Report Practical 1 – Petunia Tissue Culture Copy
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Report practical 1 – Petunia Tissue Culture Student Number 32391471 - Marcelo Vieira Da Silva Abstract: Introduction Plant tissue culture can be initiated in vitro by placing explants onto a growth medium under sterile and controlled conditions. Endogenous growth regulators or growth regulating compounds added to the medium can stimulate the metabolism of cells, and they begin active division. In this process, cell differentiation and specialisation, which may have been occurring in the intact plant, are reversed, and the explant originates a new tissue composed of undifferentiated and totipotent cell types (George et al., 2008). Some endogenous substances have a regulatory, instead of a nutritional function in plant growth and development. There are basically two classes of compounds that stimulate plant growth and development: plant hormones (endogenous) and plant growth regulators (synthetic), which are generally active at very low dosages. Auxins, abscisic acid, cytokinin, ethylene, and gibberellins are commonly recognized as the five main classes of endogenous plant hormones. Auxins, cytokinin, and the interactions of auxin-cytokinin are usually considered the most important hormones because they regulate growth and organized development in plant tissue and organ cultures (Gaspar et al, 1996). Auxins concentrations in plant tissue culture generally range from 0.01 to 10mg/L. and according to the concentrations they may regulate several process leading to organogenesis such as, and induction of embryogenesis.
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relatório de aula prática - cultura de tecido de Petunia
Transcript of Report Practical 1 – Petunia Tissue Culture Copy
Report practical 1 – Petunia Tissue Culture Student Number 32391471 - Marcelo Vieira Da Silva
Abstract:
IntroductionPlant tissue culture can be initiated in vitro by placing explants onto a growth
medium under sterile and controlled conditions. Endogenous growth regulators or growth
regulating compounds added to the medium can stimulate the metabolism of cells, and they
begin active division. In this process, cell differentiation and specialisation, which may have
been occurring in the intact plant, are reversed, and the explant originates a new tissue
composed of undifferentiated and totipotent cell types (George et al., 2008). Some
endogenous substances have a regulatory, instead of a nutritional function in plant growth
and development. There are basically two classes of compounds that stimulate plant growth
and development: plant hormones (endogenous) and plant growth regulators (synthetic),
which are generally active at very low dosages.
Auxins, abscisic acid, cytokinin, ethylene, and gibberellins are commonly recognized
as the five main classes of endogenous plant hormones. Auxins, cytokinin, and the
interactions of auxin-cytokinin are usually considered the most important hormones
because they regulate growth and organized development in plant tissue and organ cultures
(Gaspar et al, 1996). Auxins concentrations in plant tissue culture generally range from 0.01
to 10mg/L. and according to the concentrations they may regulate several process leading
to organogenesis such as, and induction of embryogenesis. Cytokinins are generally used at
a concentration range of 0.1-10 mg/L. When added in appropriate concentrations they may
regulate cell division, stimulate auxiliary and adventitious shoot proliferation, regulate
differentiation, inhibit root formation, activate RNA synthesis, and stimulate protein and
enzyme activity (Ponmurugan et al, 2005).
In these experiment was analysed if the plant growth substances promote cell
redifferentiation and development in explants of Petunia leaves when cultured under
several different treatment containing different combinations proportions of plant growth
auxin:cytokinin and different amount of sugar source.
Materials and methods: this is from the lab book but must be written in paragraphs with
subheadings for sections preparation of plant material, experiment 1, experiment 2,
MARCELO VIEIRA, 20/04/15,
Parafrasear e achar um local pra por esta informacao.
experiment 3
Results:
Current year class data
Experiment 1 Effect of auxin
After two weeks in culture the explants became slightly rough in texture, indicating
callus formation in some of the numbered media (image 1), but none of them developed
shoots or roots, all material in the media seemed to be dead, presenting a brownish colour.
For the experiment 1, the Murashige and Skoog basal medium containing different levels of
auxin did not show any effect of concentration for development of shoots or roots (Fig. 1).
The higher mean value for callus weight in experiment 1 was 0.67g where the auxin
concentration was 10uM (Fig. 2).
Figure 2. Effects of Auxin on Callus weight
Experiment 2 Effect of sugar
The effect of sugar on tissues redifferentiation was not possible, because none of
the explants survived the treatment.
Experiment 3 Relationship between auxin and cytokinin
The best combination of auxin and cytokinin proportion was observed in
___________ (Fig. 3).
Last year class data
Experiment 1 Effect of auxin
After two weeks in culture the explants became slightly rough in texture, indicating
callus formation in some of the numbered media (image 1), and others included shoot
(image 2) and root formation (image 3). The Murashige and Skoog basal medium containing
different levels of auxin showed that the effect of concentration is more pronounced when
in small amounts (Fig. 1).
Figure 1. Effects of different dosages of Auxin on Callus Development in the Medium Containing 0.5 uM of BAP
2-4 D (μM) Mean Callus weight (g)
SD Callus Weight (g)
SE Callus Weight (g)
0 0.2906 0.2086 0.06951 0.6591 0.1786 0.0565
10 0.6698 0.4091 0.129450 0.4018 0.3455 0.1093
100 0.2308 0.1218 0.0385
Experiment 2 Effect of sugar
The effective proportion of sugar was ___________, those medium that contained
Figure 3. Effects of the Interactions Auxin-Cytokinin on Tissue Development
NAA:BAP Ratio
Mean Callus
Weight (g)
SD Callus Weight (g)
SE Callus Weight (g)
Mean Shoot
Weight (g)
SD Shoot Weight (g)
SE Shoot Weight (g)
No. plants with
shoots
No. Roots
11 0.036 0.0576 0.0204 0.000 0.0000 0.0000 0 012 0.070 0.0917 0.0324 0.000 0.0000 0.0000 0 013 0.136 0.1102 0.0390 0.000 0.0000 0.0000 0 514 0.658 0.3142 0.1111 0.149 0.2692 0.0952 9 315 0.683 0.3753 0.1327 0.192 0.3614 0.1278 8 416 0.930 0.6156 0.2177 0.203 0.3815 0.1349 6 0
______levels of sugar presented the mean callus weight equal to (Fig. 2).
Figure 2. Relationship between Sugar Concentration and Tissue Development
Sucrose gl-¹Mean callus
weight
SD Mean callus
weight
SE Mean callus
weight
Mean shoot
weight no. plants
with shoots0 0.3870 0.1876 0.0593 0.0000 0
10 1.1067 0.7184 0.2272 0.5638 1920 1.4545 1.3053 0.4128 0.7624 2440 0.7656 0.3575 0.1131 0.6488 1760 0.4062 0.2232 0.0706 0.0590 6
Experiment 3 Relationship between auxin and cytokinin
the best combination of auxin and cytokinin proportion was ___________ (Fig. 3).
Figure 3. Auxin:Cytokinin Ratio and Organogenesis
NAA:BAP (uM)
Mean Callus
weight (g)
SD Callus Weight (g)
SE Callus Weight (g)
Mean Shoot
Weight (g)
SD Mean shoot
Weight (g)
No. Plants with
ShootsNo. Roots
0:0 0.177 0.086 0.0272 0.0000 0.0000 0 0 0.5:0 0.168 0.069 0.0217 0.0000 0.0000 0 0 5.0:0 0.179 0.119 0.0376 0.0000 0.0000 0 0
0:2.5 0.291 0.093 0.0295 0.0088 0.0247 3 80.5:2.5 1.036 0.898 0.2839 0.9385 1.2489 18 05.0:2.5 1.542 0.805 0.2547 0.5650 0.4555 31 0
Discussion: The first paragraph gives a summary of your results. In subsequent paragraphs write about the importance of plant hormones in tissue culture also include a paragraph on the possible reasons for the poor results in this years data. Do not refer to figures or tables in the discussion
References:
Dodds, J.H. and Roberts, L.W. 1985. Experiments in Plant Tissue Culture. 9780521315166.
https://books.google.com.au/books?id=-XlXQchGe5wC. Cambridge University Press 2 ed.
Ponmurugan, P., and Kumar, Suresh K.. Applications of Plant Tissue Culture. Daryaganj, IND:
New Age International, 2012. Accessed April 20, 2015. ProQuest ebrary.
Gaspar, T., Kevers, C.,Penel, C., Greppin, H., Reid, D.M., Thorpe, A. T. 1996. Plant Hormones
and Plant Growth Regulators in Plant Tissue Culture. In Vitro Cellular & Developmental
Biology. Plant Vol. 32, No. 4 (Oct. – Dec., 1996), pp. 272-289
Edwin F. George, Michael A. Hall, Geert-Jan De Klerk. 2008. Plant Propagation byTissue Culture. Chapter 1 Plant Tissue Culture Procedure – Background. Pp. Springer
The growth regulator requirements for most callus cultures are auxin and cytokinin. Auxins, as a class of compounds that stimulate shoot cell elongation, resemble IAA in their spectrum of activity. Cytokinins, which promote cell division in plant tissues under certain bioassay conditions, regulate growth and development in the same manner as kinetin. Auxin-cytokinin supplements are instrumental in the regulation of cell division, cell elongation, cell differentiation, and organ formation (Dodds and Roberts, 1985).
