Transgenic Fruit Research Articles

The Fat Body-Specific GST Gene SlGSTe11 Enhances the Tolerance of Spodoptera litura to Cyantraniliprole and Nicotine.

Spodoptera litura is a significant agricultural pest, and its glutathione S-transferase (GST) plays a crucial role in insecticide resistance. This study aimed to investigate the relationship between the SlGSTe11 gene of S. litura and resistance to cyantraniliprole and nicotine. Transcriptome analysis revealed that SlGSTe11 is highly expressed mainly in fat bodies, with a significant increase in SlGSTe11 gene expression under induction by cyantraniliprole and nicotine. The ectopic expression of the SlGSTe11 gene in transgenic fruit flies resulted in a 5.22-fold increase in the tolerance to cyantraniliprole. Moreover, compared to the UAS-SlGSTe11 line, the Act5C-UAS>SlGSTe11 line laid more eggs and had a lower mortality after nicotine exposure. RNAi-mediated inhibition of SlGSTe11 gene expression led to a significant increase in the mortality of S. litura under cyantraniliprole exposure. In vitro metabolism experiments demonstrated that the recombinant SlGSTe11 protein efficiently metabolizes cyantraniliprole. Molecular docking results indicated that SlGSTe11 has a strong affinity for both cyantraniliprole and nicotine. These findings suggest that SlGSTe11 is involved in the development of resistance to cyantraniliprole and nicotine in S. litura.

ORANGE gene positively regulates chromoplast formation and carotenoid accumulation in Osmanthus fragrans

Sweet osmanthus (Osmanthus fragrans) cultivar from Aurantiacus Group shows orange or orange-red flower color due to the accumulation of abundant carotenoids in the petals. Orange (Or) gene has been functionally characterized in the formation of plant chromoplasts and carotenoid accumulation in several species. However, whether Or gene from sweet osmanthus plays essential role in the formation of plant chromoplasts is not clear. In this study, we isolated Or homologous gene OfOr from sweet osmanthus by transcriptome homology search. The protein encoded by the gene has two transmembrane domains and a zinc finger domain, and is co-localized with the chloroplasts. Transmission electron microscopy analysis of the petal tissues of sweet osmanthus showed that the chromoplast volume, the plastoglobule number and volume increased significantly with petal development, accompanied by an increase in the expression level of OfOr. Correlation analysis showed that the development of chromoplasts was significantly positively correlated with the expression pattern of OfOr. In addition, after stable overexpression of OfOr in tomato (Solanum lycopersicum), it was found that the transgenic tomato fruits exhibited significantly higher redness, yellowness and chroma, as well as an increased distribution of carotenoids in the pulp cells, with total carotenoid content significantly higher than that of wild type. In summary, our study indicate that OfOr positively regulates chromoplast formation and carotenoid accumulation. This study also further reveals that OfOr can be used as a novel genetic tool to increase the accumulation of carotenoids and improve the coloring of colored organs in crops and ornamental plants.

Glycoside hydrolase PpGH28BG1 modulates benzaldehyde metabolism and enhances fruit aroma and immune responses in peach.

Benzaldehyde (BAld) is one of the most widely distributed volatiles that contributes to flavor and defense in plants. Plants regulate BAld levels through various pathways, including biosynthesis from trans-cinnamic acid (free BAld), release from hydrolysis of glycoside precursors (BAld-H) via multiple enzymatic action steps, and conversion into downstream chemicals. Here, we show that BAld-H content in peach (Prunus persica) fruit is up to 100-fold higher than that of free BAld. By integrating transcriptome, metabolomic and biochemical approaches, we identified glycoside hydrolase PpGH28BG1 as being involved in the production of BAld-H through the hydrolysis of glycoside precursors. Overexpressing and silencing of PpGH28BG1 significantly altered BAld-H content in peach fruit. Transgenic tomatoes heterologously expressing PpGH28BG1 exhibited a decrease in BAld-H content and an increase in SA accumulation, while maintaining fruit weight, pigmentation, and ethylene production. These transgenic tomato fruits displayed enhanced immunity against Botrytis cinerea compared to wild type (WT). Induced expression of PpGH28BG1 and increased SA content were also observed in peach fruit when exposed to Monilinia fructicola infection. Additionally, elevated expression of PpGH28BG1 promoted fruit softening in transgenic tomatoes, resulting in a significantly increased emission of BAld compared to WT. Most untrained taste panelists preferred the transgenic tomatoes over WT fruit. Our study suggests that it is feasible to enhance aroma and immunity in fruit through metabolic engineering of PpGH28BG1 without causing visible changes in the fruit ripening process.

ABIOTIC STRESS GENE 1 mediates aroma volatiles accumulation by activating MdLOX1a in apple.

Fruit aroma is an important organoleptic quality, which influences consumer preference and market competitiveness. Aroma compound synthesis pathways in plants have been widely identified, among the lipoxygenase pathway is crucial for fatty acid catabolism to form esters in apple. However, the regulatory mechanism of this pathway remains elusive. In this study, linear regression analysis and transgene verification revealed that the lipoxygenase MdLOX1a is involved in ester biosynthesis. Yeast one-hybrid library screening indicated that a protein, MdASG1 (ABIOTIC STRESS GENE 1), was a positive regulator of MdLOX1a and ester production based on yeast one-hybrid and dual-luciferase assays, as well as correlation analysis among eight different apple cultivars. Overexpression of MdASG1 in apple and tomato stimulated the lipoxygenase pathway and increased the fatty acid-derived volatile content, whereas the latter was decreased by MdASG1 silencing and CRISPR/Cas9 knockout. Furthermore, MdASG1 overexpression enhanced the salt-stress tolerance of tomato and apple 'Orin' calli accompanied by a higher content of fatty acid-derived volatiles compared to that of non-stressed transgenic tomato fruit, while MdASG1-Cas9 knockdown calli do not respond to salt stress and promote the biosynthesis of fatty acid-derived volatiles. Collectively, these findings indicate that MdASG1 activates MdLOX1a expression and participates in the lipoxygenase pathway, subsequently increasing the accumulation of aroma compounds, especially under moderate salt stress treatment. The results also provide insight into the theory for improving fruit aroma quality in adversity.

Ectopic overexpression of ShCBF1 and SlCBF1 in tomato suggests an alternative view of fruit responses to chilling stress postharvest.

Postharvest chilling injury (PCI) is a physiological disorder that often impairs tomato fruit ripening; this reduces fruit quality and shelf-life, and even accelerates spoilage at low temperatures. The CBF gene family confers cold tolerance in Arabidopsis thaliana, and constitutive overexpression of CBF in tomato increases vegetative chilling tolerance, in part by retarding growth, but, whether CBF increases PCI tolerance in fruit is unknown. We hypothesized that CBF1 overexpression (OE) would be induced in the cold and increase resistance to PCI. We induced high levels of CBF1 in fruit undergoing postharvest chilling by cloning it from S. lycopersicum and S. habrochaites, using the stress-inducible RD29A promoter. Harvested fruit were cold-stored (2.5°C) for up to three weeks, then rewarmed at 20°C for three days. Transgene upregulation was triggered during cold storage from 8.6- to 28.6-fold in SlCBF1-OE, and between 3.1- to 8.3-fold in ShCBF1-OE fruit, but developmental abnormalities in the absence of cold induction were visible. Remarkably, transgenic fruit displayed worsening of PCI symptoms, i.e., failure to ripen after rewarming, comparatively higher susceptibility to decay relative to wild-type (WT) fruit, lower total soluble solids, and the accumulation of volatile compounds responsible for off-odors. These symptoms correlated with CBF1 overexpression levels. Transcriptomic analysis revealed that the ripening and biotic and abiotic stress responses were altered in the cold-stored transgenic fruit. Seedlings grown from 'chilled' and 'non-chilled' WT fruit, in addition to 'non-chilled' transgenic fruit were also exposed to 0°C to test their photosynthetic response to chilling injury. Chilled WT seedlings adjusted their photosynthetic rates to reduce oxidative damage; 'non-chilled' WT seedlings did not. Photosynthetic parameters between transgenic seedlings were similar at 0°C, but SlCBF1-OE showed more severe photoinhibition than ShCBF1-OE, mirroring phenotypic observations. These results suggest that 1) CBF1 overexpression accelerated fruit deterioration in response to cold storage, and 2) Chilling acclimation in fructus can increase chilling tolerance in seedling progeny of WT tomato.

Synergistic effects of SlCBF1 and ethylene signaling on the maintenance of tomatoes quality during long-term cold storage

Refrigeration is commonly employed for postharvest storage of fruit and vegetable; however, its usage can increase the risk of chilling injury (CI). C-repeat/dehydration-responsive element binding factors (CBFs) have been identified in various plants. However, the significance of CBFs in preserving fruit quality during cold storage remains understudied, particularly in transgenic fruit. In this study, we demonstrate that the mutation of SlCBF1 gene worsen CI symptoms in tomatoes, hinder cold-induced ethylene production, and influence the expression of genes associated with ethylene signaling. Notably, our correlation analysis reveals that the severity of CI is partially correlated with ethylene production. Additionally, blocking ethylene action using 1-methylcyclopropene (1-MCP), decrease the cold resistance of tomatoes. Knocking out SlCBF1 gene impact the effect of 1-MCP-inhibited cold responses, including the suppression of antioxidant enzymes activities. These findings underscore positive regulation of cold resistance in postharvest tomatoes by SlCBF1 in collaboration with ethylene signaling.

Functional analysis of SDR112C1 associated with fenpropathrin tolerance in Tetranychus cinnabarinus (Boisduval).

Short-chain dehydrogenases/reductases (SDRs) are ubiquitously distributed across diverse organisms and play pivotal roles in the growth, as well as endogenous and exogenous metabolism of various substances, including drugs. The expression levels of SDR genes are reportedly upregulated in the fenpropathrin (FEN)-resistant (FeR) strain of Tetranychus cinnabarinus. However, the functions of these SDR genes in acaricide tolerance remain elusive. In this study, the activity of SDRs was found to be significantly higher (2.26-fold) in the FeR strain compared to the susceptible strain (SS) of T. cinnabarinus. A specific upregulated SDR gene, named SDR112C1, exhibited significant overexpression (3.13-fold) in the FeR population compared with that in the SS population. Furthermore, the expression of SDR112C1 showed a significant increase in the response to FEN induction. Additionally, knockdown of the SDR112C1 gene resulted in decreased SDR activity and reduced mite viability against FEN. Importantly, heterologous expression and in vitro incubation assays confirmed that recombinant SDR112C1 could effectively deplete FEN. Moreover, the overexpression of the SDR112C1 gene in Drosophila melanogaster significantly decreased the toxicity of FEN to transgenic fruit flies. These findings suggest that the overexpression of SDR SDR112C1 is a crucial factor contributing to FEN tolerance in T. cinnabarinus. This discovery not only enhances our understanding of SDR-mediated acaricide tolerance but also introduces a new family of detoxification enzymes to consider in practice, beyond cytochrome P450s, carboxyl/choline esterases and glutathione S-transferases.

Overexpression of Solanum lycopersicum DDB1 interacting protein 4 (SlDDI4) extends tomato shelf life by methylation-mediated suppression of softening-related genes expression

Prolonging the fruit storage period is strategically important due to the limitation of postharvest life determined by excessive softening. Efforts have been made to alleviate the issue of fruit softening in transgenic fruit by suppressing genes that encode cell wall-degrading proteins. However, individual genes related to the degradation of cell walls have limited ability, resulting in minimal success. SET domain-containing protein (SET protein) is widely expressed in most plant organs and crucial in plant growth and development. A tomato SET protein was previously identified as SlDDI4 (Solanum lycopersicum DDB1 interacting protein 4) with a H3K9 methyltransferase activity involving cell proliferation and organ size regulation. In this study, we demonstrated that SlDDI4 overexpression (SlDDI4-OE) promotes fruit firmness in addition to increased fruit size. Further experimental investigations revealed that the increased expression of SlDDI4 inhibited cellulase and pectinase activities, consequently increasing the proportion of cellulose and insoluble pectin. RNA-seq profiling indicated the genes involved in cell wall metabolism in SlDDI4-OE fruit were significantly down-regulated. DNA methylation and McrBC-PCR analysis at the SlCEL2, SlPG2, and SlEXP12 loci revealed that their down-regulation was associated with the elevated methylation level in the individual promoter regions. Furthermore, up-regulation of SlDDI4 generated fruit with increased lycopene content and prolonging shelf-life by approximately 20 d without adverse impacts on the flavor indicators. Thus, our results suggest that manipulation of SlDDI4 expression enables broad favorable effects on postharvest life as well as fruit size and quality through chromatin remodeling.

Transcriptional factor MdESE3 controls fruit acidity by activating genes regulating malic acid content in apple.

Acidity is a key factor controlling fruit flavor and quality. In a previous study, combined transcriptome and methylation analyses identified a P3A-type ATPase from apple (Malus domestica), MdMa11, which regulates vacuolar pH when expressed in Nicotiana benthamiana leaves. In this study, the role of MdMa11 in controlling fruit acidity was verified in apple calli, fruits, and plantlets. In addition, we isolated an APETALA2 domain-containing transcription factor, designated MdESE3, based on yeast one-hybrid (Y1H) screening using the MdMa11 promoter as bait. A subcellular localization assay indicated that MdESE3 localized to the nucleus. Analyses of transgenic apple calli, fruits, and plantlets, as well as tomatoes, demonstrated that MdESE3 enhances fruit acidity and organic acid accumulation. Meanwhile, chromatin immunoprecipitation quantitative PCR, luciferase (LUC) transactivation assays, and GUS reporter assays indicated that MdESE3 could bind to the ethylene-responsive element (ERE; 5'-TTTAAAAT-3') upstream of the MdMa11 transcription start site, thereby activating its expression. Furthermore, MdtDT, MdDTC2, and MdMDH12 expression increased in apple fruits and plantlets overexpressing MdESE3 and decreased in apple fruits and plantlets where MdESE3 was silenced. The ERE was found in MdtDT and MdMDH12 promoters, but not in the MdDTC2 promoter. The Y1H, LUC transactivation assays, and GUS reporter assays indicated that MdESE3 could bind to the MdtDT and MdMDH12 promoters and activate their expression. Our findings provide valuable functional validation of MdESE3 and its role in the transcriptional regulation of MdMa11, MdtDT, and MdMDH12 and malic acid accumulation in apple.

Visualization of mitochondrial dynamics in tomato based on green fluorescent protein

This study aimed to visualize the morphological features and dynamic changes of tomato mitochondria to provide a basis for the study of its mitochondrial functions. In this study, transgenic tomatoes expressing mitochondria-localized green fluorescent protein (mitochondria-GFP, Mt-GFP) were obtained by Agrobacterium-mediated genetic transformation. The color, hardness, soluble solids, acidity content, respiration rate, and ethylene production of the transgenic Mt-GFP tomato fruits were determined at the stage of mature green, breaker, and 3, 6, 9 days after breaker, while the wild-type tomato fruits were used as a control. As expected, Mt-GFP recombinant protein did not affect the ripening process, but induced the increased acidity of tomato fruits. The accumulations of Mt-GFP protein in tomato leaves and fruits were successfully verified by Western blotting. The morphological characteristics of mitochondria in flower, leaf and fruit cells as well as the dynamic changes of mitochondria in flower cells were clearly observed and studied under confocal laser microscope. The development of transgenic Mt-GFP tomato plants helps the visualization of tomato mitochondria and provides good research materials for the study of mitochondrial function during tomato development and fruit ripening.

MdWRKY45 contributes to resistance against Botryosphaeria dothidea in apple

Malus domestica “Fuji” is the predominant apple cultivar grown around the world, but it is highly susceptible to apple ring rot caused by Botryosphaeria dothidea. WRKY transcription factors (TFs) play key roles in multiple physiological responses to plant defense. We have previously reported the involvement of several MdWRKY TFs in plant defense against. In this study, we identified a new MdWRKY TF, MdWRKY45, which is induced by B. dothidea infection. MdWRKY45 was localized in the nucleus and exhibited transcriptional activity. Over-expression of MdWRKY45 enhanced the resistance of transgenic apple calli and fruits to B. dothidea infection. Conversely, suppression of MdWRKY45 decreased the disease resistance of apple calli and fruits. Further transcriptome profiling and expression analysis showed that various genes regulating the release of salicylic acid (SA), jasmonic acid (JA), and reactive oxygen species (ROS) were either up-regulated or down-regulated in MdWRKY45-RNAi transgenic apple calli compared with wild-type (WT). During B. dothidea infection, MdWRKY45 modulated the levels of SA and JA, as well as ROS burst. Moreover, MdWRKY45 could bind to the W-box cis-element in the promoters of MdPR1, MdJAZ1, and MdRbohF genes, thereby regulating their expression. In conclusion, MdWRKY45 primarily confers resistance to B. dothidea infection through pathways involving SA, JA, and ROS burst defense.

High β-carotene accumulation in transgenic eggplant fruits grown under artificial light.

Eggplant (Solanum melongena L.) fruits are known to contain few carotenoids such as β-carotene, which are abundant in congener tomato fruits. In a previous study, we introduced a fruit-specific EEF48 gene promoter-driven crtB gene encoding phytoene synthase (PSY) of Erwinia uredovora into eggplant 'Senryo No. 2'. The transgenic plants grown in a greenhouse set fruits that accumulated β-carotene (∼1.67 µg g-1FW) in the T0 and T1 generations. In the present study, we grew T1 and T2 generations of the transgenic eggplant plants in artificial climate chambers to investigate their fruit set and β-carotene accumulation. No clear difference in β-carotene accumulation was observed in the fruit of transgenic plants grown under either HID (high-intensity discharge) or LED (light-emitting diode) light, or between T1 and T2 generations. The β-carotene accumulation (8.83 µg g-1FW on average) was approximately 5 times higher than the previous results obtained from greenhouse-grown plants. However, the fruit weight and size of the T-DNA (+) plants were significantly smaller than that of their null-segregant T-DNA (-) plants derived from the same line, suggesting that β-carotene accumulation may inhibit fruit development. Considering that a part of plants grown under LED irradiation failed to set fruits or set smaller fruits than those grown under HID irradiation, the light condition in the LED chamber may not be sufficient to promote fruit development. The present results are expected to provide valuable information for the selection of transgenic eggplants with high β-carotene content in fruit under artificial lighting.

A syndromic neurodevelopmental disorder caused by rare variants in PPFIA3

PPFIA3 encodes the protein-tyrosine phosphatase, receptor-type, F-polypeptide-interacting-protein-alpha-3 (PPFIA3), which is a member of the LAR-protein-tyrosine phosphatase-interacting-protein (liprin) family involved in synapse formation and function, synaptic vesicle transport, and presynaptic active zone assembly. The protein structure and function are evolutionarily well conserved, but human diseases related to PPFIA3 dysfunction are not yet reported in OMIM. Here, we report 20 individuals with rare PPFIA3 variants (19heterozygous and 1 compound heterozygous) presenting with developmental delay, intellectual disability, hypotonia, dysmorphisms, microcephaly or macrocephaly, autistic features, and epilepsy with reduced penetrance. Seventeen unique PPFIA3 variants were detected in 18 families. To determine the pathogenicity of PPFIA3 variants invivo, we generated transgenic fruit flies producing either human wild-type (WT) PPFIA3 or five missense variants using GAL4-UAS targeted gene expression systems. In the fly overexpression assays, we found that the PPFIA3 variants in the region encoding the N-terminal coiled-coil domain exhibited stronger phenotypes compared to those affecting the C-terminal region. In the loss-of-function fly assay, we show that the homozygous loss of fly Liprin-α leads to embryonic lethality. This lethality is partially rescued by the expression of human PPFIA3 WT, suggesting human PPFIA3 function is partially conserved in the fly. However, two of the tested variants failed to rescue the lethality at the larval stage and one variant failed to rescue lethality at the adult stage. Altogether, the human and fruit fly data reveal that the rare PPFIA3 variants are dominant-negative loss-of-function alleles that perturb multiple developmental processes and synapse formation.

Transient Expression Assay and Microscopic Observation in Kumquat Fruit.

Citrus fruits encompass a diverse family, including oranges, mandarins, grapefruits, limes, kumquats, lemons, and others. In citrus, Agrobacterium tumefaciens-mediated genetic transformation of Hongkong kumquat (Fortunella hindsii Swingle) has been widely employed for gene function analysis. However, the perennial nature of woody plants results in the generation of transgenic fruits taking several years. Here, we show the procedures of Agrobacterium-mediated transient transformation and live-cell imaging in kumquat (F. crassifolia Swingle) fruit, using the actin filament marker GFP-Lifeact as an example. Fluorescence detection, western blot analysis, and live-cell imaging with confocal microscopy demonstrate the high transformation efficiency and an extended expression window of this system. Overall, Agrobacterium-mediated transient transformation of kumquat fruits provides a rapid and effective method for studying gene function in fruit, enabling the effective observation of diverse cellular processes in fruit biology.

Suppressing the rhamnogalacturonan lyase gene FaRGLyase1 preserves RGI pectin degradation and enhances strawberry fruit firmness

Plant rhamnogalacturonan lyases (RGLyases) cleave the backbone of rhamnogalacturonan I (RGI), the “hairy” pectin and polymer of the disaccharide rhamnose (Rha)-galacturonic acid (GalA) with arabinan, galactan or arabinogalactan side chains. It has been suggested that RGLyases could participate in remodeling cell walls during fruit softening, but clear evidence has not been reported. To investigate the role of RGLyases in strawberry softening, a genome-wide analysis of RGLyase genes in the genus Fragaria was performed. Seventeen genes encoding RGLyases with functional domains were identified in Fragaria × ananassa. FaRGLyase1 was the most expressed in the ripe receptacle of cv. Chandler. Transgenic strawberry plants expressing an RNAi sequence of FaRGLyase1 were obtained. Three transgenic lines yielded ripe fruits firmer than controls without other fruit quality parameters being significantly affected. The highest increase in firmness achieved was close to 32%. Cell walls were isolated from ripe fruits of two selected lines. The amount of water-soluble and chelated pectins was higher in transgenic lines than in the control. A carbohydrate microarray study showed a higher abundance of RGI epitopes in pectin fractions and in the cellulose-enriched fraction obtained from transgenic lines. Sixty-seven genes were differentially expressed in transgenic ripe fruits when compared with controls. These genes were involved in various physiological processes, including cell wall remodeling, ion homeostasis, lipid metabolism, protein degradation, stress response, and defense. The transcriptomic changes observed in FaRGLyase1 plants suggest that senescence was delayed in transgenic fruits.

Kiwifruit Monodehydroascorbate Reductase 3 Gene Negatively Regulates the Accumulation of Ascorbic Acid in Fruit of Transgenic Tomato Plants.

Ascorbic acid is a potent antioxidant and a crucial nutrient for plants and animals. The accumulation of ascorbic acid in plants is controlled by its biosynthesis, recycling, and degradation. Monodehydroascorbate reductase is deeply involved in the ascorbic acid cycle; however, the mechanism of monodehydroascorbate reductase genes in regulating kiwifruit ascorbic acid accumulation remains unclear. Here, we identified seven monodehydroascorbate reductase genes in the genome of kiwifruit (Actinidia eriantha) and they were designated as AeMDHAR1 to AeMDHAR7, following their genome identifiers. We found that the relative expression level of AeMDHAR3 in fruit continued to decline during development. The over-expression of kiwifruit AeMDHAR3 in tomato plants improved monodehydroascorbate reductase activity, and, unexpectedly, ascorbic acid content decreased significantly in the fruit of the transgenic tomato lines. Ascorbate peroxidase activity also increased significantly in the transgenic lines. In addition, a total of 1781 differentially expressed genes were identified via transcriptomic analysis. Three kinds of ontologies were identified, and 106 KEGG pathways were significantly enriched for these differently expressed genes. Expression verification via quantitative real-time PCR analysis confirmed the reliability of the RNA-seq data. Furthermore, APX3, belonging to the ascorbate and aldarate metabolism pathway, was identified as a key candidate gene that may be primarily responsible for the decrease in ascorbic acid concentration in transgenic tomato fruits. The present study provides novel evidence to support the feedback regulation of ascorbic acid accumulation in the fruit of kiwifruit.

Bioengineering of horticultural crops in Russia and in the world

Modern bioengineering technologies make it possible to create fruit and berry crops with signs that are unattainable by traditional breeding methods, while significantly reducing the time of breeding work. A review of more than 90 scientific publications in the period from 1989 to 2020 with research work on the creation of transgenic fruit and berry crops using various bioengineering technologies, which showed a wide variety of methods for modifying and identifying the resulting plants.
 For example, the modification of the genome of an apple tree using genes that accelerate flowering, the so-called “Fast breeding” technology, allows you to speed up breeding work to create valuable varieties. The developed new varieties of transgenic apple trees are resistant to the fungi, bacteria, apple moth, and also reduced ability to browning fruits, with an increased content of sucrose, etc.
 Modern methods of genetic engineering make it possible to significantly accelerate the processes of creating highly productive varieties of fruit crops with increased or complete resistance to viruses. Transgenic papaya, expressing the gene for the envelope protein of the virus, made it possible to save plantations in Hawaii. The plum varieties created with the help of bioengineering were resistant to the plum pox virus (PPV), which poses a greater danger to perennial fruit crops.
 In Russia, clonal rootstocks of apple and pear trees resistant to herbicides, strawberries and pears with improved fruit taste, rootstocks and varieties of plums and cherries resistant to the Plum Pox Virus have been created and field tested, a technology for obtaining cisgenic plants has been developed and applied using the example of apple and tomato trees that do not contain viral and bacterial sequences. Methods of genomic editing and accelerated selection of fruit crops are being developed.

WUSCHEL-related homeobox transcription factor SlWOX13 regulates tomato fruit ripening.

Fruit ripening is a complex, genetically programmed process involving the action of critical transcription factors (TFs). Despite the established importance of WUSCHEL-related homeobox (WOX) TFs in plant development, the involvement of WOX and its underlying mechanism in the regulation of fruit ripening remain unclear. Here, we demonstrate that SlWOX13 regulates fruit ripening in tomato (Solanum lycopersicum). Overexpression of SlWOX13 accelerates fruit ripening, whereas loss-of-function mutation in SlWOX13 delays this process. Moreover, ethylene synthesis and carotenoid accumulation are significantly inhibited in slwox13 mutant fruit but accelerated in SlWOX13 transgenic fruit. Integrated analyses of RNA-seq and chromatin immunoprecipitation (ChIP)-seq identified 422 direct targets of SlWOX13, of which 243 genes are negatively regulated and 179 are positively regulated by SlWOX13. Electrophoretic mobility shift assay, RT-qPCR, dual-luciferase reporter assay, and ChIP-qPCR analyses demonstrated that SlWOX13 directly activates the expression of several genes involved in ethylene synthesis and signaling and carotenoid biosynthesis. Furthermore, SlWOX13 modulates tomato fruit ripening through key ripening-related TFs, such as RIPENING INHIBITOR (RIN), NON-RIPENING (NOR), and NAM, ATAF1, 2, and CUC2 4 (NAC4). Consequently, these effects promote fruit ripening. Taken together, these results demonstrate that SlWOX13 positively regulates tomato fruit ripening via both ethylene synthesis and signaling and by transcriptional regulation of key ripening-related TFs.

The apple transcription factor MdZF-HD11 regulates fruit softening by promoting Mdβ-GAL18 expression.

Fruit ripening and the associated softening are major determinants of fruit quality and post-harvest shelf life. Although the mechanisms underlying fruit softening have been intensively studied, there are limited reports on the regulation of fruit softening in apples (Malus domestica). Here, we identified a zinc finger homeodomain transcription factor MdZF-HD11that trans-activates the promoter of Mdβ-GAL18, which encodes a pectin-degradation enzyme associated with cell wall metabolism. Both MdZF-HD11 and Mdβ-GAL18 genes were up-regulated by exogenous ethylene treatment and repressed by 1-methylcyclopropene treatment. Further experiments revealed that MdZF-HD11 binds directly to the Mdβ-GAL18 promoter and up-regulates its transcription. Moreover, using transgenic apple fruit calli, we found that overexpression of Mdβ-GAL18 or MdZF-HD11 significantly enhanced β-galactosidase activity, and overexpression of MdZF-HD11 induced the expression of Mdβ-GAL18. We also discovered that transient overexpression of Mdβ-GAL18 or MdZF-HD11 in 'Golden Delicious' apple significantly increased the release of ethylene, reduced fruit firmness, promoted the transformation of skin color from green to yellow, and accelerated ripening and softening of the fruit. Finally, the overexpression of MdZF-HD11 in tomato also promoted fruit softening. Collectively, these results indicate that ethylene-induced MdZF-HD11 interacts with Mdβ-GAL18 to promote the post-harvest softening of apple.

PusALDH1 gene confers high levels of volatile aroma accumulation in both pear and tomato fruits

Aroma is an important commercial trait that determines fruit quality and has an important influence on the overall flavor of fruits. Plant ALDH genes have been implicated in diverse pathways and play crucial roles in physiological activities. In this study, via genome resequencing we identified one gene PusALDH1 (Pbr034873.1) related to aroma biosynthesis that can respond to the induction of methyl jasmonate. Transient transformation of pear fruits and heterologous stable transformation of tomato further confirmed the function of PusALDH1 in aroma accumulation. The content of ALDH precursor substance, benzaldehyde, was reduced in the overexpressing pear and tomato fruits, and the content of ALDH product, benzoic acid and benzoic acid derivatives, was increased in the pear fruits. Meanwhile, transgenic tomato fruits with PusALDH1 overexpression exhibited a greater area of yellow placenta, indicating that the gene may be related to the growth and development of the fruit. Taken together, PusALDH1 could act as a strong candidate gene in aroma synthesis.