TABLE 2.
Fluorescence assays and PCR analysis of T1 transgenic rice plants transformed with pBDL23.
| Line no. | GFP and mCherry fluorescence assays |
PCR analysis of non-fluorescent plants |
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| G–R– | G–R+ | G+R– | G+R+ | GFP: χ2(3:1) | GFP: χ2(15:1) | GFP: χ2(63:1) | mCherry: χ2(3:1) | mCherry: χ2(15:1) | mCherry: χ2(63:1) | Plants | Pi21 RNAi+ | GFP+ | HPT+ | AcTPase+ | MF | false MF | |
| 14YD262-1 | 0 | 0 | 14 | 29 | 13.03b | 1.90 | 0.04 | 0.94 | 46.40b | 248.82b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD263-1 | 29 | 1 | 0 | 58 | 3.41 | 111.71b | 584.42b | 2.56 | 102.59b | 543.60b | 1 | 0 | 0 | 0 | 0 | 1 | 0 |
| 14YD264-2 | 24 | 0 | 0 | 71 | 0.00 | 55.41b | 331.71b | 0.00 | 55.41b | 331.71b | 1 | 0 | 0 | 0 | 0 | 1 | 0 |
| 14YD265-1 | 23 | 0 | 1 | 75 | 0.08 | 45.87b | 288.32b | 0.00 | 51.67b | 316.50b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD266-3 | 7 | 0 | 0 | 23 | 0.00 | 12.17b | 78.83b | 0.00 | 12.17b | 78.83b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD268-1 | 6 | 0 | 15 | 60 | 12.45b | 0.04 | 14.39b | 0.00 | 50.21b | 296.95b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD269-1 | 25 | 0 | 0 | 67 | 0.13 | 65.22b | 375.88b | 0.13 | 65.22b | 375.88b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD270-1 | 25 | 0 | 0 | 51 | 2.12 | 87.59b | 464.93b | 2.12 | 87.59b | 464.93b | 13 | 0 | 0 | 0 | 0 | 13 | 0 |
| 14YD271-3 | 32 | 0 | 0 | 59 | 4.49a | 124.96b | 646.37b | 4.49a | 124.96b | 646.37b | 8 | 0 | 0 | 0 | 0 | 8 | 0 |
| 14YD272-1 | 21 | 0 | 61 | 0 | 0.00 | 49.20b | 292.86b | 242.02b | 1214.05b | 5102.20b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD273-2 | 29 | 0 | 0 | 35 | 13.02b | 160.07b | 768.25b | 13.02b | 160.07b | 768.25b | 14 | 0 | 0 | 0 | 0 | 14 | 0 |
| 14YD275-1 | 24 | 0 | 0 | 71 | 0.00 | 55.41b | 331.71b | 0.00 | 55.41b | 331.71b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD276-1 | 19 | 0 | 0 | 60 | 0.00 | 39.74b | 245.33b | 0.00 | 39.74b | 245.33b | 7 | 0 | 0 | 0 | 0 | 7 | 0 |
| 14YD278-1 | 25 | 0 | 0 | 61 | 0.56 | 72.59b | 405.38b | 0.56 | 72.59b | 405.38b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD279-3 | 26 | 0 | 0 | 68 | 0.23 | 69.93b | 399.43b | 0.23 | 69.93b | 399.43b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD280-2 | 6 | 9 | 17 | 66 | 4.41a | 12.21b | 111.58b | 0.05 | 46.70b | 291.70b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD289-1 | 22 | 0 | 0 | 43 | 2.26 | 79.84b | 419.71b | 2.26 | 79.84b | 419.71b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD294-1 | 25 | 0 | 0 | 48 | 2.85 | 92.93b | 485.98b | 2.85 | 92.93b | 485.98b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD303-1 | 19 | 0 | 0 | 48 | 0.24 | 52.18b | 295.59b | 0.24 | 52.18b | 295.59b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD305-2 | 27 | 0 | 0 | 66 | 0.61 | 78.54b | 438.57b | 0.61 | 78.54b | 438.57b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD306-1 | 13 | 0 | 0 | 50 | 0.43 | 19.86b | 136.85b | 0.43 | 19.86b | 136.85b | 1 | 0 | 0 | 0 | 0 | 1 | 0 |
| 14YD307-1 | 14 | 0 | 0 | 21 | 3.44 | 62.40b | 311.67b | 3.44 | 62.40b | 311.67b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD309-1 | 5 | 0 | 73 | 0 | 13.40b | 0.03 | 8.97b | 230.02b | 1154.05b | 4850.21b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD310-1 | 18 | 0 | 0 | 58 | 0.02 | 36.51b | 227.64b | 0.02 | 36.51b | 227.64b | 3 | 0 | 0 | 0 | 0 | 3 | 0 |
| 14YD311-1 | 3 | 9 | 9 | 39 | 0.56 | 17.08b | 120.89b | 0.56 | 17.08b | 120.89b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD312-1 | 18 | 0 | 42 | 33 | 1.29 | 25.07b | 180.02b | 75.36b | 528.95b | 2355.56b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD313-1 | 23 | 0 | 0 | 48 | 1.69 | 78.42b | 418.99b | 1.69 | 78.42b | 418.99b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD315-1 | 25 | 0 | 0 | 38 | 6.48a | 114.54b | 570.68b | 6.48a | 114.54b | 570.68b | 1 | 0 | 0 | 0 | 0 | 1 | 0 |
| 14YD316-1 | 25 | 0 | 0 | 62 | 0.46 | 71.28b | 400.18b | 0.46 | 71.28b | 400.18b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD318-1 | 19 | 0 | 70 | 1 | 0.53 | 31.43b | 211.08b | 258.13b | 1302.43b | 5479.63b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD319-1 | 19 | 0 | 0 | 63 | 0.07 | 37.23b | 235.08b | 0.07 | 37.23b | 235.08b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD320-1 | 6 | 13 | 0 | 70 | 0.45 | 32.10b | 213.84b | 14.87b | 0.00 | 12.34b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD321-1 | 29 | 0 | 0 | 38 | 10.99b | 150.57b | 731.35b | 10.99b | 150.57b | 731.35b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD323-1 | 20 | 0 | 0 | 54 | 0.07 | 51.03b | 295.64b | 0.07 | 51.03b | 295.64b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD325-2 | 5 | 14 | 15 | 59 | 0.81 | 29.54b | 203.15b | 0.43 | 34.38b | 227.69b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD326-2 | 20 | 0 | 0 | 75 | 0.59 | 33.04b | 222.12b | 0.59 | 33.04b | 222.12b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD328-1 | 11 | 0 | 0 | 36 | 0.01 | 20.77b | 131.92b | 0.01 | 20.77b | 131.92b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD330-1 | 4 | 0 | 0 | 68 | 13.50b | 0.00 | 5.09a | 13.50b | 0.00 | 5.09a | 1 | 0 | 0 | 0 | 0 | 1 | 0 |
| 14YD335-1 | 27 | 0 | 0 | 53 | 2.82 | 98.61b | 518.15b | 2.82 | 98.61b | 518.15b | 16 | 0 | 0 | 0 | 0 | 16 | 0 |
| 14YD343-1 | 25 | 0 | 52 | 3 | 1.35 | 81.12b | 439.31b | 212.82b | 1090.61b | 4601.96b | 2 | 0 | 0 | 0 | 0 | 2 | 0 |
| 14YD344-2 | 32 | 0 | 50 | 0 | 7.87b | 144.78b | 724.03b | 242.02b | 1214.05b | 5102.20b | 0 | ND | ND | ND | ND | 0 | 0 |
| 14YD347-1 | 16 | 0 | 0 | 50 | 0.00 | 33.46b | 206.22b | 0.00 | 33.46b | 206.22b | 11 | 0 | 0 | 0 | 0 | 11 | 0 |
The ratio of the number of GFP- or mCherry-fluorescent T1 segregants to the number of non-fluorescent siblings in each T1 line was analyzed by a chi-squared test. Mendelian segregation patterns (3:1, 15:1, or 63:1) were accepted at the 0.05 and 0.01 probability levels when the calculated chi-square values are less than the theoretical values of 3.84 and 6.64, respectively. PCR primers for amplification of Pi21-RNAi, GFP, mCherry, HPT, and AcTPase were listed in Supplementary Table 1. G, GFP; R, mCherry; +, positive; −, negative; MF, marker-free Ds(Pi21-RNAi) plants; false MF, non-fluorescent Ds(Pi21-RNAi) plants containing residual T-DNA sequences; a, significant level (0.01 < P < 0.05); b, most significant level (P < 0.01); ND, not determined.