Floral aromas are a mixture of volatile organic compounds, essential attributes associated with the attraction of different pollinators. This investigation is the first in-depth exploration of the volatile profiles of sixteen muscadine grape genotypes, producing female and perfect flowers using the headspace solid-phase microextraction (HS-SPME)-GC-MS-based untargeted volatilomics approach. A total of one hundred fifty volatile metabolites were identified in the muscadine flower genotypes, including the functional groups of hydrocarbons, esters, alcohols, ketones, aldehydes, miscellaneous, and acids. Multivariate statistical analysis for volatile terpenes revealed eleven bio-marker terpene volatiles that primarily distinguish between female and perfect flowers. The β-elemene, β-bisabolene, and α-muurolene were the marker volatiles characterizing perfect flowers; however, α-selinene, (Z,E)-α-farnesene, and (E,E)-geranyl linalool were the typical marker terpene in the female flowers. Perfect flowers exhibited better pollinator attraction capacity associated with a higher number of flowers per inflorescence, enhanced pollinator rewards, and higher numbers and quantities of terpene volatiles than female flowers, resulting in superior pollinator attraction capacity and fruit set efficiency. The pollinator attraction mechanism of female flowers exhibited several morphological and biochemical floral defects, causing random pollinator visits and low fruit set efficiency. The controlled pollination assay could express female flowers’ full fruit set capabilities by avoiding casual insect pollination. This comprehensive study suggests that these marker terpenes might contribute to pollinator attraction in muscadine flower genotypes and should be considered an excellent reference for agroecosystem ecologists and entomologists.

A comparative study of two grapevine rootstocks with contrasting drought stress responses revealed that the drought-resilient RUG harbors an efficient antioxidant defense system, characterized by increased activities of superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT), along with elevated proline (Pro) levels compared to the drought-sensitive MGT. This robust scavenging machinery enables RUG to maintain redox balance, effectively mitigating oxidative stress and preserving cellular integrity during drought. Anatomical evaluations showed severe xylem disruptions in MGT, including extensive tylosis, leading to leaf necrosis and impaired water transport. Conversely, RUG maintained a structurally intact and functional xylem, crucial for sustaining hydraulic conductivity and water supply during drought. The pronounced rise in Pro underscores its critical role in drought resilience, working synergistically with other cellular components to facilitate osmotic adjustment while detoxifying reactive oxygen species (ROS) and minimizing oxidative damage. Transcriptome profiling suggested that RUG displays sequential gene expression during drought driven by distinct molecular processes for photosynthesis, osmotic adjustment, and structural remodeling, a dynamic notably absent in MGT. These findings emphasize the complex interplay of osmotic and oxidative homeostasis in RUG, illustrating the adaptive mechanisms that contribute to its drought resilience, potentially guiding future rootstock selection and breeding strategies.

Salinity is one of the substantial threats to plant productivity and could be escorted by other stresses such as heat and drought. It impairs critical biological processes, such as photosynthesis, energy, and water/nutrient acquisition, ultimately leading to cell death when stress intensity becomes uncured. Therefore, plants deploy several proper processes to overcome such hostile circumstances. Grapevine is one of the most important crops worldwide that is relatively salt-tolerant and preferentially cultivated in hot and semi-arid areas. One of the most applicable strategies for sustainable viticulture is using salt-tolerant rootstock such as Ruggeri (RUG). The rootstock showed efficient capacity of photosynthesis, ROS detoxification, and carbohydrate accumulation under salinity. The current study utilized the transcriptome profiling approach to identify the molecular events of RUG throughout a regime of salt stress followed by a recovery procedure. The data showed progressive changes in the transcriptome profiling throughout salinity, underpinning the involvement of a large number of genes in transcriptional reprogramming during stress. Our results established a considerable enrichment of the biological process GO-terms related to salinity adaptation, such as signaling, hormones, photosynthesis, carbohydrates, and ROS homeostasis. Among the battery of molecular/cellular responses launched upon salinity, ROS homeostasis plays the central role of salt adaptation.

Like other plant stresses, salinity is a central agricultural problem, mainly in arid or semi-arid regions. Therefore, salt-adapted plants have evolved several adaptation strategies to counteract salt-related events, such as photosynthesis inhibition, metabolic toxicity, and reactive oxygen species (ROS) formation. European grapes are usually grafted onto salt-tolerant rootstocks as a cultivation practice to alleviate salinity-dependent damage. In the current study, two grape rootstocks, 140 Ruggeri (RUG) and Millardet et de Grasset 420A (MGT), were utilized to evaluate the diversity of their salinity adaptation strategies. The results showed that RUG is able to maintain higher levels of the photosynthetic pigments (Chl-T, Chl-a, and Chl-b) under salt stress, and hence accumulates higher levels of total soluble sugars (TSS), monosaccharides, and disaccharides compared with the MGT rootstock. Moreover, it was revealed that the RUG rootstock maintains and/or increases the enzymatic activities of catalase, GPX, and SOD under salinity, giving it a more efficient ROS detoxification machinery under stress.

Anthocyanins, a major class of flavonoids, are important pigments of grape berries. Despite the recent discovery of the genetic cause underlying the loss of color, the metabolomic and molecular responses are unknown. Anthocyanin quantification among diverse berry color muscadines suggests that all genotypes could produce adequate anthocyanin quantities, irrespective of berry color. Transcriptome profiling of contrasting color muscadine genotypes proposes a potential deficiency that occurs within the anthocyanin transport and/or degradation mechanisms and might cause unpigmented berries. Genome-wide association studies highlighted a region on chromosome-4, comprising several genes encoding glutathione S-transferases involved in anthocyanin transport. Sequence comparison among genotypes reveals the presence of two GST4b alleles that differ by substituting the conserved amino acid residue Pro 171 -to-Leu. Molecular dynamics simulations demonstrate that GST4b2–Leu 171 encodes an inactive protein due to modifications within the H-binding site. Population genotyping suggests the recessive inheritance of the unpigmented trait with a GST4b2/2 homozygous. A model defining colorless muscadines’ response to the mutation stimulus, avoiding the impact of trapped anthocyanins within the cytoplasm is established.

Muscadine grapes accumulate higher amounts of bioactive phenolics compared with other grape species. To identify the molecular events associated with polyphenolic accumulation that influence antioxidant capacity, two contrasting muscadine genotypes (C5 and C6) with varied phenolic/flavonoid content and antioxidant activity were investigated via RNA-sequencing during berry development. The results showed that berry development is concomitant with transcriptome profile changes, which was more pronounced at the véraison (V) stage. Despite that the downregulation pattern of gene expression dominated the upregulation through berry development, the C5 genotype maintained higher expression levels. Comparative transcript profiling allowed the identification of 94 differentially expressed genes with potential relevance in regulating fruit secondary metabolism, including 18 transcription factors and 76 structural genes. The genes underlying the critical enzymes in the modification reactions of polyphenolics biosynthetic pathway, including hydroxylation, methylation, and glycosylation were more pronounced during the immature stages of prevéraison (PrV), V, and postvéraison (PoV) in the C5 genotype, resulting in more accumulation of biologically active phenolic/flavonoid derivatives. The results suggested that muscadine grapes, as in bunch grapes (Vitis sp.); possess a similar mechanism that organizes polyphenolics accumulation; however, the set of total flavonoids (TFs) and structural genes coordinating the pathway varies between the two species.

Vitis has two subgenera: Euvitis, which includes commercially important Vitis vinifera and interspecific hybrid cultivars, and Muscadinia. Of note, the market for Muscadinia grapes remains small, and only Muscadinia rotundifolia is cultivated as a commercial crop. To establish a basis for the study of Muscadinia species, we generated chromosome-level whole-genome sequences of Muscadinia rotundifolia cv. Noble. A total of 393.8 Mb of sequences were assembled from 20 haploid chromosomes, and 26 394 coding genes were identified from the sequences. Comparative analysis with the genome sequence of V. vinifera revealed a smaller size of the M. rotundifolia genome but highly conserved gene synteny. A genome-wide association study of 12 Muscadinia berry-related traits was performed among 356 individuals from breeding populations of M. rotundifolia. For the transferability of markers between Euvitis and Muscadinia, we used 2000 core genome rhAmpSeq markers developed to allow marker transferability across Euvitis species. A total of 1599 (80%) rhAmpSeq markers returned data in Muscadinia. From the GWAS analyses, we identified a total of 52 quantitative trait nucleotides (QTNs) associated with the 12 berry-related traits. The transferable markers enabled the direct comparison of the QTNs with previously reported results. The whole-genome sequences along with the GWAS results provide a new basis for the extensive study of Muscadinia species.

Biochemical juice attributes and color-related traits of muscadine grape genotypes have been investigated. For this study, 90 muscadine genotypes, including 21 standard cultivars, 60 breeding lines, and 9 Vitis x Muscadinia hybrids (VM), were evaluated. The biochemical properties of total soluble solids (TSS), titratable acidity, and TSS/Acid (T/A) ratio showed modest diversity among genotypes with a range of 10.3 °Brix, 2.1 mg tartaric acid/L, and 4.6, respectively. Nonetheless, the pH trait exhibited a tight range of 0.74 among the population with a minimum and maximum pH of 3.11 ± 0.12 and 3.85 ± 0.12. Color-related traits showed more deviation between individuals. Total anthocyanin content (TAC), luminosity index (L*), hue angle (h°), and chroma index (C*) displayed a range of 398 µg/g DW, 33.2, 352.1, and 24, respectively. The hierarchical clustering map classified the population into two large groups of colored and non-colored grapes based on L* and h°, suggesting the predominance of these two characters among the population. The colored berries genotypes clade was further divided into several sub-clades depending on C*, TAC, and TSS levels. The principal component analysis (PCA) separated the four-color characteristics into two groups with a negative correlation between them, L* and C* versus TAC and h°. Further, PCA suggested the positive influence of acidity in enhancing the different nutraceutical components. Despite the nature of anthocyanins as a member of phenolic compounds, a lack of significant correlation between TAC and nutraceutical-related traits was detected. The dissimilatory matrix analysis highlighted the muscadine individuals C11-2-2, E16-9-1, O21-13-1, and Noble as particular genotypes among the population due to enhanced color characteristics.

In this study, fertility-related traits of 90 muscadine grape genotypes were evaluated. Selected genotypes included 21 standard cultivars, 60 breeding lines, and nine Vitis × Muscadinia hybrids (VM hybrids). The first fruiting bud (FFB), bud fertility (BF), bud fertility coefficient (BFC), number of flowers/flower cluster (N.F/FC), fruit-set efficiency (FSE), number of clusters/vine (N.C/V), and yield/vine (Y/V) traits were evaluated. The FFB trait did not show significant differences among genotypes. The muscadine genotype O28-4-2-2 (1.6 ± 0.2) displayed the FFB closest to the base; however, O17-16-2-1, O18-2-1, and VM A12-10-2 genotypes had the most distant FFB (3.6 ± 0.3). All the other fertility-related traits varied widely among the population. The BF, BFC, N.F/FC, FSE, N.C/V, and Y/V exhibited a range estimated at 35.1%, 81.5%, 259.7, 63.3%, 177 C/V, and 22.3 kg/V, respectively. The muscadine genotypes O42-3-1 (36.7% ± 1.3) and Majesty (34% ± 1.2) exhibited the highest BF; however, the VM A12-10-2 (1.6% ± 0.1) recorded the lowest BF. The VM genotype O15-16-1 (82.8% ± 4.1) displayed the highest BFC; however, the VM A12-10-2 (1.3% ± 0.1) showed the lowest BFC. The muscadine genotypes D7-1-1 (280.3 F/FC ± 21.7) and O17-17-1 (20.7 F/FC ± 5.5) showed the highest and lowest N.F/FC, respectively. The maximum and minimum FSE was observed for the Rosa cultivar (65.7% ± 2.4) and muscadine genotype D7-1-1 (2.4% ± 0.2), respectively. The minimum N.C/V was recorded for VM genotype A12-10-2 (6 C/V ± 0.2) and maximum noted for muscadine genotypes B20-18-2 (183 C/V ± 7.5) and O44-14-1 (176 C/V ± 7.3). Muscadine genotype O23-11-2 (22.6 kg ± 1.1) produced the highest Y/V; however, the lowest yield was recorded for O15-17-1, Fry Seedless, Sugargate, and the VM genotypes and A12-10-2, with an average yield among them estimated at 0.4 kg ± 0.2.

Three muscadine grape genotypes (Muscadinia rotundifolia (Michx.) Small) were evaluated for their metabolite profiling and antioxidant activities at different berry developmental stages. A total of 329 metabolites were identified using UPLC-TOF-MS analysis (Ultimate 3000LC combined with Q Exactive MS and screened with ESI-MS) in muscadine genotypes throughout different developmental stages. Untargeted metabolomics study revealed the dominant chemical groups as amino acids, organic acids, sugars, and phenolics. Principal component analysis indicated that developmental stages rather than genotypes could explain the variations among the metabolic profiles of muscadine berries. For instance, catechin, epicatechin-3-gallate, and gallic acid were more accumulated in ripening seeds (RIP-S). However, tartaric acid and malonic acid were more abundant during the fruit-set (FS) stage, and malic acid was more abundant in the veraison (V) stage. The variable importance in the projection (VIP > 0.5) in partial least-squares–discriminant analysis described 27 biomarker compounds, representing the muscadine berry metabolome profiles. A heatmap of Pearson’s correlation analysis between the 27 biomarker compounds and antioxidant activities was able to identify nine antioxidant determinants; among them, gallic acid, 4-acetamidobutanoic acid, trehalose, catechine, and epicatechin-3-gallate displayed the highest correlations with different types of antioxidant activities. For instance, DPPH and FRAP conferred a similar antioxidant activity pattern and were highly correlated with gallic acid and 4-acetamidobutanoic acid. This comprehensive study of the metabolomics and antioxidant activities of muscadine berries at different developmental stages is of great reference value for the plant, food, pharmaceutical, and nutraceutical sectors.

In this study, biometrics assessment of flower structure, cluster-, and berry-related traits were evaluated in a population of 90 muscadine grape genotypes for three consecutive years. This population consisted of 21 standard cultivars, 60 breeding lines, and 9 Vitis x Muscadinia hybrids (VM hybrids). Cluster length (CL) and width (CWI) characteristics exhibited slight differences among the population, with a range estimated at 7.1 and 4.6 cm, respectively. However, cluster weight (CWE), number of berries/cluster (N.B/C), and cluster compactness (CC) traits showed more diversity between individuals with a calculated range of 205.6 g, 32.6 B/C, and 24.1, respectively. Interestingly, all berry-related traits greatly varied between individuals, excluding the number of seeds/berry (N.S/B) character. The N.S/B trait displayed a narrow range of 5.6 seeds within the population. However, characters of berry length (BL), width (BWI), weight (BWE), the weight of seeds/berry (W.S/B), firmness (FF), and dry scar pattern (SP) demonstrated a wide estimated range of 21.2 mm, 21.7 mm, 25.4 g, 0.71 g, 0.21 N, and 82%, respectively. Normal distribution analysis for each trait suggested different distribution patterns extended between unimodal to multimodal behavior. Hierarchical mapping analysis was able to classify the population into several clades based on physical cluster- and berry-related attributes. The PCA suggested that hermaphroditic (perfect) flower structure is associated with compact clusters exhibiting small berries in size and weight (i.e., muscadine genotypes suitable for wine production). However, female flower structure is associated with clusters displaying large berries in size and weight (i.e., muscadine genotypes appropriate for fresh consumption). These patterns occurred independently of cluster size and weight characters. This research is the first study evaluating muscadine biometrics characters at a population level, providing valuable information for market demand and muscadine breeding programs.

ABSTRACT Stilbenes are phytoalexins expressed by plants to avoid/resist certain biotic and abiotic stresses. This study was envisioned to determine the interrelationship between stilbenes producing ability of peanut genotypes and Aspergillus colonization level. Stilbenes were induced in peanut cotyledon in vitro by soaking in water, cutting them into thin slices, and subsequently challenging them with Aspergillus flavus. Fungal colonization was then measured in the cotyledon slices. The results showed major differences in fungal colonization levels between the control (seed without stilbene induction) and stilbenes-induced seeds. This finding was further validated using twenty peanut genotypes to ensure the relationship between stilbenes producing ability of the seed and fungal colonization level. The result showed that of the 20 genotypes tested, seeds of genotypes K1504, K1620 and K1632 showed minimal fungal colonization compared to control seed (without stilbenes induction), while genotypes DRT40, Kadiri-7, Narayani, DRT43 and Tirupati-3 showed no fungal colonization. The differences in stilbenes content and composition of cotyledon slices was determined by HPLC to assess genetic differences in their stilbenes producing ability. Comparative evaluation of these data showed that the genotypes that showed no fungal colonization expressed significantly higher amounts of stilbenes compared to genotypes which expressed relatively lower amounts of stilbenes. Overall, these data suggest that the genotypes expressing high amounts of stilbenes were able to mitigate fungal colonization while the genotypes expressing relatively lower amounts of stilbenes sustained fungal colonization, which indicates that this technique may be useful for screening breeding population to identify genotypes capable of avoiding Aspergillus colonization and aflatoxin contamination.