microscopic observation of in vitro aquilaria malaccensis after 1 copy (1)
TRANSCRIPT
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