植物組織培養

Introduction to Plant Tissue

Culture
 Definitions

• Plant Tissue Culture

• Plant Cell, Tissue and Organ Culture
• In Vitro Culture
• Aseptic Culture

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Advantages of Plant Tissue Culture

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 Comparisons of the Culture Among
Microorganism, Plant and Animal
Microorganism Plant Animal
Technique ＋ ＋＋ ＋＋＋
Asepticize ＋ ＋＋ ＋＋＋
Medium ＋ ＋＋ ＋＋＋
Explant ＋ ＋＋ ＋＋＋
Growth rate ＋＋＋ ＋＋ ＋
Development × 可發育成各 ×
種型態如葉
或花 5
 Key point for successful plant tissue
culture
• 培植體
• 培養基
• 培養環境
• 無菌
• 操作者

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 Explants

• Meristems （生長點）
• Leaf sections （葉）
• Bulb sections （鱗莖、球莖）
• Embryos （胚）
• Anthers （花藥）

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 Medium components
• MS
1962 年 Murashige & Skoog ： Tobacco
tissue culture
※ Development of MS medium

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 Medium preparation
• Select suitable medium formula
• Prepare stock solutions
• Properly dilute and mix of each stock
solutions
• Add adequate amount of sugar, agar,…
• Heating to melt the agar
• Adjust pH
• Dispense into culture vessels
• sterilization
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 Culture conditions

• Light
• Temperature
• Relative humidity

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 Aseptic techniques

• Dry heat : for glassware ,

metal instruments,…
ex: Oven : 160℃, 2hrs.

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 Aseptic techniques

• Wet heat : for glassware ,

metal instruments, medium,
liquid… ex: Autoclave : a
steam pressure of 15 lb/in2
at a temperature of 121℃
15 min.

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 Aseptic techniques

• Millipore membrane : pore

size, 0.22μm, 0.45μm
• HEPA filter : for air
sterilization
• Chemicals : EtOH (70-
75%), Sodium hypochlorite,
…
• Irradiation : UV
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Development of Plant Tissue Culture
Science
• 器官培養 (Culture of Isolated Organs)
• 癒合組織培養 (Callus Culture on Solidified
Medium)
• 懸浮細胞培養 (Suspension culture)
• 器官再生與胚胎形成 (Regeneration and
Embryogenesis)
• 生長點培養 (Meristem culture)
• 原生質體培養 (Protoplast Culture, Fusion)
• 基因轉殖植物培養 (Genetic Engineering//Gene
Transforation)
• 生物反應器 (Bioreator System) 14
器官培養 (Culture of Isolated Organs)

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Callus :
An amorphous mass of loosely arranged thin-
walled parenchyma cells rising from the
proliferating cells of the parent tissue cultured on
agar medium.

Carrot callus 16
Tobacco callus
Suspension Cell
Culture :
Rapidly dividing,
homogenous cell
aggregates suspended
in a liquid medium,
cultured to produce a
“cell line”.

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Organogenic process
Competence Determination
+
Explant + Organ

Dedifferentiation Induction Differentiation

Somatic embryogenesis

Plant explants Embryoids

Somatic embryos
Callus
Suspension cells
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生長點
培養
原生質體培養 (Protoplast Culture,
Fusion)

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基因轉殖植物培養 (Genetic Engineering//Gene
Transforation)

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 生物反應器 (Bioreator System)

• Small batch culture

Bioreactor
Scale up
Large volume culture

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Comparison Between in vitro and in
vivo Culture Systems
• 植物細胞具備易被誘導表現分化全能性
(Totipotency)
• 不是所有種類之植物的培養條件都可以完
全掌握
• 培養之材料與在天然環境中生長之植株的
生理 ( 和遺傳 ) 存在或多或少的差異
• 長時間的培養可能改變常態下之基因表現
的能力
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 Totipotency
1902 Father of Plant Tissue
Culture (Haberlandt)
* Concept of “Totipotency”

植物的組織或每個細胞都
含著該物種全部遺傳信息，
從而發育成完整植株（即單
一細胞可完全分化成根莖葉
之完整植株）。

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 Application

• 微體繁殖 (Micropropagation)
• 育種 (Breeding--Horticulture, Agronomy…)
• 生產二次代謝產物 (Production of
Secondary Metabolites)
• 基因轉殖植物 (Production of Transgenic
Plants)

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 微體繁殖 (Micropropagation)
• 微體繁殖：

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 生產二次代謝產物 (Production
of Secondary Metabolites)
• Application of plant Sm:
Drug, Spice, Cosmetic,
Pigment, Insecticides,
Nature latex

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Arbutin ( 熊果素 )
謝謝大家

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