EFFECT OF THE DIFFERENT AUXINS AND CYTOKININS IN CALLUS INDUCTION,

SHOOT,AND ROOT REGENERATION IN

SUGARCANE

Presented by:

Arghya Narendra Dianastya

5620000087

Departement of Agronomy

Kasetsart University, Kamphaeng Saen Campus

INTRODUCTION

Modern sugarcane varieties are breeding of species, involving crosses between the species Saccharum officinarum L. and S. spontaneum L (Heinze,1987).

Sugarcane is a tall growing monocotyledonous crop.

Sugarcane has ability to store high concentrations of sucrose, or sugar, in the internodes of the stem.

SUGARCANE PRODUCER

In 2010, Brazil was the largest producer at 719 million tonnes (FAO, 2013)

GLOBAL PRODUCTION AND CONSUMPTION FAO’s current estimate for world sugar production in

2012/13, at 180 million tonnes, (new record production)

SUGARCANE AREA HARVESTED

INTRODUCTION

What can we do to improve the quality and productivity of sugarcane

IMPROVEMENT OF SUGARCANE PRODUCTIVITY AND QUALITY

Plant improvement Conventional methode

Plant biotechnology (ex: plant tissue culture)

Cultural practice improvement

ADVANTAGES OF TISSUE CULTURE

Plant regeneration through tissue culture technique would be a

better alternative for improving the quality, production, and

improvement of crop through the rapid production of clones (Jabbot

& Atkin, 1987)

WHAT IS TISSUE CULTURE ?

Plant tissue culture can be defined as a procedure used to mantain and grow plant tissue (callus, cells, protoplasm) and organ (stem, root, embryos) in aseptic or in vitro culture.

Basic Concept is Totipotency :Totipotency ability in individual plant cells that can be regenerated to a whole plant by modulating culture conditions (Reinert, 1959).

(Hartmann et.al., 1990 )

EXPLANT

CELL SUSPENSI

ON

CALLUS

ORGANOGENESIS

EMBRYOGENESIS

PROTOPLASTS

EMBRYOGENESIS

PLANT

PATHWAY OF PLANT TISSUE AND CELL CULTURE

OBJECTIVES OF THE RESEARCH To analyze the effect of different

concentration of auxins and cytokinins in Callus Induction, Shoot,and Root Regeneration in Sugarcane.

To analyze the effect of different level of pH and sucrose in root regenenation of sugarcane.

MATERIAL

Sugarcane Cultivar CO 671 (3-5 months old)

MS Medium - Murashige and skoog medium

Cytokinin BAP Kn

Auxin IBA IAA NAA

METHODOLOGY

sterelizedLeaf

Sheath

Callus InductionMS+ CM + different concentration of : 2,4-D ; IBA ; NAA

Shoot RegenerationMS + different concentration

of :BAP; Kn ; (BAP + NAA) ;

(BAP + IBA)

Root Regeneration½ strength MS + different

concentration of :IBA ; NAA ; IAA ;

(NAA+IBA)

Different concentration ofSucrose & pH

RESULT

CALLUS INDUCTION

Effect of different concentration, auxin and 2,4-D on callus induction from leaf sheath explants of sugarcane

CALLUS INDUCTION

The best callus induction suplement 3.0mg/L of 2,4-D = 85% explant produced

callus

No callus induction IBA

SHOOT REGENERATION

Effect of the cytokinin (BAP,Kn) and the auxin (IBA,NAA) at different concentrations and combination is Ms medium on

shoot on shoot regeneration from the callus tissue of sugarcane

SHOOT REGENERATION cont..

Effect of the cytokinin (BAP,Kn) and the auxin (IBA,NAA) at different concentrations and combination is Ms medium on

shoot on shoot regeneration from the callus tissue of sugarcane

SHOOT REGENERATION

BAP+IBA The best respond 1.0+0.5

mg/L = 95 % produced shoot

BAP+NAA The best respond 1.0+0.5

mg/L = 94% produced shoot

ROOT REGENERATION

Effect of different auxin on formation of root on the invitro grown micro shoots Cultured

ROOT REGENERATION

The Best Rooting Media ½ strength of MS medium + 3mg/L NAA =

95% explant produced root

Comparison Media Used NAA and IBA were better response than

IAA for producing roots.

CONCENTRATION OF SUCROSE

Effect of sucrose concentration on rooting of invitro differentiated shoots of sugarcane

CONCENTRATION OF SUCROSE

The Best Sucrose Concentration 50 gram/L of sucrose = 96%

explant produced root

pH LEVEL

Effect of different pH level on rooting of in vitro differentiated Shoots of sugarcane

pH LEVEL

The Best pH Level pH 5.7 = 96% explant produced root

The Poorest pH Level pH 3 = 0 % explant produced root

CONCLUSION The best callus induction was observed

at 3.0 mg/L of 2,4-D

The best shoot regeneration was observed at 1.0 mg/L and 0.5 mg/L of (BAP + NAA)

The best root regeneraration was observed at 3.0 mg/L of NAA

The best sucrose concentration 50 gram/L of Sucrose

The best pH level pH 5.7

REFERENCE

Gopitha A., Lakshmi B., Senthilmanickam J. 2010. Effect of the different auxins and cytokinins in callus induction, shoot, root regeneration in sugarcane. International Journal of Pharma and Bio Science. Vol.1/Issue-3/Jul-Sep.2010.

Reinert J (1959) The control of morphogenesis and induction of adventitious embryos in cell cultures of carrots. Planta 53:318–333.

Jabbott A.J and Atkin R.A. 1987 Improving Vegetatively Propagated Crop. Academic Press Limited.

Hartmann H.T., Kester D.E., Davies F.T., and Geneve R.L. 1997. Plant Propagation: Principles and Practices, Sixth Edition. Prentice Hall International, Inc.

ACKNOWLEDGMENTS

Kasetsart University Advisor Prof. Sontichai Chanprame, Ph.D Prof. Sermsiri Chanprame, Ph.D

Jember University Advisor Dr. Kacung Haryono

CAB (Center of Agriculture Biotechnology)

And for All that have helped me to study in Kasetsart University

THANK YOU FOR

YOUR ATTENTION