Fig. 5: LAL regulated MtLHCB1s in salinity tolerance.

a Transient expression assays of MtLHCB1.4 with LAL indicated by ‘+’ and without LAL indicated by ‘-’ in N. benthamiana. Bar = 20 µm. b The relative GFP fluorescence intensity of 35 S:MtLHCB1.4-GFP with and without LAL. Values are shown by mean ± SD (grey dots). n = 5. Columns labeled with asterisks indicate significant differences from those in 35 S:MtLHCB1.4-GFP without LAL (*P < 0.05, student’s t test). The intensity in 35 S:MtLHCB1.4-GFP without LAL was set at 100%. c The expression level of MtLHCB1.4 with and without LAL by qRT-PCR. L25 was used as the internal control. Error bars represent the SD from three biological replicates (grey dots). Columns labeled with asterisks indicate significant differences from those in 35 S:MtLHCB1.4-GFP without LAL (*P < 0.05, student’s t test). The expression level in 35 S:MtLHCB1.4-GFP without LAL was set at 1.0. d The scheme of LAL and MtLHCB1.1, MtLHCB1.2, MtLHCB1.3 and MtLHCB1.4 genes structure and the complementary sequences to siRNALAL in MtLHCB1s. Empty boxes represent MtLHCB1s, and black boxes represent LAL. Four arrowheads indicate the positions of complementary sequences to siRNALAL. e Complementary sequence of MtLHCB1.4 and siRNALAL. Red vertical line indicates the cleavage site for siRNALAL validated by RLM 5’-RACE. f The expression level of siRNALAL in lal-2 by qRT-PCR. MtU6 was used as the internal control. The expression level in WT was set at 1.0. Error bars represent the SD from three biological replicates (grey dots). Columns labeled with asterisks indicate significant differences from those in WT (*P < 0.05, student’s t test). Complementary sequences of MtLHCB1.1 (g), MtLHCB1.2 (h) and MtLHCB1.3 (i) to siRNALAL. j The expression level of MtLHCB1.1, MtLHCB1.2, MtLHCB1.3 and MtLHCB1.4 in lal-2, lal-2;RNAi-MtLHCB1s#1 and lal-2;RNAi-MtLHCB1s#2 plants by qRT-PCR. MtUBIQUITIN was used as the internal control. The expression level in lal-2 was set at 1.0. Error bars represent the SD from three biological replicates (grey dots). Columns labeled with asterisks indicate significant differences from those in lal-2 (*P < 0.05, student’s t test). k The plant survival rates measured after the four-week treatment of salinity. Error bars represent the SD from three biological replicates (grey dots). Columns labeled with asterisks indicate significant differences from those in lal-2 (*P < 0.05, student’s t test). l Four-week-old plants of lal-2, lal-2; RNAi-MtLHCB1s #1 and lal-2;RNAi-MtLHCB1s#2 plants after a four-week successive exposure to 50, 100, 150 and 200 mM NaCl. Bar = 5 cm. n = 3.