How To Tissue Culture Plants

Contents

​DIY Tissue Culture – Why You Should Be Trying It

The improvements in biological and scientific research have been many throughout the years, but there have been a few noteworthy discoveries that have had a significant impact on our industrial and consumer worlds. The procedure of tissue culture is one of the most notable of them. Whether you are an at-home grower, a small-scale agriculturalist, or a large enterprise, the tissue culture procedure is relevant to your needs and objectives.

What Exactly is this Tissue Culture Process?

You can obtain extra cells, new cells, or tissue from existing plant matter through the use of the tissue culture procedure. I get what you’re saying, but isn’t that how seeds and germination work? The distinction is that the tissue culture procedure, rather of using seeds, allows you to generate new plants or plantlets by using live materials or organisms rather than seeds. A synthetic environment is used for this procedure, which involves the cultivation of cells. While this method appears to be hard at first glance, it is actually more simpler than it appears.

However, while certain plants are more difficult to produce than others, plants such as cannabis may be grown with reasonable simplicity as long as the proper procedures are followed.

Why You Should use Plant Tissue Culture

The most advantageous aspect of the plant tissue culture procedure is that it allows you to produce several plants in a short amount of time that are not only high-quality disease-free genes, but are also genetically consistent in their genetic composition as a whole. This is particularly important for medical marijuana producers who rely on a certain strain of cannabis for the precise health advantages it provides, as well as for businesses that make a profit from the sale of plant material or derived goods from the cannabis plant.

In other words, you could have a good time doing it.

DIY Tissue Culture

First and foremost, you’ll need a sufficient quantity of space in which to set up your home lab, grow room, or whatever you choose to call your new facility. Regardless of whether it is a garage or a spare room, the place must be one that can be tightly regulated and monitored. Autoclaves are pricey pieces of lab equipment that are often used by scientists to physically disinfect their specimens. They are used to physically disinfect your specimens. A pressure cooker or even a microwave are suitable options for cooking when you are at home.

Take a look at the following list of goods that you might require for your DIY Tissue Culture:

  • Cooking in a microwave or pressure cooker
  • Using disinfectants such as hydrogen peroxide or bleach
  • Using recycled glass bottles (the container for the developing plants). Some folks have reported that the size of a baby feeding bottle is the right fit. Make sure you have enough water, vinegar, baking soda, and sugar on hand in case you need them as well.

DIY Tissue Culture: The Essentials

ProtocolPreviously, we discussed a protocol for culturing cannabis; this indicates that you should conduct study about how it has been done in the past with the plant species you desire to culture, and that this protocol should be utilized as a guideline for your DIY cannabis cultivation process. Plant-Based Substances After that, you’ll need to collect the plant that will be used in the cultivation procedure. The plant you choose should be as disease-free as feasible and free of pests and illnesses.

  • You should keep in mind that the tissue culture process is dependent on a clean environment, which means that you should always ensure that you are thoroughly cleaned and wearing clean clothes before you begin the culturing procedure.
  • Air Clean air is also required by the plants.
  • The most typical handmade box is built by repurposing an old fish tank or by covering PVC pipework with clear plastic.
  • In this case, the substance on which the plant material will be put should be able to supply two things: stability as well as nutrition.
  • It is a gel-like substance that contains vital hormones and nutrients that aid in the formation of roots and the growth of shoots.
  • These are the most important components of the tissue culture process, and as you can see, they are all readily available at your local convenience shop or at your house if you are interested in carrying out a simplified procedure DIY project.
  • Articles that have been highlighted as resources

Tissue Culture Propagation of Banana

Bananas are a tropical fruit that can be eaten both raw and cooked by humans. They are widely available in supermarkets. It is said to have originated in Southeastern Asia, in nations such as India, the Philippines, Malaysia, and so on and so forth. The edi is a. click here to find out more

How PPM™ Can Save Your Tissue Culture Experiment

In plant tissue culture experiments, the Plant Preservative Mixture (PPMTM) is a stable formulation that is used as a broad-spectrum biocide. It is available in a variety of strengths. By focusing on bacteria, fungus, and other contaminations, we are able to. click here to find out more

PPM vs Antibiotics – A Comparison

Whether you are a seed-to-fruit grower or a plant cloning genius, you understand how important it is to maintain your plants free of contaminants in order to achieve success. From airborne microbial infections to airborne microbial diseases, airborne. click here to find out more

Tissue Culture Contamination and 7 Easy Steps of Prevention

Contamination has struck once more! Tissue culture is a time-consuming and labor-intensive procedure, and it may be frustrating when fungus or bacteria infect our delicate cultures. Culturing cells in the laboratory necessitates the use of. click here to find out more

Plant tissue culture – Wikipedia

Plant tissue culture is a set of procedures that are used to retain or develop plant cells, tissues, or organs under sterile circumstances on a nutrient culture medium that has a specified composition in order to produce a specific product. Micropropagation, often known as plant cloning, is a technique that is frequently used to make clones of plants. The use of different methods in plant tissue culture may provide significant benefits over traditional methods of propagation, such as the following:

  • It is the practice of creating precise replicas of plants that produce exceptionally nice flowers, fruits, or have other desired characteristics. In order to grow mature plants as rapidly as possible
  • Generation of large numbers of plants in the lack of seeds or the presence of pollinators essential for the production of seeds
  • Genetically engineered plant cells are used in the regeneration of whole plants. The production of plants in sterile containers that allows them to be moved with a greatly reduced risk of transmitting diseases, pests, and pathogens
  • The production of plants in sterile containers that allows them to be moved with a greatly reduced risk of transmitting diseases, pests, and pathogens
  • Developing plants from seeds that would normally have very poor possibilities of germination and growing, such as orchids and Nepenthes
  • To rid certain plants of viral and other diseases and to rapidly proliferate these plants for use in horticulture and agriculture as ‘cleaned stock.’

Using plant tissue culture, researchers take use of the fact that a large number of plant cells have the potential to regenerate a whole plant (Cellular totipotency). Single cells, plant cells without cell walls (protoplasts), sections of leaves, stems, or roots can all be used to grow a new plant on culture media if the medium has the necessary nutrients and plant hormones (see below).

Techniques used for plant tissue culture in vitro

The preparation of plant tissue for tissue culture is carried out under aseptic circumstances in an alaminar flow cabinet with HEPAfiltered air supplied by a dehumidifier. Following that, the tissue is cultivated in sterile containers, such as Petri dishes or flasks, in a growth environment with temperature and light intensity that are carefully regulated. The surfaces of living plant materials from the environment are naturally contaminated with microorganisms, so their surfaces are sterilized in chemical solutions (typically containing alcohol and sodium or calcium hypochlorite) before suitable samples (known as explants) can be collected and studied.

  • Solid and liquid media are primarily composed of inorganic salts, with a small amount of organic nutrients, vitamins, and plant hormones thrown in for good measure.
  • Potato explants were cultured in vitro to produce tissue.
  • For example, an excess ofauxinwill frequently result in the proliferation of roots, whereas an excess ofcytokininwill likely result in the production of shoots.
  • In order to allow for development or to modify the morphology of the culture, portions of the culture are routinely cut off and subcultured onto fresh medium as the culture matures.

As shoots emerge from a culture, they can be sliced off and rooted with auxin to produce plantlets, which can then be transplanted to potting soil for further growth in the greenhouse as normal plants once they have reached maturity.

Regeneration pathways

Plant tissue cultures are being cultivated at the National Center for Genetic Resources Preservation, which is operated by the United States Department of Agriculture. There are a variety of theories for the unique variances in the regeneration capability of different organs and explants that exist. The variations in cell cycle stage, the availability of endogenous growth regulators or the capacity to transfer them, and the metabolic capacities of the cells are the most critical elements to consider.

These tissues have rapid cell division and either concentrate or synthesize the necessary growth-regulating chemicals, including as auxins and cytokinins, that are essential for proper development.

As a result, tissue culture regeneration can become challenging, especially when several regeneration processes for various genotypes within the same species must be devised.

In most cases, the propagation of shoots or nodal segments is performed in four stages for the mass production of plantlets through in vitro vitrovegetative multiplication, but organogenesis is a common method of micropropagation that involves the tissue regeneration of adventitious organs or axillary buds, either directly or indirectly from the explants, and is a common method of micropropagation.

Non-zygotic embryogenesis is a significant developmental process that is extremely analogous to that of zygotic embryos.

Because non-zygotic embryos are formed from a single cell, they are favoured in a variety of regeneration systems, including micropropagation, ploidy modification, gene transfer, and synthetic seed synthesis, among others.

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Choice of explant

An explant is a piece of tissue taken from a plant that will be used in a culture. Single undifferentiated cells, as well as many different types of mature cells, can be used to create explants. Explants can be taken from many different parts of a plant, including portions of shoots, leaves, stems, flowers, roots, single undifferentiated cells, and many different types of mature cells, as long as they contain living cytoplasm and nuclei and are capable of de-differentiating and resuming cell division.

This, however, is not true for all cells or all plants in the same way.

The choice of explant material also impacts whether the plantlets produced during tissue culture are haploid or diploid in nature.

When compared to the other two methods, the first method, which involves the use of meristems and the induction of multiple shoots, is the preferred method for the micropropagation industry because the risks of somaclonal variation (genetic variation induced in tissue culture) are reduced to an absolute minimum.

  • Some explants, such as the root tip, are difficult to separate and are contaminated with soil microorganisms, which can cause problems throughout the tissue culture process if not handled properly.
  • Soil particles that have been entangled with roots are difficult to remove without causing damage to the roots, which then permits a microbial attack to take place.
  • Some cultured tissues develop at a snail’s pace, and this is a problem.
  • Necrosis can cause the degradation of cultured tissues.
  • As a result, it can be controlled by cultivating particularly sensitive cultivars (or tissues).
  • Although they are more readily removed from the explant by gentle washing, the remaining bacteria are typically eliminated by surface sterilization, which is a common practice.
  • Visual evaluation of the explant will frequently reveal such associations as a mosaic, de-colorization, or localized necrosis on the surface of the explant.
  • Considering that seeds have a hard surface that makes them less permeable to the penetration of severe surface sterilizing chemicals, such as hypochlorite, the allowable sterilization conditions employed for seeds can be far more strict than those used for vegetative tissues.

If the original mother plant that was used to create the first explants is vulnerable to a disease or a certain environmental situation, the entire crop will be susceptible to the same ailment. Positive characteristics, on the other hand, would remain inside the limits of the line.

Applications of plant tissue culture

Plant tissue culture is widely utilized in the plant sciences, forestry, and horticulture, among other fields. Among the applications are:

  • This refers to the commercial manufacture of plants for use as potting, landscaping and florist subjects, which employs meristem and shoot culture to generate vast numbers of identical individuals. Toconservare or endangered plant species are two words that come to mind. Instead of screening cells for desirable characteristics, a plant breeder may utilize tissue culture to screen cells for advantageous characteristics, such as herbicide resistance or tolerance. Plant cells are grown on a large scale in liquid culture in bioreactors for the synthesis of important substances such as plant-derived secondary metabolites and recombinant proteins that are utilized as biopharmaceuticals.
  • By fusing protoplasts and regenerating the novel hybrid, it is possible to bridge species that are distantly related. To swiftly investigate the molecular underpinnings of physiological, metabolic, and reproductive functions in plants, for example, by in vitro selection for stress resistant varieties. Cross-pollination between distantly related species followed by tissue culture of the resultant embryo that would otherwise perish (Embryo Rescue)
  • For the purpose of chromosomal doubling and the production of polyploidy, such as doubled haploids, tetraploids, and other kinds of polyploidy, This is often accomplished by the use of antimitotic medicines such as colchicine or oryzalin. Transformation of a tissue followed by either short-term testing of genetic constructs or regeneration of transgenic plants It is possible to grow clean plant material from virused stock using procedures such as meristem tip culture, which may be used to a variety of plant species including sugarcane, potatoes, and many types of soft fruit. It is possible to generate identical sterile hybrid species by breeding them together
  • Somatic embryogenesis is used to produce fake seeds on a large scale.
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Laboratories

A variety of independent laboratories provide specialized plant propagation services, despite the fact that several growers and nurseries have their own labs for the method of tissue culture. Many commercial tissue culture laboratories are listed on the Plant Tissue Culture Information Exchange website. Because plant tissue culture is a time-consuming and labor-intensive procedure, it would be crucial to consider this when evaluating which plants would be financially feasible to produce in a laboratory setting.

See also

  • Root culture with a lot of hair
  • Gottlieb Haberlandt was a pioneer in plant tissue culture, and he was born in Germany. Frederick Campion Steward was a pioneer in plant tissue culture and was known as the “champion.” Among the most significant plant growth mediums are Murashige and Skoog medium as well as Hoagland solution. Physiology of plants

References

  1. B.N. Sathyanarayana, B.N. Sathyanarayana, B.N. (2007). Growing Plant Tissues: Current Practices and Innovative Experimental Protocols. I. K. International, pp. 106–. ISBN 978-81-89866-11-2
  2. Bhojwani, S. S.
  3. Razdan, M. K. Bhojwani, S. S.
  4. Razdan, M. K. International, pp. 106–. ISBN 978-81-89866-11-2
  5. Razdan, M. K. (1996). Plant tissue culture: principles and applications (Revised ed.). Elsevier.ISBN978-0-444-81623-8
  6. s^ The Vasils, I.K., and Vasil, V. (1972). “Totipotency and embryogenesis in plant cell and tissue cultures” is the title of this article. In Vitro.8(3): 117–125.doi: 10.1007/BF02619487.PMID4568172.S2CID20181898
  7. In Vitro.8(3): 117–125.doi: 10.1007/BF02619487.PMID4568172.S2CID20181898 Brian James Atwell, Colin G. N. Turnbull, and Paul E. Kriedemann are among those who have contributed to this work (1999). Nature’s Adaptation and Cultivation’s Performance: Plants in Action (1st ed.). The original version of this article was published on March 27, 2018. retrieved on May 7, 2020
  8. Retrieved on May 7, 2020
  9. Indra K. Vasil and Trevor A. Thorpe have collaborated on this project (1994). Plant Cell and Tissue Culture are two different things. Springer Publishing Company, pp. 4–. ISBN 978-0-7923-2493-5
  10. AbPazuki, Arman Sohani and Mehdi are sisters (2013). Indica rice varieties were evaluated for their calluses, which were formed by scutellum-derived keratin (PDF). Mukund R. Shukla
  11. A. Maxwell P. Jones
  12. J. Alan Sullivan
  13. Chunzhao Liu
  14. Susan Gosling
  15. Praveen K. Saxena
  16. Acta Agriculturae Slovenica.101(2): 239–247.doi:10.2478/acas-2013-0020
  17. Mukund R. Shukla
  18. (April 2012). A possible function for auxin metabolism in prolonged plant proliferation has been proposed for the preservation of American elm (Ulmus americana) in vitro. Canadian Journal of Forest Research, volume 42, number 4, pages 686–697. Milen I. Georgiev, Jost Weber and Alexandre MacIuk have published a paper with the doi:10.1139/x2012-022 (2009). Plant cell culture bioprocessing for the mass manufacture of specific chemicals is described in detail in the paper. Applied Microbiology and Biotechnology, volume 83, number 5, pages 809–23. Manoj K. Rai
  19. Rajwant K. Kalia
  20. Rohtas Singh
  21. Manu P. Gangola
  22. A.K. Dhawan
  23. Doi: 10.1007/s00253-009-2049-x.PMID19488748.S2CID30677496
  24. Manoj K. Rai
  25. A.K. Dhawan (April 2011). « Developing stress tolerant plants by in vitro selection—An summary of recent developments in this field », according to the journal “Developing Stress Tolerant Plants through In Vitro Selection.” Environmental and Experimental Botany, volume 71, number 1, pages 89–98. Aina, O., Quesenberry, K., and Gallo, M. (2010). doi: 10.1016/j.envexpbot.2010.10.021 (2012). Arachis paraguariensis tetraploids are produced in vitro, according to the study. In vitro regeneration ofSaccharum officinarumvar. Co 92005 utilizing shoot tip explants. Plant Cell, Tissue, and Organ Culture.111(2): 231–238.doi: 10.1007/s11240-012-0191-0.S2CID9211804
  26. Pawar, K. R., Waghmare, S. G., Tabe, R., Patil, A., and Ambavane, A. R. 2017. In vitro regeneration ofS International Journal of Science and Nature, volume 8, number 1, pages 154-157
  27. Waghmare, S. G., Pawar, K. R., and Tabe, R. 2017. Waghmare, S. G., Pawar, K. R., and Tabe, R. 2017. Strawberry (Fragaria ananassa) var. Camarosa somatic embryogenesis was seen. Global Journal of Bioscience and Biotechnology, volume 6, number 2, pages 309-313
  • B.N. Sathyanarayana is a well-known Indian philosopher (2007). Growing Plant Tissues: Current Practices and New Experimental Protocols Bhojwani, S. S.
  • Razdan, M. K. (eds.). I. K. International, pp. 106–. ISBN 978-81-89866-11-2. (1996). Theories and applications of plant tissue culture (Revised ed.). Elsevier.ISBN978-0-444-81623-8
  • s^ I.K. Vasil and V. Vasil (1972). “Totipotency and embryogenesis in plant cell and tissue cultures” is the title of the paper that was presented at the conference. S2CID20181898
  • Doi: 10.1007/BF02619487.PMID4568172.S2CID20181898
  • In Vitro.8(3): 117–125.doi: 10.1007/BF02619487.PMID4568172.S2CID20181898 Brian James Atwell, Colin G. N. Turnbull, and Paul E. Kriedemann are among those who have contributed to this publication (1999). Nature’s Adaptation, Cultivation’s Performance: Plants in the Field (1st ed.). On March 27, 2018, an archived version of this article appeared. the 7th of May, 2020 (retrieved)
  • Vasil, Indra K., and Thorpe, Trevor A. (1994). Cultivation of Plant Cells and Tissue ISBN 978-0-7923-2493-5
  • AbPazuki, Arman (Springer, pp. 4–). Amir, Mehdi, and Sohani (2013). Rice varieties from the ‘Indica’ subcontinent were examined for their phenotypic characteristics (PDF). Agriculturae Slovenica.101(2): 239–247.doi:10.2478/acas-2013-0020
  • Mukund R. Shukla
  • A. Maxwell P. Jones
  • J. Alan Sullivan
  • Chunzhao Liu
  • Susan Gosling
  • Praveen K. Saxena
  • Mukund R. Shukla, A. Maxwell P. Jones, A. Maxwell P. Jones, A. Maxwell P. Jones, A. Maxwell P (April 2012). A possible function for auxin metabolism in prolonged plant proliferation has been proposed for the preservation of American elm (Ulmus americana) in culture. In the Canadian Journal of Forest Research, Vol. 42, No. 4, 686–697, the authors report that DOI: 10.1139/x2012-022
  • Georgiev, Milen I
  • Weber, Jost
  • MacIuk, Alexandre
  • Georgiev, Milen I (2009). Plant cell cultures are being processed in order to produce specified chemicals in large quantities. 93.5(5): 809–23 (Applied Microbiology and Biotechnology). PMID19488748.S2CID30677496
  • Manoj K. Rai, Rajwant K. Kalia, Rohtas Singh, Manu P. Gangola, and A.K. Dhawan
  • Doi: 10.1007/s00253-009-2049-x.PMID19488748.S2CID30677496
  • (April 2011). « Developing stress tolerant plants by in vitro selection—An summary of recent developments in this field », according to the journal “Developing Stress Tolerant Plants Through In Vitro Selection.” Botanical Journal of the Linnean Society.71(1): 89–98. Aina, O.
  • Quesenberry, K.
  • Gallo, M.
  • Doi: 10.1016/j.envexpbot.2010.10.021
  • (2012). Arachis paraguariensis tetraploids are produced in vitro, according to the researchers. In vitro regeneration ofSaccharum officinarumvar. Co 92005 utilizing shoot tip explants. Plant Cell, Tissue, and Organ Culture.111(2): 231–238.doi: 10.1007/s11240-012-0191-0.S2CID9211804
  • Pawar, K. R., Waghmare, S. G., Tabe, R., Patil, A., and Ambavane, A. R. 2017.In vitro regeneration ofS IJSN 8(1): 154-157
  • International Journal of Science and Nature In S. G. Waghmare et al. (2017), Pawar K. R. and Tabe R., Pawar and Tabe et al. (2017), Pawar K. R. and Tabe R. 2017. Strawberry (Fragaria ananassa var. Camarosa) somatic embryogenesis was observed. Journal of Bioscience and Biotechnology, Volume 6, Number 2, Pages 309-313

Tissue Culture Technology

Every plant, just like every individual is different and one-of-a-kind, is also unique. Some have characteristics like as improved color, yield, or insect resistance. Scientists have been searching for technologies that would allow them to create perfect replicas of these exceptional individuals for many years. Plants normally reproduce by generating seeds, which is a process known as sexual reproduction. In other words, pollen from the stamens of the plants fertilizes the egg cells in the flowers of the plants.

DNA from both parents is mixed in unforeseen and novel ways during sexual reproduction, resulting in the creation of distinct plants.

Several of us believe that all plants are propagated from seeds.

And scientists are now accomplishing this with the use of a technique known as “tissue culture.”

What is Tissue Culture?

It is the growing of plant cells, tissues, or organs on specially prepared nutritional medium that is referred to as tissue culture (TC). Under the correct circumstances, it is possible to regenerate a whole plant from a single cell. Plant tissue culture has been around for more than 30 years and is a relatively simple procedure. When it comes to the development of disease-free, high-quality planting material as well as the quick production of large numbers of homogeneous plants, tissue culture is considered a critical technique for developing nations.

  1. It is possible to create thousands of duplicates of a plant in a short period of time in this manner.
  2. They also have a shorter and more consistent production cycle, and generate larger yields.
  3. Its implementation needs simply a sterile workplace, nursery, and greenhouse, as well as highly trained personnel.
  4. Oil palm, plantain, pine, banana, date, eggplant, jojoba, pineapple, rubber tree, cassava, yam, sweet potato, and tomato are among the plants that have been grown in tissue culture and are important to developing countries.

Other plants that have been grown in tissue culture include yam, sweet potato, and tomato. This application of traditional biotechnology is the most widely used kind of traditional biotechnology in Africa.

Uses of TC technology in Asia

  • In order to meet the needs of orchid species and hybrids that are known to thrive in Southeast Asia, tissueculture has been refined over time. Thailand, Singapore, and Malaysia have all found that the ornamental and cut flower trade is a significant source of foreign exchange and additional income for small growers. In Thailand, tissue culture is used to reproduce slow-growing and environment-sensitive orchids, which are difficult to grow in their natural environment. With a production capacity of 50 million plantlets per year, Thailand is the leader in tissue culture in Southeast Asia. For the most part, these are orchids, which have helped the country to become the world’s leading exporter of whole and cutorchids. Micropagation by shoot culture technique has been developed for the mass propagation of bananas. Banana bunchy top virus (BBTV) and banana bract mosaic virus (BBrMV) are two viral diseases that are commonly spread through propagative materials in the Philippines, and this is used as a control strategy.

Benefits of TC technology for small-scale banana producersin Kenya(Source: ISAAA)

When it comes to food crops, bananas are extremely important in Kenya and many other parts of the tropical and subtropical developing world. However, during the previous 20 years, there has been a significant fall in banana output, owing to extensive soil deterioration and the infection of banana plants with pests and diseases. Adding to the difficulty was the widespread practice of cultivating new banana plants from sick suckers, which exacerbated the situation. Food security, employment, and income in banana-producing areas were jeopardized as a result of the crisis.

With proper pest and disease management and field hygiene, yield losses caused by pests and diseases have been reduced significantly at the farm level.

  • Large quantities of superior clean planting materials that are early maturing (12-16 months as opposed to the conventional banana’s 2-3 years)
  • Larger bunch weights (30-45 kg as opposed to the 10-15 kg from conventional material)
  • Higher annual yield per unit of land (40-60 tons per hectare as opposed to the 15-20 tons previously realized with conventional material
  • And higher annual yield per unit of land

Furthermore, regularity in orchard creation and simultaneous plantation expansion made it easier to coordinate marketing efforts in the field. It also provided the opportunity to turn banana farming from a purely subsistence activity into a profitable commercial enterprise, if desired. The project’s cost-benefit study revealed that transgenic banana cultivation is more profitable as an enterprise than traditional banana farming, which is a promising development. In addition, the project has mostly benefited women who tend the crop, so contributing to the reduction of the gender gap.

Benefits of TC technology for rice farmers in West Africa (Source: WARDA)

Over the course of several decades, scientists have fantasized about fusing the ruggedness of the African rice species (Oryza glaberrima) with the productivity of the Asian rice species (Oryza edulis) (Oryza sativa). However, the two are diametrically opposed. Attempts to cross them were unsuccessful since the offspring produced were all infertile. When rice breeders from the West Africa Rice Development Association (WARDA) faced infertility issues in the 1990s, they resorted to biotechnology in an attempt to address the problem.

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Scientists were able to cross these two species thanks to advancements in agricultural research.

Once the fertility of the offspring had been enhanced (typically after several cycles of back-crossing), anther cultivation was employed to double the gene complement of the male sex cells (anthers) and, as a result, generate true-breeding plants, which were then sent to the field.

Some of the new plants blended the yield characteristics of the sativa parent with the characteristics of the glaberrima parent to produce hybrids with higher yields. NERICAs are characterized by the following qualities in general:

  • Wide and droopy leaves that smother weeds in their early stages of development
  • Panicles or grain heads that are longer and have ‘forked’ branches that can hold up to 400 grains
  • Moretillers with strong stems that can support and hold tightly the heavy grainheads
  • Rice yields as high as 2.5 tons per hectare with low inputs — and 5 tons or more with only a minimal increase in fertilizer usage (amounting to a 25 percent to 250 percent increase in production)
  • Matures 30 to 50 days earlier than current varieties, allowing farmers to plant additional crops of vegetables or legumes
  • And matures 30 to 50 days earlier than current varieties, allowing farmers to plant additional crops of vegetables or legumes. It grows higher than other rice types and is more resistant to pests and drought than most others. thrives well on barren and acidic soils—which account for 70% of WestAfrica’s upland rice region
  • Contains 2% more protein per kilogram of body weight than their African or Asian ancestors

NERICAs were soon embraced by farmers as a result of their widespread success. The new rices were thought to span around 8,000 hectares in Guinea in 2000, with 5000 ha under cultivation by 20,000 farmers under the supervision of a national extension organization, according to estimates. It was anticipated that 330,000 hectares (ha) of NERICAs would be planted in 2002, which would be adequate to cover the country’s own seed requirements while also providing a surplus for sale to neighboring nations.

Glossary

Anther: The most important male reproductive structure, in which pollen is produced and preserved. Apical meristems are found at the tips of roots and stems, where new cells are produced. DNA is a molecule present in the cells of living beings that stores genetic information (also known as genetic material). In plant breeding, embryo rescue refers to a series of tissue culture procedures that are used to allow an unfertilized immature embryo arising from an interspecific cross to continue growing and developing until it may be regenerated into a full plant.

References:

  1. DANIDA.2002. Development and use of plant biotechnology in the context of plant breeding and agricultural production in underdeveloped countries are being evaluated for their potential and restrictions. This is a working document. DeVries, J., and Toenniessen, G. (2001)
  2. Ministry of Foreign Affairs, Denmark
  3. DeVries, J., and Toenniessen, G. African crop security is ensured by the use of biotechnology, breeding, and seed systems. The Rockefeller Foundation in New York, USA
  4. The Food and Agriculture Organization of the United Nations in 2002 An administrative and policy document on crop biotechnology for Sub-Saharan African administrators and policy officials. Kitch, L., Koch, M., and Sithole-Nang, I.
  5. International Service for the Acquisition of Agri-biotech Applications (ISAAA)
  6. Philippine Recommendations, 1994
  7. Ruff, Anne Marie. International Service for the Acquisition of Agri-biotech Applications (ISAAA)
  8. Philippine Recommends, 1994. As the saying goes, “We are sowing the seeds of revolution.” ()
  9. Wambugu, F., and Kiome, R. (2001)
  10. Wambugu, F., and Kiome, R. (2001). The advantages of biotechnology for small-scale banana growers in Kenya are well documented. ISAAABriefs No. 22 is now available. ISAAA is based in Ithaca, New York, while the West Africa Rice Development Association (WARDA) is based in Lagos, Nigeria.

*The month of November 2006

Next Pocket K:’Omics’ Sciences:Genomics, Proteomics, and Metabolomics

The month of November 2006*

  1. Open the flask (plastic container) in a well-ventilated, cool, and, most importantly, clean environment
  2. Carefully remove the plants and rinse off any gel that may have remained on the roots
  3. And repeat the process. If left in place, it will promote fungal development when exposed to non-sterile environments. It is critical that this is removed. Open the flask and pour some lukewarm water into it
  4. This is a good technique to utilize. Swirl the flask until the whole clump of plants, as well as the jelly, becomes loose and begins to swirl in response to the swirling movement. After that, delicately tip the plant and gel clump into a bowl of lukewarm water, and carefully separate the jelly from the roots/base of the plant. Not only does this make it easier to remove all of the gel, but it also has the added benefit of causing less harm to the roots. Remove plantlets from just one flask at a time, and plant them as soon as possible after removal. Planting should be done first thing in the morning, before the temperatures begin to rise substantially. If feasible, use a sterilised potting mix with a low nutrient concentration (i.e. one that has been decontaminated). We make use of peat moss and perlite (50:50). Check to see that the potting mix has been watered but is not overly saturated
  5. Do not plant in potting mix that is too hot (above 25 degrees Celsius). During the first few weeks of development, it is advantageous (but not necessary) to keep air temperatures in the 18-23°C range
  6. However, this is not mandatory. Each plantlet should be carefully placed in its respective seedling tray cell, firming it in gently and ensuring that the base of the plant is in excellent contact with the potting material (see illustration). If there are any existing roots, they should be buried because new ones will form if they are damaged or absent. Transfer this tray to a polyhouse, hothouse, or humidity chamber, where the humidity should be maintained at around 70%. If a humidity chamber is not accessible, a styrofoam box filled with two inches of wet sand at the bottom and a sheet of glass covering the top can suffice to accomplish the same results. With a few holes punched in it, cling wrap may also be used to cover the top and allow air circulation underneath. Place the seedling tray on the wet sand and water it down. This improvised chamber has enough space to accommodate one seedling tray (with 48 cavities). A tiny polytunnel can be used as a humidity chamber to accommodate additional seedling trays. It is lightweight and extremely simple to set up. It has the capacity to hold eleven 48-cavity seedling trays (equivalent to about 500 plants). This tunnel may be used for a variety of purposes, including herb gardening and seed cultivation. To achieve better results, you can spray the plants with a MoistureTrap solution after they have been planted. With the help of this solution, a protective coating is formed over the leaves, reducing rapid moisture loss from the newborn plants and allowing them to harden off faster. This lotion is also beneficial in the prevention and treatment of frost-burn, sunburn, salt spray, windburn, and transplanting loss. Maintain a temperature of 20-22°C in filtered light for the trays (direct sun will turn the mini-greenhouse into a mini-pressure cooker). Shade screens must be utilized to minimize the amount of light entering the building by late spring, throughout the summer, and into early fall. During the height of summer, up to 50% of the available shade may be utilized
  7. As the plants mature, a little liquid fertilizer can be sprinkled on the plants to maximize plant growth rates. Every week or fortnight, depending on the necessity, we use 20ml of 100 percent seaweed solution diluted in 10L of water

In conjunction with the purchase of citrus plants from us, we offer a discounted cost on seedling trays, micro polytunnels, and moisture traps. Please get in touch with us for pricing information. This material has been prepared solely as a guide for Australian and New Zealand circumstances, and the use of any practices mentioned below is entirely at your own risk. Please contact your local adviser if you require extra information or an application tailored to your specific circumstances.

TISSUE CULTURE OF WOODY PLANTS – Earth-Kind® Landscaping Earth-Kind® Landscaping

College Station, Texas 77843R. Daniel LinebergerProfessor of HorticultureTexas A M University, College Station, Texas 77843 [email protected] This story initially appeared in The Buckeye Nurseryman, which was published in Columbus, Ohio, in September 1980. Much has been written and spoken about the uses of plant tissue culture in the nursery business, and there is yet more to be said. The issue is still poorly understood by the majority of plant propagators, according to the latest research.

This is a misconception that is fast being dispelled.

Tissue culture propagation, according to this method, is actually applicable to species that are deemed “difficult to propagate.” Equally significant, it may provide economic benefits for those species that are regarded generally “simple to propagate.” The purpose of this communication is to provide a brief overview of present methodologies for woody plant tissue culture, as well as a discussion of the benefits and drawbacks of using tissue culture as a plant propagation method in general.

THE METHODOLOGY OF TISSUE CULTURE

Tissue culture is only one of several approaches that may be used to induce plant development in vitro (that is, using procedures such as tissue culture). The range of techniques available is vast and totally depending on the species being studied. Single cells of leaf tissue, as well as shoot tips, leaf parts, root pieces, lateral buds, and stem sections, have the ability to regenerate whole plants. Not all of these strategies are appropriate to woody plants, and in fact, not all of these methods have been successfully applied to any plants on a commercial scale.

  1. It is necessary to surface sterilize an actively developing shoot tip before placing it on a designated culture medium under sterile conditions.
  2. Depending on how the growth regulators are adjusted, the shoot tip will lengthen, lateral buds will break and begin to develop, and adventitious shoots will appear on the stem piece.
  3. From a single tip, it is possible to grow as many as one hundred shoots in as little as eight to twelve weeks.
  4. Shoots are taken from the cultures on a regular basis, and a portion of the mass is replanted on fresh medium to ensure that the organism’s growth continues uninterrupted.
  5. Shoots produced through tissue culture are generally easy to root, whereas the same cultivar may be difficult to root through cutting propagation.
  6. Techniques for rooting tissue grown shoots, as well as ways for producing these shoots in a greenhouse environment, are now gaining a significant lot of study interest.

THE TISSUE CULTURE LABORATORY

Most propagators would most likely cite the price of setting up a tissue culture laboratory as one of the primary reasons for their decision not to participate in this technique. A lab is more expensive than a mist bed, no doubt about that! A more serious barrier is, without a doubt, the scarcity of individuals who are knowledgeable in the procedures of cell culture. The minimal minimum of equipment is sufficient for an individual with prior knowledge in tissue culture methods to run a laboratory successfully.

When the need arises, the laboratory may be expanded, extra equipment can be installed, and additional employees can be engaged.

Other processes, in addition to the actual culturing process, become increasingly important when a tissue culture facility becomes operational.

Additionally, the management and maintenance of the hundreds of plantlets before they are transplanted to the field or placed in containers is critical.

Production will eventually be limited by the capacity to grow off of the plantlets, rather than by any limitations imposed by the propagation phase itself.

TISSUE CULTURE PROPAGATION OF WOODY SPECIES

Over the past 10 years, significant progress has been achieved in the field of mass multiplication of woody plants. Because of the rather varying response of species and cultivars to the in vitro environment, a great deal of study was (and continues to be) necessary to establish the cultural parameters required by ornamental species in order to thrive. As an illustration of the wide range of results obtained, we have discovered that crabapple seedling shoot tip cultures can generate as few as one or as many as sixty-four shoots per explant after three months in tissue culture, depending on the cultivar.

  • In reality, most economically significant ornamental species have been investigated, and the number of woody plant species that have been clonally reproduced using tissue culture continues to grow at an alarming rate.
  • There has been an intensive commercial scale development program for apple scion cultivars and rootstocks that has been established in England as well as numerous other European nations.
  • While it is true that cultivar variations are responsible for some plants’ failure in tissue culture, a growing amount of research demonstrates that taxonomic groups react in a similar manner in this setting.
  • A similar link exists between the trees and shrubs of the Rosaceae family, which includes the roses (including crabapples, pears, plums, and hawthorns).

It is clear from the anomalies revealed by the crabapple seedling responses to tissue culture, as well as from the emerging similarities between the requirements of rosaceous trees and shrubs, that much more research is required to define the conditions necessary for commercial scale tissue culture production programs.

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IMPACT OF THE TISSUE CULTURE INDUSTRY

As is typically the case with developing technologies, those most enthusiastic about their acceptance are likely to be individuals who were directly involved in its invention in the first place. After acknowledging one’s own prejudices, one must assess the advantages and limits of tissue culture propagation, as well as the potential economic impact of this technology on the nursery business. Not every nursery will have a tissue culture laboratory that is operational. Future nursery operations are anticipated to become more specialized, as either “propagators” or “producers,” as a result of the trend toward specialization.

A discussion of the relatively limited initial investment in physical buildings and equipment had already taken place earlier in this report, when the founding of the laboratory was being contemplated.

A tissue culture laboratory will not be required in every nursery.

The success of tissue culture in the nursery sector will be decided by economic reasons, just as it would be with any other technology.

Will the propagules be able to be integrated into the nursery industry’s regular manufacturing sequence? Are the propagules exactly as expected? Each species and cultivar must be examined individually in order to acquire the best possible responses.

The Procedure

Plant tissue culture techniques were first established and implemented at university and government-sponsored research facilities in the early 1900s. Plant propagation is now widely used in commercial organizations as a cost-effective technique for plant propagation, new variety introductions, and research, and has extended beyond the confines of research institutions in recent years. In the flower and nursery industries, plant tissue culture has transformed the industry by making lucrative novel hybrid clones available in commercial numbers very quickly after they are discovered.

Plant Propagation Methods

Plants propagated via vegetative means – such as top cuttings, root divisions, pseudobulbs, offshoots, and plantlets – are genetically identical to the mother plant and hence members of a single clone, as opposed to seeds. Plant tissue culture is an extension of existing plant propagation techniques that takes place in the laboratory. By using tissue culture, it is possible to produce very large numbers of identical plantlets from a single mother plantlet. This technology, as well as the plantlets it produces, has become the foundation of many plant nursery and flower trade industries.

The Tissue Culture Process

The mother plant that is chosen should be in good condition and devoid of any germs or pathogens. Aseptically extracted living tissue is inserted into culture flasks containing suitable nutritional medium under aseptic conditions after it has been taken from a specific section of the plant. This cell population will grow and differentiate into thousands of plantlets that will have the same features as the parent plants if the culture is deemed to be successful. The presence of bacteria and fungus in a plant tissue culture will hinder the development of the plant tissue from taking place.

Contaminant-free environment

Laboratory procedures and specialized equipment, such as a laminar flow cabinet, are used in conjunction to create an environment conducive to the handling of sterile plant tissues. It is necessary for good plant tissue culture to operate in an environment where practically all of the bacteria and fungus spores have been eliminated. We’ve created a sterile environment, and now it’s time to prepare sterile equipment for the removal of an apical branch from a plant.

Tissue Culture Media

Afterward, the excision bud is put into an empty tube containing sterile growth media. It is important to note that the effectiveness of tissue culture is highly dependent on the stage of the explant that is chosen, the sterilizing time, and the kind of culture medium that is used; different types of plants necessitate the use of different culture media.

Tissue culture media with a high concentration of nutrients is an excellent food source for bacteria and fungi; as a result, precautions against microbial contamination must be taken during all in vitro procedures.

Propagation Rate

Once a plant has been successfully transplanted into an in vitro culture and has begun to develop, it is possible to begin a multiplication program. The manipulation of the culture medium and plant tissues with care will result in a 4-10 times rise in plant numbers every 18-40 days if the culture media and plant tissues are handled with care. It is necessary to begin moving the plantlets from the culture vessel to a greenhouse once a sufficient number of plantlets have been produced in the culture vessel environment.

Take note of the number of days it took and the number of plants that resulted:

Tissue Culture Propagation
Day 1 One plant in tissue culture media
Day 40 5 plantlets cuttransplanted in media
Day 80 25 plantlets
Day 120 125 plantlets
Day 160 625 plantlets
Day 200 3,125 plantlets
Day 240 15,625 plantlets
Day 280 Transplant 15,625 plants to Greenhouse
Day 320 Transplant 15,625 plants to Field

Chapter VIII

Banana Pest and Disease Management in the Tropical Pacific: A Guidebook for Banana Producers is a resource for banana growers across the world.

Tissue culture

Tissue culture is the process of growing tissues or cells in a culture dish apart from the organism. This is often helped by the use of a liquid, semi-solid, or solid growth medium, such as broth or agar, in vitro under sterile growing conditions, such as those provided by the laboratory. When it comes to banana, it is typically propagated vegetatively; therefore, tissue culture as a propagation technique provides a reliable means of preparing disease-free planting materials that can serve as the first line of defense in the development of an integrated disease-management program for the tropical fruit.

Commercially produced tissue-cultured banana seedlings, on the other hand, are not always readily available.

It is the purpose of this book chapter to discuss a banana shoot tip cultivation technique that Damasco invented (2005).

Collection of suckers

  1. It is possible to harvest banana keikis at various stages of development (peepers, swords, or maiden suckers) that are devoid of BBTV symptoms and are around 1–3 ft (40–100 cm) tall for tissue culture. The desired keiki should be separated from the main stem without causing the corm of the keiki to be cracked. At least two suckers should be collected from each plant source, one for micropropagation and the other for a nursery farm to meet the demands of future keiki. According to Fig. 8-2, banana suckers are chosen and removed to extract roughly 4 inches (10 cm) of inner pseudostem tissue containing the banana meristem, which is then used to construct the banana meristem. A newly unfolded banana leaf from the keiki should be collected and submitted to an agriculture disease diagnostic laboratory such as the Agriculture Diagnostic Service Center (ADSC) at the College of Tropical Agriculture and Human Resources (CTAHR) for testing to confirm the plant is BBTV-free.

Disinfection of propagule

  1. Wash the pseudostem that has been retrieved from the field with running water to remove any dirt that has adhered to it. Remove the pseudostem from the vase and soak it in undiluted household bleach (5.25 percent NaOCl) for 30–45 minutes. Remove the pseudostem from the container once it has been surface-sterilized by the bleach solution

Tissue-culture medium for shoot growth

recipe adapted from Damasco and Barba (1984) recipe

Inoculation

  1. Table 1 outlines the procedure for preparing the medium. Autoclave medium scalpels, forceps, cutting plates, and Magenta boxes (Fig. 8-4) according to conventional autoclaving process
  2. Work in a laminar flow hood that has been surface-sterilized
  3. Trim the surface-sterilized banana pseudostem, taking off the outer leaf sheath that come in contact with the bleach. Then, transfer the shoot to a clean cutting dish and continue cutting until it measures 11 cm in length, with the corm tissue being as thin as possible
  4. Using a fresh cutting dish, transfer the shoot tip and cut the shoot into quarters lengthwise, through the middle of the shoot, as shown. Transplant each quadrant onto a solid culture medium

Maintenance of shoot cultures

  1. Maintain shoot cultures in an air-conditioned environment with a photoperiod of 40 E/m 2 S-1 (supplied by two 40-watt fluorescent bulbs) for 16 hours at a time. Check the cultures for signs of contamination. Contaminated cultures should be discarded as soon as contamination is detected. During the first month of culture, look for browning and bulging of corm tissue, greening of leaf tissue, and the development of new shoots. Whenever the shoot tips emerging from the apex of the leaf axis reach a height of about 2 cm, the shoot tips are ready for subculture.

Proliferation of shoots (subculture)

  1. When the propagules are about 2 cm tall, transfer the shoot or parts of shoot to a new growth medium in vitro and repeat the process. Overgrown shoots have a lower proliferative capacity. If the growth of the shoots has progressed beyond 2 cm, make a longitudinal cut through the apex of the expanding shoot. Inoculate onto a half-strength MS medium supplemented with 5 mg/l BAP and 100 mL/l coconut water, and allow to grow for 24 hours. This medium, which contains no auxins, is used to prevent the formation of nubbins at high frequencies from occurring too early. It is essential that all subculturing be done under sterile conditions. Approximately 3–4 weeks of subculture is required until the necessary number of shoots is attained. The number of proliferating shoots has reached an all-time high. Keep repeating the subculture for a maximum of 5 cycles. A greater number of subculturing cycles will result in off-type banana mutations such as dwarfism, elongation, and other abnormalities in the banana plant. When a sufficient number of shoots have multiplied to serve as a nuclear stock, rooting can begin.

Rooting

  1. Rooting medium should be prepared according to Table 2 (Damasco 2005) and used within a week after preparation for the best results. Allow for a 3–4 week (proliferation time) establishment of the last cycle of the shoot cultures in order to acquire little plantlets. Distinguish individual shoots from a group of shoots and place them in rooting media
  2. . The formation of roots will take 3–4 weeks. It is appropriate to introduce plantlets into soil when they have 3–4 fully developed leaves and are firmly rooted.

Preparing tissue-cultured banana plantlets for field planting

  1. Prior to transplanting tissue-cultured banana plantlets into soil, the seedlings must be hardened or acclimated to the external environment, which takes several weeks. For example, you may shift them to a liquid media (without agar) or expose them to partial sunlight in a tissue-culture tube under greenhouse conditions for a few days. All of the agar medium that has adhered to the tissue-cultured plantlets should be carefully removed before they are ready to be transplanted into potting media in a nursery environment
  2. Using a potting mix that has high moisture-holding and drainage qualities, such as 2 parts Sunshine Pro mix, 1 part perlite, and 3 parts medium- to coarse-grade vermiculite, is a suitable choice. Maintaining the moisture content of the media is essential for the health of the tissue-cultured seedlings. Utilize a slow-release or liquid fertilizer to feed your plants. Banana seedlings should be grown in a slightly shaded environment (50 percent shadow) for two weeks before being exposed to full sunshine. BBTV and banana aphid-free conditions should be maintained around the plants. Aphids of other species, whiteflies, and spider mites are regularly seen on banana plants in greenhouses and clustered nurseries, and when populations are large, it is necessary to use insecticide to control the pests. However, after the plants have been transplanted into the field, these pests are usually not a concern. Before field planting, the entire acclimatization process should take around 2 months, or until the seedlings are approximately 8 inches or taller, depending on the type. It is recommended that an active BBTV scouting program be in place if tissue-cultured banana is being used to replace plants in a field infected with BBTV. Young plants should be checked every 5 days since new leaves appear every 5 days
  3. This includes checking them every 5 days. The cultivar determines how long it takes to harvest bananas once they have been transplanted into their natural environment. It is possible to harvest ‘Dwarf Apple’ bananas as soon as nine to ten months after they are planted in the field.

Home Gardener’s and Farmer’s Corner

It is necessary to maintain a sterile working environment when cultivating plant tissue in order to minimize contamination of the growth media. Most commercial tissue-culture facilities are outfitted with laminar flow hoods and autoclaves, and they adhere to strict sterile procedures when conducting their operations. If tissue-cultured bananas are available, home gardeners can obtain them through plant sales. If a farmer is interested in propagating tissue-cultured bananas but does not have the necessary equipment to do it themselves, he or she can contact tissue-culture laboratories that specialize in this type of production.

To give an example, the Hawaii Agriculture Research Center (HARC) offers micropropagation services upon request (page=microprop).

Web Resources

R. Sathes’ Banana Culture was published in 2010. A.V. Jamale’s micropropagation for the creation of high-quality banana planting material was published in 2011.

References

Damasco, O.P., and colleagues (2005) published Tissue Culture of Banana, pp. 59-62. F.S. dela Cruz and colleagues (eds). The management of Musanematodes throughout Asia and the Pacific is being worked on. Laguna, Philippines is home to the International Plant Genetic Resources Institute (INIBAP). PEREZ, Ernesto A., and C.R.R. HOOKS 2008. Preparing tissue-cultured banana plantlets for field planting is a time-consuming process. BIO-8 is a CTAHR Cooperative Extension Service publication of three pages.

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