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Expression of KxhKN4 and KxhKN5 genes in Kalanchoë blossfeldiana ‘Molly’ results in novel compact plant phenotypes: towards a cisgenesis alternative to growth retardants

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Abstract

Many potted plants like Kalanchoë have an elongated natural growth habit, which has to be controlled through the application of growth regulators. These chemicals will be banned in the near future in all the EU countries. Besides their structural functions, the importance of homeotic genes to modify plant architecture appears evident. In this work, the full length cDNA of five KNOX (KN) genes were sequenced from K. x houghtonii, a viviparous hybrid. Two constructs with the coding sequence of the class I and class II homeobox KN genes, KxhKN5 and KxhKN4, respectively, were overexpressed in the commercially important ornamental Kalanchoë blossfeldiana ‘Molly’. Furthermore, a post-transcriptional gene silencing construct was made with a partial sequence of KxhKN5 and also transformed into ‘Molly’. Several transgenic plants exhibited compact phenotypes and some lines had a relative higher number of inflorescences. A positive correlation between gene expression levels and the degree of compactness was found. However, a correlation between the induced phenotypes and the number of inserted copies of the transgene were not observed, although line ‘70-10’ with a high copy number also had the highest expression level. Moreover, overexpression of KxhKN4 resulted in plants with dark green leaves due to an elevated content of chlorophyll, a highly desired property in the ornamental plant industry. These transgenic plants show that a cisgenesis approach towards production of compact plants with improved quality as an alternative to chemical growth retardants may be feasible.

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Acknowledgments

Part of the work described was funded under the Danish Ministry of Food, Agriculture and Fisheries (DFFE) and by the Italian Ministry of Agriculture, Food and Forestry in the framework of the AGRONANOTECH project. ML, CB and AA thank G. Morreale for his contribution to preliminary steps of gene cloning and C. Regis for sharing cloning of KxhKN4 and KxhKN5. Sille Høgly Petersen, Karen Rysbjerg Munk and Hanne Hasselager are acknowledged for technical assistance.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Agriculture and Ecology, Faculty of Life Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark

    Henrik Lütken, Marian Ørgaard & Søren K. Rasmussen

  2. CRA, Unit of Research for Floriculture, Corso Inglesi 508, 18038, San Remo, Italy

    Marina Laura, Cristina Borghi & Andrea Allavena

Authors
  1. Henrik Lütken
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  2. Marina Laura
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  3. Cristina Borghi
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  4. Marian Ørgaard
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  5. Andrea Allavena
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  6. Søren K. Rasmussen
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Correspondence to Henrik Lütken.

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Communicated by P. Kumar.

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299_2011_1132_MOESM1_ESM.doc

Online Resource 1. Species of origin, accession number and brief description of selected class I KNOX homeodomain proteins used for the phylogenetic analysis reported in Fig. 1. To improve results, sequence of Physcomitrella patens (fern) and Ceratopteris richardii (moss) were added; the homeodomain protein of Acetabularia acetabulum (algae) was used as out group. (DOC 70 kb)

299_2011_1132_MOESM2_ESM.doc

Online Resource 2. Species of origin, accession number and brief description of selected class II KNOX homeodomain proteins used for the phylogenetic analysis reported in Fig. 2. To improve results, sequence of Physcomitrella patens (fern), Ceratopteris richardii (moss) and Selaginella kraussiana (clubmoss) were added; the homeodomain protein of Acetabularia acetabulum (algae) was used as out group. (DOC 56 kb)

Online Resource 3. Representative chromosome pictures for selected KN lines (jpg 61 kb)

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Lütken, H., Laura, M., Borghi, C. et al. Expression of KxhKN4 and KxhKN5 genes in Kalanchoë blossfeldiana ‘Molly’ results in novel compact plant phenotypes: towards a cisgenesis alternative to growth retardants. Plant Cell Rep 30, 2267–2279 (2011). https://doi.org/10.1007/s00299-011-1132-9

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