Certain soil microflora can form tight associations with the root systems , or even grow within the root. Soil particles bound to roots are difficult to remove without injury to the roots that then allows microbial attack. These associated microflora will generally overgrow the tissue culture medium before there is significant growth of plant tissue. Some cultured tissues are slow in their growth. For them there would be two options: i Optimizing the culture medium; ii Culturing highly responsive tissues or varieties. Generally, plant varieties differ in susceptibility to tissue culture necrosis.
Thus, by culturing highly responsive varieties or tissues it can be managed. Aerial above soil explants are also rich in undesirable microflora. However, they are more easily removed from the explant by gentle rinsing, and the remainder usually can be killed by surface sterilization. Most of the surface microflora do not form tight associations with the plant tissue.
Practical Cell Culture Techniques
Such associations can usually be found by visual inspection as a mosaic, de-colorization or localized necrosis on the surface of the explant. An alternative for obtaining uncontaminated explants is to take explants from seedlings which are aseptically grown from surface-sterilized seeds. The hard surface of the seed is less permeable to penetration of harsh surface sterilizing agents, such as hypochlorite , so the acceptable conditions of sterilization used for seeds can be much more stringent than for vegetative tissues.
Tissue cultured plants are clones. If the original mother plant used to produce the first explants is susceptible to a pathogen or environmental condition, the entire crop would be susceptible to the same problem.
Conversely, any positive traits would remain within the line also. Plant tissue culture is used widely in the plant sciences, forestry, and in horticulture. Applications include:. Although some growers and nurseries have their own labs for propagating plants by the technique of tissue culture, a number of independent laboratories provide custom propagation services.
Since plant tissue culture is a very labour-intensive process, this would be an important factor in determining which plants would be commercially viable to propagate in a laboratory.
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From Wikipedia, the free encyclopedia. Different techniques in plant tissue culture may offer certain advantages over traditional methods of propagation, including: The production of exact copies of plants that produce particularly good flowers, fruits, or have other desirable traits. To quickly produce mature plants.
3 days of intensive lab experiments and lectures
The production of multiples of plants in the absence of seeds or necessary pollinators to produce seeds. The regeneration of whole plants from plant cells that have been genetically modified. The production of plants in sterile containers that allows them to be moved with greatly reduced chances of transmitting diseases, pests, and pathogens. The production of plants from seeds that otherwise have very low chances of germinating and growing, i.
To clear particular plants of viral and other infections and to quickly multiply these plants as 'cleaned stock' for horticulture and agriculture. In vitro tissue culture of potato explants.
This section does not cite any sources. Please help improve this section by adding citations to reliable sources. Unsourced material may be challenged and removed. February Learn how and when to remove this template message. Plant tissue culture: theory and practice Revised ed. In Vitro. Vasil; Trevor A. Thorpe Plant Cell and Tissue Culture. Acta Agriculturae Slovenica.
Shukla; A. Maxwell P. Jones; J. Saxena April Canadian Journal of Forest Research. Applied Microbiology and Biotechnology. Rai; Rajwant K.
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Kalia; Rohtas Singh; Manu P. Gangola; A. Dhawan April Environmental and Experimental Botany. In vitro regeneration of Saccharum officinarum var. Co using shoot tip explant. International Journal of Science and Nature 8 1 : Somatic embryogenesis in Strawberry Fragaria ananassa var.
Global Journal of Bioscience and Biotechnology 6 2 : - George, Edwin F. Plant propagation by tissue culture. The background 3rd ed. A cell culture course requires traditional laboratory space for performing routine laboratory operations, such as making media and immunochemical staining cells for further analysis. It is possible to share the traditional laboratory space with other teaching activities. However, a cell culture course also requires a properly equipped aseptic cell culture room.
It is impossible to share animal cell culture operations with bacterial operations, but it has been possible to share the animal cell culture facility with a plant tissue culture course, which precedes the animal cell culture course in the semester. Animal cell culture and plant tissue culture are never taught concurrently.
It is not an ideal situation to culture animal cells in the same laboratory space as hydroponically cultured plant cells, but it has been functional. The capital equipment required for the cell culture course are biological safety cabinets, inverted microscopes, table top centrifuges, pH meters, cell culture incubators, refrigerator, and vacuum pumps.
Generally, we have found it optimal to employ one pH meter, one biological safety cabinet, and one microscope per pair of students. Another issue is that there is a considerable recurring supply cost per student. Recurring costs arise from cell culture vessels, serum, reagents for the gene transfer laboratory, and reagents for the immunohistochemisty laboratory.
Another major challenge to teaching cell culture is the number of seats available at the biological safety cabinets because much of the training occurs at the biological safety cabinet. Initially — , the cell culture course was provided with two biological safety cabinets.
The course was taught in one section and accommodated seven students the first time it was taught in Also, a single section of nine students was taught in A second section was added to the course in , with eight students in one section and 12 students in a second laboratory session with a common lecture. Although there were no major problems, teaching 12 students in two biological safety cabinets was not very efficient for the instructor nor the student because students often began class before the scheduled laboratory time, while the last group of students tended to finish the exercise after the usually scheduled laboratory time.
Instructing eight students was manageable with two biological safety cabinets. Students are clearly presented the information to keep in the notebook during the first laboratory session, and the notebook guidelines are made available online throughout the course. Last, a class participation grade is given to the students to encourage punctual attendance and participation. Because the course is entirely techniques based, it is important that the students be evaluated for their laboratory efforts.
In general, students have been excited about the laboratory sessions, and students missing class sessions has not been a problem. Currently, there have not been a sufficient number of students completing the course to undertake a detailed analysis of the success of the graduates. However, representatives from local biotechnology companies have begun to seek graduates as employees, and they have indicated that the course provides useful skills. In the course evaluations provided by the students, they typically indicate that they had limited or no cell culture experience before enrolling in the course, but they would be comfortable performing tissue culture in unfamiliar surroundings at the conclusion of the class.
Several students have personally indicated that they have extensively used the skills learned in the course either in subsequent employment or in completing their research projects. Furthermore, teaching cell culture is labor intensive and requires close supervision during the initial phases of training. Institutions and instructors have to make a conscious investment in capital equipment, recurring supplies, and time. Although animal cell culture is a classical and straightforward technique, it is also a technique that is going to become even more prevalent in modern biology because it reduces the number of animals employed in biological studies, and it is a simple system free of the confounding effects of many different cell types found in vivo.
Related Practical Cell Culture Techniques
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