Molecular Biology and Proteomics During Somatic Embryogenesis in Dimocarpus Longan Lour.

Lai, Zhen‐Zhen; He, Yan; Chen, Y. T.; Cai, Yanfang; Lai, Chunwang; Lin, Yuling; Lin, Xiaoying; Fang, Zhenfu

doi:10.17660/actahortic.2010.863.10

Cited by 9 publications

(7 citation statements)

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“…Somatic embryogenesis, which resembles zygotic embryogenesis, is an essential component of plant cell differentiation and embryo development. The longan SE system has been used widely as a model system to investigate in vitro and in vivo regulation of embryogenesis in plants ( Lai et al, 2010 ; Lin and Lai, 2010 ). In our previous study, we found that miR160a was barely detectable during the early stages of longan SE, but highly expressed during the heart- and torpedo-shaped embryonic stages ( Lin and Lai, 2013b ).…”

Section: Introductionmentioning

confidence: 99%

Endogenous target mimics down-regulate miR160 mediation of ARF10, -16, and -17 cleavage during somatic embryogenesis in Dimocarpus longan Lour

et al. 2015

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MicroRNA160 plays a critical role in plant development by negatively regulating the auxin response factors ARF10, -16, and -17. However, the ways in which miR160 expression is regulated at the transcriptional level, and how miR160 interacts with its targets during plant embryo development, remain unknown. Here, we studied the regulatory relationships among endogenous target mimics (eTMs), and miR160 and its targets, and their involvement in hormone signaling and somatic embryogenesis (SE) in Dimocarpus longan. We identified miR160 family members and isolated the miR160 precursor, primary transcript, and promoter. The promoter contained cis-acting elements responsive to stimuli such as light, abscisic acid, salicylic acid (SA) and heat stress. The pri-miR160 was down-regulated in response to SA but up-regulated by gibberellic acid, ethylene, and methyl jasmonate treatment, suggesting that pri-miR160 was associated with hormone transduction. Dlo-miR160a, -a∗ and -d∗ reached expression peaks in torpedo-shaped embryos, globular embryos and cotyledonary embryos, respectively, but were barely detectable in friable-embryogenic callus. This suggests that they have expression-related and functional diversity, especially during the middle and later developmental stages of SE. Four potential eTMs for miR160 were identified. Two of them, glucan endo-1,3-beta- glucosidase-like protein 2-like and calpain-type cysteine protease DEK1, were confirmed to control the corresponding dlo-miR160a∗ expression level. This suggests that they may function to abolish the binding between dlo-miR160a∗ and its targets. These two eTMs also participated in 2,4-D and ABA signal transduction. DlARF10, -16, and -17 targeting by dlo-miR160a was confirmed; their expression levels were higher in friable-embryogenic callus and incomplete compact pro-embryogenic cultures and responded to 2,4-D, suggesting they may play a major role in the early stages of longan SE dependent on 2,4-D. The eTMs, miR160, and ARF10, -16, and -17 exhibited tissue specificity in ‘Sijimi’ longan vegetative and reproductive organs, but were not significant negatively correlated. These results provide insights into the possible role of the eTM-miR160-ARF10-16-17 pathway in longan somatic embryo development.

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Section: Introductionmentioning

confidence: 99%

Endogenous target mimics down-regulate miR160 mediation of ARF10, -16, and -17 cleavage during somatic embryogenesis in Dimocarpus longan Lour

et al. 2015

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“…However, the molecular mechanism of longan seed development remains unclear because of the extreme genetic heterozygosity exhibited and the difficulty of sampling the early embryos ( Liu et al, 2010 ; Lai and Lin, 2013 ; Lin and Lai, 2013a , b ; Lin et al, 2015 ). To avoid these difficulties, longan SE, which resembles zygotic embryogenesis, has previously been used widely as a model system to investigate in vitro and in vivo regulation of embryogenesis in plants ( Lai et al, 2010 ; Lin and Lai, 2010 ). Our previous work indicated that dlo-miR390 a.1 and -a ∗ .1 accumulated during heart- and TE stages ( Lin and Lai, 2013b ).…”

Section: Introductionmentioning

confidence: 99%

MicroRNA390-Directed TAS3 Cleavage Leads to the Production of tasiRNA-ARF3/4 During Somatic Embryogenesis in Dimocarpus longan Lour

Lin

Lai

et al. 2015

Front. Plant Sci.

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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.

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“…), a tropical/subtropical fruit tree in the family Sapindaceae, is of great economic importance in Southeast Asia. Because the status of embryo development determines seed size, fruit quality, percentage of fruit set, and yield in longan, efforts to improve fruit quality and yield have included studies on regulation of longan embryo development using cytological, molecular, and proteomics approaches [1,2]. Such research has been hampered, however, by the extremely high genetic heterozygosity of longan and early embryo sampling difficulties [3].…”

Section: Introductionmentioning

confidence: 99%

“…Because plant somatic embryogenesis (SE) shows close similarities on morphological and molecular levels to normal zygotic embryogeny [4-7], the longan SE system has been used as a system for investigating regulation of in vitro and in vivo embryogenesis in longan [8-10]. Studies focusing on molecular biology and proteomics of the longan SE system have been conducted using differential display reverse transcription PCR (DDRT-PCR), homology cloning, quantitative real-time PCR (qPCR), two-dimensional electrophoresis, and protein bio-mass spectrometry (MALDI-TOF, Q-TOF), resulting in the isolation and identification of hundreds of related genes and proteins [1]. …”

Section: Introductionmentioning

confidence: 99%

Analysis of the global transcriptome of longan (Dimocarpus longan Lour.) embryogenic callus using Illumina paired-end sequencing

Zhang

Lin

2013

BMC Genomics

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BackgroundLongan is a tropical/subtropical fruit tree of great economic importance in Southeast Asia. Progress in understanding molecular mechanisms of longan embryogenesis, which is the primary influence on fruit quality and yield, is slowed by lack of transcriptomic and genomic information. Illumina second generation sequencing, which is suitable for generating enormous numbers of transcript sequences that can be used for functional genomic analysis of longan.ResultsIn this study, a longan embryogenic callus (EC) cDNA library was sequenced using an Illumina HiSeq 2000 system. A total of 64,876,258 clean reads comprising 5.84 Gb of nucleotides were assembled into 68,925 unigenes of 448-bp mean length, with unigenes ≥1000 bp accounting for 8.26% of the total. Using BLASTx, 40,634 unigenes were found to have significant similarity with accessions in Nr and Swiss- Prot databases. Of these, 38,845 unigenes were assigned to 43 GO sub-categories and 17,118 unigenes were classified into 25 COG sub-groups. In addition, 17,306 unigenes mapped to 199 KEGG pathways, with the categories of Metabolic pathways, Plant-pathogen interaction, Biosynthesis of secondary metabolites, and Genetic information processing being well represented. Analyses of unigenes ≥1000 bp revealed 328 embryogenesis-related unigenes as well as numerous unigenes expressed in EC associated with functions of reproductive growth, such as flowering, gametophytogenesis, and fertility, and vegetative growth, such as root and shoot growth. Furthermore, 23 unigenes related to embryogenesis and reproductive and vegetative growth were validated by quantitative real time PCR (qPCR) in samples from different stages of longan somatic embryogenesis (SE); their differentially expressions in the various embryogenic cultures indicated their possible roles in longan SE.ConclusionsThe quantity and variety of expressed EC genes identified in this study is sufficient to serve as a global transcriptome dataset for longan EC and to provide more molecular resources for longan functional genomics.

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Molecular Biology and Proteomics During Somatic Embryogenesis in Dimocarpus Longan Lour.

Cited by 9 publications

References 0 publications

Endogenous target mimics down-regulate miR160 mediation of ARF10, -16, and -17 cleavage during somatic embryogenesis in Dimocarpus longan Lour

Endogenous target mimics down-regulate miR160 mediation of ARF10, -16, and -17 cleavage during somatic embryogenesis in Dimocarpus longan Lour

MicroRNA390-Directed TAS3 Cleavage Leads to the Production of tasiRNA-ARF3/4 During Somatic Embryogenesis in Dimocarpus longan Lour

Analysis of the global transcriptome of longan (Dimocarpus longan Lour.) embryogenic callus using Illumina paired-end sequencing

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