introduction to Convolvulaceae family, claasification, floral formula and diagram, Diagnostic features . Plant Biochemistry- estimation of protein by bradford method. M.SC BOTANY 2nd semester.

1. TAXONOMY OF ANGIOSPERMS
(CONVOLVULACEAE FAMILY)
TOPIC

2. CONTENTS
 INTRODUCTION
 CLASSIFICATION
 CHARACTERISTICS FEATURES
 FLORAL FORMULA
 DIAGNOSTIC FEATURES

3. INTRODUCTION
 Cuscuta species can be found on all continents.
 Cuscuta reflexa is a parasitic plant which belongs to family Convolvulaceae.
 It is commonly known as dodder plant, amarbel, akashabela.
 The parasitic plant sucks all the nutrients from the host plant with the help of
haustoria.

4. Kingdom Plant
Class Dicotyledonae
Subclass Gamopetalae
Series Bicarpellate
Order Solanales
Family Convolvulaceae
Genus Cascuta
Species reflexa
CLASSIFICATION

5. CHARACTERISTICS FEATURES
HABIT
Parasitic twiner
Leafless
STEM
Herbaceous
Aerial
Weak
Climbing
Branched
VEGETATIVE CHARACTERS

6. INFLORESCENCE
Flowers either solitary
or in clusters
FLOWER
BRACTEATE
PEDICELLATE
ACTINOMORPHIC
HERMEPHRODITE
HYPOGYNOUS CYCLIC
FLORAL CHARACTERS

7. CALYX
 5 SEPALS
 POLYSEPALOUS
 QUINCUNCIAL
COROLLA
 5 PETALS
 GAMOPETALOUS
 VALVATE
ANDROCEIUM
 5 STAMENS
 POLYANDROUS
 EPIPETALOUS
 DITHECOUS
 INTROSE
GYNOCEIUM
 BICARPELLARY
 SYNCARPOUS
 OVARY SUPERIOR
 AXILE PLACENTATION
 2-4 OVULES
FLORAL DIAGRAM

8. DIAGNOSTIC FEATURES
 Class. Dicotyledonae – venation reticulate
- flowers pentamerous
 Sub-class. Gamopetalae – petals fused
 Series. Bicarpellate – Carpels two, ovary superior
 Order. Paleomoniales – Leaves alternate, stipulate
- Flowers Actinomorphic
 Family. Convovulaceae – Gynoceium bicarpellary, Fruit capsule

9. Fruit:
Capsule (Convolvulus, Evolvulus, Cuscuta) or berry.
Seed:
Endospermic.
Pollination:
Entomophilous.
Floral formula:

10. ECONOMIC IMPORTANCE
STEM
 USE IN TREATMENT OF BILIOUS DISORDER.
 USED INTERNALLY IN TREATING PROTRACTED
FEVERS.
Food
Tuberous roots of Ipomoea batatus (Sweet
potato) are rich in starch and edible.

11. ECONOMIC IMPORTANCE
 Dodder helps to improve depression in people
 Bladder problems.
 Liver problems.
 Pain.
 Spleen problems
 It might cause side effects such as stomach upset
and diarrhea.

12. ESTIMATION OF PROTEIN BY BRADFORD'S
METHOD.
EXPERIMENT

13. CONTENTS
 PRINCIPLE
 MATERIALS REQUIRED
 PROCEDURE
 ADVANTAGES
 DISADVANTAGES

14. The Bradford protein assay is used to
measure the concentration of total
protein in a sample.
The principle of this assay is that the
binding of protein molecules to
Coomassie dye under acidic conditions
results in a color change from brown to
blue.
PRINCIPLE
Coomassie Brilliant Blue G-250

15. CBB
CATIONIC STATE
UNSTABLE
REDDISH COLOR
ABSORBANCE SHIFT
ANIONIC STATE
STABLE
BLUE COLOR
DYE
595nm
NEUTRAL
GREEN COLOR
NEUTRAL

17.  Sample extract (for which proteins to be estimated)
 Coomassie Brilliant Blue 1
 0.1M phosphate buffer (prepare freshly)
 Bradford reagent
 BSA Standard solution
 Spectrophotometer and cuvettes
 Micropipettes
MATERIALS REQUIRED

18. PROCEDURE
(A) PREPARATION OF SAMPLE SOLTION
 Take 1g of sample and add 5ml of 0.1M phosphate buffer to it.
 Centrifuged at 800rpm for 20 minutes.
 Collect supernatant and dilute it.
(B) Preparation of BSA (Bovine Serum Albumin)
Standard solution or protein standard solution :-
 Dissolve 5mg of BSA in 50ml of 0.1M phosphate buffer .
 It contains 100 μg protein/ml.
 This is prepared stock solution.
 For 20 μg /ml :- 0.2ml stock sol. + 0.8ml phosphate buffer.
 For 40 μg /ml :- 0.4ml stock sol. + 0.6ml phosphate buffer.
 For 60 μg /ml :- 0.6ml stock sol. + 0.4ml phosphate buffer.
 For 80 μg /ml :- 0.8ml stock sol. + 0.2ml phosphate buffer.

19. (C) PROCEDURE FOR BRADFORD REAGENT
 Take 0.1ml sample solution and add 0.9ml phosphate buffer to make volume 1ml in centrifuge
tube.
 Take 20 μg /ml protein in other 4 centrifuge tubes.
 Add 5ml Bradford reagent in all test tube and mix thoroughly.
 Observe the instant blue colour change.
 Record the absorbance of sample in all test tubes at 595nm.
 Plated a standard curve of A595 v/s μg of proteins.
 Determine the protein content in sample solution from standard curve.

20. S.NO. STANDARD BSA
SOLUTION (μg /ml)
A595
1 20 1.048
2 40 1.380
3 60 1.650
4 80 1.880
RESULT

21. ADVANTAGES
 Fast and inexpensive.
 Highly specific for proteins.
 Very sensitive.
 Compatible for wide range of substances(sodium-potassium ions, sucrose ,
carbohydrates).
 Dry reagent complex is stable for approximately 1hour.

22. DISADVANTAGES
 Basic conditions and detergents, such as SDS, can interfere
with the dye's ability to bind to the protein through its side
chains.
 The Coomassie Blue G250 dye used to bind to the proteins
in the original Bradford method readily binds to arginine
and lysine groups of proteins.