Professional Documents
Culture Documents
Robert Thangjam
Hopkins, 1994
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HOOC NH2
HCHO
Cysteine Thioproline
1.1 1.0 0.9
Formaldehyde
Curc umin + 60% methanolic ext ract Curcumin + water ext ract Curcumin
Absorbance at 420 nm
0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0 50 100 150 200 250
MULTIPURPOSE TREE
Protein content - kernel and the pods are 29% and 13-19% respectively. essential amino acid patterns is comparable to FAO/WHO/UNU (1985) amino acid requirement for preschoolers Good source of fuel wood burns slowly & completely Industrial purposes easy to saw & good finish, bark (615% tannins) for tanning & dye Oil fruit contain 19.6% oil Agro-forestry tree nitrogen fixing; produce 10-12 cubic meter of biomass in 10-15 years Ornamental tree planted as ornamental avenue tree
AS TRADITIONAL MEDICINE
.
bark or skin of fruits dysentery, diarrhoea & piles
seeds as well as tender pods - stomach disorders & regulate liver function
bark and leaves - lotions for skin diseases, ulcers & rheumatic affected parts skin of fresh - applied on scabbies
B
D
A C
D F G E
A C E
Genetic characterization
Sl. No. 1 2 3 4 5
Kangchup
Kangpokpi
Narankonjin
Kangpokpi (KP)
32.43
1.32
1.25
1.14
0.13
0.07
0.19
0.11
Narankonjin (NA) 18.92 1.18 Overall Gst (genetic differentiation) = 0.29 Overall Nm (gene flow from Gst) = 1.23
Plant material : Seeds from an 18 year old elite plus tree Explant: Cotyledonary node from 10 day old in vitro germinated plant Media: MS, WPM & Gamborg Hormones: NAA & BAP (singly or in combinations)
No. of roots
0.00 0.00 1.60 0.16 2.70 0.15 2.70 0.15 2.50 0.16 3.20 0.13 8.70 0.33
Genetic transformation
Bacterial strain
Agrobacterium tumefaciens strain EHA105 harboring the binary vector pCAMBIA2301 containing a nptII (neomycin phosphotransferase) as selectable marker and the glucuronidase (GUS) gene (uidA) with an intron as the
reporter gene within the T-DNA region driven by CaMV 35S promoter was used for the transformation.
Kanamycin Selection
Shoot induction medium (SM): MSB + 0.5 mg/l NAA + 2.5 mg/l BAP) containing different concentrations of kanamycin (0, 50, 75 and 100 mg/l) Root induction medium (RM): MSB + 2.5 mg/l IBA supplemented with different concentrations of kanamycin (0, 1, 5, 7.5 and 10 mg/l) Influence of cefotaxime on the shoot induction and subsequent growth was also checked by culturing explants on SM and RM medium containing 500 mg/l cefotaxime. Concentration (mg/l) 0 50 75 Percentage (%) Shoot regeneration per Average number survival explants explant (%) of shoots/explant SE 6.07 0.28 100 45/45 (100) 77.14 11.11 27/45 (77.14) 5/45 (11.11) 1.4 0.29 0.33 0.12
100
0.00
0/45 (0.00)
0.00 0.00
GUS analysis
(Minutes) Control 15 30 45 60
60
Percentage (%) of GUS+ plants (Mean SE) 0.00 21.94 1.00 54.72 1.21 43.61 0.73 35.64 2.18
% of GUS+ Plants
50
40
30
20
10
Period of Inoculation
3. Concentration of acetosyringone
Acetosyringone concentration (M) Control 0 50 100 200 Percentage (%) of GUS+ plants (Mean SE) 0.00 0.00 36.39 0.73 47.22 1.47 57.78 2.65 64.44 1.54
540 bp
Southern analysis
Southern blot analysis of genomic DNA of transformed and nontransformed (control) plants. Lane 1 - Hind III digested Lamda DNA , lane 2 - DNA from untransformed plant (control), lanes 3,4 - transformed plants, lane 5 plasmid DNA (pCAMBIA2301) of size 11.6 kb
Summary of the transformation of cotyledonary node explants of P. timoriana cocultured with Agrobacterium tumefaciens strain EHA105 harbouring binary vector pCAMBIA 2301
Total no. of explants inoculated with Agrobacterium Total no. of shoots Total no. of shoots Percentage of Percentage of regenerated on rooted on plants established plants positive for selection mediuma selection mediumb in soil nptII by PCR (no. of explants showing multiple shoots)
825 (125)
a
275
225
60 (135/225)
33.33 (5/15)
Selection medium for shoot regeneration: MSB + 0.5 mg/l NAA + 2.5 mg/l BAP + kanamycin (75 mg/l) + cefotaxime (500 mg/l) b Selection medium for root induction: MSB + 2.0 mg/l IBA + kanamycin (7.5 mg/l) + cefotaxime (500 mg/l)
Summary
genetic status of tree bean in Manipur established
gene pool status at the population level worked out
Thank you