Nutrient stress harms plant growth and yield. Melatonin is a biologically active, multifunctional hormone that relieves abiotic stress in plants. Although previous studies have shown that melatonin plays an important role in improving nutrient-use efficiency, the mechanism of its regulation of nutrient stress remains unclear. In this study, melatonin was applied to apple plants under nutrient stress, and morphological indices, physiological and biochemical indices, and stomatal morphology were evaluated. The response of apple plants to nutrient deficiency and the melatonin mechanism to alleviate nutrient stress were analyzed by combining ionome, transcriptome, and metabolome. The results showed that exogenous melatonin significantly alleviated the inhibitory effect of nutritional stress on the growth of apple plants by regulating stomatal morphology, improving antioxidant enzyme activity, promoting ion absorption, and utilizing and changing the absorption and distribution of minerals throughout the plant. The transcriptome results showed that melatonin alleviated nutrient stress and promoted nutrient absorption and utilization by regulating glutathione metabolism and upregulating some metal ion transport genes. The metabolome results indicated that levels of oxalic acid, L-ascorbic acid, anthocyanins (cyanidin-3-O-galactoside), lignans (lirioresinol A and syringaresinol), and melatonin significantly increased after exogenous melatonin was applied to plants under nutrient stress. These differentially expressed genes and the increase in beneficial metabolites may explain how melatonin alleviates nutrient stress in plants.

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.

Crop load has a substantial impact on growth of the aerial and belowground parts of apple trees. Here, we examined the effects of different crop loads on growth and hormone levels in apple roots. A crop load of 1.5 (T1.5) fruits per cm² trunk cross-sectional area (TCSA) treatment resulted in lower root growth vigor, while non-fruiting (T0) and T0.4 conditions showed higher root growth vigor. In all treatments, dead roots increased in length 90 days after full bloom (DAFB), whereas live roots were more abundant at about 50 and 170 DAFB, showing a bimodal curve. During each root growth peak, levels of cytokinins (CTKs), indole acetic acid (IAA), and gibberellic acid (GA₃) were higher. Moreover, hormone levels gradually decreased with increasing crop load within each peak. Root turnover tended to decrease with decreasing crop load. These findings indicate that root growth and hormone contents were positively correlated during the fruit growth phase, and that the negative impact of crop load on root growth may have been caused by hormone level decreases.

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.

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.

Dwarfing interstocks play an essential role in determining the performance of fruit trees. SH40, Jizhen 1, and Jizhen 2 are widely used dwarfing interstocks in Hebei Province, China. This study examined the influence of these three dwarfing interstocks on vegetative growth, fruit quality and yield, and leaf and fruit macro- (N, P, K, Ca, and Mg) and micro- (Fe, Zn, Cu, Mn, and B) element contents for ‘Tianhong 2’. Five-year-old ‘Fuji’ apple cultivar, ‘Tianhong 2’, on ‘Malus. Robusta’ rootstock, was cultivated with SH40, Jizhen 1, or Jizhen2 dwarfing rootstock as an interstock bridge. Jizhen 1 and 2 had more branches and a higher proportion of short branches than SH40. Jizhen 2 had a higher yield, good fruit quality, and higher leaf macro- (N, P, K, and Ca) and micro-element (Fe, Zn, Cu, Mn, and B) contents; Jizhen 1 had the highest leaf Mg content in the growing period. The fruit N, P, K, Fe, Zn, Cu, Mn, and B contents were higher in Jizhen 2. SH40 had the highest fruit Ca content. There were significant correlations in nutrient elements between leaves and fruit in June and July. Comprehensive analysis showed that Tianhong 2 had moderate tree vigor, high yield, good fruit quality, and high mineral element content in leaves and fruits when Jizhen 2 was used as an interstock.