Table 1.
Mutation frequencies for six RBCS genes in the radish mutant population revealed by two-step TILLING
| Gene ID (EMS concentration) | PCR product screened (bp) | Primer length (bp) | No. of screened lines | Total screened length (kb) | No. of TILLING positive | No. of mutations obtained | Mutation density (kb−1) | ||
|---|---|---|---|---|---|---|---|---|---|
|
| |||||||||
| Missense | Intronica | Other | |||||||
| RsRBCS1A | 1224 | 57 | 955 | 1114 | 1 | 0 | 0 | 1 (deletion) | 0 |
| RsRBCS2A | 1082 | 62 | 955 | 974 | 1 | 1 | 0 | 0 | 1/974 |
| RsRBCS4A | 1180 | 62 | 955 | 1067 | 0 | 0 | 0 | 0 | 0 |
| RsRBCS5A | 1166 | 70 | 955 | 1046 | 2 | 0 | 2 | 0 | 1/523 |
| RsRBCS3A | 1172 | 72 | 955 | 1050 | 4 | 0 | 0 | 4 (natural variation) | 0 |
| RsRBCS2B | 1105 | 72 | 955 | 986 | 5 | 3 | 0 | 2 (deletion) | 1/328 |
|
| |||||||||
| (0.25% EMS) | 6929 | 395 | 222 | 1450 | – | 1 | 2 | – | 1/483 |
|
| |||||||||
| (0.50% EMS) | 6929 | 395 | 270 | 1764 | – | 3 | 0 | – | 1/588 |
|
| |||||||||
| Total | 6929 | 395 | 5730 | 6239 | 13 | 4 | 2 | 7 | 1/1039 |
The mutation frequency was calculated as [total number of identified mutations]/[(PCR product size screened – total primer size) × (total number of screened lines)] (Lai et al. 2012). The average mutation frequency was estimated to be one mutation per 1039 kb.
a
A nucleotide was substituted out of an exon region.