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In vitro corm formation and flowering and ex vitro acclimatization Table 6.2: Percentage corm induction for Romulea minutiflora shoots cultured on medium supplemented with growth retardants. Treatment Basal swelling (%) Corm induction (%) Corms > 5 mm (%) Control 70% 0% 0% 3.4 M PP3 55% 15% 10% 17.0 M PP3 50% 35% 20% 34.0 M PP3 35% 35% 20% 4.9 M ABA 45% 20% 5% No corm formation was observed for any of the temperature and medium treatments tested for R. leipoldtii. When 100 bottles of shoots multiplied on media supplemented with BA placed at 25°C were not subcultured for 6 months no corm formation or basal thickening was observed. This was not the case with the shoot cultures placed at 25°C multiplied with media supplemented with kinetin and topolins. Here corms or basal thickening was observed for all shoots not multiplied after 6 months. Such basal thickening was observed to a certain extent for non-subcultured shoot cultures of R. minutiflora and R. sabulosa, but no corm formation was observed in any cultures placed at 25°C for these species. The percentage corm induction for R. sabulosa after 6 months was significantly higher at 10 and 20 °C with all tested sucrose concentrations compared to 15 °C (Table 6.3). Corm induction was inhibited at 25 °C in this experiment (data not shown). The largest corm mass was recorded for shoots placed at 15 °C on a MS medium supplemented with 6% sucrose (Table 6.3). The mass of these corms were however not significantly different from the mass of the corms obtained for cultures placed at the same temperature on a medium with 9% sucrose or the mass of corms produced on a medium with activated charcoal and the mass of corms at 20 °C placed on a medium with 6% sucrose. Product analysis shows that corms at 15 °C with 6% sucrose achieved the highest value (Table 6.3). 143
In vitro corm formation and flowering and ex vitro acclimatization Table 6.3: The effect of different temperatures and media composition on the in vitro formation and growth of Romulea sabulosa corms. Data shows the means ± the standard error. Letters indicate significant differences between treatments according to Duncan’s multiple range test. Culture temperature and medium Corm induction Corm mass (mg) Product treatment (%) 10°C 3% sucrose 100.0 ± 0.0 a 325.5 ± 39.0 b 325.5 6% sucrose 100.0 ± 0.0 a 346.5 ± 56.3 b 346.5 9% sucrose 100.0 ± 0.0 a 294.1 ± 49.6 b 294.1 5.0 g.l -1 activated charcoal 87.5 ± 7.2 abc 336.3 ± 50.8 b 294.2 23.2 µM kinetin 100.0 ± 0.0 a 371.1 ± 49.8 b 371.1 15°C 3% sucrose 71.3 ± 10.9 bc 325.2 ± 21.9 b 231.7 6% sucrose 81.7 ± 6.9 bc 522.7 ± 61.7 a 426.9 9% sucrose 5.0 g.l 83.8 ± 5.5 bc 439.3 ± 71.4 ab 348.9 -1 activated charcoal 68.8 ± 12.6 c 409 ± 59.4 ab 281.2 23.2 µM kinetin 20.0 ± 14.1 d 295.3 ± 39.2 b 58.5 20°C 3% sucrose 100.0 ± 0.0 a 310.9 ± 54.7 b 310.9 6% sucrose 100.0 ± 0.0 a 383.5 ± 36.6 ab 383.5 9% sucrose 5.0 g.l 100.0 ± 0.0 a 372.7 ± 57.8 b 372.7 -1 activated charcoal 91.7 ± 8.3 ab 362.6 ± 63.8 b 332.4 23.2 µM kinetin 18.8 ± 18.8 d 281.8 ± 52.9 b 52.2 Multiple corm formation from the same shoot was observed in some cultures of R. sabulosa (Table 6.4). The highest percentage of multiple corm formation was observed at 15°C on a medium supplemented with 23.2 µM kinetin, while the highest number of corms per shoot was observed at 10°C on a medium with 3% sucrose. Multiple corm formation also occurs in Romulea species under natural environmental conditions (DE VOS, 1972). Table 6.4: Cultures with multiple corm formation for Romulea sabulosa. This shows the percentage of corm formation in cultures in which corm formation observed (Total cultures with corms) and the average number of corms produced in instances of multiple corm formation. Culture temperature and medium treatment Multiple corm formation (%) Total cultures with corms Average number of multiple corms 10°C 3% sucrose 6.3% 16 7.0 9% sucrose 16.7% 18 2.3 5.0 g.l -1 activated charcoal 6.3% 16 3.0 15°C 6% sucrose 7.1% 14 3.0 9% sucrose 12.5% 16 2.0 5.0 g.l -1 activated charcoal 7.7% 13 2.0 23.2 µM kinetin 50.0% 4 2.0 20°C 3% sucrose 9.1% 11 2.0 144
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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
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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
Page 131 and 132: Germination physiology Figure 4.4:
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 and 166: In vitro corm formation and floweri
Page 167: 6.3 RESULTS 6.3.1 Corm formation In
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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