GhLBDs Promote Callus Initiation and Act as Selectable Markers to Increase Transformation Efficiency

Detached organs or differentiated tissues could form a mass of pluripotent cells termed as callus on an auxin-rich medium, the underlying molecular mechanism of which remains elusive in cotton. LATERAL ORGAN BOUNDARIES DOMAIN (LBD) transcription factor is a key regulator of plant cell totipotency/pluripotency, and a number of cotton GhLBDs with high-level differential expression during the callus induction process have been identified. Their overexpression in cotton calli fostered promotions in and callus induction without exogenous auxin. Expression analysis and histological observation using paraffin sectioning suggested that the first 72 h on culture is a key time point for callus initiation, whereby the GhLBDs showed high transcript abundance and enlarged calli that were rapidly developed from procambium and cambium. GhLBDs’ expression level could be precisely modulated by the gradient concentrations of exogenous auxin, whereas auxin transport inhibitor 2,3,5-triiodobenzoic acid could severely inhibit its expression. The LBD-mediated callus formation was also dependent on the expression levels of GhLBDs. Further, a β-estradiol-inducible promoter pER8 was used to drive GhLBD18-1 expression, which led to rapid callus proliferation, suggesting that pER8/GhLBD18-1 could be used as a selectable marker system to replace the existing antibiotic/herbicide-resistance selectable markers in plant transformation. Our study provides new insights for callus initiation regulatory mechanism and strategies for improving transformation efficiency in cotton.

[1]  W. Gruissem,et al.  Current progress and challenges in crop genetic transformation. , 2021, Journal of plant physiology.

[2]  Jun Xiao,et al.  LEAFY is a pioneer transcription factor and licenses cell reprogramming to floral fate , 2020, Nature Communications.

[3]  H. Cai,et al.  A common wild rice-derived BOC1 allele reduces callus browning in indica rice transformation , 2020, Nature Communications.

[4]  Xueyan Zhang,et al.  Extensive intraspecific gene order and gene structural variations in upland cotton cultivars , 2019, Nature Communications.

[5]  Y. Zou,et al.  Temporal and spatial expression of fgfbp genes in zebrafish. , 2018, Gene.

[6]  K. Nowak,et al.  5-Azacitidine Induces Cell Death in a Tissue Culture of Brachypodium distachyon , 2018, International journal of molecular sciences.

[7]  Yuxin Hu,et al.  The WOX11–LBD16 Pathway Promotes Pluripotency Acquisition in Callus Cells During De Novo Shoot Regeneration in Tissue Culture , 2018, Plant & cell physiology.

[8]  Yuxin Hu,et al.  Control of auxin-induced callus formation by bZIP59–LBD complex in Arabidopsis regeneration , 2018, Nature Plants.

[9]  B. Scheres,et al.  Non-canonical WOX11-mediated root branching contributes to plasticity in Arabidopsis root system architecture , 2017, Development.

[10]  Hai Huang,et al.  Divergent regeneration‐competent cells adopt a common mechanism for callus initiation in angiosperms , 2017, Regeneration.

[11]  Jungmook Kim,et al.  Dimerization in LBD16 and LBD18 Transcription Factors Is Critical for Lateral Root Formation1 , 2017, Plant Physiology.

[12]  Lin Xu,et al.  Transcription Factors WOX11/12 Directly Activate WOX5/7 to Promote Root Primordia Initiation and Organogenesis1 , 2016, Plant Physiology.

[13]  P. Testillano,et al.  Auxin Biosynthesis, Accumulation, Action and Transport are Involved in Stress-Induced Microspore Embryogenesis Initiation and Progression in Brassica napus. , 2015, Plant & cell physiology.

[14]  Lei Fang,et al.  Sequencing of allotetraploid cotton (Gossypium hirsutum L. acc. TM-1) provides a resource for fiber improvement , 2015, Nature Biotechnology.

[15]  Chao-jun Zhang,et al.  iTRAQ protein profile differential analysis between somatic globular and cotyledonary embryos reveals stress, hormone, and respiration involved in increasing plantlet regeneration of Gossypium hirsutum L. , 2015, Journal of proteome research.

[16]  Jia-bao Huang,et al.  ROP3 GTPase Contributes to Polar Auxin Transport and Auxin Responses and Is Important for Embryogenesis and Seedling Growth in Arabidopsis[C][W] , 2014, Plant Cell.

[17]  Hai Huang,et al.  WOX11 and 12 Are Involved in the First-Step Cell Fate Transition during de Novo Root Organogenesis in Arabidopsis[W] , 2014, Plant Cell.

[18]  Leonie Steinhorst,et al.  A new β-estradiol-inducible vector set that facilitates easy construction and efficient expression of transgenes reveals CBL3-dependent cytoplasm to tonoplast translocation of CIPK5. , 2013, Molecular plant.

[19]  P. Springer,et al.  Functional divergence in the Arabidopsis LOB-domain gene family , 2012, Plant signaling & behavior.

[20]  Xiyan Yang,et al.  Transcript profiling reveals complex auxin signalling pathway and transcription regulation involved in dedifferentiation and redifferentiation during somatic embryogenesis in cotton , 2012, BMC Plant Biology.

[21]  Yuxin Hu,et al.  LATERAL ORGAN BOUNDARIES DOMAIN transcription factors direct callus formation in Arabidopsis regeneration , 2012, Cell Research.

[22]  L. Burgos,et al.  Aminoglycoside antibiotics: structure, functions and effects on in vitro plant culture and genetic transformation protocols , 2010, Plant Cell Reports.

[23]  C. Forestan,et al.  ZmPIN1-Mediated Auxin Transport Is Related to Cellular Differentiation during Maize Embryogenesis and Endosperm Development1[W] , 2009, Plant Physiology.

[24]  Han Woo Lee,et al.  LBD18/ASL20 Regulates Lateral Root Formation in Combination with LBD16/ASL18 Downstream of ARF7 and ARF19 in Arabidopsis1[C][W][OA] , 2009, Plant Physiology.

[25]  L. Laurens,et al.  Pluripotency of Arabidopsis xylem pericycle underlies shoot regeneration from root and hypocotyl explants grown in vitro. , 2009, The Plant journal : for cell and molecular biology.

[26]  Jungmook Kim,et al.  LBD 18 / ASL 20 Regulates Lateral Root Formation in Combination with LBD 16 / ASL 18 Downstream of ARF 7 and ARF 19 in Arabidopsis 1 [ C ] [ W ] [ OA ] , 2009 .

[27]  S. Bridges,et al.  Comparative analysis of proteome differential regulation during cell dedifferentiation in Arabidopsis , 2008, Proteomics.

[28]  S. Howell,et al.  Developmental steps in acquiring competence for shoot development in Arabidopsis tissue culture , 2007, Planta.

[29]  T. Thorpe.  History of plant tissue culture. , 2006, Methods in molecular biology.

[30]  X. Gu,et al.  Expression divergence between duplicate genes. , 2005, Trends in genetics : TIG.

[31]  Gretchen Vogel,et al.  How Does a Single Somatic Cell Become a Whole Plant? , 2005, Science.

[32]  S. Howell,et al.  Developmental events and shoot apical meristem gene expression patterns during shoot development in Arabidopsis thaliana. , 2002, The Plant journal : for cell and molecular biology.

[33]  D. Gingerich,et al.  Global and Hormone-Induced Gene Expression Changes during Shoot Development in Arabidopsis , 2002, The Plant Cell Online.

[34]  N. Chua,et al.  Technical advance: An estrogen receptor-based transactivator XVE mediates highly inducible gene expression in transgenic plants. , 2000, The Plant journal : for cell and molecular biology.

[35]  J. Reinert,et al.  Control of Totipotency in Plant Cells growing in vitro , 1968, Nature.

[36]  C. Miller,et al.  Chemical regulation of growth and organ formation in plant tissues cultured in vitro. , 1957, Symposia of the Society for Experimental Biology.