Understanding the different physiological responses of Malus species under salt stress in the seedling stages will be useful in breeding salt-tolerant dwarfing apple rootstocks. Seedlings of Malus Zumi (Mats.) Rehd. (M. zumi), Malus sieversii (Led.) Roem. (M. sieversii), and Malus baccata (L.) Borkh. (M. baccata) were treated with solution of 0, 0.20%, 0.40%, and 0.60% salinity. Physiological parameters of their leaves and roots were measured at 0 d, 4 d, 8 d and 12 d after salinity treatments. Superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), malondialdehyde (MDA), solution protein (SP), and proline (PRO) initially increased and then decreased. The activities and contents of these parameters were higher in the 0.40% and 0.60% NaCl treatments than in the 0.20% treatment and in the 0% control. M. zumi was the most resistant to salt stress, showing the lowest content of MDA in the leaves and roots, which increased slightly under salt stress. M. baccata had the highest increase in both the content and proportion of MDA. High enzyme activity was shown to play an important role in the salt resistance of M. zumi. Moreover, it can be speculated that there are other substances that also play a major role. We found that osmotic regulation played a key role in response to salt stress for M. baccata even though it was sensitive to salt stress. For M. sieversii, both the osmotic regulation and enzymatic antioxidants were observed to play a major role in mitigating salt stress.

Apples are a major horticultural crop worldwide. Grafting techniques are widely utilized in apple production to keep the varieties pure. Interstocks are frequently used in Northern China to achieve intensive apple dwarfing cultivation. High-throughput sequencing was used to investigate differentially expressed genes in the phloem tissues of two different xenograft systems, M (‘Gala’/‘Mac 9’/Malus baccata (L.) Borkh.) and B (‘Gala’/Malus baccata (L.) Borkh.). The results showed that dwarfing interstocks could significantly reduce the height and diameters of apple trees while have few effects on the growth of annual branches. The interstocks were found to regulate the expression of genes related to hormone metabolism and tree body control (GH3.9, PIN1, CKI1, ARP1, GA2ox1 and GA20ox1), these effects may attribute the dwarf characters for apple trees with interstocks. Besides, the interstocks reduce photosynthesis-related genes (MADH-ME4 and GAPC), promote carbon (C) metabolism gene expression (AATP1, GDH and PFK3), promote the expression of nitrogen (N)-metabolism-related genes (NRT2.7, NADH and GDH) in rootstocks, and improve the expression of genes related to secondary metabolism in scions (DX5, FPS1, TPS21 and SRG1). We also concluded that the interstocks acquired early blooming traits due to promotion of the expression of flowering genes in the scion (MOF1, FTIP7, AGL12 and AGL24). This study is a valuable resource regarding the molecular mechanisms of dwarf interstocks’ influence on various biological processes and transplantation systems in both scions and rootstocks.

The breeding of salt-tolerant rootstock relies heavily on the availability of salt-tolerant Malus germplasm resources. The first step in developing salt-tolerant resources is to learn their molecular and metabolic underpinnings. Hydroponic seedlings of both ZM-4 (salt-tolerant resource) and M9T337 (salt-sensitive rootstock) were treated with a solution of 75 mM salinity. ZM-4’s fresh weight increased, then decreased, and then increased again after being treated with NaCl, whereas M9T337′s fresh weight continued to decrease. The results of transcriptome and metabolome after 0 h (CK) and 24 h of NaCl treatment showed that the leaves of ZM-4 had a higher content of flavonoids (phloretinm, naringenin-7-O-glucoside, kaempferol-3-O-galactoside, epiafzelechin, etc.) and the genes (CHI, CYP, FLS, LAR, and ANR) related to the flavonoid synthesis pathway showed up-regulation, suggesting a high antioxidant capacity. In addition to the high polyphenol content (L-phenylalanine, 5-O-p-coumaroyl quinic acid) and the high related gene expression (4CLL9 and SAT), the roots of ZM-4 exhibited a high osmotic adjustment ability. Under normal growing conditions, the roots of ZM-4 contained a higher content of some amino acids (L-proline, tran-4-hydroxy-L-prolin, L-glutamine, etc.) and sugars (D−fructose 6−phosphate, D−glucose 6−phosphate, etc.), and the genes (GLT1, BAM7, INV1, etc.) related to these two pathways were highly expressed. Furthermore, some amino acids (S-(methyl) glutathione, N-methyl-trans-4-hydroxy-L-proline, etc.) and sugars (D-sucrose, maltotriose, etc.) increased and genes (ALD1, BCAT1, AMY1.1, etc.) related to the pathways showed up-regulation under salt stress. This research provided theoretical support for the application of breeding salt-tolerant rootstocks by elucidating the molecular and metabolic mechanisms of salt tolerance during the early stages of salt treatment for ZM-4.

‘Huahong’ is a popular apple cultivar because of its anti-browning properties and appealing aroma and flavor. It is mainly planted by grafting on dwarf interstocks in Northeast China. We investigated the different aroma profiles of apple fruits grown from six dwarf interstocks (‘CG24’, ‘SH38’, ‘SH3’, ‘MD001’, ‘Mac9’, and ‘CX5’) and from no interstocks (CK). A total of 55 VOCs were detected, including esters (25), aldehydes (14), alcohols (8), ketones (3), alkane hydroxyls (3), and acids (2). Among the VOCs, 48 were detected in the skin and 21 in the pulp. The skin of ‘Huahong’ apples had a strong sweet aroma, and the pulp was green with a subtle aroma. The dominant compounds (>5% of total content) in the skin were 2-methyl butyl acetate, hexyl 2-methyl butyrate, caproic acid butyl ester, hexanal, (Z)-2-heptene aldehyde, and 6-methyl-5-heptene-2-ketone, while in the pulp, they were 2-methyl butyl acetate, methanol, 2-methyl-1-butanol, hexanol, and hexane. Compared with CK, ‘SH38’, ‘MD001’, and ‘SH3’ interstocks had increased total aroma content, and ‘CX5’ and ‘CG24’ had suppressed aroma. The effects of interstocks on aroma were mainly reflected in skin. The VOC content ranged from 3297.52 to 9895.75 µg·kg⁻¹ in skin, and from 748.62 to 1369.21 µg·kg⁻¹ in pulp. PCA revealed that use of interstock ‘SH38’ mainly affected esters. ‘MD001’ affected hexane and 4-pentene-1-acetate; ‘Mac9’ and ‘SH3’ affected octanoic acid-2-methyl butyl ester, hexyl butyrate, and 2-methyl-1-butanol; and ‘CX5’ and ‘CG24’ had a greater impact on isoamyl propionate and 1-pentene-3-ol. Finally, ‘SH38’ had the highest principal comprehensive score. ‘SH38’ and ‘SH3’ interstocks resulted in significantly increased apple VOC content.

In this study, we used genotyping to determine the genetic structure and molecular identities of apple landraces from six species of Malus Mill. in China, based on the fingerprints revealed by microsatellite markers with tailed primer M13. A total of 46 apple landraces of Malus Mill. selected from the National Apple and Pear Germplasm Repository in Xingcheng, China were genotyped with 14 SSR markers. The primers differentiated all the accessions. At least three SSR primers, CH04h02, CH01f07a and CH04g07, with a higher heterozygosity and Shannon’s information index than other combinations can distinguish all the accessions. All the alleles for these three primers were arranged in descending order, and they were assigned values beginning with 01. Character strings were constituted by combining all the codes of the three primers for every accession. By such means, separate and special molecular identities were obtained for every apple landrace, which could be expressed in the form of a bar code. Using such bar codes, trees can be labeled and scanned, which aids in the identification and tracking of genebank collections. The eight-step method for establishing the molecular identities of apple landraces reported here may serve as a reference when determining the molecular identities establishment of other apple germplasms of Malus Mill. This method might also be used for the establishment of a molecular database to aid the preservation of Malus Mill., which is in imminent danger in China. It may also be used to improve the gene bank management of Malus Mill.

Background: Wampee (Clausena lansium (Lour.) Skeels) is an understudied Rutaceae crop native to southern China, whose fruit features a complex aroma profile with simultaneous sour, sweet, bitter and astringent notes. Although bioactive compounds including flavonoids, alkaloids and volatile oils in wampee fruit have been partially characterized, the tissue-specific metabolic and transcriptional basis underlying its distinctive aroma formation remains largely unclear. Methods: We performed an integrated volatile metabolomic and transcriptomic analysis on the pericarp (peel) and flesh of wampee fruit across three cultivars (Shanyellowpi, Heijingang, Bingtangxin). Volatile metabolites were profiled via headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS), and transcriptome profiles were generated by RNA-Seq. Multi-omics integration was conducted using Procrustes analysis, gene–metabolite correlation network construction and weighted gene co-expression network analysis (WGCNA). Results: A total of 288 volatile metabolites were identified, representing the most comprehensive volatile inventory for C. lansium reported to date. Principal component analysis and partial least squares discriminant analysis revealed distinct volatile profiles between pericarp and flesh; terpenoids were the dominant chemical class, accounting for 71.56–91.48% of total volatiles in pericarp and 61.88–67.22% in flesh. Notably, organoheterocyclic compounds were significantly enriched in Shanyellowpi flesh (40.45%), forming a cultivar-specific metabolic signature absent in the other two cultivars. Transcriptomic analysis showed that phenylpropanoid biosynthesis was the most significantly enriched pathway among differentially expressed genes, followed by monoterpene biosynthesis and sesquiterpenoid biosynthesis. Procrustes analysis demonstrated a strong global concordance between the two omics layers (M² = 0.535, p < 0.001). Gene–metabolite correlation networks identified terpene synthase (TPS) genes (HP075360, HP217350) and oxidoreductase genes (SOD1, GST, 10HGO) as candidate co-regulators of terpenoid biosynthesis. WGCNA further prioritized TPS genes, cytochrome P450 genes and MYB transcription factor genes as key regulators driving volatile metabolic divergence between tissues. Conclusion: This study provides a comprehensive volatile and transcriptomic atlas of wampee fruit, and identifies tissue-specific and cultivar-specific metabolic signatures as well as their candidate regulatory genes. These findings advance our understanding of quality differentiation in Rutaceae fruits and lay a foundation for molecular breeding and flavor improvement of wampee.