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. 2021 Apr 20;12:644437. doi: 10.3389/fpls.2021.644437

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
GR GR+ 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.