Micropropagation and medicinal properties of Barleria greenii

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and preparation of stock plants would not only allow for the availability of standardized and healthy explants, but also increase the rate of explant survival during culture initiation (DEBERGH and MAENE, 1981). 2.1.3 Stage I: Initiation and establishment of aseptic culture Owing to the inherent totipotentiality of plant cells (MURASHIGE, 1978), explants obtained from different plant tissues have a potential to regenerate plantlets when given appropriate environmental and nutrient conditions. It is important that the explants be able to grow well in an aseptic in vitro environment free from any obvious infection (MURASHIGE, 1974). In order to achieve this objective, the choice of explants and their decontamination are given serious attention during this stage. MURASHIGE (1974) listed five important factors to be considered when choosing suitable explants. These include (i) the organ serving as explant source; (ii) the physiological or ontogenic age of the organ; (iii) the season in which the explant is obtained; (iv) the size of the explant; and (v) the overall quality of the stock plant. Nearly all plant organs or tissues can serve as a source of explants in micropropagation. However, explants taken from different plant parts often manifest different regeneration and morphogenic responses. KOMALAVALLI and RAO (2000), for example, reported different propagation rates and morphogenic responses in axillary node, shoot-tip, cotyledonary node, leaf, petiole, root and internode explants of Gymnema sylvestre. They observed that only axillary node, cotyledonary node and shoot-tip explants readily regenerated multiple shoots while other explants produced only callus. These observations were attributed to the variation in endogenous levels of plant growth regulators in the explants at the time of excision. Similar results have been reported by other researchers in different plant species (NHUT et al., 2004; ISLAM et al., 2005; CONDE et al., 2008). The regenerative and morphogenic capacities of explants are sometimes affected by the physiological age of the explant and the degree of differentiation among 11
their constituent cells (MURASHIGE, 1974). SUDHA et al. (1998), while investigating the regeneration potential of shoot-tip, terminal and basal node explants in Holostemma annulare, observed more rapid multiple shoot formation from basal node explants. They concluded that this variation in regenerative response might be due to the differences in endogenous growth regulator level, nutrient availability and physiological status of the explants. In general, explants obtained from juvenile plants often have a better regenerative capacity compared to those from older plants, especially in trees and shrubs (NHUT et al., 2007), perhaps due to their meristematic cell types and other endogenous factors (FENNELL, 2002). BECERRA et al. (2004) reported an inverse linear regeneration capacity in relation to the age of donor plants in Passiflora edulis. They noted that leaf explants taken from juvenile plants (1- to 6-month-old) showed a statistically significant higher shoot regeneration compared to explants taken from reinvigorated adult plants (shoots emerging next to the base after severe pruning of 1-year-old adult plants). Even among the juvenile explant sources, the same inverse linear regeneration capacity with age was observed. In Gerbera jamesonii, flower bud age was reported to significantly affect the survival rate and shoot regeneration capacity of the receptacle transverse thin cell layers, with the optimum observed in 10-day-old flower buds (NHUT et al., 2007). The authors further observed that the position of receptacle transverse thin cell layers significantly affected the receptacle morphogenic ability. They noted that shoot production and regeneration frequency decreased from the middle to exterior receptacle layers, possibly due to more nutrient reserves in the middle layers. The regenerative capacity of explants in tissue culture could also be affected by the season in which the explant is obtained due to seasonal influences on the plant developmental stages. PRAKASH and VAN STADEN (2008) observed that offshoots of Searsia dentata collected during the vegetative stage (April - May) exhibited a better morphogenic response compared to those collected during the flowering stage (October – November). Similarly, young offshoots of Hoslundia opposita collected during the months of March and April were reported to elicit a better morphogenic response compared to those collected during other months of the year (PRAKASH and VAN STADEN, 2007). Other researchers have reported 12
Page 1 and 2: MICROPROPAGATION AND MEDICINAL PROP
Page 3 and 4: 2.1.4 Stage II: Proliferation or mu
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: 2.1 Micropropagation 2.1.1 Introduc
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
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to be a slow release reservoir of a
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Percentage survival 70 60 50 40 30
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exponential population growth rates
Page 95 and 96:
Cultures were incubated under the s
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loosely closed screw cap jars (300
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Frequency of decontamination (%) 60
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Table 4.2: Frequencies of shoot, ro
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SALISBURY and ROSS (1992), a temper
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Table 4.5: Effects of temperature a
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photoperiod could therefore be due
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Titratable acidity (µM H + g -1 FW
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Figure 4.4: Huernia hystrix adventi
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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 =
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5.2.3.5 Gallotannin content The rho
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Barleria species, the EtOH extracts
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Table 5.2: Yield (% w/w) of extract
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Table 5.4: Antibacterial activity (
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Table 5.5: Antifungal activity of c
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5.3.2.3 Anti-inflammatory activity
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Prostaglandin synthesis inhibition
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The AChE inhibitory activity of ext
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(A) DPPH Scavenging activity (%) (B
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Table 5.8: DPPH radical scavenging
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Absorbance 630 nm 2.2 2.0 1.8 1.6 1
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(ORHAN et al., 2009; ABDEL-HAMEED,
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5.3.3 Phytochemical evaluation 5.3.
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5.3.3.2 Total iridoid content Figur
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5.3.3.3 Flavonoid content The flavo
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5.3.3.4 Gallotannin content The roo
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5.3.3.5 Condensed tannin (proanthoc
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Chapter 6 General conclusions An ef
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evaluating their safety may be nece
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ANAND, Y. and BANSAL, Y.K. 1998. Pl
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BASKARAN, P., VELAYUTHAM, P. and JA
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CHENG, S.H. and EDWARDS, G.E. 1991.
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DE KLERK, G.J., BRUGGE, J.T. and MA
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ETKIN, N.L. 1998. Indigenous patter
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GIRIDHAR, P., GURURAJ, H.B. and RAV
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HUANG, D., OU, B. and PRIOR, R.L. 2
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KIRANMAI, C., ARUNA, V., KARUPPUSAM
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LITZ, R.E. and CANOVER, R.A. 1981.
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MARKS, T.R. and SIMPSON, S.E. 1994.
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NDAWONDE, B.G., ZOBOLO, A.M., DLAMI
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PAREEK, A. and KOTHARI, S.L. 2003.
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RAI, M.K., AKHTAR, N. and JAISWAL,
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threatened plants - progress in the
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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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