DOTFL1 affects the floral transition in orchid Dendrobium Chao Praya Smile

2021 ◽  
Author(s):  
Yan Li ◽  
Bin Zhang ◽  
Yanwen Wang ◽  
Ximing Gong ◽  
Hao Yu
Keyword(s):  
Vegetative Growth ◽  
Apical Meristem ◽  
Functional Characterization ◽  
Ectopic Expression ◽  
Floral Transition ◽  
Inflorescence Meristem ◽  
Terminal Flower ◽  
Vegetative Traits ◽  
Carg Box

Abstract A major obstacle for orchid (Orchidaceae)breeding and production is a long juvenile phase before orchid reproductive development. The molecular basis for prolonged vegetative growth in orchids remains largely unclear despite many efforts to clarify the relevant mechanisms. In this study, we report functional characterization of Dendrobium Orchid TERMINAL FLOWER1 (DOTFL1), an ortholog of TFL1 in Arabidopsis (Arabidopsis thaliana), from the orchid Dendrobium Chao Praya Smile. DOTFL1 is highly expressed in pseudobulbs and the shoot apical meristem (SAM) before and during the floral transition, but is downregulated in inflorescence apices and open flowers. Ectopic expression of DOTFL1 rescues the early-flowering and terminal-flower phenotypes of tfl1-20 in Arabidopsis. Overexpression of DOTFL1 in Dendrobium orchids delays flowering and produces defective inflorescence meristems and flowers with vegetative traits, whereas knockdown of DOTFL1 accelerates flowering and perturbs the maintenance of the inflorescence meristem. Notably, DOTFL1 suppresses orchid flowering and associated pseudobulb formation during the floral transition. We further reveal that two orchid MADS-box transcription factors, Dendrobium Orchid SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (DOSOC1) and AGAMOUS-LIKE 24 (DOAGL24), could interact with each other and bind to the CArG-box motif at DOTFL1, implying a regulatory hierarchy similar to their counterparts in Arabidopsis. Taken together, our findings suggest that DOTFL1 promotes vegetative growth, modulates successive developmental events required for reproductive success in Dendrobium orchids, and may have evolved with a previously unknown role in controlling pseudobulb formation in the Orchidaceae family.

scholarly journals Molecular Characterization of the Interaction of Borrelia parkeri and Borrelia turicatae with Human Complement Regulators

2010 ◽  
Vol 78 (5) ◽  
pp. 2199-2208 ◽  
Author(s):  
Melanie Schott ◽  
Sonja Grosskinsky ◽  
Christiane Brenner ◽  
Peter Kraiczy ◽  
Reinhard Wallich
Keyword(s):  
Binding Capacity ◽  
Functional Characterization ◽  
Ectopic Expression ◽  
Factor H ◽  
Human Complement ◽  
Related Protein ◽  
Relapsing Fever ◽  
Extracellular Matrix Components ◽  
Complement Regulators

ABSTRACT In North America, tick-borne relapsing fever is caused by the species Borrelia hermsii, B. parkeri, and B. turicatae, which are transmitted to humans through the bite of the respective infected tick vectors. Here we describe the identification and functional characterization of a surface lipoprotein of B. parkeri, designated BpcA, that binds the human complement regulators factor H and factor H-related protein 1 and, simultaneously, the host protease plasminogen. In contrast, the homologous B. turicatae protein failed to bind human factor H and factor H-related protein 1 but retained its plasminogen binding capacity. Factor H bound to BpcA maintains its regulatory capacity to control C3b deposition and C3 convertase activity. Ectopic expression of BpcA in a serum-sensitive B. burgdorferi strain protects transformed cells from complement-mediated killing. Furthermore, bound plasminogen/plasmin endows B. parkeri and B. turicatae with the potential to degrade extracellular matrix components. These findings expand our understanding of the putative recent evolutionary separation of Borrelia parkeri and Borrelia turicatae, provide evidence that B. parkeri differs from B. turicatae in its ability to resist complement attack, and may help in understanding the pathological processes underlying tick-borne relapsing fever.

scholarly journals Characterization of the BnA10.tfl1 Gene Controls Determinate Inflorescence Trait in Brassica napus L.

Agronomy ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 722 ◽  
Author(s):  
Yongpeng Jia ◽  
Kaixiang Li ◽  
Haidong Liu ◽  
Lingxiong Zan ◽  
Dezhi Du
Keyword(s):  
Brassica Napus ◽  
Shoot Apex ◽  
Apical Meristem ◽  
Brassica Napus L ◽  
Reading Frame ◽  
Terminal Flower ◽  
Terminal Flower 1 ◽  
Oilseed Breeding ◽  
Phosphatidylethanolamine Binding Protein

Determinate inflorescences have a significant effect on the genetic improvement of rapeseed, so understanding the molecular function underlying the inflorescence trait may be beneficial to oilseed breeding. A previous study found candidate gene BnTFL1 (Terminal Flower 1) for control of the inflorescence trait on Brassica napus chromosome A10 (16,627–16,847 kb). However, little is known about the function of the BnTFL1 gene in B. napus. In this study, we firstly studied the formation of the shoot apical meristem and gene expression in indeterminate and determinate inflorescences; the results showed that the inflorescence architecture and BnA10.TFL1 expression showed significant differences in the shoot apex at the budding stage. Then, two alleles (named BnA10.TFL1 gene from indeterminate and BnA10.tfl1 gene from determinate) were cloned and sequence-analyzed; the results suggest that the open reading frame of the alleles comprises 537 bp, encodes 178 amino acids containing a conserved phosphatidylethanolamine-binding protein (PEBP) domain, and shares high similarity with Arabidopsis thaliana TFL1. To analyze the function of BnA10.TFL1, the BnA10.TFL1 gene was introduced into the determinate A. thaliana tfl1 mutant and B. napus 571 line by complementation experiment. The determinate traits were restored to indeterminate, and expression of BnA10.TFL1 was increased in the indeterminate shoot apex. These results reveal that BnA10.tfl1 is a gene controlling the determinate inflorescence trait. Moreover, the BnA10.TFL1 protein was localized to the nucleus, cytoplasm, and plasma membrane. Collectively, the results of this study help us to understand the molecular mechanism of determinate inflorescences and will provide a reliable research basis for the application of determinate inflorescences in B. napus.

scholarly journals Functional Characterization of BRASSINAZOLE-RESISTANT 1 in Panax Ginseng (PgBZR1) and Brassinosteroid Response during Storage Root Formation

2020 ◽  
Vol 21 (24) ◽  
pp. 9666
Author(s):  
Hyeona Hwang ◽  
Hwa-Yong Lee ◽  
Hojin Ryu ◽  
Hyunwoo Cho
Keyword(s):  
Panax Ginseng ◽  
Glycogen Synthase ◽  
Functional Characterization ◽  
Secondary Growth ◽  
Ectopic Expression ◽  
Floral Organ ◽  
Membrane Receptors ◽  
Storage Root ◽  
Functional Conservation

Brassinosteroids (BRs) play crucial roles in the physiology and development of plants. In the model plant Arabidopsis, BR signaling is initiated at the level of membrane receptors, BRASSINOSTEROIDS INSENSITIVE 1 (BRI1) and BRI1-ASSOCIATED RECEPTOR KINASE 1 (BAK1) complex, thus activating the transcription factors (TFs) BRASSINAZOLE RESISTANT 1/BRI1-EMS-SUPPRESSOR 1 (BZR1/BES1) to coordinate BR responsive genes. BRASSINOSTEROIDS INSENSITIVE 2 (BIN2), glycogen synthase kinase 3 (GSK3) like-kinase, negatively regulates BZR1/BES1 transcriptional activity through phosphorylation-dependent cytosolic retention and shuttling. However, it is still unknown whether this mechanism is conserved in Panax ginseng C. A. Mayer, a member of the Araliaceae family, which is a shade-tolerant perennial root crop. Despite its pharmacological and agricultural importance, the role of BR signaling in the development of P. ginseng and characterization of BR signaling components are still elusive. In this study, by utilizing the Arabidopsisbri1 mutant, we found that ectopic expression of the gain of function form of PgBZR1 (Pgbzr1-1D) restores BR deficiency. In detail, ectopic expression of Pgbzr1-1D rescues dwarfism, defects of floral organ development, and hypocotyl elongation of bri1-5, implying the functional conservation of PgBZR1 in P. ginseng. Interestingly, brassinolide (BL) and BRs biosynthesis inhibitor treatment in two-year-old P. ginseng storage root interferes with and promotes, respectively, secondary growth in terms of xylem formation. Altogether, our results provide new insight into the functional conservation and potential diversification of BR signaling and response in P. ginseng.

scholarly journals Ectopic expression and functional characterization of type III polyketide synthase mutants from Emblica officinalis Gaertn

2016 ◽  
Vol 35 (10) ◽  
pp. 2077-2090 ◽  
Author(s):  
Girija Aiswarya ◽  
Vijayanathan Mallika ◽  
Luis A. J. Mur ◽  
Eppurathu Vasudevan Soniya
Keyword(s):  
Polyketide Synthase ◽  
Functional Characterization ◽  
Ectopic Expression ◽  
Emblica Officinalis ◽  
Type Iii Polyketide Synthase ◽  
Type Iii

Anterior neurectoderm is progressively induced during gastrulation: the role of the Xenopus homeobox gene orthodenticle

Development ◽  
1995 ◽  
Vol 121 (4) ◽  
pp. 993-1004 ◽  
Author(s):  
I.L. Blitz ◽  
K.W. Cho
Keyword(s):  
Expression Patterns ◽  
Functional Characterization ◽  
Ectopic Expression ◽  
Homeobox Gene ◽  
Neural Induction ◽  
Cement Gland ◽  
Spemann's Organizer ◽  
Vertical Contact

In order to study the regional specification of neural tissue we isolated Xotx2, a Xenopus homolog of the Drosophila orthodenticle gene. Xotx2 is initially expressed in Spemann's organizer and its expression is absent in the ectoderm of early gastrulae. As gastrulation proceeds, Xotx2 expression is induced in the overlying ectoderm and this domain of expression moves anteriorly in register with underlying anterior mesoderm throughout the remainder of gastrulation. The expression pattern of Xotx2 suggests that a wave of Xotx2 expression (marking anterior neurectoderm) travels through the ectoderm of the gastrula with the movement of underlying anterior (prechordal plate) mesoderm. This expression of Xotx2 is reminiscent of the Eyal-Giladi model for neural induction. According to this model, anterior neural-inducing signals emanating from underlying anterior mesoderm transiently induce anterior neural tissues after vertical contact with the overlying ectoderm. Further patterning is achieved when the ectoderm receives caudalizing signals as it comes in contact with more posterior mesoderm during subsequent gastrulation movements. Functional characterization of the Xotx2 protein has revealed its involvement in differentiation of the anterior-most tissue, the cement gland. Ectopic expression of Xotx2 in embryos induces extra cement glands in the skin as well as inducing a cement gland marker (XAG1) in isolated animal cap ectoderm. Microinjection of RNA encoding the organizer-specific homeo-domain protein goosecoid into the ventral marginal zone results in induction of the Xotx2 gene. This result, taken in combination with the indistinguishable expression patterns of Xotx2 and goosecoid in the anterior mesoderm suggests that Xotx2 is a target of goosecoid regulation.

scholarly journals Functional Characterization of Pneumocystis carinii brl1 by Transspecies Complementation Analysis

2007 ◽  
Vol 6 (12) ◽  
pp. 2448-2452 ◽  
Author(s):  
Libera Lo Presti ◽  
Moira Cockell ◽  
Lorenzo Cerutti ◽  
Viesturs Simanis ◽  
Philippe M. Hauser
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Membrane Proteins ◽  
Nuclear Membrane ◽  
Opportunistic Infections ◽  
Functional Characterization ◽  
Pneumocystis Carinii ◽  
Ectopic Expression ◽  
Functional Complementation ◽  
Null Alleles

ABSTRACT Pneumocystis jirovecii is a fungus which causes severe opportunistic infections in immunocompromised humans. The brl1 gene of P. carinii infecting rats was identified and characterized by using bioinformatics in conjunction with functional complementation in Saccharomyces cerevisiae and Schizosaccharomyces pombe. The ectopic expression of this gene rescues null alleles of essential nuclear membrane proteins of the Brr6/Brl1 family in both yeasts.

scholarly journals Genetic tools for the stable overexpression of circular RNAs

2021 ◽  
Author(s):  
Nicol Mecozzi ◽  
Arianna Nenci ◽  
Olga Vera ◽  
Aimee Falzone ◽  
Gina M DeNicola ◽  
...  
Keyword(s):  
Mouse Model ◽  
Cultured Cells ◽  
Functional Characterization ◽  
Ectopic Expression ◽  
Circular Rnas ◽  
Genetic Tools ◽  
Mammalian Expression ◽  
Biological Studies ◽  
Genetically Engineered Mice

Circular RNAs (circRNAs) are a class of non-coding RNAs that feature a covalently closed ring structure formed through backsplicing. circRNAs are broadly expressed and contribute to biological processes through a variety of functions. Standard gain-of-function and loss-of-function approaches to study gene functions have significant limitations when studying circRNAs. Overexpression studies in particular suffer from the lack of efficient genetic tools. While mammalian expression plasmids enable transient overexpression of circRNAs in cultured cells, most cell biological studies require long-term ectopic expression. Here we report the development and characterization of genetic tools enabling stable circRNA overexpression in vitro and in vivo. We demonstrated that circRNA expression constructs can be delivered to cultured cells via transposons, whereas lentiviral vectors have limited utility for the delivery of circRNA constructs. We further showed that circRNA transposons can be supplied to mouse livers via hydrodynamic tail vein injection, resulting in ectopic circRNA expression in a hepatocellular carcinoma mouse model. Furthermore, we generated genetically engineered mice harboring circRNA expression constructs. We demonstrate that this approach enables constitutive, global circRNA overexpression as well as inducible circRNA expression directed specifically to melanocytes in a melanoma mouse model. Overall, these tools expand the genetic toolkit available for the functional characterization of circRNAs of interest.

scholarly journals Functional characterization of the ABF gene family in upland cotton (Gossypium hirsutum L.)

2017 ◽  
Author(s):  
Tyson C. C. Kerr ◽  
Haggag Abdel-Mageed ◽  
MiYoung Kang ◽  
Dakota Cryer ◽  
Randy D. Allen
Keyword(s):  
Abscisic Acid ◽  
Abiotic Stress ◽  
Gossypium Hirsutum ◽  
Water Deficit ◽  
Upland Cotton ◽  
Expression Patterns ◽  
Functional Characterization ◽  
Ectopic Expression ◽  
Differentially Expressed

AbstractThe AREB/ABF bZIP transcription factors play a pivotal role in abscisic acid-dependent abiotic stress-responsive gene expression. Despite the perennial damage and reduced productivity that result from water-deficit and unpredictable early season temperature fluctuations, these critical genes have not been previously examined in upland cotton (Gossypium hirsutum). Here, we report the isolation of the G. hirsutum ABF homologs, characterization of their expression patterns in response to abiotic stress treatments, and examination of their functions through heterologous ectopic expression in Arabidopsis. As expected for an allotetraploid, G. hirsutum ABF homologs are present in the genome as homeologous pairs. These genes are differentially expressed, both among the homologs and within the homeologous pairs, in response to exogenous abscisic acid (ABA) application, dehydration, and chilling temperatures. Furthermore, heterologous ectopic expression of many of the G. hirsutum ABF genes in Arabidopsis conferred increased tolerance to water deficit and osmotic stress, as well as cold tolerance, in a gene specific manner. These results indicate the G. hirsutum ABF homologs are functional in Arabidopsis and, as in other species, are likely to play an essential role in the abiotic stress response.HighlightThe Gossypium hirsutum ABF homeologs are differentially expressed in response to abiotic stress, and their ectopic expression in Arabidopsis can confer increased water deficit tolerance.

scholarly journals Functional Characterization of Dma1 and Dma2, the Budding Yeast Homologues of Schizosaccharomyces pombe Dma1 and Human Chfr

2004 ◽  
Vol 15 (8) ◽  
pp. 3796-3810 ◽  
Author(s):  
Roberta Fraschini ◽  
Denis Bilotta ◽  
Giovanna Lucchini ◽  
Simonetta Piatti
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Budding Yeast ◽  
Functional Characterization ◽  
Ectopic Expression ◽  
Mitotic Exit ◽  
Spindle Positioning ◽  
Septin Ring ◽  
Daughter Cells ◽  
Spindle Position

Proper transmission of genetic information requires correct assembly and positioning of the mitotic spindle, responsible for driving each set of sister chromatids to the two daughter cells, followed by cytokinesis. In case of altered spindle orientation, the spindle position checkpoint inhibits Tem1-dependent activation of the mitotic exit network (MEN), thus delaying mitotic exit and cytokinesis until errors are corrected. We report a functional analysis of two previously uncharacterized budding yeast proteins, Dma1 and Dma2, 58% identical to each other and homologous to human Chfr and Schizosaccharomyces pombe Dma1, both of which have been previously implicated in mitotic checkpoints. We show that Dma1 and Dma2 are involved in proper spindle positioning, likely regulating septin ring deposition at the bud neck. DMA2 overexpression causes defects in septin ring disassembly at the end of mitosis and in cytokinesis. The latter defects can be rescued by either eliminating the spindle position checkpoint protein Bub2 or overproducing its target, Tem1, both leading to MEN hyperactivation. In addition, dma1Δ dma2Δ cells fail to activate the spindle position checkpoint in response to the lack of dynein, whereas ectopic expression of DMA2 prevents unscheduled mitotic exit of spindle checkpoint mutants treated with microtubule-depolymerizing drugs. Although their primary functions remain to be defined, our data suggest that Dma1 and Dma2 might be required to ensure timely MEN activation in telophase.

Ectopic expression of LhMYC2 increases susceptibility to Botrytis cinerea in Arabidopsis thaliana

2020 ◽  
Author(s):  
Qi Cui ◽  
Xue Gao ◽  
Lianjuan Wang ◽  
Guixia Jia
Keyword(s):  
Functional Characterization ◽  
Ectopic Expression ◽  
Defense Responses ◽  
Gray Mold ◽  
Wild Type ◽  
Transcriptional Changes ◽  
Negative Role ◽  
Ja Signaling ◽  
Fungal Inoculation

Gray mold disease, mainly caused by Botrytis elliptica and B. cinerea, leads to severe losses in lily cut flower and bulb production. MYC2 is a critical regulator of the activation of jasmonate (JA)-mediated defense responses in plants. However, information about the lily MYC2 gene is limited. Therefore, functional characterization of MYC2 in lily, especially its role in plant immune responses, should be performed. Here, significant differences between the Botrytis-resistant Lilium hybrid ‘Sorbonne’ and the Botrytis-susceptible ‘Tresor’ were found following B. cinerea inoculation, as indicated by jasmonic acid (JA) and JA-isoleucine (JA-Ile) accumulation and related gene expression. More JA and JA-Ile were detected in ‘Sorbonne’ than in ‘Tresor’ following fungal inoculation, and higher transcript levels of JA biosynthesis genes (LhAOS, LhAOC, and LhOPR3) and a signaling gene (LhCOI1) were detected in ‘Sorbonne’ than in ‘Tresor’. In contrast, expression of the critical signaling regulator LhMYC2 was higher in ‘Tresor’ than in ‘Sorbonne’. LhMYC2 was then isolated from ‘Sorbonne’ and found to be similar to several plant MYC2 homologs that have pivotal roles in JA signaling. The expression of LhMYC2 increased significantly in response to JA and salicylic acid (SA) in ‘Sorbonne’. Ectopic expression of LhMYC2 in Arabidopsis resulted in greater susceptibility to B. cinerea than that observed in wild-type plants. This susceptibility was coupled with the transcriptional changes in SA- and JA-responsive genes. Overall, our findings indicate that LhMYC2 plays a negative role in Arabidopsis resistance to B. cinerea.

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