Pitaya is a tropical and subtropical fruit; it can produce several batches fruit in one year. To find out the fruit quality differences between various batches in the same year in Guangzhou, South China, 11 pitaya varieties were used as the materials. Comparative analysis was performed between these varieties of each batch by 14 indexes, comprehensive evaluation and ranking were evaluated by the principal component analysis (PCA). Results showed that the red-peel and red-pulp pitaya has the longer fruit period and could obtain more batches fruits. By comparing the fruit quality of these 11 varieties in different batches: Except "Guanhuahong," fruit weight is significant different between other 10 varieties. The edible rate of fruits from 2(nd) and 3(rd) batches is significantly higher than others. The hardness, total sugar, total acid, betalain, total phenol, and flavonoids were significant difference between batches. The PCA results indicated that in most varieties, the 1(st), 8(th), 9(th) batches are generally with heavier fruit, better color, harder and sweeter; more stable antioxidant compounds were shown in 6(th), 7(th), 8(th), 9(th) batches; the 3(rd), 4(th), 5(th), and 6(th) batches are smaller, softer, lower soluble sugar and higher titratable acid. Pitaya fruit quality and tastes from various batches are different in the same year, the climate may be the main factor. The fruits of 7(th), 8(th) and 9(th) batches picking from Sep to Nov has better quality and higher economic value. This research has practical application value and could provide theoretical basis for the production of pitaya.

Anthocyanin is a natural plant pigment whose biosynthesis is intricately linked to light regulation. Here, pericarps of red mango cultivars named 'Xinshiji (XSJ)' and 'Hongmang No.6 (HM6)' directly exposed to sunlight were red while they were green under shade conditions, whereas the pericarps of green mango cultivar known as 'Guiqi (GQ)' maintained green within both light and shade conditions. However, the mechanisms underlying these color changes under different light conditions remain largely unexplored. Therefore, we analyzed the contents, components, and biosynthesis of anthocyanins in the above mango pericarps through metabolome and transcriptome analyses. High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis releaved a total of 35 anthocyanin compounds were detected in the mango pericarps. Transcriptome analysis uncovered 44 differentially expressed genes among the comparison of 'XSJ', 'HM6', and 'GQ' samples. Twelve and eight candidate genes were identified in the anthocyanin biosynthesis and chlorophyll breakdown pathways, respectively. A co-expression network of the key anthocyanin biosynthetic genes revealed that twelve transcription factors may govern anthocyanin biosynthesis and chlorophyll breakdown processes under light. These candidate genes were further verified by real-time quantitative PCR (RT-qPCR) for providing a regulatory model for the formation of red-and green-pericarps in mangoes under light and shade conditions. These results deepen our understanding of light-induced anthocyanin biosynthesis in fruits.

The CONSTANS/CONSTANS-Like (CO/COL) family has been shown to play important roles in flowering, stress tolerance, fruit development and ripening in higher plants. In this study, three COL genes, MiCOL6, MiCOL7A and MiCOL7B, which each contain only one CCT domain, were isolated from mango (Mangifera indica), and their functions were investigated. MiCOL7A and MiCOL7B were expressed mainly at 20 days after flowering (DAF), and all three genes were highly expressed during the flowering induction period. The expression levels of the three genes were affected by light conditions, but only MiCOL6 exhibited a clear circadian rhythm. Overexpression of MiCOL6 promoted earlier flowering, while overexpression of MiCOL7A or MiCOL7B delayed flowering compared to that in the control lines of Arabidopsis thaliana under long-day (LD) and short-day (SD) conditions. Overexpressing MiCOL6, MiCOL7A or MiCOL7B in transgenic plants increased superoxide dismutase (SOD) and proline levels, decreased malondialdehyde (MAD) levels, and improved survival under drought and salt stress. In addition, yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) analyses showed that the MiCOL6, MiCOL7A and MiCOL7B proteins interact with several stress- and flower-related proteins. This work demonstrates the functions of MiCOL6, MiCOL7A and MiCOL7B and provides a foundation for further research on the role of mango COL genes in flowering regulation and the abiotic stress response.

Objectives Myeloblastosis (MYB) proteins, recognized as crucial transcriptional regulators, play an integral role in orchestrating the range of plant colors.Materials and Methods Pitaya (Hylocereus spp.), a distinctive commercially cultivated fruit, stands apart due to its unique betalains, water-soluble nitrogen-enriched pigments that confer its vibrant hues. Although betalain biosynthesis has been extensively explored, the potential influence of co-activators that might disrupt pigment production remains relatively unexplored.Results In this study, we found that HuMYB9, an R2R3-MYB repressor, exhibited a remarkable decreasing during pitaya maturation. Sequence alignment analyses showed conserved R2 and R3 domains within the C-terminal of HuMYB9. HuMYB9's regulatory activities were found to be nuclear localized and it interacted with specific elements within the promoters of HuDODA1, HuADH1, and HuCYP76AD1-1, thus influencing the transcriptional activities in vitro. HuMYB9 transiently downregulated the expression of key betalain biosynthetic genes with a corresponding effect on the levels of pitaya pulp betalains.Conclusions Our results suggest that HuMYB9 operates as a suppressor, specifically downregulating the expression of HuCYP76AD1-1, HuDODA1, and HuADH, thereby modulating betalain biosynthesis in pitaya. Collectively, our findings provide invaluable insights into the regulation of betalain accumulation in pitaya.

Thidiazuron (TDZ) is used for the expansion of fruits, but excessive levels of TDZ lead to a decline in fruit quality. The appropriate concentration of TDZ for mango expansion without a decline in quality is not clear. In the present study, four different concentrations (5, 10, 15, and 20 mg/L) of TDZ were applied via spraying on mango plants, and several physiological and biochemical indicators were measured. The results showed that TDZ treatment significantly increased mango fruit size and single-fruit weight. In mango fruit, TDZ treatment decreased the disease index, delayed the increase in the malondialdehyde and H2O2 content, and maintained firmness and antioxidant capacities at a relatively high level during postharvest storage. At the same time, TDZ treatment delayed the decrease in the giberellin, indoleacetic acid and jasmonic acid content in mango, and reduced the accumulation of abscisic acid and ethylene. These trends are consistent with TDZ treatment leading to extension of the shelf life of mango. Furthermore, ethylene biosynthesis, signal transduction, and cell wall dismantling-related genes were investigated. The results indicated that the expression of the MiACS, MiETR2, MiERF113, MiERF010, MiERF054, MiEXP, MiPG14, MiPG21, MiCEL, and MiPEL genes in mango was inhibited under TDZ treatment compared with the control. In summary, TDZ treatment can significantly increase the size and weight of mango fruit and can extend its shelf life. The most suitable concentration is 10-15 mg/L TDZ, which will not affect the quality of mango fruit.

为了探究不同时间采摘的火龙果果实品质之间的差异,以红皮红肉"红冠1号""红冠2号""湛江红肉""49-3""莞华红""红水晶"、红皮白肉"莞华白""红宝石""阳西白肉"、红皮粉红肉"莞华红粉"和红皮双色肉"双色1号"11个火龙果品种为材料,测定分析不同批次采摘的果实品质.结果表明,红肉品种一年内结果批次比其他品种多,产果期更长.同一品种果实单个品质指标不同批次间基本都存在显著性差异,综合比较发现,11个品种的第7、8、9批次果实的综合品质普遍较好,第3、4、5、6批次果实综合品质普遍稍差.说明火龙果一年间不同批次果实品质上存在显著性差异,可通过栽培技术调节来满足市场需求.

Three Di19-4 genes were identified in mango. Overexpression of MiDi19-4B in A. thaliana promoted earlier flowering and enhanced drought, salt, and ABA resistance. Drought-induced protein 19 (Di19) is a drought-induced protein that is mainly involved in multiple stress responses. Here, three Di19-4 genes (MiDi19-4A/B/C) in mango (Mangifera indica L.) were identified, and the coding sequences (CDS) had lengths of 684, 666, and 672 bp and encoded proteins with 228, 222, and 224 amino acids, respectively. The promoters of the MiDi19-4 genes contained phytohormone-, light-, and abiotic stress-responsive elements. The MiDi19-4 genes were expressed in every tissue and highly expressed in leaves. Moreover, MiDi19-4 genes were highly correlated with the vegetative growth period and induced by polyethylene glycol (PEG) or salt stress. MiDi19-4B displayed the highest expression during the vegetative growth period and then showed decreased expression, and MiDi19-4B was highly expressed at both the late stage of the vegetative growth period and the initial stage of the flowering induction period. The 35S::GFP–MiDi19-4B fusion protein was located in the cell nucleus. The transgenic plants ectopically expressing MiDi19-4B exhibited earlier flowering and increased expression patterns of FRUITFULL (AtFUL), APETALA1 (AtAP1), and FLOWERING LOCUS T (AtFT). The drought and salt tolerance of MiDi19-4B transgenic plants was significantly increased, and these plants showed decreased sensitivity to abscisic acid (ABA) and considerably increased expression levels of drought- and salt-related genes and ABA signalling pathway genes. Additionally, bimolecular fluorescence complementation (BiFC) experiments revealed that the MiDi19-4B protein interacted with CAULIFLOWER (MiCAL1), MiCAL2, MiAP1-1, and MiAP1-2. Taken together, these results highlighted the important regulatory roles of MiDi19-4B in tolerance to multiple abiotic stresses and in flowering.

The SQUAMOSA promoter binding protein-like (SPL) gene family is a unique family of plant-specific transcription factors (TFs), which plays vital roles in a variety of plant biological processes. Its role in betalain biosynthesis in Hylocereus undantus; however, is still unclear. Here, we report a total of 16 HuSPL genes from the pitaya genome, which were unevenly distributed among nine chromosomes. The HuSPL genes were clustered into seven groups, and most HuSPLs within the same group shared similar exon-intron structures and conserved motifs. Eight segment replication events in the HuSPL gene family were the main driving force behind the gene family expansion. Nine of the HuSPL genes had potential target sites for Hmo-miR156/157b. Hmo-miR156/157b-targeted HuSPLs exhibited differential expression patterns compared with constitutive expression patterns of most Hmo-miR156/157b-nontargeted HuSPLs. The expression of Hmo-miR156/157b gradually increased during fruit maturation, while the expression of Hmo-miR156/157b-targeted HuSPL5/11/14 gradually decreased. In addition, the lowest expression level of Hmo-miR156/157b-targeted HuSPL12 was detected 23rd day after flowering, when the middle pulps started to turn red. HuSPL5, HuSPL11, HuSPL12, and HuSPL14 were nucleus-localized proteins. HuSPL12 could inhibit the expression of HuWRKY40 by binding to its promoter. Results from yeast two-hybrid and bimolecular fluorescence complementation assays showed that HuSPL12 could interact with HuMYB1, HuMYB132, or HuWRKY42 TFs responsible for betalain biosynthesis. The results of the present study provide an essential basis for future regulation of betalain accumulation in pitaya.

SHORT VEGETATIVE PHASE (SVP) belongs to the MADS-box gene family, its main function is to regulate plant flowering, maintain normal flower development and bud dormancy. However, its function in mango (Mangifera indica L.) flowering needs further study. In this research, we obtained two SVP genes, MiSVP3 and MiSVP4, from genomic and transcriptomic data of M. indica L. cv. 'SiJiMi'. Sequence analysis showed that MiSVP3 and MiSVP4 were typical MADS-box transcription factors. Promoter element analysis suggested that MiSVP3 and MiSVP4 were associated with several hormone responses, stress responses and CArG box cis-elements. Expression analysis indicated that MiSVP3 and MiSVP4 were lowly expressed in flower/buds and highly expressed in vegetative tissue. Subcellular localization indicated that MiSVP3 and MiSVP4 were localized in the nucleus. Overexpression of MiSVP3 and MiSVP4 delays flowering time and results in abnormalities in inflorescences and floral organs in Arabidopsis. MiSVP3 was sensitive only to prohexadione-calcium (Pro-Ca) treatment, while MiSVP4 was sensitive to abscisic acid (ABA), gibberellin (GA3) and Pro-Ca treatment. The Y2H and BiFC assays showed that MiSVP3 interacted with MiSEP1-1 and MiSOC1D but not MiFLC, MiAP1-1 and MiAP1-2, MiSVP4 interacted with MiFLC, MiAP1-1, MiAP1-2 and MiSOC1D but not MiSEP1-1. The Y1H assay showed that MiSVP3 and MiSVP4 could interact with MiFLC, but only MiSVP3 could interact with MiSOC1D. These results suggest that MiSVP3 and MiSVP4 can inhibit mango flowering and participate in the regulation of mango flower formation gene network.

BACKGROUND:Multiple MYB transcription factors (TFs) are involved in the regulation of plant coloring. Betalain is a kind of natural plant pigment and its biosynthesis is regulated by a number of enzymes. Despite this, little is known about the molecular properties and roles of MYB TFs in pitaya betalain biosynthesis.RESULTS:In the present study, we identified a 1R-MYB gene, HuMYB132, which is preferentially expressed in red-pulp pitaya at the mature stage. It was clustered with Arabidopsis R-R-type genes and had two DNA-binding domains and a histidine-rich region. The expression assays in N. benthamiana and yeast indicated that HuMYB132 is a nucleus-localized protein with transcriptional activation activity. Dual luciferase reporter assay and electrophoretic mobility shift assays (EMSA) demonstrated that HuMYB132 could promote the transcriptional activities of HuADH1, HuCYP76AD1-1, and HuDODA1 by binding to their promoters. Silencing HuMYB132 reduced betalain accumulation and the expression levels of betalain biosynthetic genes in pitaya pulps.CONCLUSIONS:According to our findings, HuMYB132, a R-R type member of 1R-MYB TF subfamily, positively regulates pitaya betalain biosynthesis by regulating the expression of HuADH1, HuCYP76AD1-1, and HuDODA1. The present study provides a new theoretical reference for the management of pitaya betalain biosynthesis and also provides an essential basis for future regulation of betalain biosynthesis in Hylocereus.

Basic helix-loop-helix (bHLH) proteins are dimeric transcription factors (TFs) involved in various plant physi-ological and biological processes. Despite this, little is known about the molecular properties and roles of bHLH TFs in pitaya betalain biosynthesis. Here we report the identification of 165 HubHLH genes in H. undantus genome, their chromosomal distribution, physiochemical characteristics, conserved motifs, gene structure, phylogeny and synteny of HubHLH genes. Based on phylogenetic relationship analysis, the 165 HubHLHs were divided into 26 subfamilies and unequally distributed on the 11 chromosomes of pitaya. Based on the pitaya transcriptome data, a candidate gene HubHLH159 was obtained, and the real-time quantitative PCR analysis confirmed that HubHLH159 showed a high expression level in 'Guanhuahong' pitaya (red-pulp) at mature stage, indicating its role in betalain biosynthesis. HubHLH159 is a Group II protein and contains a bHLH domain. It is a nuclear protein with transcriptional activation activity. Dual luciferase reporter assays and virus-induced gene silencing (VIGS) experiments showed that HubHLH159 promotes betalain biosynthesis by activating the expression of HuADH1, HuCYP76AD1-1, and HuDODA1. The results of the present study lay a new theoretical reference for the regulation of pitaya betalain biosynthesis and also provides as essential basis for the future analysis of the functions of HubHLH gene family.

The WRKY gene family is a plant-specific transcription factor (TF) that regulates many physiological processes and (a) biotic stress responses. Despite this, little is known about the molecular properties and roles of WRKY TFs in pitaya betalain biosynthesis. Here we report the identification of 70 WRKY in Hylocereus undatus, their gene structure, locations on each chromosome, systematic phylogenetic analysis, conserved motif analysis, and synteny of HuWRKY genes. HmoWRKY42 is a Group IIb WRKY protein and contains a coiled-coil motif, a WRKY domain and a C2H2 zinc-finger motif (CX5CX23HXH). Results from yeast one-hybrid and transient dual-luciferase assays showed that HmoWRKY42 was a transcriptional repressor and could repress HmocDOPA5GT1 expression by binding to its promoter. Yeast two-hybrid assays showed that HmoWRKY42 could interact with itself to form homodimers. Knocking out the coiled-coil motif of HmoWRKY42 prevented its self-interaction and prevented it from binding to the HmocDOPA5GT1 promoter. Knocking out the WRKY domain and C2H2 zinc-finger motif sequence of HmoWRKY42 also prevented it from binding to the HmocDOPA5GT1 promoter. The coiled-coil motif, the WRKY domain and the C2H2 zinc finger motif are key motifs for the binding of HmoWRKY42 to the HmocDOPA5GT1 promoter. HmoWRKY42 is localized in the nucleus and possesses trans-activation ability responsible for pitaya betalain biosynthesis by repressing the transcription of HmocDOPA5GT1. As far as we know, no reports are available on the role of HmoWRKY42 in pitaya betalain biosynthesis. The results provide an important foundation for future analyses of the regulation and functions of the HuWRKY gene family.

Sugar and organic acids are important factors determining pitaya fruit quality. However, changes in sugars and acids, and expressions of metabolism-associated genes during fruit maturation of yellow-peel pitayas are not well-documented. In this study, metabolic and expression analyses in pulps of different fruit developmental stages of 'Wucihuanglong' ('WCHL', Hylocereus undatus) and 'Youcihuanglong' pitaya ('YCHL', Hylocereus megalanthus) were used to explore the sugar and organic acid metabolic process. Total phenols and flavonoids were mainly accumulated at S1 in pitaya pulps. Ascorbic acid contents of 'WCHL' pitaya were higher than that of 'YCHL' pitaya during fruit maturation. Starch was mainly accumulated at early fruit development stages while soluble sugars were rich in late stages. Sucrose, fructose, and glucose were the main sugar components of 'YCHL' pitaya while glucose was dominant in 'WCHL' pitaya. Malic and citric acids were the main organic acids in 'WCHL' and 'YCHL' pitayas, respectively. Based on the transcriptome analyses, 118 genes involved in pitaya sugar and organic acid metabolism were obtained. Results from the correlation analyses between the expression profiling of candidate genes and the contents of sugar and organic acid showed that 51 genes had a significant correlation relationship and probably perform key role in pitaya sugar and organic acid metabolism processes. The finding of the present study provides new information for quality regulation of pitayas.

NAC transcription factors are one of the largest families of transcriptional regulators in plants, and members of the gene family play vital roles in regulating plant growth and development processes including biotic/abiotic stress responses. However, little information is available about the NAC family in pitaya. In this study, we conducted a genome-wide analysis and a total of 64 NACs (named HuNAC1-HuNAC64) were identified in pitaya (Hylocereus). These genes were grouped into fifteen subgroups with diversities in gene proportions, exon–intron structures, and conserved motifs. Genome mapping analysis revealed that HuNAC genes were unevenly scattered on all eleven chromosomes. Synteny analysis indicated that the segmental duplication events played key roles in the expansion of the pitaya NAC gene family. Expression levels of these HuNAC genes were analyzed under cold treatments using qRT-PCR. Four HuNAC genes, i.e., HuNAC7, HuNAC20, HuNAC25, and HuNAC30, were highly induced by cold stress. HuNAC7, HuNAC20, HuNAC25, and HuNAC30 were localized exclusively in the nucleus. HuNAC20, HuNAC25, and HuNAC30 were transcriptional activators while HuNAC7 was a transcriptional repressor. Overexpression of HuNAC20 and HuNAC25 in Arabidopsis thaliana significantly enhanced tolerance to cold stress through decreasing ion leakage, malondialdehyde (MDA), and H2O2 and O2− accumulation, accompanied by upregulating the expression of cold-responsive genes (AtRD29A, AtCOR15A, AtCOR47, and AtKIN1). This study presents comprehensive information on the understanding of the NAC gene family and provides candidate genes to breed new pitaya cultivars with tolerance to cold conditions through genetic transformation.

Pitaya (Selenicereus) is a kind of novel fruit with a delicious taste and superior horticulture ornamental value. The potential economic impact of the pitaya lies in its diverse uses not only as agricultural produce and processed foods but also in industrial and medicinal products. It is also an excellent plant material for basic and applied biological research. A comprehensive database of pitaya would facilitate studies of pitaya and the other Cactaceae plant species. Here, we constructed pitaya genome and multiomics database, which is a collection of the most updated and high-quality pitaya genomic assemblies. The database contains various information such as genomic variation, gene expression, miRNA profiles, metabolite and proteomic data from various tissues and fruit developmental stages of different pitaya cultivars. In PGMD, we also uploaded videos on the flowering process and planting tutorials for practical usage of pitaya. Overall, these valuable data provided in the PGMD will significantly facilitate future studies on population genetics, molecular breeding and function research of pitaya.

Sugar composition not only affects fruit flavor but is also an important determinant of fruit taste and consumer preference. In this study, changes in the sugar content and sugar-metabolizing enzymes were investigated from different sections of various fruit development phases of 'Hongshuijing' pitaya (Hylocereus monacanthus). Genes related to sugar metabolism were also screened by transcriptome analyses. The results indicated that glucose was the major sugar in mature pitaya fruit, and was mainly regulated by vacuolar acid invertase (VAI) and sucrose synthase (SS) (degradative direction). Sugar accumulation varied in pulp between different sections of the pitaya fruit. VAI, neutral invertase (NI) and SS (degradative direction) are crucial enzymes for sugar accumulation in pitaya. The expression of 17 genes related to sucrose metabolism obtained from seven databases [NCBI non-redundant protein database (Nr), NCBI non-redundant nucleotide sequence database (Nt), EuKaryotic Orthologous Groups (KOG), The Protein Families (Pfam), Kyoto Encyclopedia of Genes and Genomes (KEGG), Swiss-prot, and Gene Ontology (GO)] were analyzed in different pitaya pulp sections. HpVAI1 had the highest relative expression level on the 29th day after pollination (DAP). Positive correlations were found between HpVAI1 expression and VAI activity; HpNI4 and NI activity; HpSS2, HpSS5, and SS activity (synthetic direction), indicating that HpVAI1, HpNI4, and HpSS2 and HpSS5 were involved in the regulation of VAI, NI, and SS (synthetic direction), respectively. HpVAI1 and HpNI4 regulated sucrose degradation and the accumulation of glucose and fructose, while HpSS2 and HpSS5 regulated sucrose synthesis. These results suggest that HpVAI1 plays a key role in sugar metabolism during fruit development of 'Hongshuijing' pitaya. The results of this study provide new information about sugar metabolism in pitaya fruit that could help improve fruit quality and the breeding of new cultivars.

Pitayas are rich in organic acids, especially citramalic acid, which is significantly higher than the plants. However, the mechanism of citramalic acid biosynthesis remains to be fully elucidated. In this study, organic acid compositions and contents, as well as expression patterns of key genes related to organic acid metabolism were analyzed during fruit maturation of four different pitaya cultivars i.e., ‘Guanhuabai’ (GHB), ‘Guanhuahong’ (GHH), ‘Wucihuanglong’ (WCHL), and ‘Youcihuanglong’ (YCHL). The total organic acid contents increased first and then declined during fruit maturation. The main organic acids were citramalic acid during the early stages of GHB, GHH, and WCHL pitayas, and dominated by malic acid as fruit maturation. In comparison, citric acid and malic acid were main organic acid for ‘YCHL’ pitaya. Citramalate synthase (IPMS) was involved in the synthesis of citramalic acid, and three types of HuIPMS i.e., HuIPMS1, HuIPMS2, and HuIPMS3, were obtained in our study. Highest expression levels of HuIPMS1 were detected in sepals, while HuIPMS2 and HuIPMS3 exhibited preferential expression in tender stems and ovaries. The expression levels of HuIPMS2 and HuIPMS3 were positively correlated with the content of citramalic acid in the four pitaya cultivars. HuIPMS2 was a chloroplast-localized protein, while HuIPMS3 presented a cytoplasmic-like and nuclear subcellular localization. These findings provide an important basis for further understanding of the molecular mechanism that leads to citramalic acid metabolism during pitaya fruit maturation.

Flowering is an integral part of the life cycle of flowering plants, which is essential for plant survival and crop production. Most woody fruit trees such as apples and pears bloom in spring, but loquat blooms in autumn and winter. Gibberellin (GA) plays a key role in the regulation of plant flower formation. In this study, we sprayed loquat plants with exogenous GA 3 , which resulted in vigorous vegetative growth rather than floral bud formation. We then performed a comprehensive RNA-seq analysis on GA 3 -treated and control-treated leaves and buds over three time periods to observe the effects of exogenous GA 3 application on floral initiation and development. The results showed that 111 differentially expressed genes (DEGs) and 563 DEGs were down-regulated, and 151 DEGs and 506 DEGs were up-regulated in buds and leaves, respectively, upon treatment with GA 3 . Among those that are homologs of the DELLA-mediated GA signal pathway genes, some may be involved in the positive regulation of flower development, including EjWRKY75, EjFT, EjSOC1, EjAGL24, EjSPL, EjLFY, EjFUL , and EjAP1 ; while some may be involved in the negative regulation of flower development, including EjDELLA, EjMYC3, EjWRKY12 , and EjWRKY13 . Finally, by analyzing the co-expression of DEGs and key floral genes EjSOC1s, EjLFYs, EjFULs, EjAP1s , 330 candidate genes that may be involved in the regulation of loquat flowering were screened. These genes belong to 74 gene families, including Cyclin_C, Histone, Kinesin, Lipase_GDSL, MYB, P450, Pkinase, Tubulin, and ZF-HD_dimer gene families. These findings provide new insights into the regulation mechanism of loquat flowering.

Pitaya (Hylocereus) is the most economically important fleshy-fruited tree of the Cactaceae family that is grown worldwide, and it has attracted significant attention because of its betalain-abundant fruits. Nonetheless, the lack of a pitaya reference genome significantly hinders studies focused on its evolution, as well as the potential for genetic improvement of this crop. Herein, we employed various sequencing approaches, namely, PacBio-SMRT, Illumina HiSeq paired-end, 10× Genomics, and Hi-C (high-throughput chromosome conformation capture) to provide a chromosome-level genomic assembly of 'GHB' pitaya (H. undatus, 2n = 2x = 22 chromosomes). The size of the assembled pitaya genome was 1.41 Gb, with a scaffold N50 of ~127.15 Mb. In total, 27,753 protein-coding genes and 896.31 Mb of repetitive sequences in the H. undatus genome were annotated. Pitaya has undergone a WGT (whole-genome triplication), and a recent WGD (whole-genome duplication) occurred after the gamma event, which is common to the other species in Cactaceae. A total of 29,328 intact LTR-RTs (~696.45 Mb) were obtained in H. undatus, of which two significantly expanded lineages, Ty1/copia and Ty3/gypsy, were the main drivers of the expanded genome. A high-density genetic map of F1 hybrid populations of 'GHB' × 'Dahong' pitayas (H. monacanthus) and their parents were constructed, and a total of 20,872 bin markers were identified (56,380 SNPs) for 11 linkage groups. More importantly, through transcriptomic and WGCNA (weighted gene coexpression network analysis), a global view of the gene regulatory network, including structural genes and the transcription factors involved in pitaya fruit betalain biosynthesis, was presented. Our data present a valuable resource for facilitating molecular breeding programs of pitaya and shed novel light on its genomic evolution, as well as the modulation of betalain biosynthesis in edible fruits.

Betalains are water-soluble nitrogen-containing pigments with multiple bioactivities. Pitaya is the only large-scale commercially grown fruit containing abundant betalains for consumers. However, the upstream regulators in betalain biosynthesis are still not clear. In this study, HmoWRKY40, a novel WRKY transcription factor, was obtained from the transcriptome data of pitaya (Hylocereus monacanthus). HmoWRKY40 is a member of the Group IIa WRKY family, containing a conserved WRKY motif, and it is located in the nucleus. The betalain contents and expression levels of HmoWRKY40 increased rapidly during the coloration of pitaya and reached their maximums on the 23rd day after artificial pollination (DAAP). Yeast one-hybrid and transient expression assays showed that HmoWRKY40 could bind and activate the promoter of HmoCYP76AD1. Silencing the HmoWRKY40 gene resulted in a significant reduction of betacyanin contents. These results indicate that HmoWRKY40 transcriptionally activates HmoCYP76AD, which is involved in the regulation of pitaya betalain biosynthesis. The results of the present study provide new regulatory networks related to betalain biosynthesis in pitaya.