Table 3. Cell death induction activity of MtLYK3-sYFP2 mutated variants in Nicotiana leaves.
MtLYK3-sYFP2 construct | Auto-phosphorylation activity* | Nodulation activity** | Cell death induction | |
Co-expression with MtNFP-mCherry | Separate expression | |||
WT | + | + | 28/30 | 0/12 |
P87S (Mtlyk3-3) | Not applicable | - | 15/15 | 0/9 |
T319A | – | – | 0/11 | 0/9 |
G334E (Mtlyk3-1) | – | – | 0/20 | 0/9 |
K349A | – | – | 0/16 | 0/9 |
E362A | – | – | 0/15 | 0/9 |
D441A | – | – | 0/16 | 0/9 |
T475A | – | – | 0/13 | 0/9 |
T480A | – | −(0/24) | 0/18 | 0/9 |
T285A/S286A/T300A | + | Reduced with T300A | 15/16 | 0/9 |
T433A | + | Reduced | Reduced 7/20 | 0/10 |
K464A | Reduced | Reduced (7/18) | 0/12# | 0/9 |
S471A | + | Reduced | 9/11 | 0/9 |
T472A | + | Reduced | Reduced 5/11 | 0/9 |
T512A | Reduced | – | Reduced 12/20 | 0/9 |
-see [20], except for the T480A (Fig. S2), **-see [20], except for the P87S [12], K464A and T480A (this study; number of plants nodulated/number of plants tested).
The designated constructs were expressed alone or co-expressed with MtNFP-mCherry construct in Nicotiana leaves. Macroscopic symptoms of cell death were scored 48 hai: only infiltrations that resulted in confluent death of (nearly) the entire infiltrated region were scored and are presented as a fraction of total infiltrations performed. # - despite the lack of pronounced macroscopic symptoms, the co-expression of MtLYK3[K464A]-sYFP2 and MtNFP-mCherry constructs resulted in increased staining with Evans blue in the entire infiltrated region.