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In vitro corm formation and flowering and ex vitro acclimatization watering regime at the time during which these experiments where conducted and plants were watered every second day with 500 ml of water. Corms of R. minutiflora and R. sabulosa were planted in plastic planting trays with a 1:1 ratio of potting soil and sand. For each species, 6 corms were placed in 6 trays. The trays were placed in the mist-house for 24 h, after which they were moved to the greenhouse. The trays were only placed in the mist-house for this short period of time because all corms were necrotic after only two weeks in an initial experiment were 2 trays with 6 corms each where placed in the mist-house, suggesting that these conditions are too moist. In another experiment corms of R. sabulosa were planted in clay pots with a 1:1 ratio of potting soil and sand. Because of the limited number of pots, 4 corms were placed in each pot and 30 pots were used. These pots were placed in a growth chamber with high light (190.1 µmol m –2 s –1 ) maintained at 20°C. Healthy rooted plantlets of R. minutiflora and R. sabulosa that did not produce corms during corm formation experiments were planted in plastic planting trays with a 1:1 ratio of potting soil and sand. For each species, 6 plantlets were placed in 5 trays. The trays were placed in the mist-house for 24 h, after which they were moved to the greenhouse. A small experiment was conducted with the remaining corms formed in vitro at 10 °C. Four corms were placed in a clean plastic container with autoclaved vermiculite moistened with 50% Hoagland’s nutrient solution. The containers were placed in a growth chamber set at 20°C with a 16 h photoperiod of 12.5 µmol m –2 s –1 irradiance for two months. The rest of the corms were used for a viability study. The viability of ten corms larger than 5 mm in diameter were tested according to the method of WAGNER (1984). 141
6.3 RESULTS 6.3.1 Corm formation In vitro corm formation and flowering and ex vitro acclimatization In the first corm formation experiments with shoots of R. minutiflora, no corms were observed at temperatures of 25°C and higher. The results of 15°C are not reported as the growth chamber malfunctioned and the cultures had to be discarded. No significant differences in corm induction from shoot explants were observed either when changing the temperature or altering the sucrose concentration (Table 6.1). Corm mass, however, increased with increasing sucrose concentration. This was true at both 10 and 20 °C. The addition of activated charcoal to media with 3% sucrose significantly increased corm mass under both temperature regimes (Table 6.1). The product of corm induction and corm mass indicates that placing the shoots at 10°C on a medium with 6% sucrose leads to the best combination of both proportion of induction and corm mass, followed by the shoots placed at 20°C on media with 9% and 3% sucrose respectively (Table 6.1). Table 6.1: The effect of different temperatures and media composition on the in vitro formation and growth of Romulea minutiflora corms. Data shows the means ± the standard error. Letters indicates significant differences between treatments according to Duncan’s multiple range test. Culture temperature and medium treatment Corm induction % Corm mass (mg) Product 10°C 3% sucrose 65.2 ± 9.2 a 108.7±21.9 b 70.9 6% sucrose 80.4±6.9 a 279.0±114.0 ab 224.3 9% sucrose 81.9±9.1 a 134.5±19.6 b 110.2 5.0 g.l -1 activated charcoal 56.7±5.6 a 285.2±33.7 a 161.7 20°C 3% sucrose 69.4±4.6 a 152.0±31.3 b 197.9 6% sucrose 73.9±3.9 a 227.0±61.1 b 105.5 9% sucrose 72.5±10.3 a 273.7±50.3 a 198.4 5.0 g.l -1 activated charcoal 63.8±5.9 a 239.3±45.0 a 157.7 PP3 and ABA both stimulated corm formation at 25°C in the second experiment with R. minutiflora. The addition of 17.0 and 34.0 M PP3 resulted in the largest percentage corm induction and corm size (Table 6.2). 142
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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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Literature review higher regenerati
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Subfamily Ixioideae (continued) Tri
Page 115 and 116: Literature review Table 2.8: Corm i
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
Page 143 and 144: In vitro culture initiation and mul
Page 145 and 146: 5.2.3 Explant comparison In vitro c
Page 159 and 160: 5.4 DISCUSSION In vitro culture ini
Page 163 and 164: 6 In vitro corm formation and flowe
Page 165: In vitro corm formation and floweri
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Page 177 and 178: Commercialization potential of Romu
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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