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| Table 1. The effect of different factors on the efficiency* of the transient transformation of Habanero pepper (Capsicum chinense) leaf explants. Data shown are Means ± standard error of three independent experiments. The number of leaves utilized in each experiment was 60. |
Factors evaluated |
Treatment of each factor |
Number of Gus-positive leaf explants (percentage ± standard deviation) |
ANOVA |
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Constructs |
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pCAMBIA2301 |
pCAMex |
pCAMex:PR10 |
pCAMex:Est |
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Co-cultivation conditions at dark (days, temperature) |
3, 20ºC |
0 |
0 |
0 |
0 |
|
2, 20ºC |
0 |
0 |
0 |
0 |
||
1, 20ºC |
0 |
0 |
0 |
0 |
P < 0.05 |
|
3, 28ºC ** |
11 (18.8 ± 3.0) |
8 (13.8 ± 3.2) |
9 (14.9 ± 4.0) |
7 (11.0 ± 3.5) |
||
2, 28ºC ** |
23 (38.9 ± 0.5) |
23 (38.4 ± 2.0) |
21 (34.9 ± 2.0) |
22 (36.0 ± 2.5) |
||
1, 28ºC ** |
0a |
0 |
0 |
0 |
||
Transformation method |
Vacuum (V) |
0 |
0 |
0 |
0 |
|
Infiltration (I) |
8 (12.7 ± 2.5) |
6 (10.5 ± 3.0) |
6 (10.5 ± 2.5) |
5 (8.8 ± 2.5) |
||
Micro-injection (M) |
03 (4.9 ± 2.6) |
05 (8.3 ± 3.0) |
02 (3.8 ± 2.0) |
04 (6.6 ± 3.6) |
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V+I |
12 (20.0 ± 3.0) |
10 (16.6 ± 4.0) |
8 (12.7 ± 4.1) |
6 (10.5 ± 4.0) |
P < 0.05 |
|
V+M |
04 (7.2 ± 2.0) |
06 (9.4 ± 2.5) |
03 (5.5 ± 3.0) |
05 (8.8 ± 3.0) |
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I+M |
14(23.8 ± 3.0) |
15 (24.4 ± 5.0) |
16 (27.2 ± 1.5) |
16 (26.6 ± 4.5) |
||
M+I+V |
28 (47.2 ± 2.0) |
24 (40.5 ± 2.5) |
23 (38.8 ± 6.8) |
24 (40.5 ± 3.5) |
||
Pre-incubation period (hours) |
0 |
28 (46.0 ± 3.0) |
24 (40.5 ± 2.5) |
23 (40.5 ± 3.0) |
25 (42.2 ± 5.0) |
|
1 |
42 (69.4 ± 5.0) |
40 (67.1 ± 2.5) |
43 (71.6 ± 5.5) |
40 (67.1 ± 2.5) |
||
6 |
33 (54.9 ± 4.3) |
36 (59.4 ± 2.5) |
30 (49.9 ± 3.6) |
34 (56.6 ± 4.0) |
P < 0.05 |
|
12 |
40 (66.6 ± 2.0) |
33 (54.4 ± 4.7) |
32 (53.8 ± 2.0) |
35 (58.3 ± 3.6) |
||
24 |
42 (69.9 ± 5.5) |
39 (65.5 ± 2.3) |
41 (67.7 ± 4.0) |
34 (56.6 ± 4.5) |
||
*Transient transformation efficiency
was evaluated based on the number and percentages of GUS positive (GUS+) leaf
explants produced. |
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