Table 1
Effect of embryo age on number of seeds per fruit and seeds with and without embryos in ‘Kagzi’ (acid lime) × ‘Konkan Seedless’ (lemon) crosses
| Embryo age (DAP)* | No. of seeds per fruit | No. of seeds obtained | No. of seeds containing functional embryos | Seeds with functional embryos (%) |
|---|---|---|---|---|
| 80–90 | 22 | 88 | 77 | 88 |
| 110–120 | 32 | 96 | 76 | 79 |
| 130–140 | 27 | 53 | 46 | 90 |
Figure 1
Fruit obtained from cross of ‘Kagzi’ acid lime × ‘Konkan Seedless’ lemon harvested at 80–90 DAP (A), seeds (B), embryo (C), hybrid seedling of embryo cultured on M2 medium (D), hybrid seedling obtained from embryo of 110–120 DAP cultured in M3 medium (E) and hybrid seedlings obtained from embryo of 130–140 DAP cultured in M3 medium (F)
Table 2
Effect of embryo age and media composition on embryo germination, days taken for germination and survival of ‘Kagzi’ (acid lime) × ‘Konkan Seedless’ (lemon) seedlings
| Medium composition | No. of embryo cultured | No. of germinated embryos | Days to initiate germination | Days to complete germination | Germination (%) | Seedling survival at 60 DAI** (%) |
|---|---|---|---|---|---|---|
| Embryo age (80–90 DAP)* | ||||||
| M1 | 25 | 19 | 3 | 8 | 76 cd | 84 a |
| M2 | 25 | 24 | 3 | 8 | 96 a | 71 cb |
| M3 | 27 | 22 | 3 | 8 | 81 cb | 73 cb |
| Embryo age (110–120 DAP) | ||||||
| M1 | 26 | 16 | 2 | 9 | 62 e | 75 b |
| M2 | 26 | 21 | 2 | 11 | 81 cb | 67 c |
| M3 | 26 | 26 | 2 | 7 | 100 a | 73 cb |
| Embryo age (130–140 DAP) | ||||||
| M1 | 16 | 12 | 2 | 7 | 75 d | 67 c |
| M2 | 15 | 13 | 2 | 7 | 87 b | 69 cb |
| M3 | 15 | 15 | 2 | 7 | 100 a | 53 d |
Table 3
Effect of embryo age and media composition on plantlets growth obtained from ‘Kagzi’ (acid lime) × ‘Konkan Seedless’ (lemon) crosses at 30 and 60 DAI
| Medium composition | Root length (mm) | Shoot length (mm) | Leaves per plantlet | ||||||
|---|---|---|---|---|---|---|---|---|---|
| 30 DAI | 60 DAI | Increase over 30 DAI (%) | 30 DAI | 60 DAI | Increase over 30 DAI (%) | 30 DAI | 60 DAI | Increase over 30 DAI (%) | |
| Embryo age (80–900 DAP) | |||||||||
| M1 | 2.1 b | 4.3 bc | 102 a | 2.8 ab | 3.9 cd | 37 b | 2.0 ab | 3.6 a | 80 d |
| M2 | 2.2 b | 4.3 bc | 97 b | 2.1 bc | 4.8 ab | 131 a | 1.2 b | 3.4 a | 183 a |
| M3 | 2.5 b | 4.9 a | 100 ab | 3.0 a | 5.3 a | 76 b | 1.6 ab | 3.2 a | 100 c |
| Means | 100 | 81 | 121 | ||||||
| Embryo age (110–120 DAP) | |||||||||
| M1 | 2.5 b | 4.7 ab | 88 b | 2.4 abc | 4.2 bc | 76 b | 1.6 ab | 4.0 a | 150 cd |
| M2 | 2.7 ab | 4.2 bc | 57 c | 2.5 ab | 4.2 bc | 687 b | 2.0 ab | 3.8 a | 90 abc |
| M3 | 3.2 a | 4.0 cd | 25 d | 2.2 abc | 3.7 cd | 66 b | 2.6 a | 3.6 a | 38 e |
| Means | 57 | 70 | 93 | ||||||
| Embryo age (130–140 DAP) | |||||||||
| M1 | 1.3 c | 2.5 e | 914 b | 1.5 c | 2.6 e | 73 b | 2.6 a | 3.6 a | 38 ae |
| M2 | 2.5 b | 3.5 d | 40 cd | 2.3 abc | 3.6 cd | 54 b | 2.2 ab | 1.6 a | −27 f |
| M3 | 2.7 ab | 3.8 cd | 40 cd | 2.7 ab | 3.4 d | 27 c | 2.6 a | 2.0 a | −23 f |
| Means | 57 | 51 | −4 | ||||||
| LSD (p ≤ 0.05) | |||||||||
| Embryo age (EA) | 0.21 | 0.21 | 8.03 | 0.31 | 0.24 | 9.50 | 0.41 | 0.88 | 4.11 |
| Media (M) | 0.21 | 0.21 | 8.03 | 0.31 | 0.24 | 9.50 | 0.41 | 0.88 | 4.11 |
| EA × M | 0.59 | 0.59 | 22.69 | 0.89 | 0.70 | 26.84 | 1.18 | NS | 11.61 |
Figure 2
Agarose gel profiles of DNA amplification products obtained with SSR primer CCSM-4 of in vitro raised plantlets obtained from 80–90 DAP (A), 110–120 DAP (B), 130–140 DAP (C), and seedlings raised in vivo at maturity (180–190 DAP). In Fig. 2A to 2C, the lanes are as follows: P1, female parent; H1–H12 hybrids of ‘Kagzi’ acid lime × ‘Konkan Seedless’ lemon; P2, male parent. In Fig. 2D, S1–S9 seedlings raised in vivo from crosses of ‘Kagzi’ acid lime × ‘Konkan Seedless’ lemon, P1 female parent and P2 male parent