Appendix – I Amount Conc. of stock (mg/l) solution...

Appendix – I

MS basal media composition (Murashige and Skoog, 1962)

Directions :

- to prepare 1 Lt medium take 50 ml of stock 1, and 5 ml of each of stock, II,

III and IV. - pH was adjusted before autoclaving and after the addition of plant growth

regulators and carbon sources.

Ingredients Amount (mg/l)

Conc. of stock solution (mg/l)

Major salts (Stock 1) NH4NO3 KNO3 CaCl2.2H2O MgSO4. 7H2O KH2PO4 Minor salts (Stock II) KI H3BO3 MnSO4.4H2O ZnSO4.7H2O Na2MoO4.2H2O CuSO4.5H2O CoCl2

Iron stock (Stock III) FeSO4.7H2O NA2.EDTA.2H2O B5 Vitamins (Stock – IV) Myo-inositol Nicotinic acid Pyridoxine HCl Thiamine HCl Sucrose (g) Agar (g)

1650.00 1900.00 440.00 370.00 170.00

0.83 6.20

22.30 8.60 0.25

0.025 0.025

27.80 37.30

100.00 1.00 1.00

10.00

30.00 8.00

33,000.00 38,000.00

8,800.00 7,400.00 3,400.00

166.00 1,240.00 4,460.00 1,720.00

50.00 5.00 5.00

5,560.00 7,460.00

20,000.00 200.00 200.00

2,000.00

--- ---

Table – 3. Preliminary Phytochemical screening of leaves and callus of

Abutilon indicum.

Hexane extract Chloroform extract Ethanol extract Test

leaves Callus leaves Callus leaves Callus

Alkaloid - - - - - -

Coumarin - - - - - -

Flavonoids - - + + + +

Lignin - - - - + +

Phenol + + + + + +

Protein - - - - + +

Quinone + + + + + +

Saponin - - - - + -

Steroid + - - - + +

Sugar - - - - + +

Tannin - - - - + +

Triterpene + + + + + -

Table – 4. Physico-chemical parameters of leaves and callus of Abutilon indicum

S. No. Parameters Leaves Callus

1 Total ash value (%) 1.02 0.08

2 Water-soluble ash (%) 2.05 0.31

3 Alkalinity of water-soluble ash (ml)

4.66 1.93

4 Acid insoluble ash (%) 6.72 4.03

Extractive value

1 Hexane (%) 2.56 0.53

2 Chloroform (%) 6.54 1.22

3 Methanol (%) 11.04 3.23

Table – 5. Fluorescent analysis of leaves and callus of Abutilon indicum

Visible/ Day light UV Light at 254 nm Experiments

Leaves Callus Leaves Callus

Drug powder Green Pale green Yellowish green Pale yellow

Drug powder + 1 N NaOH (aq.) Pale green Pale yellowish green Yellowish green Pale yellow

Drug powder + 1N NaOH (alc.) Pale green Pale yellowish green Yellowish green Pale yellow

Drug powder + 1 N HCl Yellowish green Pale yellow Green Pale green

Drug powder + 50% H2SO4 Olive green Pale green Fluorescent green Fluorescent green

Drug powder + 50% HNO3 Orange Pale yellow Dark green Green

Drug Powder + C6H3N3O7 Yellow Orange Dark green Green

Drug Powder + CH3COOH Olive green Yellowish green Fluorescent green Fluorescent green

Drug Powder + FeCl3 Pale green Yellowish green Green Pale green

Drug Powder + HNO3+NH3 Orange Pale yellow Green Pale green

Table – 6. Preliminary Phytochemical screening of leaves and callus of Solanum

surattense

Hexane extract Chloroform extract Ethanol extract Test

leaves Callus leaves Callus leaves Callus

Alkaloid + + + + + +

Coumarin - - - - - -

Flavonoids + + + + + +

Lignin - - - - + +

Phenol + + + + + +

Protein - - - - + +

Quinone + + + + + +

Saponin - - - - + -

Steroid + - - - + +

Sugar - - - - + +

Tannin - - - - + +

Triterpene + + + + + -

Table – 7. Physico-chemical parameters of leaves and callus of Solanum surattense.

S. No. Parameters Leaves Callus

1 Total ash value (%) 1.02 0.09

2 Water-soluble ash (%) 2.05 0.21

3 Alkalinity of water-soluble ash (ml) 3.66 1.93

4 Acid insoluble ash (%) 5.72 4.03

Extractive value

1 Hexane (%) 1.56 0.53

2 Chloroform (%) 5.54 1.20

3 Methanol (%) 10.04 3.21

Table – 8. Fluorescent analysis of leaves and callus of Solanum surattense.

Visible/ Day light UV Light at 254 nm Experiments

Leaves Callus Leaves Callus

Drug powder Green Pale green Yellowish green Pale yellow

Drug powder + 1 N NaOH (aq.) Yellowish green Pale yellowish green Pale green Pale yellow

Drug powder + 1N NaOH (alc.) Pale green Pale yellowish green Yellowish green Pale yellow

Drug powder + 1 N HCl Pale green Pale yellow Green Yellowish green

Drug powder + 50% H2SO4 Olive green Pale green Fluorescent green Fluorescent green

Drug powder + 50% HNO3 Orange Pale yellow Dark green Yellowish green

Drug Powder + C6H3N3O7 Yellow Orange Dark green Green

Drug Powder + CH3COOH Olive green Yellowish green Fluorescent green Fluorescent green

Drug Powder + FeCl3 Pale green Green Green Pale green

Drug Powder + HNO3+NH3 Orange Pale yellow Green Pale green

Table - 9. Antimicrobial Activity of Abutilon indicum Leaves

Microorganisms Control

Hexane (50,25,

12.5,6.25 mg/ml)

Control

Chloroform (50,25,

12.5,6.25 mg/ml)

Control

Ethanol (50,25,

12.5,6.25 mg/ml)

Standard

Bacillus subtilis -

13.1±0.1 12.1±0.1 11.1±0.1 11.1±0.1

-

17.2±0.1 16.0±0.1 14.1±0.1 12.2±0.1

-

18.0±0.1 9.0±0.1

- -

27(A)

Staphylococcus aureus

-

13.0±0.2 12.0±0.2 10.2±0.2

-

-

12.0±0.2 11.1±0.1 10.0±0.1

-

-

11.0±0.2 11.0±0.3 12.0±0.1 11.0±0.1

42(M)

Aeromonas hydrophila

-

13.0±0.3 13.1±0.3 11.0±0.2 12.0±0.1

-

15.1±0.2 14.0±0.2 16.0±0.2 12.0±0.1

-

22.0±0.2 20.1±0.3 18.0±0.3 16.0±0.2

18(Tr)

Escherichia coli -

14.1±0.2 12.1±0.1 12.1±0.2 10.2±0.2

17

12.0±0.1 10.2±0.1

- -

-

17.0±0.3 15.0±0.2 14.0±0.3 12.0±0.1

28(K)

Proteus vulgaris -

- - - -

-

- - - -

-

- - - -

30(T)

Pseudomonas aeruginosa

-

13.1±0.1 11.1±0.2 10.0±0.1

-

-

14.0±0.2 13.1±0.1 12.1±0.1 12.1±0.2

-

16.1±0.4 15.1±0.3 13.1±0.1 11.1±0.3

20(K)

- = No activity;

C = DMSO(dimethyl sulphoxide); Measurements are given in millimimeter (mm); Amphicillin (A); Kanamycin (K); Methicillin (M); Nalidixic acid (Na); Trimethoprien (Tr); Tetracycline (T); Gendamicin (G);

Salmonella paratyphi (A)

-

- - - -

-

13.0±0.2 12.2±0.3 11.1±0.2 10.1±0.3

-

15.2±0.1 12.1±0.2 11.1±0.2 10.1±0.1

30(G)

Salmonella typhi -

13.1±0.1 12.1±0.2 13.1±0.2 11.1±0.1

-

16.1±0.3 14.1±0.2 13.1±0.2 11.1±0.2

-

18.1±0.3 16.1±0.1 15.1±0.2 13.1±0.1

20(Na)

Fungus Aspergillus niger

-

- - - -

-

- - - -

-

15.1±0.1 13.1±0.1 12.0±0.2 10.1±0.1

(12)K

Candida albicans

-

- - - -

-

- - - -

-

18.0±0.1 16.1±0.2 14.1±0.2 12.1±0.1

(24)K

Table - 10. Antimicrobial Activity of Abutilon indicum Callus


Hexane

(50,25,

12.5,6.25

mg/ml)

Control

Chloroform

(50,25,

12.5,6.25

mg/ml)

Control

Ethanol

(50,25,

12.5,6.25

mg/ml)

Standard

Bacillus subtilis -

-

-

-

-

-

21.0±0.3

18.0±0.2

16.0±0.3

12.0±0.1

-

14.2±0.1

12.0±0.1

10.1 ±0.1

-

27(A)


-

-

-

-

-

-

22.1±0.3

20.1±0.1

19.1±0.2

17.1±0.1

-

12.0±0.2

10.0±0.3

-

-

42(M)


-

-

-

-

-

-

-

-

-

-

-

25.0±0.2

22.1±0.3

20.0±0.3

16.0±0.2

18(Tr)

Escherichia coli -

-

-

-

-

17

-

-

-

-

-

20.0±0.3

18.0±0.2

16.0±0.3

12.0±0.1

28(K)

Proteus vulgaris -

-

-

-

-

-

-

-

-

-

-

-

-

-

-

30(T)


-

-

-

-

-

-

-

-

-

-

-

16.1±0.4

15.1±0.3

13.1±0.1

11.1±0.3

20(K)


-

-

-

-

-

-

-

-

-

-

-

15.2±0.1

12.1±0.2

11.1±0.2

10.1±0.1

30(G)

Salmonella typhi -

-

-

-

-

-

-

-

-

-

-

23.1±0.3

21.1±0.1

18.1±0.2

15.1±0.1

20(Na)

Fungus

Aspergillus niger

-

-

-

-

-

-

-

-

-

-

-

16.1±0.1

14.1±0.1

13.0±0.2

11.1±0.1

(12)K

Candida albicans

-

-

-

-

-

-

-

-

-

-

-

19.0±0.1

17.1±0.2

14.1±0.2

12.1±0.1

(24)K

- = No activity;

C = DMSO (dimethyl sulphoxide); Measurements are given in millimimeter (mm); Amphicillin (A); Kanamycin (K); Methicillin (M); Nalidixic acid (Na); Trimethoprien (Tr); Tetracycline (T); Gendamicin (G);

Table - 11. Antimicrobial Activity of Solanum surattense Leaf


Hexane (50,25,

12.5,6.25 mg/ml)

Control

Chloroform (50,25,

12.5,6.25 mg/ml)

Control

Ethanol (50,25,

12.5,6.25 mg/ml)

Standard

Bacillus subtilis -

- - - -

- - - -

14.2±0.1 12.0±0.1 10.1 ±0.1

-

27(A)


-

- - - -

16.1±0.4 15.1±0.3 13.1±0.1 11.1±0.3

- - - -

42(M)


-

- - - -

- - - -

15.2±0.1 12.1±0.2 11.1±0.2 10.1±0.1

18(Tr)

Escherichia coli -

- - - -

17

19.0±0.1 17.1±0.2 14.1±0.2 12.1±0.1

- - - -

28(K)

Proteus vulgaris -

- - - -

- - - -

- - - -

30(T)


-

- - - -

- - - -

23.1±0.3 21.1±0.1 18.1±0.2 15.1±0.1

20(K)


-

- - - -

- - - -

- - - -

30(G)

Salmonella typhi -

- - - -

20.0±0.3 18.0±0.2 16.0±0.3 12.0±0.1

- - - -

20(Na)


-

- - - -

12.0±0.2 10.0±0.3

- -

16.1±0.1 14.1±0.1 13.0±0.2 11.1±0.1

(12)K

Candida albicans

-

- - - -

- - - -

23.0±0.2 22.1±0.3 20.0±0.3 16.0±0.2

(24)K

- = No activity;


Table - 12. Antimicrobial Activity of Solanum surattense hairy root.


Hexane (50,25,

12.5,6.25 mg/ml)

Control

Chloroform (50,25,

12.5,6.25 mg/ml)

Control

Ethanol (50,25,

12.5,6.25 mg/ml)

Standard

Bacillus subtilis -

- - - -

- - - -

-

14.2±0.1 12.0±0.1 10.1 ±0.1

-

27(A)


-

- - - -

- - - -

-

12.0±0.2 10.0±0.3

- -

42(M)


-

- - - -

- - -

-

22.0±0.2 21.1±0.3 20.0±0.3 16.0±0.2

18(Tr)

Escherichia coli -

- - - -

17

- - - -

-

- - - -

28(K)

Proteus vulgaris -

- - - -

- - - -

-

- - - -

30(T)


--

- - - -

20.0±0.3 18.0±0.2 16.0±0.3 12.0±0.1

-

16.1±0.4 15.1±0.3 13.1±0.1 11.1±0.3

20(K)


-

- - - -

- - - -

-

15.2±0.1 12.1±0.2 11.1±0.2 10.1±0.1

30(G)

Salmonella typhi -

- - - -

16.1±0.1 14.1±0.1 13.0±0.2 11.1±0.1

-

20.1±0.3 21.1±0.1 18.1±0.2 15.1±0.1

20(Na)


-

- - - -

- - - -

-

- - - -

(12)K

Candida albicans

-

- - - -

- - - -

-

19.0±0.1 17.1±0.2 14.1±0.2 12.1±0.1

(24)K

- = No activity;


Table - 13. Histochemical Localization of Abutilon indicum

Stem Root

S.No. Histochemical Methods Localized Substance

Colour

Indication Outer zone

Middle Zone

Inner zone

Outer zone

Inner zone

1. Toludine Blue O

a.Carboxylated Polysaccharide b.Lignin

Blue Purple to Pink

+++

++

++

+

+++

++

++++

++

+

++++

2. Periodic acid schiffs reagent

Total insoluble Polysaccharides

Purple ++ (-)

+ +++ +

3. Ruthenium Red Pectin Pink ++++ +++ (-) ++++ ++ 4. Iodine Pottasium Iodine Starch Black ++ (-) (-) ++++ + 5. Coomassie Brilliant Blue Total Protein Blue ++ + (-) ++++ (-) 6. Fast Green Histones and basic proteins Green +++ + + ++ (-) 7. Nile Blue Sulphate a. Neutral lipids

b. Acidic lipids Red Blue

++ ++

(-) (-)

(-)

++++ (-)

8. Sudan red - III Lipids Red +++ (-) ++ +++ (-) 9. Fast blue BB Phenols Red to dark

brown +++ (-) ++ ++ (-)

10. Wagner reagent Alkaloids Reddish brown ++ (-) + +++ ++

(-)

11. AlCl3 method Flavonoids Yellowish red +++ (-) ++ ++ (-) 12. Nadi reagent Terpenoids ++ ++ (-) ++++ (-) 13. Benzidine Method Peroxidase Black +++ ++ +++ +++ +++ 14. DOPA method Polyphenol oxidase Blue to Black ++++ ++ (-) ++++ (-)

Outer zone (Epidermis, Hypodermis, Cortex), Middle zone (Cambium, Xylem), Inner zone (Pith).

Stem outer zone (cork cells, old cortical cells) Inner zone (Cambium and Sec. Xylem cells)

Table - 14. Histochemical Localization of Abutilon indicum

Leaf Callus S.

No.

Histochemical Methods

Localized Substance

Colour

Indication Outer region

Inner zone Outer zone

Inner zone

1. Toludine Blue O

a. Carboxylated Polysaccharide b.Lignin

Blue Purple to Pink

++ +

++ ++

+++ ++

+++

++++



Purple ++ + +++ +

3. Ruthenium Red Pectin Pink + +++ ++++ ++ 4. Iodine Pottasium Iodine Starch Black (-) + (-) ++++ 5. Coomassie Brilliant Blue Total Protein Blue (-) ++ ++++ +++ 6. Fast Green Histones and basic proteins Green (-) + ++ +++ 7. Nile Blue Sulphate a. Neutral lipids


++ ++

+

(-) (-)

++

8. Sudan red - III Lipids Red ++ ++ +++ ++ 9. Fast blue BB Phenols +++ + ++ ++

10. Wagner reagent Alkaloids Reddish brown (-) + +++ ++ 11. AlCl3 method Flavonoids Yellowish red ++ ++ ++ ++ 12. Nadi reagent Terpenoids (-) + + ++ 13. Benzidine Method Peroxidase Black + +++ +++ ++++ 14. DOPA mehtod Polyphenol oxidase Blue to Black + ++ +++ +++

Leaf: Outer (Lower and Upper epidermis), Inner (Mesophyll cells, Vascular elements)

Table - 15. Histochemical Localization of Solanum surattense

Stem Root S.

No. Histochemical Methods

Localized Substance

Colour Indication Outer

region Middle Zone

Inner zone

Outer zone

Inner zone

1. Toludine Blue O


Blue Purple to Pink

+++ ++

++ +

+++ ++

++++

++

++ +



Purple ++ +

+ +++ ++

3. Ruthenium Red Pectin Pink ++++ ++ (-) ++++ ++ 4. Iodine Pottasium Iodine Starch Black + (-)

+++ +++ +

5. Coomassie Brilliant Blue Total Protein Blue ++ + ++ ++++ + 6. Fast Green Histones and basic proteins Green +++ + ++ ++ (-) 7. Nile Blue Sulphate a. Neutral lipids


++ ++

(-) (-)

(-)

++++ (-)

8. Sudan red - III Lipids +++ (-) ++ +++ + 9. Polyphenol oxidase Polyphenol oxidase +++ (-) ++ ++ (-)

10. Wagner reagent Alkaloids Reddish brown

++ (-) + ++++ ++

11. AlCl3 method Flavonoids Yellowish red +++ (-) +++ ++ ++ 12. Nadi reagent Terpenoids Red ++ ++ (-) +++ + 13. Benzidine Method Peroxidase Black +++ ++ +++ +++ +++ 14. DOPA method Polyphenol oxidase Blue to Black ++++ (-) (-) ++++ (-)

Outer region (Epidermis, Hypodermis, Cortex), Middle zone (Cambium, Xylem), Inner region (Pith).

Stem outer zone (cork cells, old cortical cells) Inner zone (Cambium and Sec. Xylem cells)

Table - 16. Histochemical Localization of Solanum surattense

Leaf Callus S.

No.

Histochemical Methods

Localized Substance

Colour

Indication Outer region

Inner zone Outer zone

Inner zone

1. Toludine Blue O


Blue Purple to Pink

++ +

+++ ++

+

++

++++

(-)



Purple ++ + + +

3. Ruthenium Red Pectin Pink ++ ++ +++ ++ 4. Iodine Pottasium Iodine Starch Black (-) + (-) ++ 5. Coomassie Brilliant Blue Total Protein Blue + ++ ++ ++ 6. Fast Green Histones and basic proteins Green (-) + ++ ++ 7. Nile Blue Sulphate a. Neutral lipids


++ ++

+ (-)

(-) (-)

(-) (-)

8. Sudan red - III Lipids +++ ++ +++ ++

9. Sudan red - III Lipids ++ ++ ++ ++

10. Wagner reagent Alkaloids Reddish brown ++ + (-) + 11. AlCl3 method Flavonoids Yellowish red ++ ++ (-) ++ 12. Nadi reagent Terpenoids Red + + (-) ++ 13. Benzidine Method Peroxidase Black +++ +++ +++ +++ 14. DOPA method Polyphenol oxidase Blue to Black ++ ++ + ++

Leaf: Outer (Lower and Upper epidermis), Inner (Mesophyll cells, Vascular elements)

Table - 17. Multiple shoot induction response of nodal and Internodal explants of

Abutilon indicum grown on MS medium supplemented with different concentrations of auxins and cytokinin after 25 days.

Values are mean of 20 replicates per treatment and repeated thrice. Values with the same superscript are not significantly different at 5% probability level according to DMRT.

Nodal explant Internodal explant Plant growth

regulators (mg/l)

Shoot bud induction

(%)

Shoot number

Shoot length (cm)

Shoot bud induction

(%)

Shoot number

Shoot length (cm)

BAP 0.5 1.0 1.5 2.0 2.5 KIN 0.5 1.0 1.5 2.0 2.5 BAP KIN 1.5 0.1 1.5 0.5 1.5 1.0 1.5 1.5 1.5 2.0 BAP KIN NAA 2.0 0.5 1.0 2.0 0.5 1.5 2.0 0.5 2.0 2.0 0.5 2.5 2.0 0.5 3.0

80.0e

83.3d e 88.3c

81.6d e 78.3e f

68.3i 71.6h 66.6i j 60.0k l 58.3l

75.0g 80.0e 68.3i 65.0j 61.6k

90.0b 96.6a 90.0b 85.0d 78.3e f

5.7k l 6.0k 10.2f 9.0h 8.6i

4.2o 6.5k 5.0m

4.8m n 4.0o p

9.8f g 13.1e 10.0f g 8.7h i 8.0j

13.6d 16.0a 15.5b 14.0c 10.5e f

1.9j 1.5n 1.3o 1.1o 1.0p

1.6m 2.1h 1.8k 1.7l 1.5n

2.8c 3.5a 3.1b 2.6d 2.5e

2.0i 2.8c 2.5e 2.3f 2.2g

70.0i

76.6f g 80.0d e 73.3g h 68.3i j

65.0j k 68.3i j 60.0l 56.6n 51.6o

75.0g 78.3f 70.0i 66.6j

58.3l m

88.3b 93.3a 85.0c 81.6d 70.0i

2.4l m 5.0k 8.4d 6.2h i 5.2j k

2.8l 5.3j

2.4l m 2.3m

2.0m n

7.1f g 8.0d e 7.8e 6.5h 6.0d

9.0c 12.0a 10.2b 8.0d e 7.2f

3.2m 3.5l 4.5h 4.4h i 4.0j

5.3d 6.2a 5.7b 5.2d e 4.9f g

4.8g 5.6b c 5.0f 4.3i 3.8k

2.7o 3.2m 2.9n 2.5p

2.4p q

Table - 18. Shoot elongation response of multiple shoots raised from node and

internodal explants of A.indicum on MS medium supplemented with different concentrations of GA3 after 30 days.

Nodal Internodal Plant

growth regulators

(mg/l)

% of cultures with elongated

shoots

Shoot length (cm)


shoots

Shoot length (cm)

GA3

1.0

1.5

2.0

2.5

3.0

70.0c

78.3a

75.0ab

68.3cd

61.6e

7.1c

8.3a

7.9ab

6.5d

6.0de

86.6b

90.0a

85.0bc

80.0d

76.6de

8.6ab

9.0a

8.2b

7.8bc

7.0d


Table - 19. Effect of different forms and concentrations of auxins on rooting in A.

indicum after 30 days.


regulators (mg/l)

% of response

No. of Roots

Root length (cm)

% of response

No. of Roots

Root length (cm)

IAA

0.1

0.5

1.0

2.0

3.0

IBA

0.1

0.5

1.0

1.5

2.0

NAA

0.1

0.5

1.0

2.0

3.0

70.0h

78.3fg

81.6f

90.0bc

85.0de

86.6d

90.0bc

93.3b

95.0ab

98.3a

50.0k

58.3j

63.0i

75.0g

70.0h

2.7i

3.0h

3.4g

4.2d

3.9e

3.0h

3.7ef

4.8bc

5.0b

5.9a

1.5 m

1.8l

2.1

3.0h

2.6ij

1.9h

2.3f

2.6de

3.0c

2.5e

2.1g

2.5c

3.0b

3.8b

4.3a

1.3j

1.5i

2.0gh

2.7d

2.3f

61.6i

65.0h

70.0g

78.0e

73.3f

80.0de

81.6d

85.0c

88.3b

91.6a

45.0m

48.3l

55.0k

63.3hi

60.0ij

2.0ij

2.1i

2.7j

3.7d

3.2l

2.6gh

3.0ef

4.3bc

4.5b

4.8a

1.2l

1.5k

2.0ij

2.4h

1.9j

1.4j

1.6i

2.0g

2.8d

2.5l

2.0g

2.4ef

3.0c

3.7b

4.0a

1.4j

1.5ij

1.8h

2.5l

2.0g

Values are mean of 20 replicates per treatment and repeated thrice. Values with the same superscript are not significantly different at 5% probability level according to DMRT

Table - 20. Callus induction from nodal and internodal explants of Abutilon indicum.

Nodal explant Internodal

Plant growth regulators (mg/l) % of

response Callus nature

% of

response No. of Roots

2,4-D

0.5 1.0 2.0 3.0 4.0

NAA 0.5 1.0 2.0 3.0 4.0

NAA KIN 3.0 0.5 3.0 1.0 3.0 1.5 3.0 2.0 3.0 2.5

NAA BAP 3.0 0.5 3.0 1.0 3.0 1.5 3.0 2.0 3.0 2.5

70.0h

78.3fg

81.6f

90.0bc

85.0de

86.6d 90.0bc

93.3b

95.0ab

94.3a

50.0k

58.3j

63.0i

75.0g

70.0h

86.6e 91.0b 96.6a 90.0bc 85.0f

YF YF YF

GYF GYF

YF YF GC GC GC

GC

GCN GCN GCN GC

GC GC GC GF GF

61.6i

65.0h

70.0g

78.0e

73.3f

80.0de

81.6d

85.0c

88.3b 87.6a

45.0m

48.3l

55.0k

63.3hi

60.0ij

65.0gh

70.0f

91.3h

95.3ij

51.6l

YF YF

GYF GYF GYF

GYF GC GC

GCN GCN

GC

GCN GCN GCN GC

GC GC

GCN GCN GC

YF – yellow friable; GYF – Green yellow friable; GC – Green compact; GCN – Green compact nodular; GF – Green friable. Values are mean of 20 replicates per treatment and repeated thrice. Values with the same superscript are not significantly different at 5% probability level according to DMRT

Table - 21. Shoot regeneration response of Node and Internodal callus of A.indicum

grown on MS medium containing cytokinin, auxin and tyrosine after 30 days



response

No. of Shoots/ Explant

Shoot length (cm)

% of response


Soot length (cm)

BAP 0.5 1.0 1.5 2.0 2.5

KIN 0.5 1.0 1.5 2.0 2.5

BAP NAA 2.0 0.1 2.0 0.5 2.0 1.0 2.0 1.5 2.0 2.0

BAP NAA Tyr 2.0 1.0 0.5 2.0 1.0 1.0 2.0 1.0 1.5 2.0 1.0 2.0 2.0 1.0 2.5

78.3fg

85.0d

86.6cd 90.0bc

75.0gh

70.0ij

71.6i

78.3fg

66.6jk

60.0m

70.0ij

76.6g

80.0f

68.3j

63.0l

88.3de

91.6b

95.0a

90.0bc

83.3d

2.4ij

3.2g

4.1ef

5.0c

3.0gh

1.5m

2.4ij

3.0gh

2.0k

1.8kl

4.9i

5.2f

7.0e

6.0g

5.5k

7.5ef

8.0b

11.6a

9.4c

9.0b

1.6n

2.4lm

3.0i

3.5h

2.5l

2.7k

3.0i

4.1de

3.6gh

2.9ij

3.7g

3.9f

4.5bc

3.8fg

3.0i

4.2d

4.6b

5.4a

4.5bc

3.9f

61.6i

65.0h

70.0g

78.0e

73.3f

80.0de

81.6d

85.0c

88.3b 91.6a

45.0m

48.3l

55.0k

63.3hi

60.0ij

2.6gh

3.0ef

4.3bc

4.5b

4.8a

1.2l

1.5k

2.0ij

2.4h

1.9j

2.0ij

2.1i

2.7j

3.7d

3.2l

78.3fg

85.0d

86.6cd 90.0bc

75.0gh

1.4j

1.6i

2.0g

2.8d

2.5l

2.0g

2.4ef

3.0c

3.7b

4.0a

1.4j

1.5ij

1.8h

2.5l

2.0g

5.9i

6.2f

7.0e

6.0g

5.5k Values are mean of 20 replicates per treatment and repeated thrice. Values with the same superscript are not significantly different at 5% probability level according to DMRT

Table - 22. Multiple shoot induction response of nodal and internodal explants of

S.surattense grown on MS medium supplemented with different concentrations of auxins and cytokinin after 45 days.


regulators (mg/l)

Shoot bud induction

(%)

Shoot number

Shoot length (cm)

Shoot bud induction

(%)

Shoot number

Shoot length (cm)

BAP 0.5 1.0 1.5 2.0 2.5 3.0

KIN 0.5 1.0 1.5 2.0 2.5 3.0

BAP KIN 2.5 0.1 2.5 0.5 2.5 1.0 2.5 1.5 2.5 2.0

BAP KIN NAA 2.5 0.5 0.5 2.5 0.5 1.0 2.5 0.5 1.5 2.5 0.5 2.0 2.5 0.5 2.5

58.3l

61.6jk

68.3hi

73.0fg

78.3de 75.0f

66.6i 63.0j 55.0m 51.6n 48.3o 40.0p

83.3c 86.0b 78.3de 70.0h 65.0ij

81.6cd 90.0a

85.0bc 80.0d 75.0f

5.3n 6.0lm 6.7jk 7.9i

9.4fg 8.1hi

5.0o 6.8j 6.1l 5.2no 4.8op 3.9q

9.0g

12.5cd 10.9e 9.7f

8.5h

13.5c 16.2a 14.0b 13.8bc 12.1d

3.9lm 3.5no 2.9p 2.6q 2.4qr

2.0s

5.0i 5.4h 4.9ij 4.3k 4.0l

3.6n

7.0fg 8.5b 7.9de

6.8g 5.0i

8.0d 9.2a 8.3bc 7.8e

7.2f

68.3ij 70.0i 76.6gh 80.0fg 85.0e 83.3ef

66.6j 70.0i 65.0jk 63.3k 60.0l

58.3lm

86.6de 88.3ef 81.6f 78.3g 75.0h

95.0ab 96.6a 91.0c 88.3d 80.0fg

8.0m

8.5l

9.8k

12.3h 13.0g 11.9hi

8.0m 8.3lm 7.0mn 6.2o 5.8op

5.5p

13.8ef 14.5e 12.9gh

11.5i

10.7j

19.2b 21.0a 18.5bc 16.0d

13.8ef

2.9lm 2.5n 2.0op 1.8p 1.6pq 1.0r

3.5k 4.3i 3.8j 3.0l 2.8m 2.1o

5.5f 6.0e 5.1g 4.8h 4.3i

7.8c 8.5a

8.0b

7.9bc 7.6cd


Table- 23. Shoot elongation response of multiple shoots raised from node and

internodal explants of S.surattense on MS medium supplemented with BAP (1.5 mg/l) and GA3 after 30 days.

Nodal Internodal Plant

growth regulators

(mg/l)


shoots

Shoot length (cm)


shoots

Shoot length (cm)

GA3

0.5

1.0

1.5

2.0

2.5

68.0c

76.3a

72.0ab

64.3cd

60.6e

7.1c

8.3a

7.9ab

6.5d

6.0de

84.6b

87.0a

85.0bc

80.0d

76.6de

8.6ab

9.0a

8.2b

7.8bc

7.0d


Table -24. Rooting response of in vitro raised shoots of S.surattense grown on MS

medium supplemented with auxins, after 30 days.


regulators (mg/l)

% of response

No. of Roots

Root length (cm)

% of response

No. of Roots

Root length (cm)

IBA 0.1 0.5 1.0 1.5 2.0 2.5

NAA 0.1 0.5 1.0 1.5 2.0 2.5

IAA 0.1 0.5 1.0 1.5 2.0 2.5

56.6ij

60.0hi

68.3f

71.6e

65.0g

61.6h

88.3ab

90.0a

85.0c

78.3d

70.0ef

66.6fg

48.3kl

50.0k

56.6ij

60.0hi

65.0g

58.3i

4.9jk

5.0j

5.5i 6.8g

6.0h

5.2ij

9.0b

9.8a

8.7bc

8.0d

7.6e

7.2f

2.8no

3.0n

3.6m

4.0l

4.8k

4.0l

3.0j

3.6hi

3.9g

4.5e

4.1f

3.8gh

5.1b

5.5a

5.0bc

4.8d

4.0fg

3.7h

2.1l

2.8k

3.0j

3.5i

4.0fg

3.9g

48.3ij

50.0i

56.6g

60.0f

58.3fg

53.3h

75.0b

78.3a

76.6ab

70.0c

66.0d

65.0de

31.6o

35.0mn

36.6m

43.3kl

50.0i

45.0k

3.2j

3.8hi

4.1g

5.0ef

4.8f

4.0gh

7.0cd

8.0a

7.5b

7.1c

6.8d

6.0e

2.5l

3.0k

3.1jk

3.9h

4.0gh

3.9h

2.4m

2.7kl

2.8k

3.4h

3.7fg

3.0lj

4.8b

5.2a

4.7bc

4.3d

4.0e

3.5gh

2.4m

2.6l

3.1i

3.6g

3.8f

3.5gh


Table - 25. Callus induction from nodal and intermodal explants of Solanum surattense.

Nodal explant Internodal Plant growth

regulators (mg/l) % of response

Callus nature % of

response No. of Roots

2,4-D 0.5 1.0 2.0 3.0 4.0

NAA 0.5 1.0 2.0 3.0 4.0

NAA BAP 3.0 0.5 3.0 1.0 3.0 1.5 3.0 2.0 3.0 2.5

BAP KIN NAA 3.0 0.5 0.5 3.0 0.5 1.0 3.0 0.5 1.5 3.0 0.5 2.0 3.0 0.5 2.5

72.0h

76.3fg

82.6f

89.0bc

84.0de

84.6d 92.0bc

93.3b

95.0ab

90.3a

50.0k

58.3j

63.0i

75.0g

70.0h

86.6e 91.0b 96.6a 90.0bc 85.0f

YF YF YF

GYF GYF

YF YF GC GC GC

GC

GCN GCN GCN GC

GC GC GC GF GF

60.6i

65.0h

72.0g

76.0e

75.3f

45.0m

48.3l

55.0k

63.3hi

60.0ij

65.0gh

70.0f

63.3h

58.3ij

51.6l

80.0de

81.6d

85.0c

84.3b 75.6a

YF YF

GYF GYF GYF

GYF GC GC

GCN GCN

GC

GCN GCN GCN GC

GC GC

GCN GCN GC

YF – yellow friable; GYF – Green yellow friable; GC – Green compact; GCN – Green compact nodular; GF – Green friable. Values are mean of 20 replicates per treatment and repeated thrice. Values with the same superscript are not significantly different at 5% probability level according to DMRT

Table - 26. Shoot regeneration response of Node and Internodal callus of S.surattense

grown on MS medium containing cytokinin, auxin and tyrosine after 30 days



response


Shoot length (cm)

% of response

No. of Shoots / Explant

Soot length (cm)

BAP 0.5 1.0 1.5 2.0 2.5

KIN 0.5 1.0 1.5 2.0 2.5

BAP NAA 2.0 0.1 2.0 0.5 2.0 1.0 2.0 1.5 2.0 2.0

BAP NAA Tyr 2.5 1.0 0.5 2.5 1.0 1.0 2.5 1.0 1.5 2.5 1.0 2.0 2.5 1.0 2.5

76.3fg

85.0d

83.6cd 91.0bc

75.0gh

70.0ij

71.6i

78.3fg

66.6jk

60.0m

70.0ij

76.6g

80.0f

68.3j

63.0l

88.3de

91.6b

96.0a

90.0bc

83.3d

2.4ij

3.2g

4.1ef

5.0c

3.0gh

1.5m

2.4ij

3.0gh

2.0k

1.8kl

4.9i

5.2f

7.0e

6.0g

5.5k

7.5ef

8.0b

11.6a

9.4c

9.0b

1.6n

2.4lm

3.0i

3.5h

2.5l

2.7k

3.0i

4.1de

3.6gh

2.9ij

3.7g

3.9f

4.5bc

3.8fg

3.0i

4.2d

4.6b

5.4a

4.5bc

3.9f

60.6i

65.0h

69.0g

78.0e

73.3f

80.0de

81.6d

85.0c

86.3b 90.6a

44.0m

49.3l

55.0k

63.3hi

60.0ij

78.3fg

85.0d

89.6cd 87.0bc

75.0gh

2.6gh

3.0ef

3.3bc

3.5b

3.8a

1.2l

1.5k

2.0ij

2.4h

1.9j

2.0ij

2.1i

2.7j

3.7d

3.2l

1.6gh

3.2ef

3.9bc

3.5b

3.8a

1.4j

1.6i

2.0g

2.8d

2.5l

2.0g

2.4ef

3.0c

3.7b

4.0a

1.4j

1.5ij

1.8h

2.5l

2.0g

5.9i

6.2f

7.0e

6.0g

5.5k Values are mean of 20 replicates per treatment and repeated thrice. Values with the same superscript are not significantly different at 5% probability level according to DMRT

Table – 27: In vitro Callus induction response (%) of Abutilon indicum

c, control; nt, not tried. Data represent mean ± SE based on 24 replicates each with one explant. The experiment was repeated twice.

Concentration ( mg-1)

Adenine Kn BA Zeatin TDZ

0.0

1.0

2.0

3.0

4.0

5.0

10.0

20.0

30.0

40.0

50.0

70.0

90.0

100.0

c

nt

nt

nt

nt

nt

90 + 0.5

95 ± 0.3

78 ± 0.8

75 ± 0.5

70 ± 0.4

70 ± 0.3

60 ± 0.3

60 ± 0.5

c

80 ± 0.5

85 ± 1.2

88 ± 0.6

90 ± 0.2

100 ± 0.8

nt

nt

nt

nt

nt

nt

nt

nt

c

95 ± 0.5

92 ± 0.2

88 ± 0.4

75 ± 0.1

74 ± 0.2

nt

nt

nt

nt

nt

nt

nt

nt

c

6 ± 0.1

92 ± 0.3

80 ± 0.3

78 ± 0.5

70 ± 0.2

nt

nt

nt

nt

nt

nt

nt

nt

c

0 ± 0.2

95 ± 0.5

74 ± 0.2

70 ± 0.4

50 ± 0.2

nt

nt

nt

nt

nt

nt

nt

nt

Table – 28: Effect of CuSo4 on fresh (FW) and dry (DW) weight (g) of callus formed

on leaf explants of Abutilon indicm Cultured on WB containing 5mg1-1 kn.

Week after culture Cuso4

(µM) Biomass

4 8 12

0

50

100

150

200

250

300

FW

DW

FW

DW

FW

DW

FW

DW

FW

DW

FW

DW

FW

DW

1.40 ± 0.12

0.18 ± 0.04

1.45 ± 0.12

0.20 ± 0.04

1.49 ± 0.12

0.20 ± 0.02

1.52 ± 0.13

0.21 ± 0.02

1.58 ± 0.13

0.21 ± 0.01

1.62 ± 0.13

0.22 ± 0.02

1.43 ± 0.14

0.14 ± 0.02

1.48 ± 0.11

0.19 ± 0.03

1.51 ± 0.12

0.22 ± 0.02

0.22 ± 0.04

1.62 ± 0.10

1.62 ± 0.10

0.22 ± 0.02

1.67 ± 0.10

0.23 ± 0.01

1.50 ± 0.12

0.15 ± 0.01

1.38 ± 0.11

0.13 ± 0.02

1.59 ± 0.12

0.21 ± 0.05

1.58 ± 0.14

0.20 ± 0.02

1.65 ± 0.14

0.22 ± 0.02

1.71 ± 0.15

0.23 ± 0.03

1.64 ± 0.11

0.22 ± 0.02

1.45 ± 0.12

0.14 ± 0.01

1.30 ± 0.11

0.12 ± 0.01

Data represent mean ± SE based on 24 replicates each with one explant. The experiment was repeated twice.

Table – 29: Effect of ZnSo4 on fresh (FW) and dry (DW) weight (g) of callus formed

on leaf explants of Abutilon indicum Cultured on WB supplemented containing 5mg1-1 kn.

Week after culture ZnSo4(µM) Biomass

4 8 12

0

50

100

150

200

250

300

FW

DW

FW

DW

FW

DW

FW

DW

FW

DW

FW

DW

FW

DW

1.30 ± 0.10

0.26 ± 0.00

1.35 ± 0.10

0.23 ± 0.00

1.75 ± 0.20

0.30 ± 0.02

1.88 ± 0.13

0.36 ± 0.02

1.22 ± 0.13

0.27 ± 0.01

1.20 ± 0.13

0.20 ± 0.02

1.18 ± 0.14

0.18 ± 0.02

1.50 ± 0.12

0.29 ± 0.00

1.58 ± 0.12

0.26 ± 0.01

1.70 ± 0.04

0.31 ± 0.10

1.82 ± 0.10

0.36 ± 0.02

1.48 ± 0.10

0.26 ± 0.01

1.43 ± 0.12

0.29 ± 0.01

1.31 ± 0.11

0.21 ± 0.02

1.62 ± 0.21

0.31 ± 0.05

1.63 ± 0.15

0.29 ± 0.01

1.75 ± 0.14

0.32 ± 0.02

1.90 ± 0.15

0.37 ± 0.03

1.31 ± 0.11

0.29 ± 0.02

1.30 ± 0.12

0.24 ± 0.01

1.12 ± 0.11

0.22 ± 0.01

Data represent mean ± SE based on 24 replicates each with one explant. The experiment was repeated twice.

Table-31: Effect of plant sample extract and silymarin pre treatment on bio-chemical parameters of the mice, intoxicated with CCl4

Values are expressed as mean ±S.E; n=5; *p<0.05 Vs. Normal, **p<0.05 Vs. Control

S. No

Groups SGOT SGPT ALP Total

Protein Total

Bilirubin Direct

Bilirubin Indirect Bilirubin

1 Normal 565.80±0.86 274.60±0.92 723.20±0.58 5.82±0.04 .24±0.02 .14±0.02 .10±.00

2 Control (CCl4) 1055.40±0.67*,** 766.00±0.70*,** 1166.40±0.60*,** 9.10±0.03*,** .86±0.02*,** .46±0.02* .40±.00*.**

3 1-A (A. indicum callus) 757.00±0.70*,** 294.60±0.50*,** 1133.20±0.58*,** 7.18±0.04*,** .66±0.02*,** .46±0.02* .20±.00*.**

4 1-B (A. indicum Leaf) 1045.00±0.70*,** 332.80±0.66*,** 1031.40±0.50*,** 8.22±0.06*,** .64±0.02*,** .44±0.02* .20±.00*.**

5 2-A (S. surattense Leaf) 985.00±0.70*,** 451.60±0.50*,** 822.00±0.44*,** 8.28±0.04*,** .46±0.02*,** .26±0.02*,** .20±.00*.**

6 2-B (S. surattense hairy root) 857.40±0.81*,** 456.80±0.58*,** 1045.60±0.51*,** 6.42±0.04*,** .44±0.02*,** .24±0.02*,** .20±.00*.**

7 Quercetin 675.40±0.51*,** 305.20±0.58*,** 663.60±0.74*,** 7.82±0.04*,** .54±0.02*,** .34±0.02*,** .20±.00*.**

8 Silymarin 989.20±0.58*,** 545.40±0.67*,** 754.20±0.37*,** 6.38±0.06*,** .30±0.03** .18±0.02** .12±.02**

Plate - I

Fig 1: Abutilon indicum – A population in uyyakondan wet land area of

Tiruchirapanni. In a favorable rainy season it dominates the entire area of

bund vegetative. Almost all other herbaceous forms inhabited in the area are

completely suppressed by this

Fig 2: A portion of twig showing young and matured fruits

Fig 3: A portion of mature root showing the limited number of secondary root lets.

Fig 4: A view of Solanum surattense in a small population. Most of the sites

approximately 4 or 5 plants are found together over the soil surface.

Fig 5: An individual twig showing the spines and flowers. The violet petals attract

honey bees at the time of anthesis.

Fig 6: The Solanum surattense root showing secondary rootlets.

Plate -II

Figs 7-61: Histochemical and Anatomical studies of Abutilon indicum and Solanum

surattense.

Fig 7: T. S. of Abutilon indicum stem stained in TBO (Toludine Blue O) showing

the anatomy and Histochemistry in the middle portion of a section. The

cortex and secondary vascular tissues are well distinguished. The inner most

cortical cells and phloem parenchyma cells contained secondary

metabolites. The cambial zone becomes activated to form secondary xylem

and phloem cells.

Fig 8: A portion of stem T.S. showing the cortical cell stored phenolic compound

in their chytoplasm. The upper border is made by collenchymatous cells.

Fig 9: In Abutilon indicum few cells in pith region also contained phenolic

compounds.

Fig 10&11: Showing the lignified secondary wall in xylem and phloem cells

respectively.

Plate -III

Fig 12: T.S. of stem stained in Ruthenium Red showing the tissue distribution

pattern. The outer starts with single layered epidermis, chlorenchymatous,

hypodermis, collenchymas, parenchymatous cortex, secondary phloem

(predominated by) fibers, phloem parenchyma, cambium, secondary xylem

and ending with pith.

Fig 13: A view of stem T.S. showing few secretary cells in the hypodermal zone

above to the collenchymas cells. Note: The pectination pattern of cell wall

varied in various layers collenchymas cells.

Fig 14: A portion of stem, T.S. showing the secondary phloem region. Where newly

formed cells contained the calcium oxalate crystals. The ray cells in phloem

are uniseriate and multiseriate in nature.

Plate - IV

Fig 15: The secondary chemical compounds are found in cortical and phloem cells.

Fig 16&17: Showing the cambial cells and their derivatives. The cambial cells only

made by pectinized walls.

Plate - V

Fig 18: T.S. of matured root in Abutilon indicum showing the outer cortex,

secondary phloem, cambium and secondary xylem. The parenchyma cells in

both secondary vascular elements stored starch, lipids and phenols.

Fig 19: T.S. of root showing the outer region, with cork and few older cortical cells.

The flavonoid compounds were found in the older cortical cells.

Fig 20: T.S. of root showing the innermost core with secondary xylem elements.

The lipids and starch are profusely found in the ray parenchyma cells.

Plate - VI

Fig 21: T.S. of mature root-enlarged at secondary xylem showing the vessels,

trachieds fibres and parenchyma cells.

Fig 22: Similar to stem, in the toot the secondary phloem also contained calcium

oxalate crystals.

Fig 23: T.S. of root stained in Ruthenium Red showing the tissue distribution

pattern.

Fig 24: The cork cells of root showing synthesis and storage of phenolic

compounds.

Fig 25&26: T.S. of leaf (Fig 25) the upper epidermis covered by many hairs. (Fig 26)

the epidermises and mesophyll cells – In between the mesophyll cells a

continued layer of non chlorophyll cells present and participated in

synthesis and storage of phytochemicals, in leaf.

Plate - VII

Fig 27: T.S. of callus showing the vascular nodulation by forming tracheid

elements. The outermost region cells are meristematic producing new cells.

Fig 28&29: The T.S. of callus cells showing differential augmentation in only cell wall

layers for the purpose to store the phenolic compounds.

Plate - VIII

Fig 30: T.S. of callus showing uniquely enlarged cell and involved in secondary

compound synthesis.

Fig 31: Formation of vascular nodulation by the differentiating treached elements in

the callus.

Fig 32&34: T.S. of callus stained in Ruthenium Red.

Fig 32: A callus cell population showing different stages of development.

Fig 33: The inner vascular elements completely encircled by the phloem like cells.

The cells in outmost region are actively divided to produce new cells.

Fig 34: A portion of callus cells showing few vacuolated cells at regular intervals

and that are modified into secretory cells.

Plate - IX

Fig 35-37: An enlarged view of outer zone cells showing the cell division process

during intensive proliferation.

Fig 38: A view of T.S. of callus showing the vascular differentiation. This complex

resembles the intact vascular elements of trachied, fibres, and parenchyma

(except vessel) of in vivo condition.

Plate - X

Fig 39 - 41: T.S. of callus showing accumulation of starch grains. The MS medium

contained all essential components for the growth, however, callus

synthesis starch and accumulate more amount in the central region (Fig

41).

Plate - XI

Fig 42: Solanum surattense T.S. of stem stained in TBO showing the cortical and

vascular regions. The cambium layer is compressed by the outer cortical and

xylem parenchyma cells. Many secretory cells were found in the cortical

region.

Fig 43: A portion of stem in an outer region showing the epidermis with thick

cuticle,the hypodermis made by chlorenchymatous cells. There is

differential pectinization in the cell wall of collenchyma.

Fig 44: T.S. of stem showing the vascular elements. Few cells in this region are

modified into the secretory cells.

Plate - XII

Fig 45: T.S. of stem stained in Ruthenium Red showing the epidermis, sub

epidermis, collenchyma layer, and parenchymatous cortex.

Fig 46: An enlarged view of stem showing epidermis sub-epidermis and

collenchymatous layer. Many of the sub-epidermis cells modified into

secretory cells and to produce secondary chemical compounds.

Fig 47&48: T.S. of stem showing cells in pith.

Fig 47 : An enlarged parenchymatous pith cell showing the pectin accumulation only

in corners of polygonal cells.

Fig 48 : The protoxylem in the pith region.

Plate - XIII

Fig 49: T.S. of root stained in TBO showing cortex and vascular tissues. Note: The

epidermis is replaced by cork cells.

Fig 50: The newly formed xylem cells in root with accumulation of phenolic

compounds.

Fig 51: An enlarged view of outer cortex showing thick walled parenchymatous

cells.

Fig 52: T.S. of root showing vessels, tracheid and fibers. The parenchyma cells in

this region shire lipid and starch.

Plate - XIV

Fig 53: T.S. of root stained in Ruthenium Red showing the tissue distribution

pattern.

Fig 54: An enlarged view of vascular elements composed of vessels and

parenchyma cells.

Fig 55: T.S. of root showing the secondary xylem.

Plate - XV

Fig 56: The outer cortical cells slowly replaced by the newly formed cork cells.

Many phenol containing cells are found in this region.

Fig 57: The active cambium and its xylem derivative cells.

Fig 58: T.S. of root showing the formation of vessel member from the cambial

initial.

Plate - XVI

Fig 59: T.S. of callus showing cell proliferation.

Fig 60: The leaf explants produces the callus in Solanum.

Fig 61: T.S. of callus showing the accumulation of secondary compound in the

inner zone.

Plate - XVII

Fig 62: Rhizogenesis from epidermal layer of leaf explants.

Fig 63&64: Profusely branched root hairs.

Plate – XVIII

Multiple shoot induction from nodal explant of Abutilon indicum

Fig .65: Shoot growth from nodal explant (after 12 days of inoculation).

Fig .66: Initiation of multiple shoots (after 17 days).

Fig .67: Elongation of multiple shoots (after 25 days).

Fig .68: Hardened plantlet after rooting in plastic pots

Plate – XIX

Shoot induction from Internodal explant of A.indicum

Fig. 69: Shoot induction of internodal explant (after 13 days of inoculation)

Fig .70: Initiation of multiple shoots (after 16 days).

Fig .71: Elongation of multiple shoots after 20 days

Fig .72: Hardened plantlet in pots

Plate – XX

Organogenic callus induction and plant regeneration from internodal explants of

A.indicum

Fig.73 : One weeks old callus obtained from internodal explants

Fig.74 : Two weeks old organogenic callus.

Fig.75: Green compact nodular calls after 3 week culture.

Fig.76: Multiple shoot proliferation from callus after two weeks of subculture.

Figs.77: Elongated shootlets after 2 weeks of subculture.

Fig.78: Hardened plantlet on a plastic pot.

Plate – XXI

Organogenic callus induction and plant regeneration from nodal explants of

A.indicum

Fig. 79: Two weeks old callus obtained from nodal explants

Fig.80: Green compact nodular calls after 18 days culture.

Fig. 81: Multiple shoots proliferation from callus after 20 days of subculture.

Fig. 82: Elongated shootlets after 2 weeks of subculture.

Fig.83: Hardened plantlet on a plastic pot.

Plate –XXII

Shoot proliferation in Solanum surattense from node explants

Fig.84: One week old multiple shoots

Fig.85: Two weeks old multiple shoots proliferated from nodal explant.

Fig.86: Four weeks old multiple shoots.

Fig. 87: Elongated shoot with root on elongation medium.

Fig. 88: Rooted shoot lets on rooting medium.

Plate –XXIII

Direct regeneration of Solanum surattense from internodal explant

Fig.89: 14 days old multiple shoot buds

Fig.90: Regeneration of shoot from cut surface.

Fig.91: 25days old multiple shoots.

Fig.92: Elongation of shoot with root on elongation medium.

Fig.93: Hardened plantlet on pot.

Plate – XXIV

Organogenic callus induction and plant regeneration from nodal explants of

Solanum surattense

Fig.94 : Two weeks old callus obtained from nodal explants

Fig.95: Shoot bud derived from compact nodular callus (after 18 days culture).

Fig.96: Multiple shoots proliferation from callus (after 25 days of subculture).

Fig.97: Elongated shootlets after 2 weeks of subculture.

Fig. 98: Elongated shoots on elongation medium

Fig.99: Elongated shoots with root on rooting medium.

Plate – XXV

Organogenic callus induction and plant regeneration from internodal explants of

Solanum surattense

Fig.100: Two weeks old green compact nodular callus obtained from internodal

explants

Fig.101: Shoot bud multiplication from compact nodular calls after 20 days culture.

Fig. 102: Shoot bud formation from callus after three weeks of subculture.

Fig. 103: Elongated shoots on elongation medium

Fig.104: Elongated shoots with root on rooting medium.

Plate – XXVI

A.indicum leaf callus used for Quercetin enhancement

Fig. 105 - 107: Three weeks old quercetin enhanced callus

Fig. 108: Normal green compact Callus

Plate – XXVII

Fig. 109: HPLC chromatogram of quercetin (Used as the standard and purchased

from Himedia RM6191).

Fig. 110: HPLC chromatogram of Abutilon indicum leaf sample.

Fig. 111: HPLC chromatogram of A.indicum callus sample.

Fig.112: HPLC chromatogram of S.surattense leaf sample.

Fig.113: HPLC chromatogram S. surattense hairy root.

Plate – XXVIII

Hepatoprotective activity

Fig. 114: Liver section of normal Albino mice.

Fig. 115: Liver section of mice treated with CCl4.

Fig. 116: Liver section of mice treated with aqueous extracts of Abutilon

indicum callus.

Fig. 117: Liver section of mice treated with aqueous extract of A.indicum leaf.

Fig. 118: Liver section of mice liver treated with S.surattense leaf.

Fig. 119: Liver section of mice treated with S.surattense hairy root.

Fig. 120: Liver section of mice treated with quercetin.

Fig. 121: Liver section of mice treated with silymarin.

Appendix – I Amount Conc. of stock (mg/l) solution...

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