FIGURE 3.

Water deficit treatments in white clover induce Tr-NCED1 transcript upregulation and proline accumulation. Accumulation of L-Proline in wild type plant under PS and NPS treatments (A) and transcription of Tr-NCED1 (B), as indicated, in the first fully expanded (FFE) leaves in response to NPS and PS treatments against the moisture content of the media, as indicated. For L-proline accumulation, each value represents the mean from two biological replications each consisting of pooled tissues from four stolons. For Tr-NCED1, relative transcript abundance was determined by qRT-PCR using at least three biological replicates, with pooled tissues collected from at least three stolons comprising a biological replicate, and was normalized using two internal reference genes, Tr-β-actin and Tr-GAPDH. The moisture content represent the mean value of the biological replicates and each data point represents mean value ± SE, of the biological replicates. Statistical analysis was performed using Student’s t-test. ^∗Indicate statistically significant (P < 0.05) differential expression in comparison with initial moisture content for NPS and PS treatment, respectively, and ‘^†’ indicates statistically significant (P < 0.05) expression in the PS treatment in comparison with the NPS treatments using Student’s t-test.