Table 1.
Complementation analysis of symbiosis-defective phenotypes using the pEpi expression system
| Lotus line | Construct | Root hair phenotype (%) | Nodulation phenotype | Total number of plants | ||||||
|---|---|---|---|---|---|---|---|---|---|---|
| No colonization | Micro-colony | Short ITs | ITs | (%) | Nodules per nodulated plant (mean ± SE) | |||||
| Nod− | Bump | Nod | ||||||||
| Wild-type | Empty | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 17.5 ± 1.1 | 31 |
| Wild-type | pEpi::GUS+ | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 22.7 ± 1.8 | 35 |
| Wild-type | p35S::GUS+ | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 20.9 ± 1.5 | 30 |
| nfr1–4 | Empty | 100 | 0 | 0 | 0 | 100 | 0 | 0 | – | 20 |
| nfr1–4 | pEpi::NFR1 | 0.9 | 34.9 | 44.9 | 19.3 | 7.3 | 67.9 | 24.8 | 1.1 ± 0.1 | 109 |
| nfr1–4 | p35S::NFR1 | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 18.3 ± 0.8 | 65 |
| nfr5–2 | Empty | 100 | 0 | 0 | 0 | 100 | 0 | 0 | – | 51 |
| nfr5–2 | pEpi::NFR5 | 0 | 12.2 | 50 | 37.8 | 4.9 | 18.3 | 76.8 | 6.0 ± 0.6 | 82 |
| nfr5–2 | p35S::NFR5 | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 13.5 ± 1.0 | 44 |
| nup85–2 | Empty | 100 | 0 | 0 | 0 | 100 | 0 | 0 | – | 59 |
| nup85–2 | pEpi::NUP85 | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 15.3 ± 0.6 | 102 |
| nup85–2 | p35S::NUP85 | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 18.0 ± 0.8 | 93 |
| nup133–3 | Empty | 100 | 0 | 0 | 0 | 100 | 0 | 0 | – | 67 |
| nup133–3 | pEpi::NUP133 | 0 | 0 | 0 | 100 | 2.3 | 0 | 97.7 | 9.8 ± 0.6 | 87 |
| nup133–3 | p35S::NUP133 | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 11.2 ± 0.7 | 60 |
| castor–4 | Empty | 100 | 0 | 0 | 0 | 100 | 0 | 0 | – | 21 |
| castor–4 | pEpi::CASTOR | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 23.0 ± 1.8 | 54 |
| castor–4 | p35S::CASTOR | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 19.4 ± 1.2 | 48 |
| pollux–3 | Empty | 100 | 0 | 0 | 0 | 100 | 0 | 0 | – | 50 |
| pollux–3 | pEpi::POLLUX | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 20.5 ± 1.3 | 55 |
| pollux–3 | p35S::POLLUX | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 18.7 ± 1.3 | 64 |
| ccamk–3 | Empty | 100 | 0 | 0 | 0 | 100 | 0 | 0 | – | 88 |
| ccamk–3 | pEpi::CCaMK | 0 | 3.7 | 0.9 | 95.4 | 58.3 | 13.9 | 27.8 | 2.2 ± 0.4 | 108 |
| ccamk–3 | p35S::CCaMK | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 11.5 ± 0.9 | 55 |
| cyclops–3 | Empty | 0 | 86.1 | 13.9 | 0 | 0 | 100 | 0 | – | 36 |
| cyclops–3 | pEpi::CYCLOPS | 0 | 0 | 0 | 100 | 0 | 100 | 0 | – | 90 |
| cyclops–3 | p35S::CYCLOPS | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 12.5 ± 1.0 | 78 |
| nsp1–1 | Empty | 100 | 0 | 0 | 0 | 100 | 0 | 0 | – | 61 |
| nsp1–1 | pEpi::NSP1 | 0 | 0 | 0 | 100 | 58.1 | 17.7 | 24.2 | 1.7 ± 0.4 | 62 |
| nsp1–1 | p35S::NSP1 | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 16.1 ± 1.0 | 67 |
| nsp2–1 | Empty | 100 | 0 | 0 | 0 | 100 | 0 | 0 | – | 61 |
| nsp2–1 | pEpi::NSP2 | 5.3 | 11.6 | 8.4 | 74.7 | 96.8 | 2.1 | 1.1 | 1.0 | 95 |
| nsp2–1 | p35S::NSP2 | 0 | 0 | 0 | 100 | 0 | 0 | 100 | 16.4 ± 1.9 | 39 |
The numbers indicate the percentage of plants that showed infection events, i.e. formation of micro-colonies or infection threads (ITs) in the epidermis, and nodulation events, i.e. formation of empty nodules (bump) or infected nodules (Nod) in the cortex, 4 weeks after inoculation of DsRed-labeled Mesorhizobium loti. Data were compiled from more than two independent experiments. Wild-type, Gifu B–129; Empty, empty vector; pEpi, Epi308 promoter; p35S, CaMV 35S promoter.