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In vitro culture initiation and multiplication Significantly more shoots were produced on a medium supplemented with 23.2 M kinetin than a medium supplemented with 2.3 or 4.7 µM kinetin (Figure 5.6). Tukey’s HSD test revealed that the number of shoots formed on a medium supplemented with 23.2 M kinetin were significantly higher than on a medium supplemented with 2.3 M kinetin (N = 14, 18; Mean difference = -3.42857; SE = 0.97832, p = 0.003). It also revealed that the number of shoots generated on a medium supplemented with 23.2 M kinetin was significantly higher than on 4.7 M kinetin (N = 16, 18; Mean difference = -3.12500; SE = 0.94330, p = 0.005). Figure 5.6: Effect of kinetin concentration on shoot production of Romulea sabulosa after 2 months. Error bars indicate standard error of the mean. Letters shows significance differences between treatments according to Tukey’s HSD test. Although it appeared that some cultures were embryogenic (Figure 5.7), sections of tissue presumed to exhibit direct and indirect embryogenesis did unfortunately not reveal any embryo initials and it was concluded that these cultures were not embryogenic, but formed abnormal shoot-like structures lacking chlorophyll. 129
In vitro culture initiation and multiplication Figure 5.7: Visual observations of Romulea sabulosa cultures. Cultures including both kinetin and 2,4-D appears to exhibit embryo-like structures. 5.3.3 Explant comparison Despite the apparent larger number of shoots produced between cultures in which embryos were used as explants compared to seedling organs, there is no significant difference between the numbers of shoots produced according to a paired t-test at 95% confidence limits. There was also no significant difference between the number of shoots formed on media supplemented with kinetin and the number of shoots formed on media supplemented with other plant growth regulator treatments (Figure 5.8). These results show that a medium supplemented with 23.2 µM mTR and 0.5 µM NAA significantly increases the amount of shoots formed per embryo compared to the control. This is not the case with seedling hypocotyls. Here cytokinins did not increase the number of shoots formed per explant (Figure 5.8). The number of shoots produced on a medium with 23.2 µM mTR and 0.5 µM NAA was not significantly different from the number of shoots observed on a medium supplemented with 2.3 µM kinetin or 2.3 µM mTR and 0.5 µM NAA. Explant response data is not shown, as there were no significant differences in explant response between the two explant types. Although these differences are not 130
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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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Page 107 and 108: Literature review environmental str
Page 109 and 110: 2.11 IN VITRO FLOWERING Literature
Page 111 and 112: Literature review higher regenerati
Page 113 and 114: 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
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
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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 and 168: 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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