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Factors Affecting Flower Development and Quality in Rhododendron Simsii

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Factors affecting flower development and quality in Rhododendron simsii Annelies Christiaens Promoters Prof. dr. ir. Marie-Christine Van Labeke Ghent University, Faculty of Bioscience Engineering, Department of Plant Production Dr. Bruno Gobin PCS Ornamental Plant Research Dean Prof. dr. ir. Guido Van Huylenbroeck Rector Prof. dr. Anne De Paepe Factors affecting flower development and quality in Rhododendron simsii Annelies Christiaens Thesis submitted in fulfillment of the requirements for the degree of Doctor (PhD) in Applied Biological Sciences Nederlandse titel: Factoren die de bloemontwikkeling en bloeikwaliteit van azalea (Rhododendron simsii) beïnvloeden Christiaens, A. (2014) Factors affecting flower development and quality in Rhododendron simsii. PhD Thesis, Ghent University, Ghent, Belgium. ISBN-number: 978-90-5989-712-0 The authors and promoters give the authorization to consult and to copy parts of the work for personal use only. Every other use is subject to the copyright laws. Permission to reproduce any material contained in the work should be obtained from the author. Table of contents Summary v Samenvatting ix List of abbreviations xiii CHAPTER 1 GENERAL INTRODUCTION 1 1.1 Taxonomy, morphology and origin 1 1.2 Azalea breeding 4 1.3 Economic importance of azalea for the Ghent region 5 1.4 Cultivation of azalea 5 1.5 Flowering: a complex process 7 1.5.1 Floral induction 8 1.5.2 Flower initiation and flower development 12 1.5.3 Dormancy 14 1.5.4 Anthesis 15 1.6 Research objectives and thesis outline 15 CHAPTER 2 FLOWER DIFFERENTIATION OF AZALEA DEPENDS ON GENOTYPE AND NOT ON THE USE OF PLANT GROWTH REGULATORS 19 2.1 Introduction 19 2.2 Materials and methods 21 2.2.1 Plant material 21 2.2.2 Experiment 1 Effect of PGR application 21 2.2.3 Experiment 2 Seasonal variations 21 2.2.4 Experiment 3 Genotypic variation in flower development 22 2.2.5 Assessment of flower bud development 22 2.2.6 Climatic registration 22 2.2.7 Statistical analysis 22 2.3 Results 23 i Table of contents 2.3.1 Effect of PGR on flower development 23 2.3.2 Seasonal effects 24 2.3.3 Genotypic variation in flower development 25 2.4 Discussion 26 2.4.1 Effect of PGR on flower initiation and differentiation 26 2.4.2 Seasonal effects on flower bud development 27 2.4.3 Genotypic variation 28 2.5 Conclusions 28 CHAPTER 3 FLOWER BUD DORMANCY OF RHODODENDRON SIMSII IS INFLUENCED BY GENOTYPE, PLANT GROWTH REGULATORS AND FLOWER BUD DEVELOPMENTAL STAGE 29 3.1 Introduction 30 3.2 Materials and methods 32 3.2.1 Experiment 1 Effect of dormancy-breaking cold treatment on endogenous ABA content, ABA sensitivity and quality of flowering 32 3.2.2 Experiment 2 Effect of plant growth regulators (PGR) on endogenous ABA content, ABA sensitivity and quality of flowering 32 3.2.3 Experiment 3 Optimum flower bud stage to respond to cold treatment 33 3.2.4 Assessment of flower bud development 33 3.2.5 ABA bio-assay 33 3.2.6 Endogenous ABA concentrations 34 3.2.7 Quality of flowering 34 3.2.8 Statistical analysis 35 3.3 Results 35 3.3.1 Experiment 1 Effect of dormancy-breaking cold treatment on endogenous ABA content, ABA sensitivity and quality of flowering 35 3.3.2 Experiment 2 Effect of plant growth regulators (PGR) on endogenous ABA content, ABA sensitivity and quality of flowering 37 3.3.3 Experiment 3 Optimum flower bud stage to respond to cold treatment 40 3.4 Discussion 41 3.4.1 Effect of dormancy-breaking cold treatment on endogenous ABA content, ABA sensitivity and quality of flowering 41 3.4.2 Effect of plant growth regulators (PGR) on endogenous ABA content, ABA sensitivity and quality of flowering 43 3.4.3 Floral developmental stage determines the start of dormancy breaking treatments 44 3.5 Conclusions 45 CHAPTER 4 DETERMINING THE MINIMUM DAILY LIGHT INTEGRAL FOR FORCING OF AZALEA (RHODODENDRON SIMSII) 47 ii Table of contents 4.1 Introduction 47 4.2 Materials and methods 49 4.2.1 Plant material 49 4.2.2 Whole-plant gas exchange measuring technique 50 4.2.3 Determining the minimum DLI 52 4.2.4 Effect of temperature on minimum DLI 53 4.2.5 Effect of developmental stage on minimum DLI 53 4.2.6 Effect of developmental stage on carbohydrate content 53 4.2.7 Forcing experiments 54 4.2.8 Statistical analysis 54 4.3 Results 55 4.3.1 Daily carbon assimilation 55 4.3.2 Determining the minimum DLI 56 4.3.3 Effect of temperature on photosynthesis and DLI 57 4.3.4 Effect of developmental stage on DLI 58 4.3.5 Effect of developmental stage on carbohydrate content 58 4.3.6 Flowering quality under different DLIs 59 4.4 Discussion 60 4.4.1 Method to determine the minimum DLI 60 4.4.2 Minimum DLI is cultivar dependent 62 4.4.3 Effect of temperature on DLI 62 4.4.4 Effect of developmental stage on DLI 63 4.4.5 Minimum DLI and plant quality 64 4.5 Conclusions 65 CHAPTER 5 COLD STORAGE TO OVERCOME DORMANCY AFFECTS THE CARBOHYDRATE STATUS AND PHOTOSYNTHETIC CAPACITY OF RHODODENDRON SIMSII 67 5.1 Introduction 67 5.2 Materials and methods 69 5.2.1 Plant material and experimental set-up 69 5.2.2 Determination of soluble sugars and starch 70 5.2.3 Gene expression analysis of carbohydrate metabolism genes 71 5.2.4 Photosynthesis measurements 71 5.2.5 Statistical analysis 72 5.3 Results 72 5.3.1 Carbohydrate metabolism during cold storage 72 5.3.2 Gene expression profiling during cold treatment 75 5.3.3 Effect of cold treatment on photosynthesis and carbohydrate metabolism 78 5.4 Discussion 79 iii Table of contents 5.4.1 Carbohydrate metabolism during cold storage 79 5.4.2 Photosynthetic recovery after cold treatment 80 5.4.3 Effect of dormancy status of the flower buds on photosynthesis 81 5.4.4 Effect of cultivar on photosynthesis 82 5.5 Conclusions 82 CHAPTER 6 SOURCE-SINK METABOLISM DURING FLOWERING OF AZALEA IS INFLUENCED BY LIGHT CONDITIONS 83 6.1 Introduction 83 6.2 Materials and methods 85 6.2.1 Plant material 85 6.2.2 Experimental set-up 1: manipulating photosynthate level 86 6.2.3 Experimental set-up 2: influence of PGR 88 6.2.4 Soluble carbohydrates and starch content 88 6.2.5 Enzyme activities 89 6.2.6 Assessment of the quality of flowering 89 6.2.7 Statistical analysis 89 6.3 Results 90 6.3.1 Carbohydrate content before forcing 90 6.3.2 Carbohydrate content during forcing 93 6.3.3 Enzymatic activity during forcing 97 6.3.4 Post-production carbohydrate content 99 6.3.5 Enzymatic activity during post-production 103 6.3.6 Quality of flowering 103 6.3.7 Effect of a high dose of paclobutrazol 105 6.4 Discussion 109 6.4.1 Sucrose metabolism during flowering under optimal conditions 109 6.4.2 Effect of forcing conditions on sucrose metabolism 110 6.4.3 Post-production sucrose metabolism and flowering quality 111 6.4.4 Effect of a high dose of paclobutrazol on sucrose metabolism and flowering 111 6.5 Conclusions 112 CHAPTER 7 CONCLUSIONS AND FUTURE PERSPECTIVES 113 References 123 Additional data to chapter 5 137 Curriculum vitae 141 Dankwoord iv Summary As part of the genus Rhododendron, azalea is well known for its luxuriant flowering. Each year, 30 million plants are produced with a production value of 48.2 million euro in East Flanders (Belgium). High-quality flowering is essential and is considered to be a homogeneous bud burst in the forcing greenhouse and a continuous development to fully open flowers at indoor conditions. Growers continuously strive to improve the quality of their products and a simple question arose: “Why do azalea flowers not always open at indoor conditions?” We tried to answer this question by an in-depth physiological research of the complex flowering process. Analysing the production methods, we see drastic changes over the last decade. The complete production process is forced to a higher productivity, but as a result it becomes more challenging to keep flowering quality up to standards. A year-round production of flowering plants is established through a shorter and different timing of production, an increased use of plant growth regulators, the use of supplemental light during forcing and a new range of cultivars. These cultivars can be divided into three groups according to their natural flowering time: early flowering, semi-early flowering and late flowering. In a first part of this research, we focus on flower differentiation and flower bud dormancy. First, we investigated the effects of the altered production processes on flower initiation and differentiation. As we expected large genotypic differences between cultivars of different flowering groups, the experiments are performed using early-, semi-early- and late-flowering cultivars. We hypothesized that the increased use of the plant growth regulators (chlormequat and paclobutrazol; inhibitors of the gibberellin biosynthesis pathway) to control the vegetative growth and initiate flowering would affect flower differentiation rate. Our results show that although flower initiation is triggered by chlormequat, subsequent flower differentiation is not influenced v Summary by the application of plant growth regulators (PGR). Nevertheless, plants treated with PGR, have an evenly distributed flower bud formation at plant level compared to control plants. A different timing of the production cycle means that climatic conditions (temperature, light intensity, day length) are different during flower differentiation.
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    The Rhododendron Genome and Chromosomal Organization Provide Insight Into Shared Whole-Genome Duplications Across the Heath Family (Ericaceae)

    GBE The Rhododendron Genome and Chromosomal Organization Provide Insight into Shared Whole-Genome Duplications across the Heath Family (Ericaceae) Valerie L. Soza 1,*, Dale Lindsley1,†, Adam Waalkes1,5, Elizabeth Ramage1, Rupali P. Patwardhan2, Joshua N. Burton2,6, Andrew Adey2,7,AkashKumar2,8, Ruolan Qiu2,†, Jay Shendure2,3,4, and Benjamin Hall1 1Department of Biology, University of Washington, Seattle, WA 2Department of Genome Sciences, University of Washington, Seattle, WA 3Brotman Baty Institute for Precision Medicine, Seattle, WA 4Howard Hughes Medical Institute, University of Washington, Seattle, WA 5Present address: Department of Laboratory Medicine, University of Washington, Seattle, WA 6Present address: Adaptive Biotechnologies, Seattle, WA 7Present address: Department of Molecular and Medical Genetics, Oregon Health and Science University, Portland, OR 8Present address: Department of Pediatrics, Stanford University, Palo Alto, CA †Retired. *Corresponding author: E-mail: [email protected]. Accepted: November 4, 2019 Data deposition: DNA sequencing data and final genome assembly for Rhododendron williamsianum have been deposited at NCBI BioProject under the accession PRJNA432092 and at CoGe under genome ID 51679. In the genome assembly, within each linkage group, all ordered scaffolds are given in their LACHESIS order, followed by all unordered scaffolds listed in descending order of length. Unclustered scaffolds are listed after linkage group 13 in descending order of length. The scripts used for genome assembly, annotation, and comparative genomics are available in the GitHub repository (https://github.com/vsoza/rhododendron-genome/). Synteny analyses within the Comparative Genomics (CoGe) platform can be obtained from the links given. Rhododendron delavayi: https://genomevolution.org/r/14kzh (whole-genome); Rhododendron williamsia- num: https://genomevolution.org/r/12blb, https://genomevolution.org/r/17qc6 (ordered scaffolds); Vaccinum corymbosum: https://genomevolu- tion.org/r/14oui.