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Abstract Background The mechanisms by which the apple MdPYL9 gene mediates the response to drought stress remain unclear. Here, transcriptome and metabolome analyses of apple plants under drought were used to investigate the mechanisms by which MdPYL9 regulates the response to drought stress in apple. MdPYL9-overexpressed transgenic and non-transgenic apple histoculture seedlings were rooted, transplanted, and subjected to drought treatments to clarify the mechanisms underlying the responses of apples to drought stress through phenotypic observations, physiological and biochemical index measurements, and transcriptomic and metabolomic analyses. Results Under drought stress treatment, transgenic plants were less affected by drought stress than non-transgenic plants. Decreases in the net photosynthetic rate, stomatal conductance, and transpiration rate of transgenic apple plants were less pronounced in transgenic plants than in non-transgenic plants, and increases in the intercellular CO2 concentration were less pronounced in transgenic plants than in non-transgenic plants. The relative electrical conductivity and content of malondialdehyde, superoxide anion, and hydrogen peroxide were significantly lower in transgenic plants than in non-transgenic plants, and the chlorophyll content and activities of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase) were significantly higher in transgenic plants than in non-transgenic plants. The number of differentially expressed genes (DEGs) involved in the response to drought stress was lower in transgenic plants than in non-transgenic plants, and the most significant and highly annotated DEGs in the transgenic plants were involved in the flavonoid biosynthesis pathway, and the most significant and highly annotated DEGs in control plants were involved in the phytohormone signal transduction pathway. The number of differentially accumulated metabolites involved in the response to drought stress was lower in transgenic plants than in non-transgenic plants, and up-regulated metabolites were significantly enriched in apigenin-7-O-glucoside in transgenic plants and in abscisic acid in non-transgenic plants. In the flavonoid biosynthetic pathway, the expression of genes encoding chalcone synthase (CHS) and chalcone isomerase (CHI) was more significantly down-regulated in non-transgenic plants than in transgenic plants, and the expression of the gene encoding 4-coumarate-CoA ligase (4CL) was more significantly up-regulated in transgenic plants than in non-transgenic plants, which resulted in the significant up-regulation of apigenin-7-O-glucoside in transgenic plants. Conclusions The above results indicated that the over-expression of MdPYL9 increased the drought resistance of plants under drought stress by attenuating the down-regulation of the expression of genes encoding CHS and CHI and enhancing the up-regulated expression of the gene encoding 4CL, which enhanced the content of apigenin-7-O-glucoside.

Abstract Background Drought is considered the main environmental factor restricting apple production and thus the development of the apple industry. Rootstocks play an important role in enhancing the drought tolerance of apple plants. Studies of the physiology have demonstrated that ‘ZC9-3’ is a strong drought-resistant rootstock, whereas ‘Jizhen-2’ is a weak drought-resistant rootstock. However, the metabolites in these two apple rootstock varieties that respond to drought stress have not yet been characterized, and the molecular mechanisms underlying their responses to drought stress remain unclear. Results In this study, the physiological and molecular mechanisms underlying differences in the drought resistance of ‘Jizhen-2’ (drought-sensitive) and ‘ZC9-3’ (drought-resistant) apple rootstocks were explored. Under drought stress, the relative water content of the leaves was maintained at higher levels in ‘ZC9-3’ than in ‘Jizhen-2’, and the photosynthetic, antioxidant, and osmoregulatory capacities of ‘ZC9-3’ were stronger than those of ‘Jizhen-2’. Metabolome analysis revealed a total of 95 and 156 differentially accumulated metabolites in ‘Jizhen-2’ and ‘ZC9-3’ under drought stress, respectively. The up-regulated metabolites in the two cultivars were mainly amino acids and derivatives. Transcriptome analysis revealed that there were more differentially expressed genes and transcription factors in ‘ZC9-3’ than in ‘Jizhen-2’ throughout the drought treatment. Metabolomic and transcriptomic analysis revealed that amino acid biosynthesis pathways play key roles in mediating drought resistance in apple rootstocks. A total of 13 metabolites, including L-α-aminoadipate, L-homoserine, L-threonine, L-isoleucine, L-valine, L-leucine, (2S)-2-isopropylmalate, anthranilate, L-tryptophan, L-phenylalanine, L-tyrosine, L-glutamate, and L-proline, play an important role in the difference in drought resistance between ‘ZC9-3’ and ‘Jizhen-2’. In addition, 13 genes encoding O-acetylserine-(thiol)-lyase, S-adenosylmethionine synthetase, ketol-acid isomeroreductase, dihydroxyacid dehydratase, isopropylmalate isomerase, branched-chain aminotransferase, pyruvate kinase, 3-dehydroquinate dehydratase/shikimate 5-dehydrogenase, N-acetylglutamate-5-P-reductase, and pyrroline-5-carboxylate synthetase positively regulate the response of ‘ZC9-3’ to drought stress. Conclusions This study enhances our understanding of the response of apple rootstocks to drought stress at the physiological, metabolic, and transcriptional levels and provides key insights that will aid the cultivation of drought-resistant apple rootstock cultivars. Especially, it identifies key metabolites and genes underlying the drought resistance of apple rootstocks.

The regulatory role of apple MdAGO4.1 gene in plant drought and salt resistance is unclear. In this study, transgenic A. thaliana in which the apple MdAGO4.1 gene was over-expressed was used to analyze the regulatory effects of the MdAGO4.1 gene on plant drought and salt resistance, to verify the function of the apple MdAGO4.1 gene. The seed germination rate, seedling fresh weight and root length of transgenic Arabidopsis strains in MS medium containing different concentrations of NaCl and mannitol were better than those of the wild type. The transgenic A. thaliana seedlings were more resistant to drought than wild type under drought stress. The transgenic strains were less affected by salt stress than thewild type. Exposure to drought and salt stress reduced the relative elektrolyte leakage, malondialdehyde (MDA), superoxide anion (O2-), and hydrogen peroxide (H2O2) levels of the transgenic strain significantly compared with the levels in the wild type. The levels of proline, protective enzyme activities, and the expression of genes related to drought and salt stress resistance were significantly higher than those of the wild type. These results indicate that MdAGO4.1 overexpression improved drought and salt tolerance in transgenic Arabidopsis. This study can provide a theoretical basis for future research on stress tolerance mechanisms and breeding new varieties of fruit trees resistant to drought and salt.

This study explored the contributions of melatonin and dopamine to the uptake and utilization of nitrogen and the formation of rhizosphere microbial communities in ‘Tianhong 2’/M. hupehensis, with the goal improving plant resistance to drought stress. Drought stress was formed by artificially controlling soil moisture content. And melatonin or dopamine solutions were applied to the soil at regular intervals for experimental treatment. After 60 days of treatment, plant indices were determined and the structure of the rhizosphere microbial community was evaluated using high-throughput sequencing technology. The findings revealed two ways through which melatonin and dopamine alleviate the inhibition of growth and development caused by drought stress by promoting nitrogen uptake and utilization in plants. First, melatonin and dopamine promote the absorption and utilization of nitrogen under drought stress by directly activating nitrogen transporters and nitrogen metabolism-related enzymes in the plant. Second, they promote the absorption of nitrogen by regulating the abundances of specific microbial populations, thereby accelerating the transformation of the soil nitrogen pool to available nitrogen that can be absorbed directly by plant roots and utilized by plants. These findings provide a new framework for understanding how melatonin and dopamine regulate the uptake and utilization of nitrogen in plants and improve their ability to cope with environmental disturbances.

The lack of dwarf stock with good cold resistance has affected the production of apples in northern China. Annual dormant branches of ‘Tianhong 2’ apple were grafted on 10 different dwarf interstocks (the rootstocks were the seedlings of Malus hupehensis var. Pingyiensis) as test materials. Among these 10 interstocks, Huang 6, 244, NO.1, 53, 24-5, ZC9-3, Jizhen 1 were newly developed by us (Apple Research Group of Hebei Agricultural University), and three interstocks with different degrees of cold resistance (GM256–with strongest cold resistance, SH40–with stronger cold resistance, M9–with poor cold resistance) were used as controls. The semi-lethal temperature (LT50) and related physiological indexes of the branches in the overwintering process were studied. Based on the comprehensive physiological indexes, the effects of 10 interstocks on cold resistance of the ‘Tianhong 2’ apple were analyzed. The results showed that the effects of 10 kinds of interstocks on the cold resistance of ‘Tianhong 2’ apple were quite different. The order of effects on cold resistance from strong to weak was as follow: GM256 > Huang 6 > 244 >NO.1> 53 > 24-5 > ZC9-3 >Jizhen 1 > SH40 > M9. The purpose of this study was to screen out the interstocks with strong cold resistance, in order to provide some basis for the selection and utilization of interstocks with strong cold resistance in apple cultivation to further promote the development of the apple industry in China.

1-Methylcyclopropene (1-MCP) can reduce the rate of fruit softening and prolong storage time. In this study, the fruit of a new early-maturing apple variety, ‘Taihangzaohong’, was treated with air (control), 2 μL/L 1-MCP, 100 μL/L ethylene (C 2H4) or 2 μL/L 1-MCP +100 μL/L C2H4 for 24 hours and then stored at 4 °C for 70 days. The postharvest physiological indices and the expression of 13 genes related to ethylene biosynthesis and signal transduction were monitored every 10 days. The results showed that 1-MCP can delay the softening rate and maintain the fruit quality of this early-maturing apple variety by reducing ethylene production by reducing the expression of MdACO1, MdACO2, and MdACS1, as well as by preventing ethylene signal transduction by decreasing the expression of MdETR2 and MdERS1 and increasing the expression of MdCTR1. Understanding the significant changes in these genes and their functions may help us explore the mechanisms controlling apple fruit softening and its response to exogenous 1-MCP and ethylene stimuli, as well as inhibition at the receptor level during ripening and senescence.

Malus hupehensis, as an apple rootstock, is an economically important tree species popular due to its excellent fruit yield and stress resistance. Nitrogen is one of the critical limiting factors of plant growth and fruit yield, so it is crucial to explore new methods to improve nitrogen use efficiency. Melatonin and dopamine, as multifunctional metabolites, play numerous physiological roles in plants. We analyzed the effects of exogenous melatonin and dopamine treatments on the growth, root system architecture, nitrogen absorption, and metabolism of M. hupehensis when seedlings were exposed to nitrate-deficient conditions. Under low nitrate stress, plant growth slowed, and chlorophyll contents and 15NO3– accumulation decreased significantly. However, the application of 0.1 μmol/L melatonin or 100 μmol/L exogenous dopamine significantly reduced the inhibition attributable to low nitrate levels during the ensuing period of stress treatment, and the effect of dopamine was more obvious. In addition to modifying the root system architecture of nitrate-deficient plants, exogenous melatonin and dopamine also changed the uptake, transport, and distribution of 15NO3–. Furthermore, both exogenous melatonin and dopamine enhanced tolerance to low nitrate stress by maintaining the activity of enzymes (NR, NiR, GS, Fd-GOGAT, and NADH-GOGAT) and the transcription levels of related genes involved in leaf and root nitrogen metabolism. We also found that exogenous melatonin and dopamine promoted the expression of nitrate transporter genes (NRT1.1, NRT2.4, NRT2.5, and NRT2.7) in nitrate-deficient plant leaves and roots. Our results suggest that both exogenous melatonin and dopamine can mitigate low nitrate stress by changing the root system architecture, promoting the absorption of nitrate, and regulating the expression of genes related to nitrogen transport and metabolism. However, according to a comprehensive analysis of the results, exogenous dopamine plays a more significant role than melatonin in improving plant nitrogen use efficiency.

There is no report on the effect of the length of Jizhen 2 interstock on the growth and fruit quality of Tianhong 2 apple trees, which are usually grown in Baoding, Hebei Province, China. We surveyed the tree size, branch types, fruit set, fruit quality and root parameters of 3–5-year-old ‘Tianhong 2/Jizhen 2/Malus × robusta Rehder’ apple trees, to study the effects of dwarfing interstock length on the growth of the tree’s aboveground parts and roots, as well as fruit yield and quality. The tree height and the stem girths of the interstock and scion decreased as interstock length increased, and the dwarfing degree of the apple trees gradually increased. Trees with an interstock length of 30 cm had the fewest long branches, the most short branches, and the greatest proportion of short branches. An interstock length of 30 cm provided the highest fruit-set rate, the highest yield per tree and per unit cross-sectional area, the highest single fruit weight, the highest soluble: acid ratio, the highest color brightness (L*), and better red skin coloration (higher a*) of the fruit skin. The root length density, root surface area density, and root volume density exhibited two growth peaks in a year, during the slow growth period when spring and autumn shoots are stopped. Root length density, root surface area density, and root volume density decreased with interstock length. Root death peaked during the growth peak period of the autumn shoots, and root length density of dead roots and root turnover frequencyincreased with the interstock length. A 30 cm length was the most suitable for the Tianhong 2 apple trees when the Jizhen 2 was used as the interstock.