Glycinebetaine (betaine) affords osmoprotection in bacteria, plants and animals, and protects cell components against harsh conditions in vitro. This and a compelling body of other evidence have encouraged the engineering of betaine production in plants lacking it. We have installed the metabolic step for oxidation of choline, a ubiquitous substance, to betaine in three diverse species, Arabidopsis, Brassica napus, and tobacco (Nicotiana tabacum), by constitutive expression of a bacterial choline oxidase gene. The highest levels of betaine in independent transgenics were 18.6, 12.8, and 13 mol g ؊1 dry weight, respectively, values 10-to 20-fold lower than the levels found in natural betaine producers. However, choline-fed transgenic plants synthesized substantially more betaine. Increasing the choline supplementation further enhanced betaine synthesis, up to 613 mol g ؊1 dry weight in Arabidopsis, 250 mol g ؊1 dry weight in B. napus, and 80 mol g ؊1 dry weight in tobacco. These studies demonstrate the need to enhance the endogenous choline supply to support accumulation of physiologically relevant amounts of betaine. A moderate stress tolerance was noted in some but not all betaine-producing transgenic lines based on relative shoot growth. Furthermore, the responses to stresses such as salinity, drought, and freezing were variable among the three species.

The deleterious effects of salinity on plant growth are attributed to a decrease in osmotic potential of the growing medium, specific ion toxicity and nutrient ion deficiency (Greenway and Munns 1980). The extent of plant growth depression under saline conditions varies with salt composition, salt concentration, the physiological stage of the plant when it is exposed to salinity, and the plant species (Sharma 1983).Inhibition of seed germination by salinity has been attributed to osmotic effects in barley cultivars (Bliss et al. 1986). Other studies with cereals suggest that salinityinduced reductions in germination and seedling growth are cau$ed by specific ion toxicity (Sharma 1983; Hampson and Simpson 1990a,b). The interaction of osmotic and specific ion effects may be significant, depending on plant species and the combination of cations and anions in a given salt (Redmann 1974;Ryan et al. 1975;Hardegree and Emmerich 1990

Salinity inhibits plant growth due to osmotic and ionic effects. However, little is known about the impact of genotype and salinity on biochemical and molecular processes in the leafy vegetable lettuce. We report here evaluations of two lettuce types, Verte (NaCl tolerant) and Romaine (NaCl sensitive), under iso-osmotic 100 mM NaCl and 77 mM Na(2)SO(4) treatments. As compared to Romaine, NaCl-treated Verte displayed better growth, contained lower levels of inorganic cations in leaves, and possessed superior antioxidative capacity due to enhanced carotenoid and phenolics biosynthesis and more active antioxidative enzymes resulting in reduced membrane damage. Both genotypes had relatively similar growth patterns under Na(2)SO(4) treatment, but Romaine showed enhanced root lignification, greater malondialdehyde formation, and suppressed Fe-superoxide dismutase expression in roots as compared with Verte.

Genomic DNA replication is a universal and essential process for all herpesvirus including human cytomegalovirus (HCMV). HCMV UL70 protein, which is believed to encode the primase activity of the viral DNA replication machinery and is highly conserved among herpesviruses, needs to be localized in the nucleus, the site of viral DNA synthesis. No host factors that facilitate the nuclear import of UL70 have been reported. In this study, we provided the first direct evidence that UL70 specifically interacts with a highly conserved and ubiquitously expressed member of the heat shock protein Hsp40/DNAJ family, DNAJB6, which is expressed as two isoforms, a and b, as a result of alternative splicing. The interaction of UL70 with a common region of DNAJB6a and b was identified by both a two hybrid screen in yeast and coimmunoprecipitation in human cells. In transfected cells, UL70 was primarily co-localized with DNAJB6a in the nuclei and with DNAJB6b in the cytoplasm, respectively. The nuclear import of UL70 was increased in cells in which DNAJB6a was up-regulated or DNAJB6b was down-regulated, and was reduced in cells in which DNAJB6a was down-regulated or DNAJB6b was up-regulated. Furthermore, the level of viral DNA synthesis and progeny production was increased in cells in which DNAJB6a was up-regulated or DNAJB6b was down-regulated, and was reduced in cells in which DNAJB6a was down-regulated or DNAJB6b was up-regulated. Thus, DNAJB6a and b appear to enhance the nuclear import and cytoplasmic accumulation of UL70, respectively. Our results also suggest that the relative expression levels of DNAJB6 isoforms may play a key role in regulating the cellular localization of UL70, leading to modulation of HCMV DNA synthesis and lytic infection.

Epigenetic regulation of gene expression is critical for controlling embryonic properties during the embryo-to-seedling phase transition. Here we report that a HISTONE DEACETYLASE19 (HDA19)-associated regulator, SCARECROW-LIKE15 (SCL15), is essential for repressing the seed maturation programme in vegetative tissues. SCL15 is expressed in and GFP-tagged SCL15 predominantly localizes to, the vascular bundles particularly in the phloem companion cells and neighbouring specialized cells. Mutation of SCL15 leads to a global shift in gene expression in seedlings to a profile resembling late embryogenesis in seeds. In scl15 seedlings, many genes involved in seed maturation are markedly derepressed with concomitant accumulation of seed 12S globulin; this is correlated with elevated levels of histone acetylation at a subset of seed-specific loci. SCL15 physically interacts with HDA19 and direct targets of HDA19–SCL15 association are identified. These studies reveal that SCL15 acts as an HDA19-associated regulator to repress embryonic traits in seedlings.