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The abscisic acid (ABA) has a key role in the regulation of grapevine fruit ripening, but the cellular and molecular biological mechanism of the hormone action in the fruit ripening remains unknown. By means of differential centrifugation, microsomes were prepared from Kyoho grapevine (Vitis vinifera L. × V. Labrusca L.) berries, and using the microsomes, we have obtained evidence for the occurrence of specific ABA-binding sites on the membranes with the microvolume radio-ligand binding assay. The binding sites had saturability, high affinity, and low capacity. The results of trypsin and dithiothreitol treatments to the microsomes suggested that ABA binding sites had the properties of proteins that might have disulfide group located at or near the binding site. The binding maximum amount of ABA in the microsomes was at pH 6.0 and the activity of ABA binding proteins was higher at 25 than at 0°C (temperature). The amount of ABA bound reached 54% of the ABA binding maximum (Bmax) for 10 minutes of incubation and Bmax reached for 30 minutes. The dissociation constant (Ka) and Bmax of ABA binding proteins in the microsomes were 17.5 nmol/L and 98.4 fmol/mg protein, respectively.
By using the micro-volume radio-ligand binding essay, the changes in the kinetic characteristics of the abscisic acid (ABA)-binding protein(s) of the Kyhoh grapevine (Vitis vinifera × V. labrusca) fruit during the different stages of fruit development have been studied. The changes in the berry volume growth, concentration of sugar, organic acids, and ABA in fruit mesocarp have been surveyed, especially for studies of ABA-binding protein. The dissociation constant (Kd) and ABA binding maximum (Bmax) were determined by the Scatchard plots for ABA binding in microsomes of the fruit. They are Kd = 17.5, 50.0, 6.3, 13.3 nmol·L–1; Bmax = 98.6, 523.0, 41.6, 85.4 μmol·mg–1 protein, respectively, for the fruit developmental phase I, II, veraison, and phase III. The Scatchard plots showed a rectilinear function for all of the developmental phases including veraison, which suggests the sole ABA-binding site of high affinity for ABA in the fruit microsomes, but this site could either be only one kind of the same protein or consist of more kinds of different proteins for different developmental stages. The binding affinity of ABA-binding protein(s) for ABA was shown to be higher at veraison time than during other developmental phases; this binding affinity increased nearly by 10 times from phase II to veraison, while the concentration (Bmax) of the ABA-binding protein(s) decreased to the minimum at veraison. The very low concentration of ABA at veraison may be able to trigger the onset of fruit ripening due to the increase of the binding affinity of ABA-binding protein(s) for ABA at this time. The possible functions of the ABA-binding protein(s) for fruit development during the different developmental stages were discussed, and it is suggested that the protein(s) detected could be the putative ABA receptor(s) or transporter(s) for the action of this plant hormone in grapevine.
Root systems of pecan trees are usually dominated by a single taproot with few lateral roots, which are commonly thought to inhibit successful transplanting. This study aimed to evaluate early growth and root/shoot development of pecan seedlings in response to taproot pruning. Taproots of ‘Shaoxing’ seedling pecan trees were mildly (1/3 of the total length of the radicle removed) and severely (2/3 of the total length of the radicle removed) pruned at different seedling development stages shortly after germination. At the end of the first growing season, top growth was measured and then trees were uprooted so that root system regrowth could be evaluated. The results showed that root pruning had no impact on increases in stem height or stem diameter. However, pruning the taproot could stimulate primary growth in taproot branches. Root weight and the number of taproot branches per tree increased with decreasing taproot length. This study indicated that severe root pruning when three to five leaves had emerged resulted in root systems with more taproot branches and the greatest root dry weight after one growth season, which may increase survival and reduce transplanting shock.
MicroRNAs (miRNAs) related to phytohormone signal transduction and self-incompatibility may play an important role in the xenia effect. However, associated research in this area is still lacking in rabbiteye blueberry (Vaccinium ashei). In this study, we identified miRNAs, predicted their target genes, performed functional enrichment analysis of the target genes, and screened for miRNAs related to phytohormone signaling and self-incompatibility. A total of 491 miRNAs were identified, of which 27 and 67 known miRNAs as well as 274 and 416 new miRNAs were found in the rabbiteye blueberry cultivars Brightwell and Premier, respectively. Compared with ‘Premier’, 31 miRNAs were upregulated and 62 miRNAs were downregulated in ‘Brightwell’. Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis indicated that the 4985 target genes predicted were involved in biosynthesis of amino acids, plant–pathogen interaction, and spliceosome pathways. A total of 10, one, one, five, two, five, and two candidate miRNAs related to auxin, cytokinin, gibberellin, abscisic acid, ethylene, brassinosteroid, and salicylic acid signaling, respectively, in rabbiteye blueberry pollen were identified. Further analysis indicated that novel_miR_49 was a candidate miRNA related to self-incompatibility, and their target gene was maker-VaccDscaff21-snap-gene-21.37. In addition, the KEGG enrichment analysis of the target genes of novel_miR_49 showed that they were involved in the ribosome, aminoacyl-tRNA biosynthesis, and glycosylphosphatidylinositol-anchor biosynthesis pathways. The results revealed that the microRNAs of rabbiteye blueberry pollen regulated to phytohormone signal transduction and self-incompatibility signal transduction based on related to auxin, cytokinin, gibberellin, abscisic acid, ethylene, brassinosteroid, and salicylic acid signaling. Results suggest that more research of the effects of miRNAs on regulation of hormone signal transduction and self-incompatibility is necessary for elucidating the molecular mechanism of the xenia effect.
Crabapples (Malus sp.) are ornamental woody plants that belong to the Rosaceae family. Flooding has severely hampered the growth and development of crabapple, and little is known about the molecular responses of crabapple to waterlogging tolerance. Cuttings of waterlogging-tolerant Malus hupehensis and waterlogging-intolerant Malus halliana received flooding treatment of 30 days and regular planting, respectively. Using transcriptome sequencing, we isolated 5703 and 2735 waterlogging-responsive genes from waterlogging-treated M. hupehensis and M. halliana leaves. Among these differentially expressed genes (DEGs), only 746 were shared by both. Several variables may explain the greater waterlogging tolerance of M. hupehensis: there were more waterlogging response genes related to carbohydrate and energy metabolism; signal transduction; antioxidation; lipid metabolism; protein and amino acid metabolism; and polysaccharide, cell wall, and cytoskeleton metabolism pathway in the waterlogged leaves of M. hupehensis than in M. halliana. In particular, the number of DEGs related to anaerobic metabolism, fatty acid metabolism, protein phosphorylation and dephosphorylation, γ-aminobutyric acid metabolism and cellulase, pectinase metabolism pathway in the flooded leaves of M. hupehensis was more than that in M. halliana. The alterations in gene expression patterns of the two crabapple species induced by waterlogging varied substantially. These outcomes pave the way for further studies into the functions of genes that may be involved in waterlogging tolerance in crabapples.
Kiwifruit (Actinidia chinensis Planchon) is an economically important fruit, and its flowering and production are affected by the chill accumulation in winter. In this study, the chilling requirements of nine kiwifruit cultivars with three ploidy levels (diploid, tetraploid, and hexaploid) were analyzed by using the Dynamic Model, Utah Model, and chilling hours (CH) Model. The chilling requirements for vegetative budbreak of these kiwifruit cultivars were 24–55 chill portions (CP), 316–991 chill units (CU), and 222–853 CH, and the chilling requirements for floral emergence were 45–69 CP, 825–1336 CU, and 655–1138 CH. The chilling requirements for vegetative budbreak and floral emergence were significantly lower for diploid than hexaploid cultivars with tetraploid cultivars intermediate. Pearson correlation analysis indicated that ploidy levels were positively correlated with chilling requirement, with the cv of 0.74 and 0.82 for vegetative budbreak and floral emergence chilling requirements, respectively. In conclusion, these results provide some novel insights of kiwifruit varieties of various chilling requirements, which is beneficial for kiwifruit cultivar selection for different climates and environments.
The plant compact and dwarf growth habit is an important agronomic trait when breeding watermelon (Citrullus lanatus) cultivars because of their reduced vine length, high-density planting, and better land utilization; however, the genetic basis of the dwarf growth habit is not well-known. In this study, the plant population of six generations, P1, P2, F1, F2, BC1P1, and BC1P2, were studied. A genetic segregation analysis demonstrated that dwarfism is mainly controlled by a single recessive Cldw gene. Furthermore, whole-genome sequencing of two distinct watermelon cultivars, W1-1 (P1) and 812 (P2), was performed and preliminarily mapped through a bulked segregant analysis of F2 individuals that revealed the Cldw gene locus on chromosome 9. Two candidate genes, Cla015407 and Cla015408, were discovered at the delimited region of 43.2 kb by fine mapping, and gene annotation exposed that the Cla015407 gene encodes gibberellic acid 3β-hydroxylase protein. In addition, a comparative analysis of gene sequence and cultivars sequences across the reference genome of watermelon revealed the splice site mutation in the intron region of the Cldw gene in dwarf-type cultivar 812. The quantitative real-time polymerase chain reaction exhibited a significantly higher expression of the Cla015407 gene in cultivar W1-1 compared with 812. There was no significant difference in the vine length of both cultivars after gibberellic acid treatment. In brief, our fine mapping demonstrated that Cla015407 is a candidate gene controlling dwarfism of watermelon plants.
To find the characteristics of somatic embryogenesis of orchids and elucidate the mechanism, we had previously established an efficient plant regeneration system via somatic embryogenesis in Dendrobium candidum Wall ex Lindl. In this study, a detailed cytological investigation was carried out on the initiation and developmental process of somatic embryogenesis. Based on our observations, the somatic embryogenesis in D. candidum originated from the transition of an embryonic callus cell to the initial somatic embryo cell, and the somatic embryos initiated from those cells. During the transition process, condensation and devacuolation successively occurred in the cytoplasm of the embryonic callus cells, giving rise to the formation of a typical initial somatic embryo cell with dense cytoplasm and a clear nucleus. One of the two pathways in somatic embryogenesis is the single-cell-derived somatic embryo which is generated from an inner initial somatic embryo cell in embryonic callus and develops into a globular somatic embryo in a way similar to zygotic embryogenesis and then keeps developing into a protocorm-like body (PLB). The other is a multiple-cell-derived somatic embryo which is generated from peripheral grouped initial somatic cells in embryonic calli and directly forms globular embryo or multicellular somatic proembryo, lacking the typical early stages of embryogenesis. Both pathways were observed in the somatic embryogenesis system, indicating that the culture system in D. candidum can be a useful tool for investigating the mechanisms underlying orchid embryogenesis.