5. AS A RESULT OF THIS REMARKABLE
DISCOVERY…………
CYCAS WAS EXCLUDED FROM
DICOTYLEDONS &………
CAME TO RECOGNISED AS ..
GYMNOSPERM
6. ……….However the first use of the Greek
word gymnosperm have been already
made as early as 300 b.c by the Greek
philospher ……
THEOPHRASTUS in his “ENQUIRY
INTO PLANTS”
8. A group of vascular plants whose
seeds are not enclosed by a ripened
ovary (fruit).
A group of plant which lack double
fertilisation in the formation of
endosperm
9. They are true land plants………..
Because they show……….
•Protection of gametophyte and young
developing sporophyte from dessication
and injury .
•Provision of nourishment for their proper
development.
•Freedom from external water for the
transference of the gametes
10. Gymnosperm achieved all these
conditions by mustering 3 succesive
generation in a single organ…….
SEED……..
•The parent sporophyte as …integuments &
nucellus
•The gametophyte as ….endosperm
•The daughter sporophyte as……. embryo
14. CYCAS…….
– They are evergreen, gymnospermous
and dioceious plants having large
pinnately compound leaves.
About 95 species are currently accepted. The
best-known species is Cycas revoluta widely
cultivated under the name "Sago Palm" or
"King Sago Palm" due to its palm-like
appearance although it is not a true palm
21. INSTRUMENTS……
.AN OVEN (set at 52 degree celcius )
this temperature depends on
environmental condition.
.AN INCUBATOR (set at 37 degree celsius)
we used orbital shaker
.A DIGITAL CAMERA(for the pics)
22. Theory ……
Cycads produce several secondry compounds
that are
carcinogenic and neurotoxic. Because of their
unique
placement in terrestrial plant evolution,
molecular
studies should help to define the origins of
structures
that led to the rise of seed plants and the role
of neurotoxic
compounds that are found in cycads.
23. The subsequent search for a neurotoxic
compound in cycads lead to the identification
of a
•Novel non-protein amino acid called BMAA
(beta-methylamino L-alanine)
•methylazoxymethanol(MAM)-glycocides
( cycasin macrozamin and neocycasin )
•sterol beta-D glucoside
25. CYCASIN N MACROZAMIN
The toxic azoxyglycosides from the seeds of several
species of Cycas.
Carcinogenic in mice, rats, hamsters, guinea-pigs,
rabbits and fish.
Azoxyglycosides may have played an important
ecological role as antiherbivore defenses.
26. ELECTRICAL
WEIGHING
WEIGHING LIMIT: MACHINE
FOR MICROGRAM AND
GRAM
27. Cycasin
1.3. Molecular formula, weight and structure:
2.C8H16N2O7; 253.23
3.4. Density: No data.
4.5. Absorption spectroscopy: UV absorption
spectra have been reported (Riggs, 1956); λ (log
ε)= 2187.
5. Solubility: Very soluble in water and dilute ethanol; less
soluble in absolute ethanol; insoluble in
6. organic solvents.
7. 8. Description, appearance: Colorless, long, needle-like
crystals.
28. METABOLISM OF CYCASIN AND
MACROZAMIN
The active component of all theses compounds is MAM
(methylazoxyglycoside),
which is metabolised to a toxic intermediate by the
p450 enzyme present in the liver and in the olfactory
epithelium in the nose and inner membrane of
mitochondria
30. P450 ENZYME
Cytochrome P450 (abbreviated CYP, P450,
infrequently CYP450) is a very large and
diverse superfamily of hemoproteins found in all
domains of life…
ncluding mammals,birds, fish, insects, worms, sea
squirts, sea urchins, plants, fungi slime molds, bacteria
and archaea
the most common reaction catalysed by cytochrome
P450 is a monooxygenase reaction :
RH + O2 + 2H+ + 2e– → ROH + H2O
31. PROCEDURE….
our first aim to isolate cycasin from cycas seeds..!
steps :
1 we were provided with 7 seeds of cycas pectinata
2 seed coat of cycas seed was quite hard so we first
isolated seed form seed coat with the help of cutter.
3 we placed the naked cycas seeds in petridish
32. MAIN CYCAS SEED
NAKED SEED
CYCAS SEED COVERED WITH
HARD SEED COAT
33. 4 since before extraction of cycasin we need to
quantitise the cycasin in cycas seeds, therefore we
measured them in electrical weighing machine one by
one and recorded them.
RECORD :
SEEDS WET WEIGHT(grams)
1 12.41
2 12.08
3 10.16
4 9.84
5 10.42
6 8.32
7 7.31
35. 5 since seeds were wet so we kept them in
oven at 54 degree celcious to dry them.
6 our next step will be the crushing of dried
seeds and soaking in water for extraction of
cycasin.........
37. After drying cycas seeds for 4 days in oven at
54degree celcius,
it was crushed in mortar and pistil untill it become
fined powedered.
the powdered form of seeds were again
weighed....,and it was found that their dry weight
were almost 50% less than there originall weight..!
42. steps:
1 firstly we fixed the embryo in appropriate size
beaker in such a way that narrow portion of head
facing downward and broder portion facing ourself.
2 then with the help of needle a hole was made in
the broder portion of egg.
3 then widened this hole with the help of scissor ,
this should be done very carefully without disturbing
the vitelline membrane.
55. A 9 DAY OLD CHICK EMBRYO
TREATED WITH CYCAS SOLUTION ..
REMAINED ALIVE AFTER 6 HOURS WITH ALL THE CONCENTRATIONS
HOWEVER DIED AFTER 24 HOURS WHEN TREATED WITH SOLUTION A
56. A 9 DAYS OLD CHICK EMBRYO
DIED AFTER 24 HOURS WHEN TREATED WITH SOL
A(HIGHLY CONCENTRATED ,1ST WASH)
57. A 9 DAYS OLD CHICK EMBRYO( ALIVE) NORMAL I.E
NOT TREATED WITH CYCAS SOLUTION
58. A 18 DAYS OLD CHICK EMBRYO
SURVIVE WITH ALL THE CONCENTRATION AND
WASHS
59. OBSERVATION &DESCUSSION
EGG EXPERIMENT OBSERVATION INFERENCE
2 SOL A ALBUMEN TURNED MAY BE BECAUSE
DAYS WASH 1 GREENISH IN BOTH OH HIGH
CASE CONCENTRATION
OLD WASH 2 TOXIC IN CYCAS
SOLUTION
SOL B FIRST WASH
CONTAIN
WASH 1 ALBUMEN TURNED RELATIVELY HIGH
WASH 2 YELLOW
AMOUNT OF TOXIN
NO CHANGE THAN 2ND WASH
SOL C EXTREMLY SMALL
WASH 1 NO CHANGE QUANTITY OF
NO CHANGE TOXIN IN BOTH
WASH 2
WASH
60. EGG EXPERIMENT OBSERVATION INFERENCE
4 SOL A NERVE NETWORK HIGH
CONCENTRATION OF
DAYS WASH 1 – DEVELOPMENT TOXIN MIGHT KILLED
OLD 6HOURS RETARTED IN E.E IN SOME OF THE
COMPARISON TO N.E SOMATOCYTES
EGG SPOILED AND A HIGH
24 HOURS VERY FOUL SMELL CONCENTRATION
MIGHT KILLED ALL
CAME & NERVE THE SOMATOCYTES IN
DEVELOPMENT WAS E.E & DESTROYED THE
VERY LITTLE IN EGG IN SUCH A LONG
COMPARISON 2 N.E TIME
NERVE NETWORK
WASH 2- REMAINS ALMOST COMPARETIVELY LOW
6 HOURS CON. AND VERY LESS
SAME IN BOTH TIME
NERVE RETARTED BUT LOW CON. AND LONG
24 HOURS FOUL SMELL DIDN’T ACTION TIME
COME
61. EGG EXPERIMENT OBSERVATION INFERENCE
4 SOL B LOW
DAYS WASH 1 – NOT DISHTINGUISHABLE CONCENTRATION
&LESS ACTION
OLD 6HOURS OBSERVATION TIME
24 HOURS NERVE NETWORK SHOW LOW
RETARDATION IN CONCENTRATION
DEVELOPMENT &BUT RELATIVELY
LONG ACTION
TIME
WASH 2-
NO CHANGE ALMOST EXTREMLY LOW
6 HOURS
SAME CONCENTRATION
24 HOURS
NO CHANGE ,E.E DIDN’T
SPOIL
62. EGG EXPERIMENT OBSERVATION INFERENCE
4 SOL C
DAYS WASH 1 – NO CHANGE
6HOURS
OLD
24 HOURS MIGHT BE
NO CHANGE DUE TO
EXTREMLY
LOW OR
ALMOST
NEGLIGIBLE
CONCENTRA
WASH 2- NO CHANGE TION OF
6 HOURS TOXIN
24 HOURS
NO CHANGE
63. EGG EXPERIMENT OBSERVATION INFERENCE
18 SOL A HIGH RESISTANCE
DAYS WASH 1 – NO EXTERNAL TO TOXIN DUE TO
OBSERVATION WELL DEVELOP
OLD 6HOURS IMMUNE SYSTEM
24 HOURS
MIGHT BE THERE
WERE CHANGES
CHICK REMAINED ALIVE IN NERVOUS
IN ALL THE CONDITIONS SYSTEM
ACCORDING TO
WASH 2- STUDIES BUT WE
6 HOURS WERE UNABLE TO
FIND THAT
24 HOURS
64. THANX……..
DR.RITA MAM
2. JIBAN SIR
3. B.T DEPARTMENT
4.SAKSHI MAM
5.SWATI MAM
6.INTERNET SOURCES,
7. INTRODUCTION TO GYMNOSPERM BY DIVYA
DARSHAN PANT.
8 PLANT ENCYCLOPEADIA
9CHICK EMBRYOLOGY..
