the way for the large-scale application of BMGs

Exposure to low-level metalaxyl impacts the cardiac development and function of zebrafish embryos.

Metalaxyl is an anilide pesticide that is widely used to control plant diseases caused by Peronosporales species. In order to study the toxic effects, zebrafish embryos were exposed to metalaxyl at nominal concentrations of 5, 50 and 500 ng/L for 72 hr, and the cardiac development and functioning of larvae were observed. The results showed that metalaxyl exposure resulted in increased rates of pericardial edema, heart hemorrhage and cardiac malformation. The distance between the sinus venosus and bulbus arteriosus, stroke volume, cardiac output and heart rate were significantly increased in larvae exposed to 50 and 500 ng/L metalaxyl compared to solvent control larvae. Significant upregulation in the transcription of tbx5, gata4 and myh6 was observed in the 50 and 500 ng/L treatments, and that of nkx2.5 and myl7 was observed in the 5, 50 and 500 ng/L groups. These disturbances may be related to cardiac developmental and functional defects in the larvae. The activity of Na+/K+-ATPase and Ca2+-ATPase was significantly increased in zebrafish embryos exposed to 500 ng/L metalaxyl, and the mRNA levels of genes related to ATPase (atp2a11, atp1b2b, and atp1a3b) (in the 50 and 500 ng/L groups) and calcium channels (cacna1ab) (in the 500 ng/L group) were significantly downregulated; these changes might be associated with heart arrhythmia and functional failure

Causes and consequences of pronounced variation in the isotope composition of plant xylem water

Stable isotopologues of water are widely used to derive relative root water uptake (RWU) profiles and average RWU depth in lignified plants. Uniform isotope composition of plant xylem water (delta(xyl)) along the stem length of woody plants is a central assumption of the isotope tracing approach which has never been properly evaluated.Here we evaluate whether strong variation in delta(xyl) within woody plants exists using empirical field observations from French Guiana, northwestern China, and Germany. In addition, supported by a mechanistic plant hydraulic model, we test hypotheses on how variation in delta(xyl) can develop through the effects of diurnal variation in RWU, sap flux density, diffusion, and various other soil and plant parameters on the delta(xyl) of woody plants.The hydrogen and oxygen isotope composition of plant xylem water shows strong temporal (i.e., sub-daily) and spatial (i.e., along the stem) variation ranging up to 25.2 parts per thousand and 6.8 parts per thousand for delta H-2 and delta O-18, respectively, greatly exceeding the measurement error range in all evaluated datasets. Model explorations predict that significant delta(xyl) variation could arise from diurnal RWU fluctuations and vertical soil water heterogeneity. Moreover, significant differences in delta(xyl) emerge between individuals that differ only in sap flux densities or are monitored at different times or heights.This work shows a complex pattern of delta(xyl) transport in the soil-root-xylem system which can be related to the dynamics of RWU by plants. These dynamics complicate the assessment of RWU when using stable water isotopologues but also open new opportunities to study drought responses to environmental drivers. We propose including the monitoring of sap flow and soil matric potential for more robust estimates of average RWU depth and expansion of attainable insights in plant drought strategies and responses

Nitrogen rather than streamflow regulates the growth of riparian trees

In arid and semiarid regions, riparian forests are crucial for maintaining ecological biodiversity and sustainability, and supporting social and economic development. For the typical arid and semiarid ecosystem, streamflow variability is thought to be the dominant factor influencing the vulnerability and evolution of the riparian forests, which often leads to the neglect of other potentially important factors such as nutrient availability and transport. Here, we measured annual stable nitrogen isotopes (δ15N) and nitrogen concentrations (N%) in the tree rings of Populus euphratica Oliv. (Euphrates poplar) over a 90 year period (1920–2012), collected from the lower researches of the inland Heihe River, northwestern China. Coupling with our previous dual-isotope (δ13C and δ18O) chronologies and estimated intrinsic water-use efficiency (iWUE), we examined the linkages between tree-ring δ15N and δ18O, iWUE, streamflow, and then explored the contributions of each to tree growth during the study period. Our results show that after 1975, a statistically significant correlation between tree-ring δ15N and river streamflow appears, indicating the river as a potential carrier of nitrogen from the upper and middle reaches to the lower research trees. In addition, the linkage between tree-ring δ15N and iWUE suggests substantial influence of carbon and nitrogen together on photosynthesis and transpiration of trees, although this connection become decoupled since AD 1986. The commonality analysis revealed that the nitrogen impacts indicated by tree-ring δ15N on tree growth cannot be ignored when evaluating riparian forest development. The fertilization effects caused by rising CO2 concentration complicate the nitrogen constraints on tree growth during the later part of the past century. Our results have potentially broad implications for identifying the limited factors for dryland forest ecosystems that are susceptible to natural water resource variations and human activities

Do 2H and 18O in leaf water reflect environmental drivers differently?

We compiled hydrogen and oxygen stable isotope compositions (δ H and δ O) of leaf water from multiple biomes to examine variations with environmental drivers. Leaf water δ H was more closely correlated with δ H of xylem water or atmospheric vapour, whereas leaf water δ O was more closely correlated with air relative humidity. This resulted from the larger proportional range for δ H of meteoric waters relative to the extent of leaf water evaporative enrichment compared with δ O. We next expressed leaf water as isotopic enrichment above xylem water (Δ H and Δ O) to remove the impact of xylem water isotopic variation. For Δ H, leaf water still correlated with atmospheric vapour, whereas Δ O showed no such correlation. This was explained by covariance between air relative humidity and the Δ O of atmospheric vapour. This is consistent with a previously observed diurnal correlation between air relative humidity and the deuterium excess of atmospheric vapour across a range of ecosystems. We conclude that H and O in leaf water do indeed reflect the balance of environmental drivers differently; our results have implications for understanding isotopic effects associated with water cycling in terrestrial ecosystems and for inferring environmental change from isotopic biomarkers that act as proxies for leaf water

Study on the Mechanism of Exogenous 5-Aminolevulinic Acid (ALA) in Regulating the Photosynthetic Efficiency of Pear Leaves

To provide a theoretical basis for the application of ALA in pear production, the effects of exogenous 5-aminolevulinic acid (ALA) treatment on leaf photosynthetic gas exchange parameters, chlorophyll fast fluorescence properties, and relative expression of the related genes were investigated using pear (Pyrus pyrifolia Nakai cv. ‘Whasan’) as a material in the study. The results show that exogenous ALA treatment improved the photosynthetic gas exchange parameters of pear leaves, upregulated the expression of multiple key genes which are related to ALA biosynthesis, metabolism, and transformation into chlorophylls. GUS staining in tobacco leaves showed that exogenous ALA activated the promoter activity of PypHEMA and PypCHLH genes, implying that the synthesis of endogenous ALA and chlorophylls was promoted by exogenous ALA. Furthermore, ALA promoted the expression of the genes encoding photosystem II (PSII) reaction center proteins, such as core protein D1, inner light-harvesting pigment proteins CP43 and CP47, and cytochrome b559. This led to increased PSII reaction center activity. In addition, ALA alleviated the donor side oxygen-evolving complex inhibition and reduced the closure rate on the receptor side, allowing for increased photochemical electron transfer and reduced heat dissipation while improving the photosynthetic performance index PIabs and PItotal. The findings of this study contribute to a better understanding of ALA’s promotion of plant photosynthetic efficiency, providing valuable insights for further research and potential applications in pear production