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In vitro culture initiation and multiplication • Shoot cultures was initiated on media supplemented with 2.3 to 23.2 M kinetin for R. flava, R. leipoldtii, R. minutiflora, R. monadelpha and R. sabulosa, with the most suitable concentration depending on the species used. • A medium supplemented with 2.5 M mTR is suitable for R. sabulosa shoot multiplication. • BA caused vitrification of shoots in all the experiments in which it was used and is not a suitable cytokinin for the micropropagation of these species. 137
6 In vitro corm formation and flowering and ex vitro acclimatization 6.1 INTRODUCTION In vitro formation of storage organs increases the ex vitro survival rate during acclimatization and these organs serve as a more attractive product than seeds, as flowering of the plant can be enjoyed at a much earlier stage (ASCOUGH et al., 2009). Commercialising in vitro produced corms of R. sabulosa will also reduce the pressures on the vulnerable populations from which seeds are harvested (RAIMONDO et al., 2009). In vitro flowering is an important tool in ornamental plant breeding, as it enables the breeder to see the floral traits in a much shorter time than in ex vitro conditions. Some factors that influence in vitro flowering and ex vitro survival is discussed in section 2.10 and 2.8.9 of Chapter 2 respectively. The aims of this Chapter were to establish an in vitro corm induction protocol for R. minutiflora, R. leipoldtii and R. sabulosa, to investigate the effect of some physical and chemical stimuli on in vitro flowering of R. minutiflora and R. sabulosa corms and to establish an ex vitro acclimatization protocol for R. minutiflora and R. sabulosa corms and plantlets. 6.2 MATERIALS AND METHODS 6.2.1 Corm formation In all experiments, the in vitro generated shoots were separated from each other, trimmed to 25 mm and all roots were removed for uniformity. If not stated otherwise a MS medium supplemented with 100 mg.l -1 myo-inositol and 3% sucrose, with pH adjusted to 5.7 and solidified with 0.8% agar was used. All experiments were conducted in a laminar flow hood and cultures were placed in a growth chamber under 3.4 µmol m –2 s –1 light using Osram ® 75 W cool white fluorescent tubes with a 16/8 light/dark photoperiod. The duration of all experiments was 6 months, as no 138
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Propagation of Romulea species Pier
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Contents 2.7 GERMINATION PHYSIOLOGY
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Contents 5.3.1 Explants from seedli
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Abstract The suitability of various
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Declarations I Pierre André Swart,
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Declarations DETAILS OF CONTRIBUTIO
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Publications from this thesis ASCOU
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List of Figures Figure 2.13: Suther
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was significantly different from th
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List of Tables Table 2.1: Names of
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List of Abbreviations 2,4-D 2,4-Dic
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Die vlakhaas spits sy oor hy het di
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As die mens dan ook so sy dankbaarh
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Introduction where a great number o
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Introduction The subgenera Romulea
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2 Literature review Leef van daad e
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Literature review Figure 2.2: Life
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Literature review Figure 2.4: Life
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Literature review The plants are sh
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Literature review flowers (DE VOS,
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Literature review often found on cl
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Literature review flowers of R. lei
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Literature review flattened upper s
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2.2.16 Romulea tabularis Literature
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Literature review extinct, R. papyr
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Literature review R. leipoldtii occ
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Literature review Figure 2.8: Calvi
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Literature review Figure 2.14: Fras
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Figure 2.20: Nieuwoudtville weather
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Literature review Figure 2.26: Grah
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Literature review Figure 2.29: Diag
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Literature review Figure 2.30: A te
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Literature review Macronutrients ca
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Literature review The soils of the
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Literature review The embryo is the
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Literature review Phase 2 is seen a
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Literature review endogenous gibber
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Literature review tolerable level (
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Literature review these channels pe
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Literature review 2.8.7 Seed dorman
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Literature review the embryo (COPEL
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Literature review germinated under
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Literature review (COPELAND, 1976).
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2.9 BRIEF REVIEW OF IN VITRO CULTUR
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Literature review (DEBERGH, 1994).
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Literature review PIERIK (1997) sta
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Literature review The distinguishin
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Literature review The essential fun
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2.9.3.4 Abscisic acid Literature re
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Table 2.6: Example of a matrix to e
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Literature review Young embryos req
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Literature review and the addition
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Literature review organ is then pro
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Literature review environmental str
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2.11 IN VITRO FLOWERING Literature
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Page 113 and 114: Subfamily Ixioideae (continued) Tri
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Page 117 and 118: 3 Investigation into the habitat of
Page 119 and 120: Soil sampling and analysis The firs
Page 121 and 122: Soil sampling and analysis Table 3.
Page 123 and 124: 4 Germination physiology 4.1 INTROD
Page 125 and 126: 4.2.2 Water content and imbibition
Page 127 and 128: Germination physiology (-N), phosph
Page 129 and 130: Germination physiology rosea seeds
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Page 133 and 134: Germination (%) 50 40 30 20 10 0 77
Page 135 and 136: Germination (%) 100 80 60 40 20 0 c
Page 137 and 138: Germination physiology The relative
Page 139 and 140: Germination physiology seeds of R.
Page 141 and 142: 5.2 MATERIALS AND METHODS In vitro
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Page 145 and 146: 5.2.3 Explant comparison In vitro c
Page 159 and 160: 5.4 DISCUSSION In vitro culture ini
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Page 167 and 168: 6.3 RESULTS 6.3.1 Corm formation In
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Page 179 and 180: Commercialization potential of Romu
Page 181 and 182: BASKIN, C. C., and BASKIN, J. M. (1
Page 183 and 184: Literature cited DOWLING, P. M., CL
Page 185 and 186: Literature cited HILTON-TAYLOR, C.
Page 187 and 188: Literature cited KUMAR, A., SOOD, A
Page 189 and 190: Literature cited PIERIK, R. L. M. (
Page 191 and 192: Literature cited STEINITZ, B., COHE
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