Transcriptional repression of GTL1 under water-deficit stress promotes anthocyanin biosynthesis to enhance drought tolerance.

The transcription factor GT2-LIKE 1 (GTL1) has been implicated in orchestrating a transcriptional network of diverse physiological, biochemical, and developmental processes. In response to water-limiting conditions, GTL1 is a negative regulator of stomatal development, but its potential rolein other water-deficit responses is unknown. We hypothesized that GTL1 regulates transcriptome changes associated with drought tolerance over leaf developmental stages. To test the hypothesis, gene expression was profiled by RNA-seq analysis in emerging and expanding leaves of wild-type and a drought-tolerant gtl1-4 knockout mutant under well-watered and water-deficit conditions. Our comparative analysis of genotype-treatment combinations within leaf developmental age identified 459 and 1073 differentially expressed genes in emerging and expanding leaves, respectively, as water-deficit responsive GTL1-regulated genes. Transcriptional profiling identified a potential role of GTL1 in two important pathways previously linked to drought tolerance: flavonoid and polyamine biosynthesis. In expanding leaves, negative regulation of GTL1 under water-deficit conditions promotes biosynthesis of flavonoids and anthocyanins that may contribute to drought tolerance. Quantification of polyamines did not support a role for GTL1 in these drought-responsive pathways, but this is likely due to the complex nature of polyamine synthesis and turnover. Our global transcriptome analysis suggests that transcriptional repression of GTL1 by water deficit allows plants to activate diverse pathways that collectively contribute to drought tolerance.

Different Leaf Anatomical Responses to Water Deficit in Maize and Soybean.

The stomata on leaf surfaces control gas exchange and water loss, closing during dry periods to conserve water. The distribution and size of stomatal complexes is determined by epidermal cell differentiation and expansion during leaf growth. Regulation of these processes in response to water deficit may result in stomatal anatomical plasticity as part of the plant acclimation to drought. We quantified the leaf anatomical plasticity under water-deficit conditions in maize and soybean over two experiments. Both species produced smaller leaves in response to the water deficit, partly due to the reductions in the stomata and pavement cell size, although this response was greater in soybean, which also produced thicker leaves under severe stress, whereas the maize leaf thickness did not change. The stomata and pavement cells were smaller with the reduced water availability in both species, resulting in higher stomatal densities. Stomatal development (measured as stomatal index, SI) was suppressed in both species at the lowest water availability, but to a greater extent in maize than in soybean. The result of these responses is that in maize leaves, the stomatal area fraction (fgc) was consistently reduced in the plants grown under severe but not moderate water deficit, whereas the fgc did not decrease in the water-stressed soybean leaves. The water deficit resulted in the reduced expression of one of two (maize) or three (soybean) SPEECHLESS orthologs, and the expression patterns were correlated with SI. The vein density (VD) increased in both species in response to the water deficit, although the effect was greater in soybean. This study establishes a mechanism of stomatal development plasticity that can be applied to other species and genotypes to develop or investigate stomatal development plasticity.