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      Comprehensive characterization of a floral mutant reveals the mechanism of hooked petal morphogenesis in Chrysanthemum morifolium

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          Summary

          The diversity of form of the chrysanthemum flower makes this species an ideal model for studying petal morphogenesis, but as yet, the molecular mechanisms underlying petal shape development remain largely unexplored. Here, a floral mutant, which arose as a bud sport in a plant of the variety ‘Anastasia Dark Green’, and formed straight, rather than hooked petals, was subjected to both comparative morphological analysis and transcriptome profiling. The hooked petals only became discernible during a late stage of flower development. At the late stage of ‘Anastasia Dark Green’, genes related to chloroplast, hormone metabolism, cell wall and microtubules were active, as were cell division‐promoting factors. Auxin concentration was significantly reduced, and a positive regulator of cell expansion was down‐regulated. Two types of critical candidates, boundary genes and adaxial–abaxial regulators, were identified from 7937 differentially expressed genes in pairwise comparisons, which were up‐regulated at the late stage in ‘Anastasia Dark Green’ and another two hooked varieties. Ectopic expression of a candidate abaxial gene, Cm YAB1 , in chrysanthemum led to changes in petal curvature and inflorescence morphology. Our findings provide new insights into the regulatory networks underlying chrysanthemum petal morphogenesis.

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          The no apical meristem gene of Petunia is required for pattern formation in embryos and flowers and is expressed at meristem and primordia boundaries.

          Petunia embryos carrying the no apical meristem (nam) mutation fail to develop a shoot apical meristem. Occasional shoots on nam- seedlings bear flowers that develop ten instead of five primordia in the second whorl. Double mutants with the homeotic gene green petals show that nam acts independently of organ identify in whorl 2 and now also affects primordium number in whorl 3. The nam gene was isolated by transposon tagging. The encoded protein shares a conserved N-terminal domain with several other proteins of unknown function and thus represents a novel class of proteins. Strikingly, nam mRNA accumulates in cells at the boundaries of meristems and primordia. These data indicate a role for nam in determining positions of meristems and primordia.
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            Radial patterning of Arabidopsis shoots by class III HD-ZIP and KANADI genes.

            Shoots of all land plants have a radial pattern that can be considered to have an adaxial (central)-abaxial (peripheral) polarity. In Arabidopsis, gain-of-function alleles of PHAVOLUTA and PHABULOSA, members of the class III HD-ZIP gene family, result in adaxialization of lateral organs. Conversely, loss-of-function alleles of the KANADI genes cause an adaxialization of lateral organs. Thus, the class III HD-ZIP and KANADI genes comprise a genetic system that patterns abaxial-adaxial polarity in lateral organs produced from the apical meristem. We show that gain-of-function alleles of REVOLUTA, another member of the class III HD-ZIP gene family, are characterized by adaxialized lateral organs and alterations in the radial patterning of vascular bundles in the stem. The gain-of-function phenotype can be obtained by changing only the REVOLUTA mRNA sequence and without changing the protein sequence; this finding indicates that this phenotype is likely mediated through an interference with microRNA binding. Loss of KANADI activity results in similar alterations in vascular patterning as compared to REVOLUTA gain-of-function alleles. Simultaneous loss-of-function of PHABULOSA, PHAVOLUTA, and REVOLUTA abaxializes cotyledons, abolishes the formation of the primary apical meristem, and in severe cases, eliminates bilateral symmetry; these phenotypes implicate these three genes in radial patterning of both embryonic and postembryonic growth. Based on complementary vascular and leaf phenotypes of class III HD-ZIP and KANADI mutants, we propose that a common genetic program dependent upon miRNAs governs adaxial-abaxial patterning of leaves and radial patterning of stems in the angiosperm shoot. This finding implies that a common patterning mechanism is shared between apical and vascular meristems.
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              Role of PHABULOSA and PHAVOLUTA in determining radial patterning in shoots.

              The upper side of the angiosperm leaf is specialized for efficient capture of sunlight whereas the lower side is specialized for gas exchange. In Arabidopsis, the establishment of polarity in the leaf probably requires the generation and perception of positional information along the radial (adaxial versus abaxial or central versus peripheral) dimension of the plant. This is because the future upper (adaxial) side of the leaf develops from cells closer to the centre of the shoot, whereas the future under (abaxial) side develops from cells located more peripherally. Here we implicate the Arabidopsis PHABULOSA and PHAVOLUTA genes in the perception of radial positional information in the leaf primordium. Dominant phabulosa (phb) and phavoluta (phv) mutations cause a dramatic transformation of abaxial leaf fates into adaxial leaf fates. They do so by altering the predicted sterol/lipid-binding domains of ATHB14 and ATHB9, proteins of previously unknown function that also contain DNA-binding motifs. This change probably renders the protein constitutively active, implicating this domain as a central regulator of protein function and the PHB and PHV proteins as receptors for an adaxializing signal.
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                Author and article information

                Contributors
                chenfd@njau.edu.cn
                Journal
                Plant Biotechnol J
                Plant Biotechnol. J
                10.1111/(ISSN)1467-7652
                PBI
                Plant Biotechnology Journal
                John Wiley and Sons Inc. (Hoboken )
                1467-7644
                1467-7652
                26 May 2019
                December 2019
                : 17
                : 12 ( doiID: 10.1111/pbi.v17.12 )
                : 2325-2340
                Affiliations
                [ 1 ] State Key Laboratory of Crop Genetics and Germplasm Enhancement Key Laboratory of Landscaping Ministry of Agriculture and Rural Affairs College of Horticulture Nanjing Agricultural University Nanjing China
                Author notes
                [*] [* ] Correspondence (Tel +86‐25‐84396579; fax +86‐25‐84396579; email chenfd@ 123456njau.edu.cn )
                [†]

                These authors contributed equally to this work.

                Author information
                https://orcid.org/0000-0001-9134-2353
                Article
                PBI13143
                10.1111/pbi.13143
                6835125
                31050173
                ccd86d8b-7506-4475-89a2-de51b9e4afff
                © 2019 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

                History
                : 07 February 2019
                : 25 April 2019
                : 26 April 2019
                Page count
                Figures: 10, Tables: 0, Pages: 16, Words: 10896
                Funding
                Funded by: National Science Fund for Distinguished Young Scholars
                Award ID: 31425022
                Funded by: the National Natural Science Foundation of China , open-funder-registry 10.13039/501100001809;
                Award ID: 31701959
                Funded by: the Natural Science Fund of Jiangsu Province , open-funder-registry 10.13039/501100004608;
                Award ID: BK20170717
                Funded by: the China Postdoctoral Science Foundation , open-funder-registry 10.13039/501100002858;
                Award ID: 2017M611843
                Funded by: the Fundamental Research Funds for the Central Universities
                Award ID: KJSY201705
                Award ID: KJQN201815
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                December 2019
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.7.1 mode:remove_FC converted:07.11.2019

                Biotechnology
                chrysanthemum,petal morphogenesis,adaxial–abaxial genes,boundary genes,transcriptomic analysis

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