Important Vegetables Research Articles

Integrated Analysis of the lncRNA-miRNA-mRNA Expression Profiles in Response to Meloidogyne incognita in Radish (Raphanus sativus L.)

Radish is an important root vegetable that is widely grown in Asia. The root-knot nematode (RKN) Meloidogyne incognita seriously affects the growth and development of radish root and causes poor appearance quality. However, the molecular mechanism of radish response to RKNs remains poorly understood. In this study, a total of 220 lncRNAs, 1144 mRNAs, 20 miRNAs and 153 proteins were differential expressed between the RKN-infected and WT samples. Correlation analysis of all DEPs compared with all DGEs showed that 8 mRNAs-DEPs showed a changed abundance. The results showed that 18 DEmiRNAs have 167 target DEGs in 220 miRNA-target modules and 29 DElncRNAs were predicted as putative targets of 16 DEmiRNAs in 37 miRNA-target modules. In all, 6 DGEs in the ABA pathway and 2 DGEs in the JA pathway were identified under RKN infection, respectively. The four regulatory networks of lncRNA-miRNA-mRNA were constructed in response to RKN infection. qRT-PCR analysis found that the expression pattern of 6 DElncRNAs, 6 DEmRNAs, 6 DEmiRNAs and 6 DEPs were consistent with sequencing results. These results provide a theoretical basis for studying the molecular mechanism of radish in response to M. incognita and breeding resistant varieties to this nematode.

Field longevity of methyl eugenol and cue-lure plugs and associated insecticidal strips: captures of Bactrocera dorsalis and Zeugodacus cucurbitae (Diptera: Tephritidae) in Hawaii.

Certain species of true fruit flies (Diptera: Tephritidae) cause tremendous damage to commercially important fruits and vegetables, and many countries operate continuous trapping programs which rely on male-specific lures such as trimedlure (TML), methyl eugenol (ME), and cue-lure (CL). Traditionally, these attractants have been applied as liquids to cotton wicks inside traps, although this results in high evaporative loss of the lure. Slow-release, polymeric plugs have been widely adopted for TML, but such devices are not widely used for ME or CL. Recent data, however, suggest that ME and CL plugs may be attractive for as long as 12 wk in the field. The present study investigates whether ME and CL plugs weathered for 18 or 24 wk are effective in capturing males of Bactrocera dorsalis (Hendel) and Zeugodacus cucurbitae (Coquillett), respectively. For B. dorsalis, 6 g ME plugs were as effective as the control treatment (fresh liquid on a wick) after 12 wk of weathering but not after 18 or 24 wk. For Z. cucurbitae, 3 g CL plugs were as effective as the control treatment (fresh CL plugs) after 12 and 18 wk of weathering but not after 24 wk. The residual content and release rate of the 2 lures were also measured over time, but, with the exception of the residual content of ME, we did not find a direct correlation between these parameters and numbers of flies captured.

Physicochemical characteristics and bacterial communities of meigan cai and zao cai, two home-made fermented vegetables

Meigan cai and zao cai are two important traditional fermented vegetables in Fujian Province, China. However, there is little information on the bacterial community of these two fermented vegetables. In the present work, we evaluated and compared the physicochemical characteristics and bacterial communities of meigan cai and zao cai. Results revealed that Firmicutes and Proteobacteria were the main dominant phyla, and Lactobacillus and Serratia were the main genera. Principal coordinate and significance analysis showed that the bacterial communities were similar between meigan cai and zao cai. Except for glucose, there was no significant difference in the physicochemical characteristics between meigan cai and zao cai. Our study provided information on the bacterial communities of meigan cai and zao cai, and a guide for the production of these two fermented vegetables.

Attractiveness, longevity, and release rates of multilure wafers for trapping males of the oriental fruit fly and melon fly (Diptera: Tephritidae).

Invasive fruit flies (Diptera: Tephritidae) pose a serious threat to the production and export of many commercially important fruits and vegetables. Detection of the agricultural pests Bactrocera dorsalis (Hendel) and Zeugodacus cucurbitae (Coquillett) relies heavily on traps baited with male-specific attractants. For B. dorsalis, traps are typically baited with liquid methyl eugenol (ME), and for Z. cucurbitae, traps are baited with liquid cue-lure (CL). Operating large-scale trapping networks is costly, consequently, there is much interest in identifying ways to maintain network sensitivity while reducing costs. One cost-cutting approach is the possibility of combining different male lures in the same dispenser, thus reducing the number of traps requiring servicing. The chief objective of this study was to compare captures of B. dorsalis and Z. cucurbitae males in Jackson traps baited with polymeric wafers impregnated with both ME and raspberry ketone (RK, a hydrolyzed form of CL) versus traps baited with liquid ME or CL freshly applied to cotton wicks. Captures were measured when the ME/RK wafers had been weathered for 12, 18, or 24wk. Captures of B. dorsalis and Z. cucurbitae males were similar between fresh lure and weathered wafers over all trapping periods, with a single exception apparently due to the lessened potency of the associated killing agent. The residual amount and release rate of ME and RK from the wafers were also measured to examine possible relationships between wafer chemistry and trap catch. The possible implications of the present results to area-wide trapping programs are discussed.

Studies on Genetic Divergence in Cabbage Genotypes in the Mid Hills of Solan District, Himachal Pradesh, India

Cabbage (Brassica oleracea var. capitata L.) is one of the most important vegetables of the family Brassicaceae and is grown under temperate to tropical climatic conditions for its head in more than ninety countries throughout the world. The diversity in cabbage is immense due to the presence of variation in shape, size, weight and maturity of heads. The greater the variability in the available germplasm, more would be the chances for selecting superior genotypes for use in breeding programmes or direct use as cultivars. Twenty three genotypes of cabbage including check variety, Golden Acre were evaluated in a Randomized Complete Block Design (RCBD) with three replications. The results concluded that twenty-three diverse genotypes of cabbage were grouped into four clusters. The resultant four clusters showed genetic diversity and maximum number of genotypes were accommodated in cluster I (12) followed by II (7), IV (3) and III (1). The maximum inter cluster distance was recorded between cluster II and III, therefore, the hybridization between the genotypes of cluster II and III can be performed to get superior hybrids or recombinants in segregating populations for further breeding programmes. Based on the mean performance, genetic distance and clustering pattern, inter-varietal crosses can be made which may be advantageous in creating wider variability for better heterotic combinations and transgressive segregants in cabbage.

Effect of integrated nutrient management on the cauliflower yield and soil properties

Cauliflower is one of the most important and preferred vegetables and effective nutrient management is essential to improve production as well as soil properties. Thus, the study was conducted to examine the impact of integrated nutrient management on cauliflower and soil at Ghorahi, Dang district of Nepal. The single-factor experiment was organized employing a Randomized Complete Block Design with three replications. In this study, nitrogen (N) was applied through different sources viz 100% N via urea, 50% N via urea + 50% N via farmyard manure, 50% N via urea + 50% N via pig manure, 50% N via urea + 50% N via vermicompost, 50% N via urea + 25% N via farmyard manure + 25% N via pig manure, 50% N via urea + 25% N via pig manure + 25% N via vermicompost, 50% N via urea + 25% N via farm yard manure + 25% N via vermicompost and 50% N via urea + 50% N via (farmyard manure + pig manure + vermicmpost in 1:1:1). Among different treatments, 50% N via urea + 50% N via vermicompost recorded the highest curd diameter (17.72 cm), curd height (11.57 cm), curd weight (0.76 kg), curd yield per hectare (31.67 t ha-1), soil organic matter (3.31%), total nitrogen (0.157%), available phosphorus (76.78 kg ha-1) as well as available potassium (860.20 kg ha-1). Therefore, it is suggested to apply 50% N via urea and 50% N via vermicompost for better cauliflower production and soil properties.

Behind the Loss of Salinity Resistance during Domestication: Alternative Eco-Physiological Strategies Are Revealed in Tomato Clade

Salinity stress impairs growth and physiological performance in tomato, which is one of the most economically important vegetables and is widely cultivated in arid and semi-arid areas of the world. Plant landraces, which are heterogeneous, local adaptations of domesticated species, offer a unique opportunity to valorize available germplasm, underpinning the productivity, resilience, and adaptive capacity of staple crops in vulnerable environments. Here, we investigated the response of fully mature tomato plants from a commercial variety, an ancestral wild relative, and a landrace under short-term salinity exposure, as well as their ability to recover upon cessation of stress. The heterogeneous panel evaluated in this study revealed different adaptative strategies to cope the stress. Our data highlighted the ability of the tomato clade to handle low and intermediate salinity stress for short-term exposure time, as well as its capacity to recover after the cessation of stress, although inter- and intraspecific variations in morphological and physiological responses to salinity were observed. Overall, the landrace and the wild type performed similarly to control conditions under low salinity, demonstrating an improved ability to maintain ionic balance. In contrast, the commercial genotype showed susceptibility and severe symptoms even under low salinity, with pronounced reductions in K+/Na+ ratio, PSII photochemical efficiency, and photosynthetic pigments. This research confirmed that improved salt tolerant genotypes can lead to substantial, positive impacts on horticultural production. While the salt tolerance mechanism of domesticated tomato was efficient under mild stress conditions, it failed at higher salinity levels.

The interaction on the horticultural characteristics of economically important vegetables in simple nutrient addition production (SNAP) hydroponics and conventional production system

The interaction on the horticultural characteristics of economically important vegetables in simple nutrient addition production (SNAP) hydroponics and conventional production system

Development of a rapid and efficient system for CR genes identification based on hairy root transformation in Brassicaceae

Many economically important crops and vegetables belonging to the cruciferous family are heavily endangered by clubroot disease caused by Plasmodiophora brassicae infection. Breeding of clubroot resistant cultivars based on mapping and cloning of resistant genes is commonly regarded as the most cost-effective and efficient way to fight against this disease. The traditional way of R gene functional validation requires stable transformation that is both time- and labor-consuming. In this study, a rapid and efficient hairy-root transgenic protocol mediated by Agrobacterium rhizogenes was developed. The transformation positive rate was over 80% in Brassica napus showed by GUS reporter gene and this transformation only took 1/6 of the time compared with stable transformation. The system was applicable to different B. napus varieties and other cruciferous crops including Brassica rapa and Brassica oleracea. In particular, two known CR genes, CRA3.7.1 and CRA8.2.4 were used respectively, as example to show that the system works well for CR gene study combined with subsequent P. brassicae infection in B. napus. Most importantly, it works both in over-expression that led to disease resistance, as well as in RNAi which led to disease susceptible phenotype. Therefore, this system can be used in batch-wise identification of CR genes, and also offered the possibility of manipulating key genes within the P. brassicae genome that could improve our knowledge on host–pathogen interaction.

Gene expression, transcription factor binding and histone modification predict leaf adaxial–abaxial polarity related genes

Leaf adaxial–abaxial (ad–abaxial) polarity is crucial for leaf morphology and function, but the genetic machinery governing this process remains unclear. To uncover critical genes involved in leaf ad–abaxial patterning, we applied a combination of in silico prediction using machine learning (ML) and experimental analysis. A Random Forest model was trained using genes known to influence ad–abaxial polarity as ground truth. Gene expression data from various tissues and conditions as well as promoter regulation data derived from transcription factor chromatin immunoprecipitation sequencing (ChIP-seq) was used as input, enabling the prediction of novel ad–abaxial polarity-related genes and additional transcription factors. Parallel to this, available and newly-obtained transcriptome data enabled us to identify genes differentially expressed across leaf ad–abaxial sides. Based on these analyses, we obtained a set of 111 novel genes which are involved in leaf ad–abaxial specialization. To explore implications for vegetable crop breeding, we examined the conservation of expression patterns between Arabidopsis and Brassica rapa using single-cell transcriptomics. The results demonstrated the utility of our computational approach for predicting candidate genes in crop species. Our findings expand the understanding of the genetic networks governing leaf ad–abaxial differentiation in agriculturally important vegetables, enhancing comprehension of natural variation impacting leaf morphology and development, with demonstrable breeding applications.

Valuation of different organic and inorganic fertilizer combinations on the growth and yield of cabbage

abbage (Brassica oleracea var. capitata) is the most important winter leafy vegetable in the world and belongs to the Cruciferae family. An experiment was conducted at the Horticultural farm of Gauradaha Agriculture Campus, Tribhuwan University, Jhapa, Nepal from 26th December 2022 to 2nd April 2023 to evaluate the different organic and inorganic fertilizer combinations on the growth and yield of cabbage (Brassica oleracea var. capitata L. cv. Green coronet). The experiment was conducted as a randomized block design with 7 treatments replicated 3 times and the treatments are T1: 25-ton FYM/ha, T2: 25-ton PM/ha, T3: 25-ton FYM/ha + NPK (RDF), T4: 25-ton PM/ha + NPK (RDF), T5: 20-ton FYM/ha + 10-ton PM/ha + NPK (RDF), T6: 10-ton FYM/ha + 20-ton PM/ha + NPK (RDF) and T7: control. Significant variation was found among the treatments. The result showed treatment had a significant role in different parameters. T3- The highest number of leaves, T5- the highest (leaf area, weight with roots, net head yield, horizontal head diameter), T4-highest (plant height, gross yield), T6- highest (stem girth, vertical head diameter) was obtained. On the result basis T5 (20-ton FYM/ha + 10-ton PM/ha + NPK (RDF) can be adopted as sustainable crop production. This study not only provides valuable insights into optimizing cabbage production but also underscores the importance of tailored fertilizer combinations for sustainable agriculture. Furthermore, the findings contribute to the broader discourse on enhancing crop yield while minimizing environmental impact. By elucidating the efficacy of specific fertilizer blends, this research paves the way for more efficient and eco-friendly farming practices, thereby promoting food security and environmental sustainability on a global scale.

Advances in and Perspectives on Transgenic Technology and CRISPR-Cas9 Gene Editing in Broccoli.

Broccoli, a popular international Brassica oleracea crop, is an important export vegetable in China. Broccoli is not only rich in protein, vitamins, and minerals but also has anticancer and antiviral activities. Recently, an Agrobacterium-mediated transformation system has been established and optimized in broccoli, and transgenic transformation and CRISPR-Cas9 gene editing techniques have been applied to improve broccoli quality, postharvest shelf life, glucoraphanin accumulation, and disease and stress resistance, among other factors. The construction and application of genetic transformation technology systems have led to rapid development in broccoli worldwide, which is also good for functional gene identification of some potential traits in broccoli. This review comprehensively summarizes the progress in transgenic technology and CRISPR-Cas9 gene editing for broccoli over the past four decades. Moreover, it explores the potential for future integration of digital and smart technologies into genetic transformation processes, thus demonstrating the promise of even more sophisticated and targeted crop improvements. As the field continues to evolve, these innovations are expected to play a pivotal role in the sustainable production of broccoli and the enhancement of its nutritional and health benefits.

PE (Prickly Eggplant) encoding a cytokinin-activating enzyme responsible for the formation of prickles in eggplant.

Eggplant is one of the most important vegetables worldwide, with some varieties displaying prickles. These prickles, present on the leaves, stems, and fruit calyxes, posing challenges during cultivation, harvesting, and transportation, making them an undesirable agronomic trait. However, the genetic mechanisms underlying prickle morphogenesis in eggplant remain poorly understood, impeding genetic improvements. In this study, genetic analyses revealed that prickle morphogenesis is governed by a single dominant nuclear gene, termed PE (Prickly Eggplant). Subsequent bulk segregant RNA-sequencing (BSR-seq) and linkage analysis preliminarily mapped PE to chromosome 6. This locus was then fine mapped to a 9233 bp interval in a segregating population of 1109 plants, harboring only one candidate gene, SmLOG1, which encodes a LONELY GUY (LOG)-family cytokinin biosynthetic enzyme. Expression analyses via transcriptome and qRT-PCR demonstrate that SmLOG1 is predominantly expressed in immature prickles. CRISPR-Cas9 knockout experiments targeting SmLOG1 in prickly parental line 'PI 381159' abolished prickles across all tissues, confirming its critical role in prickle morphogenesis. Sequence analysis of SmLOG1 pinpointed variations solely within the non-coding region. We developed a cleaved amplified polymorphic sequences (CAPS) marker from a distinct SNP located at -735-bp within the SmLOG1 promoter, finding significant association with prickle variation in 190 eggplant germplasms. These findings enhance our understanding of the molecular mechanisms governing prickle development in eggplant and facilitate the use of marker-assisted selection (MAS) for breeding prickleless cultivars.

Physiological and molecular bases of the nickel toxicity responses in tomato

Nickel (Ni), a component of urease, is a micronutrient essential for plant growth and development, but excess Ni is toxic to plants. Tomato (Solanum lycopersicum L.) is one of the important vegetables worldwide. Excessive use of fertilizers and pesticides led to Ni contamination in agricultural soils, thus reducing yield and quality of tomatoes. However, the molecular regulatory mechanisms of Ni toxicity responses in tomato plants have largely not been elucidated. Here, we investigated the molecular mechanisms underlying the Ni toxicity response in tomato plants by physio-biochemical, transcriptomic and molecular regulatory network analyses. Ni toxicity repressed photosynthesis, induced the formation of brush-like lateral roots and interfered with micronutrient accumulation in tomato seedlings. Ni toxicity also induced reactive oxygen species accumulation and oxidative stress responses in plants. Furthermore, Ni toxicity reduced the phytohormone concentrations, including auxin, cytokinin and gibberellic acid, thereby retarding plant growth. Transcriptome analysis revealed that Ni toxicity altered the expression of genes involved in carbon/nitrogen metabolism pathways. Taken together, these results provide a theoretical basis for identifying key genes that could reduce excess Ni accumulation in tomato plants and are helpful for ensuring food safety and sustainable agricultural development.

Optimum flow rate enhances the performance of lettuce (Lactuca sativa L.) in hydroponic culture

Poor substate flow rate affects the production of crops under hydroponic systems as contact and collision times of ions (nutrients) with roots is affected deciding nutrients uptake rate and quantity required for crop growth. Keeping in view, a study was conducted under hydroponic culture to obtain optimal flow rate for higher production of lettuce, which is an important leafy vegetable. Two lettuce varieties (Boston and Lollo Bionda) were subjected to 4 different flow rates (0.5, 1.0, 1.5, 2.0 L min-1) under completely randomized design (CRD) with factorial arrangement and replicated four times. The outcomes of the study suggest that performance of Lollo Bionda variety was better than Boston regarding root and shoot fresh and dry biomass, leaf number, area, length and width and head diameter and plant height. The same parameters kept on increasing with increase of flow rate and achieved maximum values at 1.5 L min-1. Beyond that flow rate the values for these parameters declined which indicated that the optimal flow rate for these 2 lettuce verities is 1.5 L min-1. Hence, it was concluded that for higher production of lettuce, the flow rate should be kept at 1.5 L min-1 in Nutrient Film Technique (NFT) using hydroponic system with environmental and substrate conditions mentioned in materials and method portion of this article.

Desempenho morfofisiologico e qualidade pós-colheita de cultivares de alface hidropônica em condições de temperatura elevada

ABSTRACT Lettuce (Lactuca sativa) is the most economically important leafy vegetable in Brazil, with the crisp type being the most popular among producers. However, in regions with hot climates, lettuce has production limitations due to bolting, requiring knowledge on the behavior of cultivars available on the market. Thus, the objective of the present study was to characterize the performance of crisp lettuce cultivars in terms of their productive, physiological, and postharvest quality aspects when grown under hydroponic cultivation and high temperature conditions. The assay was performed in a greenhouse and the plants were grown in a hydroponic system of DFT type. The average temperature during the experiment was 30.33 °C. The experimental design used was completely randomized with nine treatments (Veneranda, Cristina, Olinda, Mônica, Cinderela, Grand Rapids, Itapuã Super, Crocantela, and Simpson Black Seed), with four replications. The cultivars were analyzed in terms of their productive, physiological, and postharvest quality performances. The Olinda and Cristina cultivars showed the best performance in growth, physiological, and postharvest variables. However, the cultivars had distinct sensory characteristics. Bolting negatively affected gas exchange, production, and postharvest quality of lettuce cultivars Veneranda, Mônica, Grand Rapids, and Cinderela. In addition, it was observed that the pigment content was not determinant for the expression of bolting. Therefore, this study recommends the use of cultivars Olinda and Cristina for the hydroponic cultivation under high temperature conditions.

LIFE HISTORY PARAMETERS OF MELON FRUIT FLY, BACTROCERA CUCURBITAE (COQUILLETT) (DIPTERA: TEPHRITIDAE) ON FOUR ECONOMICALLY IMPORTANT VEGETABLES

The majority of plant species that are used for consumption and medicinal purposes belongs to Cucurbitaceae. These vegetables are attacked by several insect pests including Tephritid flies. Keeping in view, four economically important cucurbit vegetables such as sponge gourd; Luffa aegyptiaca, bitter gourd; Momordica charantia, musk melon; Cucumis melo, and bottle gourd; Lagenaria siceraria were selected to investigate the life-history parameters of Bactrocera cucurbitae (Coq.) (Diptera: Tephritidae) under laboratory conditions. For this purpose, B. cucurbitae was reared in the laboratory on each host vegetable and egg hatchability %, egg incubation period, larval survival (%), duration (days), length and width of larvae, pupal parameters, male-female ratio, and age-specific fecundity were recorded. The results showedno statistical difference in hatchability percentages, larval duration, and pupal survival (%) in all tested vegetables. However, greater length and width of larvae were recorded when the B. cucurbitae larvae were provided with bottle gourd. Moreover, maximum pupal recovery was recorded in bottle and sponge gourd among all the tested vegetables. Maximum females were also recorded in bottle gourd. Adult longevity and fecundity were significantly affected by the host. Maximum female longevity (52 days) was recorded when B. cucurbitae were provided with bottle gourd and minimum (41 days) on sponge gourd. Similarly, maximum fecundity (823 eggs) was also recorded when maggots were fed on bottle gourd and the lowest (611 eggs) on a sponge gourd. In conclusion, Bottle gourd is the most preferable and nutritious host among all the tested vegetables and needs effective management strategies for B. cucurbitae. The output of the present studies would be helpful for the development of an effective integrated pest management (IPM) program for B. cucurbitae in a variety of vegetables.

Expanding the targeting scope of CRISPR/Cas9-mediated genome editing by Cas9 variants in Brassica

AbstractCRISPR/Cas9, presently the most widely used genome editing technology, has provided great potential for functional studies and plant breeding. However, the strict requirement for a protospacer adjacent motif (PAM) has hindered the application of the CRISPR/Cas9 system because the number of targetable genomic sites is limited. Recently, the engineered variants Cas9-NG, SpG, and SpRY, which recognize non-canonical PAMs, have been successfully tested in plants (mainly in rice, a monocot). In this study, we evaluated the targeted mutagenesis capabilities of these Cas9 variants in two important Brassica vegetables, Chinese cabbage (Brassica rapa spp. pekinensis) and cabbage (Brassica oleracea var. capitata). Both Cas9-NG and SpG induced efficient mutagenesis at NGN PAMs, while SpG outperformed Cas9-NG at NGC and NGT PAMs. SpRY achieved efficient editing at almost all PAMs (NRN > NYN), albeit with some self-targeting activity at transfer (T)-DNA sequences. And SpRY-induced mutants were detected in cabbage plants in a PAM-less fashion. Moreover, an adenine base editor was developed using SpRY and TadA8e deaminase that induced A-to-G conversions within target sites using non-canonical PAMs. Together, the toolboxes developed here induced successful genome editing in Chinese cabbage and cabbage. Our work further expands the targeting scope of genome editing and paves the way for future basic research and genetic improvement in Brassica.

Effect of Maturity Stages on the Quality of Cold Storage Iceberg Lettuce (Lactuca sativa var. capitate) for Export

Iceberg lettuce is one of the most important vegetables economically and nutritionally, but its perishable nature poses challenges for storage and long-distance transportation. This study investigated the effect of maturity stages on the quality of iceberg lettuce for export under cold storage conditions. A completely randomized design with three replications was used to evaluate four maturity stages based on days after planting (DAP): 52, 55, 58, and 61 DAP. The lettuce heads were stored at 3±2oC and a 95% relative humidity, and quality parameters were analyzed every three days until the end of storage. Sensory properties, weight loss, color change, total soluble solids, total phenolics, and decay rates were evaluated. The results showed that maturity stages significantly (P <0.05) affected the quality attributes of iceberg lettuce and that harvesting at the optimal stage is very important. Findings from this study confirmed that the quality parameters were preserved for the 58 DAP samples throughout the storage time. They indicated that at three weeks of storage time, the lowest weight loss (5.9%), color change (14.1), and decay rate (8.3%), and the highest sensory analysis (5 scores) and appearance were shown by the 58 DAP maturity stage lettuce. Therefore, harvesting iceberg lettuce at 58 DAP is recommended for long-distance exportation.

Developing carotenoids-enhanced tomato fruit with multi-transgene stacking strategies

As natural dominant pigments, carotenoids and their derivatives not only contribute to fruit color and flavor quality but are regarded as phytochemicals beneficial to human health because of various bioactivities. Tomato is one of the most important vegetables as well as a main dietary source of carotenoids. So, it's of great importance to generate carotenoid-biofortified tomatoes. The carotenoid biosynthesis pathway is a network co-regulated by multiple enzymes and regulatory genes. Here, we assembled four binary constructs containing different combinations of four endogenous carotenoids metabolic-related genes, including SlORHis, SlDXS, SlPSY, and SlBHY by using a high efficiency multi-transgene stacking system and a series of fruit-specific promotors. Transgenic lines overexpression SlORHis alone, three genes (SlORHis/SlDXS/SlPSY), two genes (SlORHis/SlBHY), and all these four genes (SlORHis/SlDXS/SlPSY/SlBHY) were enriched with carotenoids to varying degrees. Notably, overexpressing SlORHis alone showed comparable effects with simultaneous overexpression of the key regulatory enzyme coding genes SlDXS, SlPSY, and SlORHis in promoting carotenoid accumulation. Downstream carotenoid derivatives zeaxanthin and violaxanthin were detected only in lines containing SlBHY. In addition, the sugar content and total antioxidant capacity of these carotenoids-enhanced tomatoes was also increased. These data provided useful information for the future developing of biofortified tomatoes with different carotenoid profiles, and confirmed a promising system for generation of nutrients biofortified tomatoes by multiple engineering genes stacking strategy.