Sivaramakrishnan Muthuswamy

Callose (beta-1,3-glucan) is produced at different locations in response to biotic and abiotic cues. Arabidopsis contains 12 genes encoding callose synthase (CalS). We demonstrate that one of these genes, CalS5, encodes a callose synthase which is responsible for the synthesis of callose deposited at the primary cell wall of meiocytes, tetrads and microspores, and the expression of this gene is essential for exine formation in pollen wall. CalS5 encodes a transmembrane protein of 1923 amino acid residues with a molecular mass of 220 kDa. Knockout mutations of the CalS5 gene by T-DNA insertion resulted in a severe reduction in fertility. The reduced fertility in the cals5 mutants is attributed to the degeneration of microspores. However, megagametogenesis is not affected and the female gametes are completely fertile in cals5 mutants. The CalS5 gene is also expressed in other organs with the highest expression in meiocytes, tetrads, microspores and mature pollen. Callose deposition in the cals5 mutant was nearly completely lacking, suggesting that this gene is essential for the synthesis of callose in these tissues. As a result, the pollen exine wall was not formed properly, affecting the baculae and tectum structure and tryphine was deposited randomly as globular structures. These data suggest that callose synthesis has a vital function in building a properly sculpted exine, the integrity of which is essential for pollen viability.

Vertebrate Tpr and its yeast homologs Mlp1/Mlp2, long coiled-coil proteins of nuclear pore inner basket filaments, are involved in mRNA export, telomere organization, spindle pole assembly, and unspliced RNA retention. We identified Arabidopsis thaliana NUCLEAR PORE ANCHOR (NUA) encoding a 237-kD protein with similarity to Tpr. NUA is located at the inner surface of the nuclear envelope in interphase and in the vicinity of the spindle in prometaphase. Four T-DNA insertion lines were characterized, which comprise an allelic series of increasing severity for several correlating phenotypes, such as early flowering under short days and long days, increased abundance of SUMO conjugates, altered expression of several flowering regulators, and nuclear accumulation of poly(A)+ RNA. nua mutants phenocopy mutants of EARLY IN SHORT DAYS4 (ESD4), an Arabidopsis SUMO protease concentrated at the nuclear periphery. nua esd4 double mutants resemble nua and esd4 single mutants, suggesting that the two proteins act in the same pathway or complex, supported by yeast two-hybrid interaction. Our data indicate that NUA is a component of nuclear pore-associated steps of sumoylation and mRNA export in plants and that defects in these processes affect the signaling events of flowering time regulation and additional developmental processes.

Plants exposed to abiotic stress show a range of morphogenetic responses, sometimes termed the Stress-Induced Morphogenetic Response (SIMR). SIMR is principally composed of three components: inhibition of cell elongation, alterations in cell differentiation, and stimulus of cell division in localized areas. An explanation proposed for SIMR has been increased accumulation of reactive oxygen species (ROS) and alterations in hormone signalling. Mutations in the Arabidopsis thaliana RADICAL-INDUCED CELL DEATH1 (RCD1) gene have altered abiotic stress responses and ROS accumulation. Even in the absence of exogenous stress, these plants show many morphological changes also seen in SIMR. In the September Issue of Plant Physiology we reported an in depth analysis of the phenotype of rcd1-3 plants as well as the phenotype of a mutations in the previously uncharacterized paralog of RCD1, SIMILAR TO RCD ONE1 (SRO1). sro1-1 plants have mild morphological changes and abiotic stress response defects while rcd1-3; sro1-1 double mutant plants have severe developmental defects, including less cell elongation. In this Addendum, we hypothesize that rcd1, sro1, and rcd1; sro1 mutant plants are under constitutive stress, and that this stress is responsible for at least some of the developmental defects seen in these plants.