• Join over 1.2 million students every month
  • Accelerate your learning by 29%
  • Unlimited access from just £6.99 per month

Work shope on plant Cryopreservation

Extracts from this document...


Name- Venkatesh Kolluru Student number- 0803568 Course- MSc. BIOTECHNOLOGY Module- BI1102A Title of the work shop- PLANT CRYOPRESERVATION Workshop tutor- Irene Tierney AIM: To learn about the practical aspects of plant cryopreservation and its current developments in the field of plant biotechnology. INTRODUCTION: Cryopreservation is a process where the cells or the whole tissues are preserved by cooling the temperature to -196C (BP of liquid nitrogen). In these extreme conditions the biological activity even the biochemical reactions which would lead to cell death is effectively stopped. But this technique consists of problems like damage to the cell due to solution effect (used in the solution), dehydration and intra cellular ice formation. At this temperature the salts present in the cell may become crystal which can ultimately lead to fatal conditions. To overcome this problem vitrification solutions (sucrose, DMSO i.e. Di-Methyl sulphoxide, glycerol, etc.) are used. Vitrification solutions prevent the formation of ice crystals. They do it by lowering down the freezing point and increasing the viscosity. The time of exposure of cryoprotectants (i.e. vitrification solutions) is very important because the long exposure of chemicals towards the plant cells may result in to phytotoxicity. Cryopreservation is a technique which is carried out frequently for the conservation of plant genetic resources. This was initially developed by Withers and King in 1980, but the cryopreservation techniques which are used now a day have several modifications in the protocol which was used by the initial developers. ...read more.


This Petri dish was labelled as control, 2hrs -LN. 13. Another 3 embryos were taken and placed on cryovial labelled as +LN, this is attached to the cryocane and was given for storage in LN for 5min. 14. Another 3 embryos were taken into the glass bottle provided for moisture content (MC) determination after 2 hrs desiccation. 15. The weight of the empty weighing bottle without cap was determined and was recorded as W1. 16. The encapsulated embryos were placed in the weighing bottle and weight was recorded as W2. 17. This bottle was labelled with cryo marker and was given for oven drying at 103oC for 17 hrs. Oven drying was overnight so dry weight of the samples was recorded as W3 after drying. Calculation of beads moisture content: % Bead MC = W2-W1 / W3-W1 x 100 18. Rapid warming will be carried out for cryoreserved embryos. 19. After rapid warming the beads were placed onto recovery medium, sealed and labelled. EXERCISE 2 AIM: cryopreservation of shoot tips by vitrification. MATERIALS: METHODS: 1. The solanum shoot tips were placed in a cryovial containing the lowest concentration of PVS2 solution for about 1-10 min. 2. Almost all the vitrification solution were removed from the tissues and were replaced with a step wise higher concentration of PVS2 solution (50% 60% 70% 80% 90% 100%). All these were performed on ice. ...read more.


The expected reason behind this could be experimental error that could be related to personal error, mechanical error and other unobservatory defects. EXCERSISE 2: In this case there was no contamination. But there was no growth of the test samples on MS media with respect to both the cases i.e. -LN and +LN. This indicates towards unknown experimental error during experiment. But in second case there was a growth in +LN and no growth in -LN. This showed success as per the expected results because liquid nitrogen has allowed the survival of the tested sample. This indicates towards the efficiency of vitrification. Vitrification solution initially dehydrated the sample. The liquid nitrogen at -196oC is very effective in preserving the vital plant genotypes and resources. Vitification solutions aid in this processes by removing the risk of crystal formation. Ice crystal formation can rupture the cell membrane of the plant cells which ultimately result in the failure in the survival of stored samples. Thus the cryopreservation represents a effective and potential technique for storing the plant tissue resources. CONCLUSION: REFERANCE: * Cryopreservation. 2009. In Encyclop´┐Żdia Britannica. [online]. Available from :http://www.britannica.com/EBchecked/topic/1452607/cryopreservation [Accessed 5 April 2009] * Lambardi, M. and Panis, B. 2005. STATUS OF CRYOPRESERVATION TECHNOLOGIES IN PLANTS (CROPS AND FOREST TREES). [online]. Available from: www.fao.org/biotech/docs/panis.pdf [Accessed 5 April 2009] * Reed, B. M. 2004. Technical guidelines for the management of field and in vitro germplasm collections. [online] Bioversity International. Available from: http://books.google.co.uk/books?id=QNWf4HW1EnMC [Accessed 7 April 2009] ?? ?? ?? ?? ...read more.

The above preview is unformatted text

This student written piece of work is one of many that can be found in our University Degree Botany section.

Found what you're looking for?

  • Start learning 29% faster today
  • 150,000+ documents available
  • Just £6.99 a month

Not the one? Search for your essay title...
  • Join over 1.2 million students every month
  • Accelerate your learning by 29%
  • Unlimited access from just £6.99 per month

See related essaysSee related essays

Related University Degree Botany essays

  1. To investigate the effects of abiotic factors specifically pH on the abundance of marram ...

    Use distilled water This would make sure that the water had a solute potential of 0 allowing osmosis and the infiltration rate to occur at an accurate speed. Rank order Error in procedure Effect of error on result/ trend Improvement Justification of improvement 1 Ranging pole distance The distance between the ranging poles was 10m.

  2. Demonstrate the separation of plant pigments using chromatography and the rate of photosynthesis in ...

    DPIP replaces the NADP+ found in chloroplasts. 3. Light is the source of electrons that will reduce DPIP. 4. The spectrophotometer was used to determine the change in light transmittance at a wavelength of 605 nm. As more light was absorbed by the chloroplasts, there was a change in the amount of light that passed through the DPIP.

  1. An investigation to find the effect of bile salts on the digestion of fats.

    I will collect the initial rate of reaction, when the chances of all the active sites being occupied are high. This ensures I am measuring the breakdown of fats while the pH of the overall mixture is alkaline, and so I can be sure that the acidity of the forming

  2. To what extent does plasticity of dipterocarp seedlings affect growth and survival in the ...

    How vital is plasticity to the growth and survival of dipterocarp seedlings, and do other factors, such as herbivory play a role? Methods STUDY SITE AND SPECIES The study was carried out in primary lowland dipterocarp rainforest surrounding Danum Valley Conservation Area, Sabah, East Malaysia, Borneo (4"58'N, 117'48'E).

  1. A review of the development, production and post harvest requirements of Gerberas

    Peat is also tricky to irrigate and can either become too wet, or dry out and then take a long time to absorb water (Mathias, 2006). Coir: Coir or coco substrate has become more popular in Gerbera cultivation because of its good stability and air/water ratio (Florist de Kwakel (no date [b]).

  2. The combination of constant warmth and abundant moisture of the tropical regions demonstrates a ...

    A few groups, such as marine infauna and fresh water invertebrates, appear not to follow this pattern but many plant and animal taxa display latitudinal gradients. *Approaches to the study of the diversity gradient have so far been mainly of two types-the method of gross geographical lumping with comparison of

  1. Induced defence responses against herbivores. The aim of the project was to study ...

    Other proteins which deter insect herbivores are amylase inhibitors and lectins. Figure 1. (Roberts M. R., et al., 2001) Showing a basic scheme for a plant response to wounding. Mechanical wounding, simulating herbivore grazing elicits the expression of local and systemic genes.

  2. Mechanisms of insect resistance induced by treatment of Lycopersicon esculentum seeds by jasmonic ...

    genes in tomato plants. JA has been found to be present in many plant species and is involved in regulating diverse plant functions including plant resistance and senescence (Creelman et al., 1997) Studies have shown that although MeJA is not elicited in sufficient quantities or over sufficient distanced to induce Pin genes in neighbouring

  • Over 160,000 pieces
    of student written work
  • Annotated by
    experienced teachers
  • Ideas and feedback to
    improve your own work