Micropropagation and medicinal properties of Barleria greenii

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TABLE OF CONTENTS STUDENT DECLARATION ................................................................................... vii DECLARATION BY SUPERVISORS ................................................................... viii FACULTY OF SCIENCE & AGRICULTURE DECLARATION 1 - PLAGIARISM.... ix FACULTY OF SCIENCE & AGRICULTURE DECLARATION 2 - PUBLICATIONS x ACKNOWLEDGEMENTS ..................................................................................... xii LIST OF FIGURES ............................................................................................... xiii LIST OF TABLES .................................................................................................xvii LIST OF ABBREVIATIONS .................................................................................. xix ABSTRACT….. ....................................................................................................xxii Chapter 1 General introduction ........................................................................ 1 1.1 Use of plants in horticulture and traditional medicine .......................... 1 1.2 The need for conservation of plant species ........................................... 1 1.3 Distribution, morphology, uses and conservation status of the studied plant species ............................................................................... 3 1.3.1 Barleria greenii .................................................................................... 3 1.3.2. Huernia hystrix..................................................................................... 5 1.4 Value of tissue culture ............................................................................. 7 1.5 Aims and objectives ................................................................................. 8 1.6 General overview of the thesis ................................................................ 8 Chapter 2 Literature review ............................................................................ 10 2.1 Micropropagation ................................................................................... 10 2.1.1 Introduction ........................................................................................ 10 2.1.2 Stage 0: Selection and preparation of mother plants ......................... 10 2.1.3 Stage I: Initiation and establishment of aseptic culture ...................... 11 ii
2.1.4 Stage II: Proliferation or multiplication of propagules ......................... 15 2.1.5 Stage III: Preparation of propagules for transfer to soil ..................... 17 2.1.6 Stage IV: In vivo rooting and acclimatization for soil establishment... 19 2.1.7 Effects of auxins and cytokinins on plant regeneration in micropropagation ............................................................................. 22 2.1.8 Environmental factors affecting micropropagation ............................. 29 2.1.8.1 Light ............................................................................................... 29 2.1.8.2 Temperature ................................................................................... 31 2.1.9 Tissue culture of the families: Acanthaceae and Asclepiadaceae ..... 32 2.2 Pharmacological and phytochemical investigation of plant extracts 36 2.2.1 Introduction ........................................................................................ 36 2.2.2 Antimicrobial activity .......................................................................... 37 2.2.3 Anti-inflammatory activity ................................................................... 39 2.2.4 Acetylcholinesterase inhibition .......................................................... 42 2.2.5 Antioxidant activity ............................................................................. 43 2.2.6 Phytochemical property ..................................................................... 45 Chapter 3 In vitro propagation of Barleria greenii ........................................ 48 3.1 Introduction ............................................................................................ 48 3.2 Materials and methods ........................................................................... 49 3.2.1 Explant decontamination, selection and bulking ................................ 49 3.2.2 Effects of BA and NAA on shoot multiplication .................................. 50 3.2.3 Effects of types and concentrations of cytokinins on shoot multiplication .................................................................................... 50 3.2.4 Effects of photoperiod on shoot multiplication ................................... 51 3.2.5 In vitro rooting of regenerated shoots ................................................ 51 3.2.6 Ex vitro rooting and acclimatization ................................................... 52 3.2.7 Data analyses .................................................................................... 52 iii
Page 1: MICROPROPAGATION AND MEDICINAL PROP
Page 5 and 6: Chapter 5 Pharmacological and phyto
Page 7 and 8: STUDENT DECLARATION Micropropagatio
Page 9 and 10: FACULTY OF SCIENCE & AGRICULTURE DE
Page 11 and 12: AMOO, S.O., FINNIE, J.F. and VAN ST
Page 13 and 14: LIST OF FIGURES Chapter 1 Figure 1.
Page 15 and 16: Figure 5.2: Anti-inflammatory activ
Page 17 and 18: LIST OF TABLES Chapter 2 Table 2.1:
Page 19 and 20: LIST OF ABBREVIATIONS [3G]BA 6-Benz
Page 21 and 22: PE Petroleum ether PEPC Phosphoenol
Page 23 and 24: status. Being a dwarf species, very
Page 25 and 26: activities shown by H. hystrix extr
Page 27 and 28: During this same period, these auth
Page 29 and 30: dark pink with magenta streaks on t
Page 31 and 32: traditional medicine (muthi) among
Page 33 and 34: 1.5 Aims and objectives Endemic and
Page 35 and 36: 2.1 Micropropagation 2.1.1 Introduc
Page 37 and 38: their constituent cells (MURASHIGE,
Page 39 and 40: media are sterile, the explants mai
Page 41 and 42: plants is higher (MURASHIGE, 1978)
Page 43 and 44: adventitious roots are produced wit
Page 45 and 46: DEBERGH and MAENE (1981) therefore
Page 47 and 48: cytokinins as well as these major e
Page 49 and 50: Auxins are known to play a crucial
Page 51 and 52: 6-Benzyladenine (BA) 6-Furfurylamin
Page 53 and 54:
STADEN and DREWES, 1992), the relat
Page 55 and 56:
growth and development throughout t
Page 57 and 58:
HASEGAWA et al (1973) reported a co
Page 59 and 60:
Table 2.1 continued Plant species E
Page 61 and 62:
2.2 Pharmacological and phytochemic
Page 63 and 64:
filamentous fungi (such as Fusarium
Page 65 and 66:
PGI2, PGF2α, are known to sensitiz
Page 67 and 68:
Many researchers have reported anti
Page 69 and 70:
inclusion of antioxidant and free r
Page 71 and 72:
activity of gallotannins from Mangi
Page 73 and 74:
3.1 Introduction Chapter 3 In vitro
Page 75 and 76:
with 30 g l -1 sucrose, 0.1 g l -1
Page 77 and 78:
andomised and maintained in a growt
Page 79 and 80:
3.3.1 Explant decontamination Figur
Page 81 and 82:
Adventitious shoots per explant (n)
Page 83 and 84:
3.3.3 Effects of types and concentr
Page 85 and 86:
Table 3.1: Effects of types and con
Page 87 and 88:
3.3.4 Effects of photoperiod on sho
Page 89 and 90:
to be a slow release reservoir of a
Page 91 and 92:
Percentage survival 70 60 50 40 30
Page 93 and 94:
exponential population growth rates
Page 95 and 96:
Cultures were incubated under the s
Page 97 and 98:
loosely closed screw cap jars (300
Page 99 and 100:
Frequency of decontamination (%) 60
Page 101 and 102:
Table 4.2: Frequencies of shoot, ro
Page 103 and 104:
SALISBURY and ROSS (1992), a temper
Page 105 and 106:
Table 4.5: Effects of temperature a
Page 107 and 108:
photoperiod could therefore be due
Page 109 and 110:
Titratable acidity (µM H + g -1 FW
Page 111 and 112:
Figure 4.4: Huernia hystrix adventi
Page 113 and 114:
strength MS medium with or without
Page 115 and 116:
Chapter 5 Pharmacological and phyto
Page 117 and 118:
Garden, Pietermaritzburg, South Afr
Page 119 and 120:
2004) respectively, were also deter
Page 121 and 122:
each of the plant extracts, the bla
Page 123 and 124:
ORR =
Page 125 and 126:
5.2.3.5 Gallotannin content The rho
Page 127 and 128:
Barleria species, the EtOH extracts
Page 129 and 130:
Table 5.2: Yield (% w/w) of extract
Page 131 and 132:
Table 5.4: Antibacterial activity (
Page 133 and 134:
Table 5.5: Antifungal activity of c
Page 135 and 136:
5.3.2.3 Anti-inflammatory activity
Page 137 and 138:
Prostaglandin synthesis inhibition
Page 139 and 140:
The AChE inhibitory activity of ext
Page 141 and 142:
(A) DPPH Scavenging activity (%) (B
Page 143 and 144:
Table 5.8: DPPH radical scavenging
Page 145 and 146:
Absorbance 630 nm 2.2 2.0 1.8 1.6 1
Page 147 and 148:
(ORHAN et al., 2009; ABDEL-HAMEED,
Page 149 and 150:
5.3.3 Phytochemical evaluation 5.3.
Page 151 and 152:
5.3.3.2 Total iridoid content Figur
Page 153 and 154:
5.3.3.3 Flavonoid content The flavo
Page 155 and 156:
5.3.3.4 Gallotannin content The roo
Page 157 and 158:
5.3.3.5 Condensed tannin (proanthoc
Page 159 and 160:
Chapter 6 General conclusions An ef
Page 161 and 162:
evaluating their safety may be nece
Page 163 and 164:
ANAND, Y. and BANSAL, Y.K. 1998. Pl
Page 165 and 166:
BASKARAN, P., VELAYUTHAM, P. and JA
Page 167 and 168:
CHENG, S.H. and EDWARDS, G.E. 1991.
Page 169 and 170:
DE KLERK, G.J., BRUGGE, J.T. and MA
Page 171 and 172:
ETKIN, N.L. 1998. Indigenous patter
Page 173 and 174:
GIRIDHAR, P., GURURAJ, H.B. and RAV
Page 175 and 176:
HUANG, D., OU, B. and PRIOR, R.L. 2
Page 177 and 178:
KIRANMAI, C., ARUNA, V., KARUPPUSAM
Page 179 and 180:
LITZ, R.E. and CANOVER, R.A. 1981.
Page 181 and 182:
MARKS, T.R. and SIMPSON, S.E. 1994.
Page 183 and 184:
NDAWONDE, B.G., ZOBOLO, A.M., DLAMI
Page 185 and 186:
PAREEK, A. and KOTHARI, S.L. 2003.
Page 187 and 188:
RAI, M.K., AKHTAR, N. and JAISWAL,
Page 189 and 190:
threatened plants - progress in the
Page 191 and 192:
SRIVASTAVA, L.M. 2002. Plant Growth
Page 193 and 194:
TURNER, S. 2001. Witwatersrand Nati
Page 195:
WILLIAMSON, E., OKPAKO, D.T. and EV
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