Commonly known as Ghritakumari, a xerophytic medicinal plant

Family: Liliaceae
Active principles: Aloin, Aloesone, Emodin, Aloesi
Medicinal Uses:
•Healing properties
•Effects on skin exposure to UV and gamma radiation
•Anti-inflammatory action
•Laxative effects
•Antiseptic effect etc
The technique to tissue and organ culture is used for rapid
multiplication of plants, and for preservation valuable germplasm.
To create a large no. of plant in short period of time plant tissue
culture is indispensible.
The tissue cultured plants often show variation. To study the level
of variation cytological studies were made in vivo and iv vitro.
Also phytochemical analysis was done from plant raised in vivo and
in vitro to compare the variation of its content.
Propagation of Aloe vera (sweet variety-A common market
variety) plant in vitro.
Chromosomal analysis of the plant grown in vivo and in vitro.
Comparative phytochemical study of the in vivo and in vitro
grown plants.
Phytochemical
analysis
 B5(Gamborg,1968)
IAA + BA , 2,4D + Kn , NAA+ BA
Callusing M.S.(Murashige and
Skoog,1962)
IAA + BA , 2,4D + Kn , NAA+ BA

B5(Gamborg,1968)
BA and Kn
Shoot tip
multiplication M.S.(Murashige and Skoog,1962)

BA and Kn

Rooting M.S.(Murashige and Skoog,1962)

BA+IAA and B A+ IBA
Pre treatment
Saturated P.D.B. (Para dichlorobenzene) for 2 1/2- 3 hrs.
Fixation
Acetic alcohol (45% glacial acetic acid : 95%ethanol :: 1:3 ).
Staining and making squash preparation
Stain use 2% Acetoorcein solution.
Acetoorcein and 1(N) Hcl in 9:1 ratio for 1 hr.
Squash preparation
Squashing was done with 45% Glacial acetic acid.
Observation
Calculate Mitotic index
M.I.=Total number of dividing cell/Total number of cell in field × 100
 Standardization of sterilization
Concentration of Duration Total Fresh & Fresh &
Infected Intolerant
HgCl2 (%) (min.) inoculated Healthy Healthy (%)

5
10 10 0 0 0
7 10 10 0 0 0
0.05 10 10 10 0 0 0
12 10 8 1 1 10
15 10 6 1 3 30
5 10 10 0 0 0
7 10 9 0 1 10
0.1 10 10 0 1 8 80
12 10 0 6 4 40
15 10 0 10 0 0
 Establishment of explant
Sl no. Basal PGR (mg/L) Inititation of % Response Types of Number of Length of
media Response response microshoot microshoot
(cm)
(Days)
1. MS Control(No PGR) 23 30 Shoot 2.3 1.5cm
elongation
2. MS NAA(0.5)+BA(0.5) 15.5 80 Shoot 2.7 3.8 cm
elongation
3. MS IAA(0.5)+BA(0.5) 17 60 Shoot 2.5 2.25cm
elongation
4. MS 2,4D(0.5)+Kn(0.5) 20 40 Shoot 2.5 1.8cm
elongation
5 B5 Control (No PGR) No response - - -
6. B5 NAA(0.5)+BA(0.5) 25 40 Shoot 2.25 1.5cm
elongation
7. B5 IAA(0.5)+BA(0.5) 24 30 Shoot 2.3 1.8cm
elongation
8. B5 2,4D(0.5)+Kn(0.5) 25.5 30 Shoot 2 1.5cm
elongation
After 15 days of After 25 days of After 35 days of
inoculation inoculation inoculation
 Multiplication of explants
Sl Basal PGR (mg/L) % Response Types of response Number of Length of
no. media microshoot microshoot(cm)
1. MS Control (No 30 Shoot elongation 1.30 1.50
PGR)
2. MS BA(2) 80 Shoot multiplication 2.80 3.37
3. MS BA(5) 70 Shoot multiplication 2.70 3.07
4. MS Kn(2) 90 Shoot multiplication 3.10 4.10
5. MS Kn(5) 70 Shoot multiplication 2.42 2.70
8 B5 Control No response - - -
9. B5 BA(2) 50 Shoot multiplication 2.4 2.5
10. B5 BA(5) 50 Shoot multiplication 2.33 2.08
11 B5 Kn(2) 40 Shoot multiplication 2.25 2.25
12 B5 Kn(5) 30 Shoot multiplication 2.33 2.5
Inoculation After 20 days of After 60 days of After 90 days of
Inoculation Inoculation subculture
 Root development
Sl no. Basal media PGR added Response initiation Types of Percentage of
(mg/l) (in days) response response (%)
1. MS Control (no PGR) No response No root 0%
formation
2. MS BA(0.5)+IBA (0.2) 12 1-2White roots 80%
develop
3. MS BA(0.5)+IAA(0.5) 15 Root formation 65%
initiate
Rooting After 20 days Rooting After 35 days
of subculture of subculture
 In vitro In vivo
No. of observation No. of cell in field No. of dividing cell No. of cell in field No. of dividing cell
1 154 13 144 17
2 146 11 112 11
3 174 17 128 11
4 164 9 92 10
5 153 23 120 10
6 123 18 162 8
7 137 17 157 11
8 118 11 185 19
9 97 5 200 20
10 134 13 145 19
Total 1400 137 1445 117
Mitotic index =9.87 Mitotic index =8.09
Dividing and non dividing Chromosome 2n = 14
field at 45X view at 45 X
It was not possible to induce callus through any of the media used.
Thus phytochemical analysis of callus was not possible.
Phytochemical analysis of in vitro and in vivo plant in regard of Aloin
is in progress.
0.1 % Hgcl2 and 10 min duration appears to be best for
sterilization
A combination of MS with NAA + BAP(0.5mg/l) best for
establishment
MS with Kn (2mg/l) best for multiplication
MS with BA(0.5 mg/l)+ IAA(0.2 mg/l) best for rooting.
Mitotic index of in vivo plant is 8.09 and in vitro plant is 9.87