Microscopic observation of in

vitro Aquilariamalaccensis after

wounding treatmentSubasini Anamulai

168246

Forest Production,Faculty of Forestry, UPM

Supervisor

Assoc. Prof. Dr. Rozi Mohamed

Aquilaria malaccensis is a species of plant in the

Thymelaeaceae family.

It is found in Bangladesh, Bhutan, India,

Indonesia, Laos, Malaysia, Myanmar, the

Philippines, Singapore, and Thailand. It is

threatened by habitat loss (Angela, 2000)

Aquilaria malaccensis is the major source of

agarwood, a resinous heartwood, used for perfume

and incense (Chang, 2005)

Introduction

Problem and justification

Previous study showed that there was no differences in anatomical features between juvenile and matured trees after wounding treatment in natural environment led to gaharuinduction.

In this study, anatomical characteristics of in vitro plants will be investigated

So, this experiment to find the anatomical changes of in vitro of Aquilaria malaccensis under microscopic observation

To characterize anatomical features of in vitro Aquilaria malaccensis under microscopic observation

To determine any changes in the anatomical features after wounding treatment

Objectives

• Samples of in vitro plantlets

• Fungal inoculum – Fusarium solani

• Chemical elicitor – Methyl jasmonate (MeJa)

• Epoxy embedding kit

Epoxy Embedding kit, contains DDSA,NMA, DMP -30

Materials and method

Wounding treatment

Wounding+ funguswounding + chemical elicitorNon-wounding + fungusnon-wounding + chemical elicitor

Wounded 24 hours

Embedding (Epoxy Embedding)

Sectioning

Microscopic study

In vitro plantlets after treatments for 2 months.

The anatomical were observed under light microscopy

Samples are embedded using Epoxy

Results

CP

VC

Controlled Aquilaria malaccensis (Thymelaceae) plantlets. PF= Phloem Fibre, P= Phloem, C=Cortex, VC= Vascular Cambium, Scale =100µm.

C

PX

cpx

ph

hb

ca

c

scl

Traverse section of stem of plantlets with F.solani inoculation and wounding treatment showing the formation of haustorium bridge. Abbreviations: c=cortex,ph=phloem,hb= haustarium bridge, sph=secondary phloem, x=xylem, ca=vascular cambium. Scale =100µm

Ip

Psx

hb

sphc

ph

Traverse section of stem of plantlets with F.solani inoculation and non wounding treatment showing the formation of haustorium bridge. Abbreviations: c=cortex,ph=phloem,hb= haustarium bridge, sph=secondary phloem, sx= secondry xylem, p=pith, ip=included phloem, Scale =100µm

Ip

Showing the extended parts of plantlets with F.solani inoculation and non wounding treatment. Scale =100µm

Ep

psx

sp

cp

Ip

TS of plantlets treated with MeJa and non wounded. Ip= Included phloem, sp= secondry phloem. Sx=secondry xylem, p=pith, cp=

Lenticels

Deposit in pores

Showing the lenticels and deposit in pore under scale=100µm

Ip

TS of plantlets treated with MeJa and wounded. Shows the changes in distribution of phloem compare to controlled and the higher number of deposit in pores Ip= Included phloem. Scale =100µm

Ip

TS of plantlets treated with MeJa and wounded. The distribution of included phloem. Scale =30µm

SX

P

VC

RP

pe

TS of plantlets wounded for 24 hours. Pe= Pholem elements, rp=ray parencyhma, sx=secondry xylem, p=pith, vc=vascular cambium. Scale =100µm

Ip

Ts of wounded 24 hours plantlets included phloem. Scale =30µm

• It shows, when Tissue culture of A.malaccensis interact withMeJa as a chemical elicitor, the deposit in pores become highand showing that potential to induce production fragnantcompound.

• The results shows similarly with study by Ito in wild Aquilariasp. (Ito et al. 200)

• There is haustorium bridge development in xylem thatextended to new growth in plants tissue, assumed that thetissue culture’s defense of these agarwood producing speciesis triggerd by F. solani attack, as Turjaman,2011 mention thesame statement in his journal.

• Treatments done chemically, fungus inoculation andtechnically, the reaction of the tissues occur in chemicaltreatment and fungus inoculation are faster.

Discussion

The anatomical changes in in vitro A.malaccensiscan be seen clearly in xylem and phloem tissues after undergo treatments chemically, fungus inoculation and technically.

Methyl Jasmonate can be potential chemical solution that can trigger the induction of agarwood.

F. solani induce the reaction by enchance the plantlets defense and change the tissues shape and structure.

Conclusion

• Rahman, M. A., & Khisa, S. K. (1984). Agar production inagar tree by artificial inoculation and wounding II. BanoBiggyan Patrika, 13,57-63.

• Yaacob, S. (1999). Agarwood: Trade and CITESImplementation in Malaysia. Unpublished report preparedfor TRAFFIC Southeast Asia, Malaysia

• Mohamed, R.1*, Wong, M. T.1 and Halis, R.2 1ForestBiotech Laboratory, Department of Forest Management,Faculty of Forestry, Universiti Putra Malaysia, 43400Serdang, Selangor, Malaysia

References

Thank you