The initiation of stomata, microscopic valves in the epidermis of higher plants that control of gas exchange, requires a co-ordinated sequence of asymmetric and symmetric divisions, which is under tight environmental and developmental control. Arabidopsis leaves grown under elevated photosynthetic photon flux density have a higher density of stomata. STOMAGEN encodes an epidermal patterning factor produced in the mesophyll, and our observations indicated that elevated photosynthetic irradiation stimulates STOMAGEN expression. Our analysis of gain and loss of function of STOMAGEN further detailed its function as a positive regulator of stomatal formation on both sides of the leaf, not only in terms of stomatal density across the leaf surface but also in terms of their stomatal index. STOMAGEN function was rate limiting for the light response of the stomatal lineage in the adaxial epidermis. Mutants in pathways that regulate stomatal spacing in the epidermis and have elevated stomatal density, such as stomatal density and distribution (sdd1) and too many mouth alleles, displayed elevated STOMAGEN expression, suggesting that STOMAGEN is either under the direct control of these pathways or is indirectly affected by stomatal patterning, suggestive of a feedback mechanism. These observations support a model in which changes in levels of light irradiation are perceived in the mesophyll and control the production of stomata in the epidermis by mesophyll-produced STOMAGEN, and whereby, conversely, stomatal patterning, either directly or indirectly, influences STOMAGEN levels.
The aim of this study was to select the best arrangements of MP–PCR (microsatellite-primed PCR) for routine large-scale fingerprinting of flax cultivars. We found optimum PCR conditions for the application of five previously published primers (PCT1–PCT5) to flax cultivar fingerprinting. We modified to optimum MP–PCR which was targeted to flax tetrameric [GATA] microsatellite loci specified by primer PCT6. We found that after a reamplification PCR step was involved we were able to generate highly discriminating fingerprinting patterns, which distinguished all eight flax cultivars individually. In particular primers 3PCT1 and 3PCT2 were promising for future large-scale fingerprinting due to the production of most polymorphic bands. Increasing annealing temperature within a temperature profile helped to generate new polymorphisms within flax microsatellite patterns especially with primer 3PCT2. Using this primer we succeeded in generating new polymorphic bands after increasing annealing temperature from 55 °C to 60 °C, and to 65 °C. A cluster analysis of flax cultivars was performed based on microsatellite data. The core group of eight flax cultivars was clustered into two homogeneous subclusters. A lower level of cultivar clustering within subclusters was not detected.
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