Magnesium alleviates growth inhibition under low potassium by enhancing photosynthesis and carbon-nitrogen metabolism in apple plants.

Potassium (K) and magnesium (Mg) play analogous roles in regulating plant photosynthesis and carbon and nitrogen (C-N) metabolism. Based on this consensus, we hypothesize that appropriate Mg supplementation may alleviate growth inhibition under low K stress. We monitored morphological, physiological, and molecular changes in G935 apple plants under different K (0.1 and 6 mmol L-1) and Mg supply (3 and 6 mmol L-1) conditions. Low K stress caused changes in root and leaf structure, inhibited photosynthesis, and limited the root growth of the apple rootstock. Further study on Mg supplementation showed that it could promote the uptake of K+ and NO3- by upregulating the expression of K+ transporter proteins such as Arabidopsis K+ transporter 1 (MdAKT1), high-affinity K+ transporter 1 (MdHKT1), and potassium transporter 5 (MdPT5) and nitrate transporters such as nitrate transporter 1.1/1.2/2.1/2.4 (MdNRT 1.1/1.2/2.1/2.4). Mg promoted the translocation of 15N from roots to leaves and enhanced photosynthetic N utilization efficiency (PNUE) by increasing the proportion of photosynthetic N and alleviating photosynthetic restrictions. Furthermore, Mg supplementation improved the synthesis of photosynthates by enhancing the activities of sugar-metabolizing enzymes (Rubisco, SS, SPS, S6PDH). Mg also facilitated the transport of sucrose and sorbitol from leaves to roots by upregulating the expression of sucrose transporter 1.1/1.2/4.1/4.2 (MdSUT 1.1/1.2/4.1/4.2) and sorbitol transporter 1.1/1.2 (MdSOT 1.1/1.2). Overall, Mg effectively alleviated growth inhibition in apple rootstock plants under low K stress by facilitating the uptake of N and K uptake, optimizing nitrogen partitioning, enhancing nitrogen use efficiency (NUE) and PNUE, and promoting the photosynthate synthesis and translocation.

Nitrogen-potassium balance improves leaf photosynthetic capacity by regulating leaf nitrogen allocation in apple.

Nitrogen (N) and potassium (K) are two important mineral nutrients in regulating leaf photosynthesis. However, the influence of N and K interaction on photosynthesis is still not fully understood. Using a hydroponics approach, we studied the effects of different N and K conditions on the physiological characteristics, N allocation and photosynthetic capacity of apple rootstock M9T337. The results showed that high N and low K conditions significantly reduced K content in roots and leaves, resulting in N/K imbalance, and allocated more N in leaves to non-photosynthetic N. Low K conditions increased biochemical limitation (BL), mesophyll limitation (MCL), and stomatal limitation (SL). By setting different N supplies, lowering N levels under low K conditions increased the proportion of water-soluble protein N (Nw) and sodium dodecyl sulfate-soluble proteins (Ns) by balancing N/K and increased the proportion of carboxylation N and electron transfer N. This increased the maximum carboxylation rate and mesophyll conductance, which reduced MCL and BL and alleviated the low K limitation of photosynthesis in apple rootstocks. In general, our results provide new insights into the regulation of photosynthetic capacity by N/K balance, which is conducive to the coordinated supply of N and K nutrients.

MdBT2 regulates nitrogen-mediated cuticular wax biosynthesis via a MdMYB106-MdCER2L1 signalling pathway in apple.

Cuticular waxes play important roles in plant development and the interaction between plants and their environment. Researches on wax biosynthetic pathways have been reported in several plant species. Also, wax formation is closely related to environmental condition. However, the regulatory mechanism between wax and environmental factors, especially essential mineral elements, is less studied. Here we found that nitrogen (N) played a negative role in the regulation of wax synthesis in apple. We therefore analysed wax content, composition and crystals in BTB-TAZ domain protein 2 (MdBT2) overexpressing and antisense transgenic apple seedlings and found that MdBT2 could downregulate wax biosynthesis. Furthermore, R2R3-MYB transcription factor 16-like protein (MdMYB106) interacted with MdBT2, and MdBT2 mediated its ubiquitination and degradation through the 26S proteasome pathway. Finally, HXXXD-type acyl-transferase ECERIFERUM 2-like1 (MdCER2L1) was confirmed as a downstream target gene of MdMYB106. Our findings reveal an N-mediated apple wax biosynthesis pathway and lay a foundation for further study of the environmental factors associated with wax regulatory networks in apple.

Metabolomics combined with physiology and transcriptomics reveal key metabolic pathway responses in apple plants exposure to different selenium concentrations.

Selenium (Se) can be absorbed by plants, thereby affects plant physiological activity, interferes gene expression, alters metabolite content and influences plant growth. However, the molecular mechanism underlying the plant response to Se remains unclear. In this study, apple plants were exposed to Se at concentrations of 0, 3, 6, 9, 12, 24, and 48 µM. Low concentrations of Se promoted plant growth, while high Se concentrations (≥24 µM) reduced photosynthesis, disturbed carbon and nitrogen metabolism, damaged the antioxidant system, and ultimately inhibited plant growth. The transcriptome and metabolome revealed that Se mainly affected three pathways, namely the 'biosynthesis of amino acids', 'starch and sucrose metabolism', and 'phenylpropanoid biosynthesis' pathways. 9 µM Se improved the synthesis, catabolism and utilization of amino acids and sugars, ultimately promoted plant growth. However, 24 µM Se up-regulated the related genes expression of PK, GPT, P5CS, SUS, SPS and CYP98A, and accumulated a large number of osmoregulation substances, such as citric acid, L-proline, D-sucrose and chlorogenic acid in the roots, ultimately affected the balance between plant growth and defense. In conclusion, this study reveals new insights into the key metabolic pathway in apple plants responses to Se.

Integrated soil-fruit-climate management system to improve apple production stability.

Unstable agricultural systems, such as unreliable fruit production, threaten global food security and could negatively impact human nutrition. However, methods of maintaining a stable fruit supply have received little attention. Therefore, this study explored the impact of a systematic service model on the establishment of an integrated soil-fruit-climate management (ISM) approach and stabilization of apple yield based on a fixed-effects model using sample data from Fuji apple farmers in China for six consecutive years (2016-2021). The results showed that the systematic service model dramatically improved the adoption of ISM-based technology. By 2021, the rate of ISM technology adoption among farmers reached 84.4 %, especially pendulous branch-fruiting technology and Osmia pollination, which increased by 82.2 % and 37.8 %, respectively, compared to that in 2016. With the adoption of the ISM approach, apple yields and tree loadings stabilized over time. For example, the yield change rate and tree loading change rate were 14.8 % and 7.3 % lower, respectively, in 2021 than in 2016. We demonstrated through a fixed-effects analysis that the ISM approach exerts a fully mediated effect on the mechanism of action of service measure quantity disclosure, which positively impacted the rate of change in yield and tree loadings. This finding indicated that the six-in-one systematic service model of soil testing, pest and disease diagnosis, course training, meteorological monitoring, on-site guidance, and experimental demonstration established by the Science and Technology Backyard model resolved the information, goal, and hardware gaps that limited the adoption of this technology by farmers and promoted the construction of an integrated soil-fruit-climate management approach, which in turn has stabilized the apple production system.

Magnesium improved fruit quality by regulating photosynthetic nitrogen use efficiency, carbon-nitrogen metabolism, and anthocyanin biosynthesis in 'Red Fuji' apple.

Introduction: Both nitrogen (N) and magnesium (Mg) play important roles in biochemical and physiological processes in plants. However, the application of excessive N and insufficient Mg may be the factor leading to low nitrogen utilization rate (NUE) and fruit quality degradation in apple production. Methods: In this study, we analyzed the effects of different application rates of Mg (0, 50, 100, 150, 200 kg/ha) on the photosynthetic nitrogen use efficiency (PNUE), the accumulation and distribution of carbon (C), N metabolism, anthocyanin biosynthesis and fruit quality of the 'Red Fuji' apple in 2018 and 2019. Results: The results showed that the application of Mg significantly increased the 15NUE and increased the allocation rate of 15N in the leaves whereas the 15N allocation rate in the perennial organs and fruits was decreased. With the increase in Mg supply, the activities of N metabolism enzymes (NiR, GS, and GOGAT) were significantly promoted and the content of intermediate products in N metabolism ( NO 2 - , NH 4 + , and free amino acid) was significantly decreased. Furthermore, an appropriate rate of Mg significantly promoted the net photosynthetic rate (Pn) and photosynthetic nitrogen use efficiency (PNUE), enhanced the enzyme activities of C metabolism (SS, SPS, S6PDH), and increased the contents of sorbitol and sucrose in leaves. In addition, Mg upregulated the gene expression of sugar transporters (MdSOT1, MdSOT3, MdSUT1, and MdSUT4) in fruit stalk and fruit fresh; 13C isotope tracer technology also showed that Mg significantly increased the 13C allocation in the fruits. Mg also significantly increased the expression of anthocyanin biosynthesis genes (MdCHS and MdF3H) and transcription factors (MdMYB1 and MdbZIP44) and the content of anthocyanin in apple peel. Conclusion: The comprehensive analysis showed that the appropriate application of Mg (150 kg/ha) promoted PNUE, C-N metabolism, and anthocyanin biosynthesis in apple trees.

Effects of Magnesium on nitrate uptake and sorbitol synthesis and translocation in apple seedlings.

Both magnesium (Mg) and nitrogen (N) play many important roles in plant physiological and biochemical processes. Plants usually exhibit low nitrogen utilization efficiency (NUE) under Mg deficiency conditions, but the mechanisms by which Mg regulates NUE are not well understood. Herein, we investigated biomass, nutrient uptake, sorbitol and sucrose transport, and relative gene expression in apple seedlings under various concentrations of Mg and N treatments in hydroponic cultures. We first observed that low Mg supply significantly limited plant growth and N, Mg concentrations. Increasing the supply of N, but not Mg, partially alleviated the inhibition of plant growth under low Mg stress, which indicated that Mg deficiency had a negative impact on plant growth because it inhibits N absorption. Moreover, we found that the expression of nitrate transporter genes MdNRT2.1 and MdNRT2.4 was significantly downregulated by low Mg stress, and sufficient Mg significantly promoted sucrose and sorbitol synthesis and transport from leaves to roots by regulating relevant enzyme activity and genes expression. Further experiments showed that exogenous sorbitol could rapidly restore MdNRT2.1/2.4 expression and nitrate uptake under low Mg availability without increasing internal Mg level, suggesting that Mg may regulate MdNRT2.1/2.4 expression by regulating more sorbitol transport to roots, the effect of Mg on N was indirect, sorbitol played a key role during this process. Taken together, Mg promoted sorbitol synthesis and transport into roots, thus upregulating the expression of MdNRT2.1/2.4 and increasing the absorption of nitrate.

Magnesium alleviates aluminum-induced growth inhibition by enhancing antioxidant enzyme activity and carbon-nitrogen metabolism in apple seedlings.

Previous studies have determined that magnesium (Mg) in appropriate concentrations prevents plants from suffering from abiotic stress. To better understand the mechanism of Mg alleviation of aluminum (Al) stress in apple, we investigated the effect of Mg on plant growth, photosynthetic fluorescence, antioxidant system, and carbon (C) and nitrogen (N) metabolism of apple seedlings under Al toxicity (1.5 mmol/L) via a hydroponic experiment. Al stress induced the production of reactive oxygen in the leaves and roots and reduced the total dry weight (DW) by 52.37 % after 20 days of treatment relative to plants grown without Al, due to hindered photosynthesis and alterations in C and N metabolism. By contrast, total DW decreased by only 11.07 % in the Mg-treated plants under Al stress. Supplementation with 3.0 mmol/L Mg in the Al treatment decreased Al accumulation in the apple plants and reduced Al-induced oxidative damage by enhancing the activity of antioxidant enzymes (superoxide dismutase, catalase, and peroxidase) and reducing the production of H2O2 and malondialdehyde (MDA). Under Al stress, the Mg-treated plants showed a 46.17 % higher photosynthetic rate than the non-treated plants. Supplementation with Mg significantly increased the sucrose content by increasing sucrose synthase (SS) and sucrose-phosphate synthase (SPS) activities. Moreover, Mg facilitated the transport of 13C-carbohydrates from the leaves to roots. Regarding N metabolism, the nitrate reductase (NR), glutamine synthase (GS), and glutamate synthase (GOGAT) activities in the roots and leaves of the Mg-treated plants were significantly higher than those of the non-treated plants under Al stress. Compared with the non-treated plants under Al stress, the Mg-treated plants exhibited a significantly high level of NO3- and soluble protein content in the leaves, roots, and stems, but a low level of free amino acids. Furthermore, Mg significantly improved nitrogen accumulation and enhanced the transport of 15N from the roots to leaves. Overall, our results revealed that Mg alleviates Al-induced growth inhibition by enhancing antioxidant capacity and C-N metabolism in apple seedlings.

Transcriptome and Metabolome Analysis Reveals the Effect of Nitrogen-Potassium on Anthocyanin Biosynthesis in "Fuji" Apple.

Nitrogen (N) and potassium (K) have significant effects on apple peel color. To further understand the molecular mechanism of N-K regulation of apple color, we analyzed the apple peel under different N and K treatments using isotope labeling, transcriptomics, and metabolomics. Under high N treatments, fruit red color and anthocyanin content decreased significantly. High N decreased the 13C distribution rate and increased the Ndff values of fruits, while K increased the expression of MdSUTs and MdSOTs and promoted 13C transportation to fruits. Anthocyanin-targeted metabonomics and transcriptome analysis revealed that high N downregulated the expression of structural genes related to the anthocyanin synthesis pathway (MdPAL, Md4CL, MdF3H, MdANS, and MdUFGT) and their regulators (MdMYBs and MdbHLHs), and also decreased some metabolites contents. K alleviated this inhibition and seven anthocyanins were regulated by N-K. Our results improve the understanding of the synergistic regulation of apple fruit coloring by N-K.

Exogenous sucrose promotes the growth of apple rootstocks under high nitrate supply by modulating carbon and nitrogen metabolism.

Excessive nitrogen (N) supply often leads to an imbalance of carbon (C) and N metabolism and inhibits plant growth. Sucrose, an important source and signaling substance of C in plants, is closely linked to N metabolism. However, it is not clear whether exogenous sucrose can mitigate the inhibitory effect of high N on plant growth by regulating C and N metabolism. In this study, we investigated the effects of exogenous sucrose on the growth, N metabolism, and C assimilation in the apple rootstock M26 seedlings under normal (5 mM NO3－, NN) and high (30 mM NO3－, HN) NO3－ concentrations. Our results showed that high NO3－ supply reduced plant growth, photosynthesis, and chlorophyll fluorescence, but spraying with 1% sucrose (HN + 1% Sucrose) significantly alleviated this inhibition. Application of 1% sucrose increased sucrose and sorbitol contents as well as sucrose-phosphate synthase and sucrose synthase activities in the plants under HN treatment and promoted the distribution of 13C photoassimilation products to the root. In addition, spraying with 1% sucrose alleviated the inhibition of N metabolizing enzyme activities by high NO3－ supply, reduced NO3－ accumulation and N content, increased free amino acid content, and promoted 15N distribution to the aboveground parts. However, spraying with 1% sucrose under the NN treatment negatively affected plant photosynthesis and carbon assimilation. In conclusion, exogenous sucrose increased the C level in plants in the presence of excess N, promoted the balance of C and N metabolism, and alleviated the inhibitory effect of high N on the apple plant growth.

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The new large-grained activated humic acid fertilizer (LAF) can significantly reduce the amount of chemical fertilizer application and stable fruit yield. Understanding its impacts on soil aggregates and organic carbon is an important basis for revealing its role in driving soil structure of apple orchard. There were four LAF treatments: LAF1 (full fertilization, fertilization period and mass ratio (the same below), germination stage: fruit expansion stage: maturity stage=3:4:3), LAF2 (full fertilization, germination stage: fruit expansion stage: maturity stage=2:3:5), LAF3 (fertilizer application reduction by 1/4, germination stage: fruit expansion stage: maturity stage=2:3:5), LAF4 (fertilizer application reduction by 1/3, germination stage: fruit expansion stage: maturity stage=2:3:5); with no fertilization as control (CK). In a four-year pot experiment, we examined the composition, stabi-lity and organic carbon content of soil aggregates under different fertilization treatments. The results showed that: 1) compared with CK, each treatment of LAF increased the content of >2 mm and 2-0.25 mm aggregate by 53.4%-77.5% and 12.3%-17.0%, respectively. The application of LAF significantly increased the content of soil water stable aggregates, and such effects were positively related with application amount. The content of soil water stable aggregate was the highest in the LAF1 treatment. 2) There was no significant difference in aggregate content of each particle size among LAF treatments, with the proportion of aggregate content of 2-0.25 mm particle size being the highest. 3) Compared with CK, all LAF treatments significantly increased the average weight diameter (MWD) and geometric mean diameter (GMD), and reduced the fractal dimension (D). LAF1 treatment had the highest MWD and GMD values, and had the strongest effect on the stability of soil aggregates. 4) Except for LAF4 treatment, the content of soil organic carbon in other LAF treatments was significantly higher than that in CK, and the content of soil organic carbon in LAF2 treatment was the highest. All LAF treatments increased the organic carbon content of soil aggregates with each particle size. LAF1, LAF2, and LAF3 treatments significantly increased the organic carbon of aggregates with particle size >2 mm. Particle size >2 mm had the highest contribution to the total organic carbon. The contribution rate of water stable large aggregate organic carbon to total organic carbon of LAF treatment was significantly higher than that of CK, which was all higher than 66.0%, and that of LAF1 treatment was the highest. In conclusion, the application of LAF enhanced the formation and stability of water stable aggregates and increased organic carbon content of aggregates in apple orchard soil, with the best performance of the full application. The application of LAF could be used as an effective measure to improve soil structure and fertility in apple orchard.

Inversion of Nitrogen Concentration in Apple Canopy Based on UAV Hyperspectral Images.

As the major nutrient affecting crop growth, accurate assessing of nitrogen (N) is crucial to precise agricultural management. Although improvements based on ground and satellite data nitrogen in monitoring crops have been made, the application of these technologies is limited by expensive costs, covering small spatial scales and low spatiotemporal resolution. This study strived to explore an effective approach for inversing and mapping the distributions of the canopy nitrogen concentration (CNC) based on Unmanned Aerial Vehicle (UAV) hyperspectral image data in a typical apple orchard area of China. A Cubert UHD185 imaging spectrometer mounted on a UAV was used to obtain the hyperspectral images of the apple canopy. The range of the apple canopy was determined by the threshold method to eliminate the effect of the background spectrum from bare soil and shadow. We analyzed and screened out the spectral parameters sensitive to CNC, including vegetation indices (VIs), random two-band spectral indices, and red-edge parameters. The partial least squares regression (PLSR) and backpropagation neural network (BPNN) were constructed to inverse CNC based on a single spectral parameter or a combination of multiple spectral parameters. The results show that when the thresholds of normalized difference vegetation index (NDVI) and normalized difference canopy shadow index (NDCSI) were set to 0.65 and 0.45, respectively, the canopy's CNC range could be effectively identified and extracted, which was more refined than random forest classifier (RFC); the correlation between random two-band spectral indices and nitrogen concentration was stronger than that of other spectral parameters; and the BPNN model based on the combination of random two-band spectral indices and red-edge parameters was the optimal model for accurately retrieving CNC. Its modeling determination coefficient (R2) and root mean square error (RMSE) were 0.77 and 0.16, respectively; and the validation R2 and residual predictive deviation (RPD) were 0.75 and 1.92. The findings of this study can provide a theoretical basis and technical support for the large-scale, rapid, and non-destructive monitoring of apple nutritional status.

Appropriate and Constant Potassium Supply Promotes the Growth of M9T337 Apple Rootstocks by Regulating Endogenous Hormones and Carbon and Nitrogen Metabolism.

Potassium (K) is an indispensable nutrient element in the development of fruit trees in terms of yield and quality. It is unclear how a stable or unstable supply of K affects plant growth. We studied the root morphology and physiological and molecular changes in the carbon and nitrogen metabolism of M9T337 apple rootstock under different K levels and supply methods using hydroponics. Five K supply treatments were implemented: continuous low K (KL), initial low and then high K (KLH), appropriate and constant K (KAC), initial high and then low K (KHL), and continuous high K (KH). The results showed that the biomass, root activity, photosynthesis, and carbon and nitrogen metabolism of the M9T337 rootstocks were inhibited under KL, KH, KLH and KHL conditions. The KAC treatment promoted root growth by optimizing endogenous hormone content, enhancing carbon and nitrogen metabolism enzyme activities, improving photosynthesis, optimizing the distribution of carbon and nitrogen, and upregulating the transcription levels of nitrogen assimilation-related genes (nitrate reductase, glutamine synthetase, glutamate synthase, MdNRT1.1, MdNRT1.2, MdNRT1.5, MdNRT2.4). These results suggest that an appropriate and constant K supply ensures the efficient assimilation and utilization of nitrogen and carbon.

DMPP reduces nitrogen fertilizer application rate, improves fruit quality, and reduces environmental cost of intensive apple production in China.

In China, excessive application of nitrogen (N) fertilizer is common in intensive apple production. To resolve issues of benefit reduction and environmental pollution caused by excessive N, a two-year trial was conducted in an apple orchard with a split-plot design, in which the main factor was the N level (500, 400, 300, and 200 kg N ha-1 year-1, expressed as TN, TN80%, TN60%, and TN40%, respectively) and the deputy factor was whether or not to add 3,4-dimethylpyrazole phosphate (DMPP, expressed as +D). The effects of N reduction combined with DMPP on soil N transformation, fruit quality, economic benefits, and environmental effects were investigated. The results showed that DMPP reduced the production of nitrate and its vertical migration by inhibiting the abundance of AOB amoA and decreased N2O emission by reducing nirKC1 levels. Moreover, N reduction combined with DMPP improved N use efficiency (26.67-49.35%) and reduced N loss rate (15.25-38.76%). Compared with TN, TN60% + D increased the content of anthocyanin and soluble sugar by 21.15% and 13.09%, respectively, and decreased environmental costs caused by NH3 volatilization and N2O emission by 33.84%, while maintaining yield and N utilization rate at relatively high levels. Considering the agronomic, economic and environmental benefits, on the basis of traditional N application rate, 40% N reduction combined with DMPP (TN60% + D) could ensure target yield, corresponding quality and economic benefits, maintain soil N fertility, and reduce the risk of N losses to the environment. The present research could provide references for green, efficient, and sustainable development of apple production.

Multiomics Analysis Reveals New Insights into the Apple Fruit Quality Decline under High Nitrogen Conditions.

Excessive application of nitrogen (N) fertilizer is common in Chinese apple production. High N reduced the contents of soluble sugar and total flavonoids by 16.05 and 19.01%, respectively, resulting in poor fruit quality. Moreover, high N increased the total N and decreased the total C and C/N ratio of apple fruits. On the basis of the transcriptomic, proteomic, and metabolomic analyses, the global network was revealed. High N inhibited the accumulation of carbohydrates (sucrose, glucose, and trehalose) and flavonoids (rhamnetin-3-O-rutinoside, rutin, and trihydroxyisoflavone-7-O-galactoside) in fruits, and more C skeletons were used to synthesize amino acids and their derivatives (especially low C/N ratio, e.g., arginine) to be transferred to N metabolism. This study revealed new insights into the decline in soluble sugar and flavonoids caused by high N, and hub genes (MD07G1172700, MD05G1222800, MD16G1227200, MD01G1174400, and MD02G1207200) and hub proteins (PFK, gapN, and HK) were obtained.

Inversion reflectance by apple tree canopy ground and unmanned aerial vehicle integrated remote sensing data.

To obtain accurate spatially continuous reflectance from Unmanned Aerial Vehicle (UAV) remote sensing, UAV data needs to be integrated with the data on the ground. Here, we tested accuracy of two methods to inverse reflectance, Ground-UAV-Linear Spectral Mixture Model (G-UAV-LSMM) and Minimum Noise Fraction-Pixel Purity Index-Linear Spectral Mixture Model (MNF-PPI-LSMM). At wavelengths of 550, 660, 735 and 790 nm, which were obtained by UAV multispectral observations, we calculated the canopy abundance based on the two methods to acquire the inversion reflectance. The correlation of the inversion and measured reflectance values was stronger in G-UAV-LSMM than MNF-PPI-LSMM. We conclude that G-UAV-LSMM is the better model to obtain the canopy inversion reflectance.

ITRAQ-based quantitative proteomic analysis of Fusarium moniliforme (Fusarium verticillioides) in response to Phloridzin inducers.

BACKGROUND: Apple replant disease (ARD) has been reported from all major fruit-growing regions of the world, and is often caused by biotic factors (pathogen fungi) and abiotic factors (phenolic compounds). In order to clarify the proteomic differences of Fusarium moniliforme under the action of phloridzin, and to explore the potential mechanism of F. moniliforme as the pathogen of ARD, the role of Fusarium spp. in ARD was further clarified. METHODS: In this paper, the quantitative proteomics method iTRAQ analysis technology was used to analyze the proteomic differences of F. moniliforme before and after phloridzin treatment. The differentially expressed protein was validated by qRT-PCR analysis. RESULTS: A total of 4535 proteins were detected, and 293 proteins were found with more than 1.2 times (P< 0.05) differences. In-depth data analysis revealed that 59 proteins were found with more than 1.5 times (P< 0.05) differences, and most proteins were consistent with the result of qRT-PCR. Differentially expressed proteins were influenced a variety of cellular processes, particularly metabolic processes. Among these metabolic pathways, a total of 8 significantly enriched KEGG pathways were identified with at least 2 affiliated proteins with different abundance in conidia and mycelium. Functional pathway analysis indicated that up-regulated proteins were mainly distributed in amino sugar, nucleotide sugar metabolism, glycolysis/ gluconeogenesis and phagosome pathways. CONCLUSIONS: This study is the first to perform quantitative proteomic investigation by iTRAQ labeling and LC-MS/MS to identify differentially expressed proteins in F. moniliforme under phloridzin conditions. The results confirmed that F. moniliforme presented a unique protein profile that indicated the adaptive mechanisms of this species to phloridzin environments. The results deepened our understanding of the proteome in F. moniliforme in response to phloridzin inducers and provide a basis for further exploration for improving the efficiency of the fungi as biocontrol agents to control ARD.

Studies on the translocation characteristics of 13C-photoassimilates to fruit during the fruit development stage in 'Fuji' apple.

In order to define translocation characteristics of 13C-photoassimilates to fruit during the fruit development stage in 'Fuji' apple, the 13C labeled tracer method was used in whole five-year-old 'Fuji'3/M26/Malus hupehensis (Pamp.) Rehder apple trees at different days after flowering (DAF). The changes in 13C translocation to the fruit, source strength of the leaves, and sink strength of the fruits were assessed. The results indicated that the δ13C value and 13C distribution rate of the fruit increased first and then decreased with the increase in the fruit development period, being higher from 120 to 135 DAF. The leaves appeared to moderately senesce in an attempt to maintain high photosynthesis during 120-135 DAF, which promoted the outward transport of photoassimilates. The single fruit weight and longitudinal and transverse diameter of the fruit increased rapidly during 120-150 DAF, which increased the sink zone for the unloading of photoassimilates in the fruit. The activity of sorbitol dehydrogenase (SDH) and amylase (AM), the content of indole-3-acetic acid (IAA), the gibberellin (GA3) and abscisic acid (ABA) in the fruit flesh, and the gene expression levels of MdSOT1, MdSOT2, MdSOT3, MdSUT1, and MdSUT4 in the fruit stalk tissue were higher during 120-135 DAF. At this point, the difference in the sorbitol content between the fruit stalk and fruit flesh was also at the highest level of the entire year. These factors together increased the sink activity of the fruit, thus improving the photoassimilate transport efficiency to the fruit.

Effects of Potassium Levels on Plant Growth, Accumulation and Distribution of Carbon, and Nitrate Metabolism in Apple Dwarf Rootstock Seedlings.

Nitrogen (N) is one of the most required mineral elements for plant growth, and potassium (K) plays a vital role in nitrogen metabolism, both elements being widely applied as fertilizers in agricultural production. However, the exact relationship between K and nitrogen use efficiency (NUE) remains unclear. Apple dwarf rootstock seedlings (M9T337) were used to study the impacts of different K levels on plant growth, nitrogen metabolism, and carbon (C) assimilation in water culture experiments for 2 years. The results showed that both deficiency and excess K inhibited the growth and root development of M9T337 seedlings. When the K supply concentration was 0 mM and 12 mM, the biomass of each organ, root-shoot ratio, root activity and NO3 - ion flow rate decreased significantly, net photosynthetic rate (P n) and photochemical efficiency (F v/F m) being lower. Meanwhile, seedlings treated with 6 mM K+ had higher N and C metabolizing enzyme activities and higher nitrate transporter gene expression levels (NRT1.1; NRT2.1). 13C and 15N labeling results showed that deficiency and excess K could not only reduce 15N absorption and 13C assimilation accumulation of M9T337 seedlings, but also reduced the 15N distribution ratio in leaves and 13C distribution ratio in roots. These results suggest that appropriate K supply (6 mM) was optimal as it enhanced photoassimilate transport from leaves to roots and increased NUE by influencing photosynthesis, C and N metabolizing enzyme activities, nitrate assimilation gene activities, and nitrate transport.

Nitrification Inhibitor 3,4-Dimethylpyrazole Phosphate Application During the Later Stage of Apple Fruit Expansion Regulates Soil Mineral Nitrogen and Tree Carbon-Nitrogen Nutrition, and Improves Fruit Quality.

In order to solve the problems of nitrogen (N) losses and fruit quality degradation caused by excessive N fertilizer application, different dosages of the nitrification inhibitor, 3,4-dimethylpyrazole phosphate (DMPP) (0, 0.5, 1, 2, and 4 mg kg-1 soil), were applied during the later stage of 'Red Fuji' apple (Malus domestica Borkh.) fruit expansion in 2017 and 2018. The effects of DMPP on soil N transformation, carbon (C)-N nutrition of tree, and fruit quality were investigated. Results revealed that DMPP decreased the abundance of ammonia-oxidizing bacteria (AOB) amoA gene, increased the retention of NH4 +-N, and decreased NO3 --N concentration and its vertical migration in soil. DMPP reduced 15N loss rates and increased 15N residual and recovery rates compared to the control. 13C and 15N double isotope labeling results revealed that DMPP reduced the capacity of 15N absorption and regulation in fruits, decreased 15N accumulation in fruits and whole plant, and increased the distribution of 13C from vegetative organs to fruits. DMPP increased fruit anthocyanin and soluble sugar contents, and had no significant effect on fruit yield. The comprehensive analysis revealed that the application of 1 mg DMPP kg-1 soil during the later stage of fruit expansion effectively reduced losses due to N and alleviated quality degradation caused by excessive N fertilizer application.