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Strigolactones (SLs), a new class of plant hormones, are implicated in the regulation of various biological processes. However, the related family members and functions are not identified in longan (Dimocarpus longan Lour.). In this study, 23 genes in the CCD, D27, and SMXL family were identified in the longan genome. The phylogenetic relationships, gene structure, conserved motifs, promoter elements, and transcription factor-binding site predictions were comprehensively analysed. The expression profiles indicated that these genes may play important roles in longan organ development and abiotic stress responses, especially during early somatic embryogenesis (SE). Furthermore, GR24 (synthetic SL analogue) and Tis108 (SL biosynthesis inhibitor) could affect longan early SE by regulating the levels of endogenous IAA (indole-3-acetic acid), JA (jasmonic acid), GA (gibberellin), and ABA (abscisic acid). Overexpression of SMXL6 resulted in inhibition of longan SE by regulating the synthesis of SLs, carotenoids, and IAA levels. This study establishes a foundation for further investigation of SL genes and provides novel insights into their biological functions.

ispartof: EUROPEAN JOURNAL OF HORTICULTURAL SCIENCE vol:84 issue:4 pages:226-236 status: published

Longan (Dimocarpus longan L.) is one of the most important tropical and subtropical fruits in the world. Longan fruit has high nutritional and medical value, and is regarded as a treasure among fruits. Since it was first reported that potassium chlorate (KClO3) could be successfully applied to promote flowering in longan, this compound has been widely used in the production of on-season and off-season longan fruits. KClO3 has thus played a great role in promoting the development of the longan industry. In this review, we summarize the application methods, influencing factors, and physiological and molecular mechanisms associated with KClO3-mediated induction of longan flowering. It can be deduced that leaves may play a crucial role in the transport of and response to KClO3. Leaves supply carbon and nitrogen nutrition, and hormone and signaling molecules needed for the differentiation of apical buds. Moreover, cytokinins may be crucial for KClO3-mediated induction of longan flowering. More effort should be focused on studying the molecular mechanisms underlying this process. This will not only help us to better understand floral induction by KClO3 in longan but also enrich our understanding of flowering regulation mechanisms in woody plants.

Longan (Dimocarpus longan) is a typical southern subtropical fruit tree species that is sensitive to cold stress. C-repeat binding factors (CBFs), as transcription factors, are crucial components involved in the molecular regulation of the plant response to cold stress. However, the role of CBF homologs in the cold response regulation of longan remains largely unknown. Here, three novel CBF genes, DlCBF1, DlCBF2, and DlCBF3, were cloned from longan. DlCBF1 and DlCBF2 contain an AP2 domain and PKKPAGR and DSAWR CBF signature motifs, while DlCBF3 has mutations within these conserved signature motifs. DlCBF1/2/3 were mainly localized in the nucleus and specifically bound to CRT/DRE cis-elements, resulting in strong transcriptional activation. DlCBF1/2 exhibited tissue expression specificity, and their expression was induced by low temperature, while DlCBF3 had no tissue specificity and barely responded to low temperature. DlCBF1, DlCBF2, and DlCBF3 overexpression in Arabidopsis-enhanced cold tolerance by increasing proline accumulation and reducing reactive oxygen species (ROS) content, accompanied by upregulated expression of cold-responsive genes (AtRD29A, AtCOR15A, AtCOR47, and AtKIN1) in the CBF cold stress response signaling pathway. In conclusion, the biological functions of DlCBF1/2/3 were somewhat conserved, but slow expression of DlCBF1/2 and low expression of DlCBF3 may partly cause the cold sensitivity of longan. Collectively, these results indicated that differences exist in the expression and function of CBF orthologs in the cold-sensitive plant species longan, and these findings may help to improve the understanding of the cold response regulation mechanism and provide important theoretical support for cold-tolerant breeding of longan.

Longan (Dimocarpus longan) is a typical southern subtropical fruit tree species that is sensitive to cold stress. C-repeat binding factors (CBFs), as transcription factors, are crucial components involved in the molecular regulation of the plant response to cold stress. However, the role of CBF homologs in the cold response regulation of longan remains largely unknown. Here, three novel CBF genes, DlCBF1, DlCBF2, and DlCBF3, were cloned from longan. DlCBF1 and DlCBF2 contain an AP2 domain and PKKPAGR and DSAWR CBF signature motifs, while DlCBF3 has mutations within these conserved signature motifs. DlCBF1/2/3 were mainly localized in the nucleus and specifically bound to CRT/DRE cis-elements, resulting in strong transcriptional activation. DlCBF1/2 exhibited tissue expression specificity, and their expression was induced by low temperature, while DlCBF3 had no tissue specificity and barely responded to low temperature. DlCBF1, DlCBF2, and DlCBF3 overexpression in Arabidopsis-enhanced cold tolerance by increasing proline accumulation and reducing reactive oxygen species (ROS) content, accompanied by upregulated expression of cold-responsive genes (AtRD29A, AtCOR15A, AtCOR47, and AtKIN1) in the CBF cold stress response signaling pathway. In conclusion, the biological functions of DlCBF1/2/3 were somewhat conserved, but slow expression of DlCBF1/2 and low expression of DlCBF3 may partly cause the cold sensitivity of longan. Collectively, these results indicated that differences exist in the expression and function of CBF orthologs in the cold-sensitive plant species longan, and these findings may help to improve the understanding of the cold response regulation mechanism and provide important theoretical support for cold-tolerant breeding of longan.

Carbohydrates are nutrients and important signal molecules in higher plants. Sugar transporters (ST) play important role not only in long-distance transport of sugar, but also in sugar accumulations in sink cells. Longan (Dimocarpus longan L.) is one of the most important commercial tropical/subtropical evergreen fruit species in Southeast Asia. In this study, a total of 52 longan sugar transporter (DlST) genes were identified and they were divided into eight clades according to phylogenetic analysis. Out of these 52 DlST genes, many plant hormones (e.g., MeJA and gibberellin), abiotic (e.g., cold and drought), and biotic stress responsive element exist in their promoter region. Gene structure analysis exhibited that each of the clades have closely associated gene architectural features based on similar number or length of exons. The numbers of DlSTs, which exhibited alternative splicing (AS) events, in flower bud is more than that in other tissues. Expression profile analysis revealed that ten DlST members may regulate longan flowerbud differentiation. In silico expression profiles in nine longan organs indicated that some DlST genes were tissue specificity and further qRT-PCR analysis suggested that the transcript level of seven DlSTs (DlINT3, DlpGlcT1, DlpGlcT2, DlPLT4, DlSTP1, DlVGT1 and DlVGT2) was consistent with sugar accumulation in fruit, indicating that they might be involved in sugar accumulations during longan fruit development. Our findings will contribute to a better understanding of sugar transporters in woody plant.

Background The APETALA2/ethylene responsive factor (AP2/ERF) superfamily members are transcription factors that regulate diverse developmental processes and stress responses in plants. They have been identified in many plants. However, little is known about the AP2/ERF superfamily in longan (Dimocarpus longan Lour.), which is an important tropical/subtropical evergreen fruit tree that produces a variety of bioactive compounds with rich nutritional and medicinal value. We conducted a genome-wide analysis of the AP2/ERF superfamily and its roles in somatic embryogenesis (SE) and developmental processes in longan. Results A genome-wide survey of the AP2/ERF superfamily was carried out to discover its evolution and function in longan. We identified 125 longan AP2/ERF genes and classified them into the ERF (101 members), AP2 (19 members), RAV (four members) families, and one Soloist. The AP2 and Soloist genes contained one to ten introns, whereas 87 genes in the ERF and RAV families had no introns. Hormone signaling molecules such as methyl jasmonate (MeJA), abscisic acid (ABA), gibberellin, auxin, and salicylic acid (SA), and stress response cis-acting element low-temperature (55) and defense (49) boxes also were identified. We detected diverse single nucleotide polymorphisms (SNPs) between the ‘Hong He Zi’ (HHZ) and ‘SI JI MI’ (SJM) cultivars. The number of insertions and deletions (InDels) was far fewer than SNPs. The AP2 family members exhibited more alternative splicing (AS) events in different developmental processes of longan than members of the other families. Expression pattern analysis revealed that some AP2/ERF members regulated early SE and developmental processes in longan seed, root, and flower, and responded to exogenous hormones such as MeJA, SA, and ABA, and 2,4-D, a synthetic auxin. Protein interaction predictions indicated that the Baby Boom (BBM) transcription factor, which was up-regulated at the transcriptional level in early SE, may interact with the LALF/AGL15 network. Conclusions The comprehensive analysis of molecular evolution and expression patterns suggested that the AP2/ERF superfamily may plays an important role in longan, especially in early SE, and in seed, root, flower, and young fruit. This systematic analysis provides a foundation for further functional characterization of the AP2/ERF superfamily with the aim of longan improvement.

BackgroundSomatic embryogenesis (SE) is a process of somatic cells that dedifferentiate to totipotent embryonic stem cells and generate embryos in vitro. Longan SE has been established and wildly used as model system for studying embryogenesis in woody plants, SE-related genes had been characterized. In spite of that, a comprehensive overview of SE at a molecular level is still absent. To understand the molecular mechanisms during longan SE, we examined the transcriptome changes by using Illumina HiSeq from the four distinct developmental stages, including non-embryogenic callus (NEC), embryogenic callus (EC), incomplete compact pro-embryogenic cultures (ICpEC), globular embryos (GE).ResultsRNA-seq of the four samples generated a total of 243.78 million high quality reads, approximately 81.5% of the data were mapped to longan genome. The cDNA libraries of NEC, EC, ICpEC and GE, generated 22,743, 19,745, 21,144, 21,102 expressed transcripts, 1935, 1710, 1816, 1732 novel transcripts, 2645, 366, 505, 588 unique genes, respectively. Comparative transcriptome analysis showed that a total of 10,642, 4180, 5846 and 1785 genes were differentially expressed in the pairwise comparisons of NEC_vs_EC, EC_vs_ICpEC, EC_vs_GE, ICpEC_vs_GE, respectively. Among them, plant hormones signalling related genes were significantly enriched, especially the auxin and cytokinin signalling components. The transcripts of flavonoid biosynthesis related genes were mainly expressed in NEC, while fatty acid biosynthesis related genes mainly accumulated in early SE. In addition, the extracelluar protein encoding genesLTP,CHI,GLP,AGP,EP1were related to longan SE. Combined with the FPKM value of longan nine tissues transcription, 27 SE specific or preferential genes (LEC1,LEC1-like,PDF1.3,GH3.6,AGL80,PIN1,BBM,WOX9,WOX2,ABI3, et al.) and 28 NEC preferential genes (LEA5,CNOT3,DC2.15,PR1–1,NsLTP2,DIR1,PIP1,PIP2.1,TIP2–1,POD-P7andPOD5et al.) were characterized as molecular markers for longan early SE. qRT-PCR validation of SE-related genes showed a high correlation between RNA-seq and qRT-PCR data.ConclusionThis study provides new insights into the role of the transcriptome during early SE in longan. Differentially expressed genes reveal that plant hormones signalling, flavonoid and fatty acid biosynthesis, and extracelluar protein related genes were involved in longan early SE. It could serve as a valuable platform resource for further functional studies addressing embryogenesis in woody plants.

Longan (Dimocarpus longan Lour.), as a kind of commercial fruit tree in the family Sapindaceae, is widely cultivated in South Asia. In this study, we obtained the complete chloroplast genome sequence of longan using Illumina paired-end sequencing. It has 160,833 bp in length, containing a pair of IR regions (28,428 bp) separated by a small single-copy region (18,270 bp) and a large single-copy region (85,707 bp). The overall GC contents of the chloroplast genome were 37.8%. This circular genome contains 130 annotated genes, including 85 protein-coding genes, 37 tRNAs and 8 rRNAs. The phylogenetic analysis using maximum–likelihood (ML) and neighbour-joining (NJ) method showed that longan has the closest relationship with Litchi chinensis, Sapindus mukorossi and Dodonaea viscosa. This complete chloroplast genomes can be subsequently used for the genetic breeding of this valuable species.

Carbohydrates are nutrients and important signal molecules in higher plants. Sugar transporters (ST) play important role not only in long-distance transport of sugar, but also in sugar accumulations in sink cells. Longan (Dimocarpus longan L.) is one of the most important commercial tropical/subtropical evergreen fruit species in Southeast Asia. In this study, a total of 52 longan sugar transporter (DlST) genes were identified and they were divided into eight clades according to phylogenetic analysis. Out of these 52 DlST genes, many plant hormones (e.g., MeJA and gibberellin), abiotic (e.g., cold and drought), and biotic stress responsive element exist in their promoter region. Gene structure analysis exhibited that each of the clades have closely associated gene architectural features based on similar number or length of exons. The numbers of DlSTs, which exhibited alternative splicing (AS) events, in flower bud is more than that in other tissues. Expression profile analysis revealed that ten DlST members may regulate longan flowerbud differentiation. In silico expression profiles in nine longan organs indicated that some DlST genes were tissue specificity and further qRT-PCR analysis suggested that the transcript level of seven DlSTs (DlINT3, DlpGlcT1, DlpGlcT2, DlPLT4, DlSTP1, DlVGT1 and DlVGT2) was consistent with sugar accumulation in fruit, indicating that they might be involved in sugar accumulations during longan fruit development. Our findings will contribute to a better understanding of sugar transporters in woody plant.

EnvironmentAsia, 11, 3, 161-169

To clone and examine expression profiles of DlRan genes during somatic embryogenesis in Dimocarpus longan Lour. Thirty cDNA sequences and two genomic sequences encoding DlRan proteins were isolated from longan embryogenic cultures. Structural analysis of DlRan genes revealed that the longan Ran gene family is more expanded than that of Arabidopsis. Expression analysis of DlRan genes during somatic embryogenesis uncovered a high abundance of DlRan genes in early embryogenic cultures and heart- and torpedo-shaped embryos. The expression of DlRan genes in embryogenic calli was affected by exogenous 2,4-dichlorophenoxyacetic acid treatment. DlRan is involved in 2,4-D induced somatic embryogenesis and development of somatic embryos in longan. Electronic supplementary material The online version of this article (doi:10.1186/s40064-016-1887-0) contains supplementary material, which is available to authorized users.

Trans-acting short-interfering RNAs (tasiRNAs) originate from TAS3 families through microRNA (miRNA) 390-guided cleavage of primary transcripts and target auxin response factors (ARF3/-4), which are involved in the normal development of lateral roots and flowers in plants. However, their roles in embryo development are still unclear. Here, the pathway miR390-TAS3-ARF3/-4 was identified systematically for the first time during somatic embryo development in Dimocarpus longan. We identified the miR390 primary transcript and promoter. The promoter contained cis-acting elements responsive to stimuli such as light, salicylic acid, anaerobic induction, fungal elicitor, circadian control, and heat stress. The longan TAS3 transcript, containing two miR390-binding sites, was isolated; the miR390- guided cleavage site located near the 3′ end of the TAS3 transcript was verified. Eight TAS3-tasiRNAs with the 21-nucleotides phase were found among longan small RNA data, further confirming that miR390-directed TAS3 cleavage leads to the production of tasiRNA in longan. Among them, TAS3_5′D5+ and 5′D6+ tasiRNAs were highly abundant, and verified to target ARF3 and -4, implying that miR390-guided TAS3 cleavage with 21-nucleotides phase leading to the production of tasiRNA-ARF is conserved in plants. Pri-miR390 was highly expressed in friable-embryogenic callus (EC), and less expressed in incomplete compact pro-embryogenic cultures, while miR390 showed its lowest expression in EC and highest expression in torpedo-shaped embryos (TEs). DlTAS3 and DlARF4 both exhibited their lowest expressions in EC, and reached their peaks in the globular embryos stage, which were mainly inversely proportional to the expression of miR390, especially at the globular embryos to cotyledonary embryos (CEs) stages. While DlARF3 showed little variation from the EC to TEs stages, and exhibited its lowest expression in the CEs stage. There was a general lack of correlation between the expressions of DlARF3 and miR390. In addition, pri-miR390, DlTAS3, DlARF3 and -4 were up-regulated by 2,4-D in a concentration-dependent manner. They were also preferentially expressed in roots, pulp, and seeds of ‘Sijimi’ longan, implying their extended roles in the development of longan roots and fruit. This study provided insights into a possible role of miR390-tasiRNAs-ARF in plant somatic embryo development.

22 Pags.- 1 Tabl.- 4 Figs. The definitive version is available at: http://www.sciencedirect.com/science/journal/03044238, Longan (Dimocarpus longan) is an important evergreen fruit crop grown in tropical and subtropical climates, with a clear expanding potential, but with a poorly described phenology. In this work, the different phenological growth stages of longan are characterized according to the BBCH (Biologische Bundesanstalt, Bundessortenamt und Chemische industrie) scale. From vegetative bud dormancy to fruit harvest, seven main growth stages are described for bud, leaf and shoot development, inflorescence emergence, flowering, fruit development and fruit maturity. Within them 41 secondary growth stages are described. The BBCH code will be an important tool to assist the development and implementation of longan management protocols and to standardize observations made in different edaphoclimatic and experimental conditions., This work was supported by the Spanish Ministerio de Economía y Competitividad—European Regional Development Fund, European Union (no. AGL2013-43732-R, and AGL2012-40239), Gobierno de Aragon A-43, and Junta de Andalucía (no. FEDERP08-AGR-3694). VTP was supported by a PhD scholarship in the frame of an agreement between the Consejo Superior de Investigaciones Científicas (CSIC), Spain and the Vietnamese Academy of Science and Technology (VAST).

6 Pags.- 5 Figs., Longan (Dimocarpus longan) is a subtropical fruit tree with a long cultivation tradition in South East Asia, and also with a clear potential in other areas of the world. In other plant species, the reproductive phase is most vulnerable to temperature conditions, but information on the effect of temperature on pollen germination and tube growth in longan is still lacking. In order to fill this gap, in this work the effect of temperature on longan pollen germination was assessed both in vivo and in vitro. For this purpose first pollen germination was evaluated in vivo at three temperature regimes, showing the best performance at 23/24 °C. Then an in vitro pollen germination medium was optimized for longan and pollen storage conditions were evaluated, showing good pollen germination in vitro after storage for one year at −20 °C. Finally, the effect of temperature was evaluated in vitro in four cultivars and for two male flower types M1 and M2, differing on the degree of pistil differentiation. Both pollen types behave similarly in all conditions, and all the cultivars studied had good pollen germination in vitro at 30 °C, a temperature higher than that reported as optimum in vivo reflecting a higher sensitivity of the stigma than pollen to high temperatures. However, clear differences between cultivars in their performance at 20 °C and 40 °C were recorded. These results set the basis for reproductive biology studies and breeding in longan, opening a window to select the best cultivars for different temperature conditions., This work was supported by the Spanish Ministerio de Economía y Competitividad—European Regional Development Fund, European Union (AGL2013-43732-R, and AGL2012-40239 and Recupera2020) and Gobierno de Aragón A-43. VTP was supported by a Ph.D. scholarship from the Consejo Superior de Investigaciones Científicas (CSIC), Spaings and Vietnam Academy of Science and Technology (VAST).

Dimocarpus longan Lour. subsp. longan GRIN Nomen number: 403166 Family: Sapindaceae Common Name: longan (English). Native: ASIA-TROPICAL - Indo-China: Cambodia; Laos; Thailand; Malesia: Indonesia; Malaysia. Field Infestation: Takeishi 1992 (“ Euphoria longan ”): Two fruits, confiscated at Narita Airport (Tokyo, Japan) from airline passengers from Thailand, were infested with B. latifrons., Published as part of McQuate, Grant T. & Liquido, Nicanor J., 2013, Annotated World Bibliography of Host Fruits of Bactrocera latifrons (Hendel) (Diptera: Tephritidae), pp. 1-61 in Insecta Mundi 2013 (289) on page 15, DOI: 10.5281/zenodo.5175793, {"references":["Takeishi, H. 1992. A study on the fruit flies (Diptera: Tephritidae) found in the fresh fruits carried by passengers from Thailand to Narita airport, Japan. Research Bulletin of the Plant Protection Service, Japan 28: 75 - 78."]}