Plant Biotechnology- PLANT TISSUE CULTURE

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Shri Tuljabhavani Sewabhawi Shaikshanik Va
Samagik Shikshan Sanstha, Kothari. HI TECH
COLLEGE OF PHARMACY, CHANDRAPUR Padoli Phata
Nagpur Highway, chandrapur-442401 M-Pharmacy
(1st year, IInd Semester) 2021-2022 Department -
Pharmacognosy(MPG) Name - Maroti Madhukar
Jeurkar Roll No - 02 SUB- Plant
Biotechnology Topic- PLANT TISSUE CULTURE
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Plant tIssue culture technIquesTools In plant
mIcropropagatIon
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WHAT IT IS?

• In vitro propagation? Or
• Micropropagation ? Or
• In vitro culture?
• THE ASEPTIC CULTURE OF PLANT
• Implies - regeneration
• - multiplication

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IMPORTANCE OF PLANT TISSUE CULTURE TECHNIQUES

• True-to-type clones
• A single explant can be multiplied into several
  thousand
• Year-round production
• Rare and endangered plants can be cloned safely
• To produce virus free plants
• Long-term germplasm storage with tissue banks
• Plant cultures easier to export than are
  soil-grown plants
• Production of difficult-to-propagate species
• Worldwide industry multibillion Euros

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FUNDAMENTAL ABILITIES OF PLANTSHOW CAN A PLANT
CELL OR TISSUE DEVELOP?

• Totipotency
• Dedifferentiation
• Competency
• Therefore, tissue can be regenerated from
  explants such as cotyledons, hypocotyls, leaf,
  ovary, protoplast, roots, anthers, etc.

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WHATS THE BACKGROUND
1902 Haberlandt The Concept 1920s Knudson
Simple Orchid Germination First commercial
use 1930s Thimann Went Auxin 1930s
White/Gautheret/Nobecourt Root Cultures 1950s
Skoogs group Cytokinins, The
discovery of the structure of DNA by Crick and
Watson 1960s Morel-Orchid micropropagation,
thermotherapy 1970s Genetic engineering took
off 1990s by Calgene genetically engineered
potatoes
Gottleib Haberlandt
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WHAT IS NEEDED?

• Appropriate tissue
• A suitable growth medium
• Aseptic (sterile) conditions
• Growth regulatorsThe ratio of auxins and
  cytokinins
• Frequent subculturing

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TYPES OF PLANT TISSUE CULTURE TECHNIQUES

• Culture of intact plants (Seed orchid culture)
• Embryo culture (embryo rescue)
• Organ cultureMicropropagation
• A. Organogenesis in solid or semi solid medium
• 1. Meristem and shoot tip culture
• 2. Bud culture
• 3. Root culture
• 4. Leaf culture
• 5. Anther culture
• B. Somatic embryogenesis
• C. Organogenesis and somatic embryogenesis in
  bioreactors
• D. In vitro micrografting
• E. Thin cell layer technology (TCLs)
• F. Photoautotrophic culture
• 4. Callus culture
• 5. Cell suspension and single cell culture
• 6. Protoplast culture, somatic hybridization

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Culturing (micropropagating) Plant Tissue - the
steps

• Stage 0 Selection preparation of the mother
  plant
• sterilization of the plant tissue takes place
• Stage I  - Initiation of culture
• explant placed into growth media
• Stage II - Multiplication
• explant transferred to shoot media shoots can be
  constantly divided
• Stage III - Rooting
• explant transferred to root media
• Stage IV - Transfer to soil
• explant returned to soil hardened off

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ORGANOGENESIS OF PISTACHIO
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ADVENTITIOUS ORGANOGENESIS IN PISTACHIO
Adventitious Buds
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Elongation and
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2
3
from Single Leaflet
Regeneration of
Plantlets
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SOMATIC EMBRYOGENESIS IN PISTACHIO
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MICROPROPAGATION IN BIOREACTORS

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WHAT IS MICROGRAFTING?
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The thin cell layer (TCL) system consists of
explants of small size excised from different
plant organs either longitudinally (lTCL) or
transversally (tTCL)
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PHOTOAUTOTROPHIC CULTURE
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APPLICATIONS OF MICROPROPAGATION

• Through micropropagation, it is now possible to
  provide clean and uniform planting materials in
  plantations for several plant species such as
  oil palm, plantain, pine, banana, abaca, date,
  rubber tree field crops eggplant, jojoba,
  pineapple, tomato root crops cassava, yam,
  sweet potato and many ornamental plants such as
  orchids and anthuriums (Alfonso, A. 2007 Singh
  et al. 2011).
• Bioreactor cultures are being established in
  several commercial laboratories for
  micropropagation of
• ferns, spathiphylum, philodendron, banana,
  potato, lilies, poinsettia, sugar-cane, and some
  forest tree species such as eucalyptus, poplar,
  and early stages of conifer somatic embryos
  (Aitkin-Christie, 1991 Mehrotra et al 2007Gross
  and Levin, 1999 Cervelli and Senaratna 1995).
  And plant products, pharmaceuticals, food
  ingredients and cosmetics (Perulllini et al.,
  2007 Vongpaseuth and Roberts 2007 Pavlov et al.
  2007)
• Micrografted seedlings are commercialised to
  avoid the serious crop loss caused by infection
  of soil-borne diseases for fruit trees and
  several vegetables (Navarro et al., 1975Navarro
  1981Jung-Myung Lee et al. 2010).
• Transverse thin layer section technology may be
  ideal for large scale micropropagation of
  ornamental plants (Jain et al. 1998).
• Photoautotrophic flow-through systems for
  enhanced micropropagation for Gerbera Hypericum,
  Myrtus communis, Momordica grosvenori, Eucalyptus
  (Nguyen and Kozai 2005 Xiao et al.2011)
• The development of transgenic methods and the
  growth of agricultural biotechnology started
  during the 1980s and the global biotech crophas
  increased hundred million of hectares area
• Palmer et al. 2005 Thomas et al. 2003
• Efficient doubled haploid technology enables
  breeders to reduce the time and the cost of new
  cultivar development relative to conventional
  breeding practices.

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WHATS IN THE FUTURE

• Adaptation of tissue culture technology to more
  species
• Fast and mass propagation of transformed plants
  with designer genes
• Chloroplast transformation methods
• Efficient, computer-controlled flow-through
  systems to cut down the labor cost
• Mass production of plant constituents
• New technologies

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ACKNOWLEDGEMENTS
All photos in this talk were taken by Ahmet Onay
and Engin Tilkat, except the ones on the slayt 12
and 15 which was obtained from the WEB pages.
TUBITAK TOVAG - 3355 has granted part of these
studies And the technical assistance of all
co-authors are gratefully acknowledged.
Ahmet Onay, Department of Biology, Faculty of
Science, University of Dicle, 21280 Diyarbakir,
Turkey
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THANK YOU