Classical arabinogalactan proteins (AGPs) are an abundant class of cell surface proteoglycans widely distributed in flowering plants. We have used a combination of enhancer detection tagging and RNA interference (RNAi)–induced posttrancriptional silencing to demonstrate that AGP18, a gene encoding a classical arabinogalactan protein, is essential for female gametogenesis in Arabidopsis thaliana. AGP18 is expressed in cells that spatially and temporally define the sporophytic to gametophytic transition and during early stages of seed development. More than 75% of the T1 transformants resulted in T2 lines showing reduced seed set during at least three consecutive generations but no additional developmental defects. AGP18-silenced T2 lines showed reduced AGP18 transcript levels in female reproductive organs, the presence of 21-bp RNA fragments specific to the AGP18 gene, and the absence of in situ AGP18 mRNA localization in developing ovules. Reciprocal crosses to wild-type plants indicate that the defect is female specific. The genetic and molecular analysis of AGP18-silenced plants containing a single T-DNA RNAi insertion suggests that posttranscriptional silencing of AGP18 is acting both at the sporophytic and gametophytic levels. A cytological analysis of all defective AGP18-RNAi lines, combined with the analysis of molecular markers acting at key stages of female gametogenesis, showed that the functional megaspore fails to enlarge and mitotically divide, indicating that AGP18 is essential to initiate female gametogenesis in Arabidopsis. Our results assign a specific function in plant development to a gene encoding a classical AGP.
16 November 2016 in Journal of Food Science and Technology
Mucilage extracted from the parenchymatous and chlorenchymatous tissues of Opuntia robusta were obtained using water or ethanol as the extraction solvent. The changes in the different tissues by using different extraction solvents were evaluated via scanning electron microscopy (SEM) and Fourier transform infrared (FT-IR) and Raman spectroscopy; in addition, the effect of mucilage coating on the various quality characteristics of tomato (Lycopersicum sculentum) was evaluated. The SEM results showed that the mucilage extracted from the parenchyma had a higher aggregation level that the mucilage extracted from the chlorenchyma. The presence of three characteristic bands of pectic substances in the FT-IR spectra between 1050 and 1120 cm−1 indicated that the mucilage extracted from the parenchymatous tissue had a higher content of pectic compounds than the mucilage extracted from the chlorenchymatous tissue. It was also observed in the Raman spectra that the level of pectic substances in the mucilage extracted from the parenchymatous was higher than that in the mucilage extracted from the chlorenchymatous tissue. The mucilage extracted from the parenchymatous tissue was more effective as an edible coating than the mucilage extracted from the chlorenchymatous tissue. Tomatoes covered with mucilage showed significantly enhanced firmness and reduced weight loss. The uncoated tomatoes showed higher lycopene content than the coated tomatoes on the 21st day. This study showed that the Opuntia robusta tissue and extraction solvent influence mucilage characteristics and that Opuntia robusta mucilage is a promising edible coating.
Cysteine-rich receptor-like protein kinases (CRKs) are transmembrane proteins containing two domains of unknown function 26 (DUF26) RLKs in their ectodomain. Despite that CRKs control important aspects of plant development, only few proteins have functionally been characterized. In this work, we analyzed the function of CRK33 by characterizing two insertional lines. The stomatal density and stomatal index were decreased in crk33-2 and crk33-3 plants in comparison to wild-type plants, correlating with a decreased transpiration in transgenic plants and a higher drought tolerance. Furthermore, photosynthesis and stomatal conductance changed. Finally, all four stomata cell fate genes were upregulated, especially the expression of TMM and SPCH in the mutant background, suggesting a role for CRK33 in stomatal spacing.
Late embryogenesis abundant (LEA) proteins are a large protein family that mainly function in protecting cells from abiotic stress, but these proteins are also involved in regulating plant growth and development. In this study, we performed a functional analysis of LEA13 and LEA30 from Arabidopsis thaliana. The results showed that the expression of both genes increased when plants were subjected to drought-stressed conditions. The insertional lines lea13 and lea30 were identified for each gene, and both had a T-DNA element in the regulatory region, which caused the genes to be downregulated. Moreover, lea13 and lea30 were more sensitive to drought stress due to their higher transpiration and stomatal spacing. Microarray analysis of the lea13 background showed that genes involved in hormone signaling, stomatal development, and abiotic stress responses were misregulated. Our results showed that LEA proteins are involved in drought tolerance and participate in stomatal density.