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In vitro culture initiation and multiplication observed when it was placed on media with activated charcoal. These cultures did not multiply well, or at all, and many became necrotic. What appeared to be indirectly formed embryogenic tissue was observed on all media supplemented with both kinetin and 2,4-D and its induction percentage was highest on a medium supplemented with 2.3 M kinetin and 2.3 M 2,4-D (Table 5.1). Cultures that appeared to have direct embryogenesis was observed on media supplemented with 4.7 M kinetin and 2.3 M 2,4-D, and 23.2 M kinetin, 4.5 M 2,4-D. Most of the cultures that appeared to have initiated embryogenesis were observed on the latter medium. The results of experiments that tested the effect of media supplemented with 5.4 M NAA and 2.3 M kinetin and media with two different concentrations of BA is not included. This is because of the vitrification observed in cultures placed on this media and the lack of subsequent explant response by these cultures when placed on media with no plant growth regulators. 127
In vitro culture initiation and multiplication Table 5.1: Effect of kinetin and 2,4-D on excised embryos of Romulea sabulosa. Mean values in a column followed by different letters that indicates significance differences between treatments according to Duncan’s multiple range test (P 0.05). S = swelling of embryo; SR = swelling of embryo with rooting; SSI = swelling of embryo with shoot initials; SRF = shoot and root formation; SC = shoot cluster; SCR = shoot cluster with roots; CSCI = callus with shoot cluster initials; CIS = corm-like structure (< 10 mm); CAI = callus appearing incompetent; CPE = callus with potential embryogenesis; PDE = potential direct embryogenesis; CSGR = cultures showed growth response. Potential embryogenesis refers to cultures that appeared to develop embryo-like structures (Figure 5.7). Treatment ( M) Observed response (%) y S SR SSI SRF SC SCR CSCI CIS CAI CPE PDE CSGR control (no PGR’s) 43 ± 6 a 20 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 e 0 ± 0 b 53 ± 8 abc kinetin (2.3) 24 ± 3 bc 0 ± 0 b 24 ± 3 a 20 ± 0 a 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 e 0 ± 0 b 50 ± 10 abc kinetin (4.7) 30 ± 4 b 40 ± 0 a 20 ± 0 b 20 ± 0 a 20 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 e 0 ± 0 b 50 ± 7 abc kinetin (23.2) 20 ± 0 cd 0 ± 0 b 20 ± 0 b 0 ± 0 b 36 ± 8 a 20 ± 0 a 20 ± 0 a 0 ± 0 b 0 ± 0 b 0 ± 0 e 0 ± 0 b 63 ± 8 ab 2,4-D (2.3) 20 ± 0 cd 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 20 ± 0 a 20 ± 0 de 0 ± 0 b 27 ± 7 c kinetin (2.3) + 2,4-D (2.3) 20 ± 0 cd 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 40 ± 7 ab 0 ± 0 b 47 ± 7 abc kinetin (4.7) + 2,4-D (2.3) 20 ± 0 cd 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 30 ± 6 a 0 ± 0 b 43 ± 6 a 27 ± 5 a 70 ± 7 a kinetin (23.2) + 2,4-D (2.3) 20 ± 0 bcd 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 20 ± 0 ab 0 ± 0 b 30 ± 5 abc 0 ± 0 b 50 ± 5 abc 2,4-D (4.5) 30 ± 4 bc 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 20 ± 0 ab 20 ± 0 b 26 ± 5 cde 0 ± 0 b 37 ± 13 bc kinetin (2.3) + 2,4-D (4.5) 20 ± 0 cd 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 20 ± 0 ab 20 ± 0 a 35 ± 8 bcd 0 ± 0 b 47 ± 13 abc kinetin (4.7) + 2,4-D (4.5) 20 ± 0 cd 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 20 ± 0 ab 0 ± 0 b 33 ± 7 ab 0 ± 0 b 47 ± 7 abc kinetin (23.2) + 2,4-D (4.5) 20 ± 0 d 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 0 ± 0 b 20 ± 0 ab 20 ± 0 a 27 ± 5 cde 32 ± 5 a 57 ± 13 ab 128
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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
Page 101 and 102: Literature review Young embryos req
Page 103 and 104: Literature review and the addition
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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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