These authors made equal contributions to this study. SummaryThe¯oral transition in Arabidopsis is regulated by at least four¯owering pathways: the long-day, autonomous, vernalization, and gibberellin (GA)-dependent pathways. Previously, we reported that the MADSbox transcription factor SUPPRESSOR OF OVEREXPRESSION OF CO 1 (SOC1) integrates the long-day and vernalization/autonomous pathways. Here, we present evidences that SOC1 also integrates signaling from the GA-dependent pathway, a major¯owering pathway under non-inductive short days. Under short days, the¯owering time of GA-biosynthetic and -signaling mutants was well correlated with the level of SOC1 expression; overexpression of SOC1 rescued the non-¯owering phenotype of ga1-3, and the soc1 null mutant showed reduced sensitivity to GA for¯owering. In addition, we show that vernalization-induced repression of FLOWERING LOCUS C (FLC ), an upstream negative regulator of SOC1, is not suf®cient to activate SOC1; positive factors are also required. Under short days, the GA pathway provides a positive factor for SOC1 activation. In contrast to SOC1, the GA pathway does not regulate expression of other owering integrators FLC and FT. Our results explain why the GA pathway has a strong effect on¯owering under short days and how vernalization and GA interact at the molecular level.

Posttranscriptional RNA metabolism plays versatile roles in the regulation of gene expression during eukaryotic growth and development. It is mediated by a group of RNA binding proteins with distinct conserved motifs. In this study, an Arabidopsis (Arabidopsis thaliana) gene, designated FLK, was identified and shown to encode a putative RNA binding protein with K homology motifs. A mutant in which FLK was inactivated by T-DNA insertion exhibited a severe late flowering phenotype both in long and short days. The late flowering phenotype was reversed by gibberellin and vernalization treatments. The FLOWERING LOCUS C (FLC) transcription was greatly upregulated, whereas those of FLOWERING LOCUS T and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 decreased in the mutant. These observations demonstrate that FLK regulates the autonomous flowering pathway via FLC. It is now evident that a battery of different RNA binding proteins are involved in the posttranscriptional regulation of flowering time in Arabidopsis.

Profilin is a small actin-binding protein that regulates cellular dynamics of the actin cytoskeleton. In Arabidopsis (Arabidopsis thaliana), five profilins were identified. The vegetative class profilins, PRF1, PRF2, and PRF3, are expressed in vegetative organs. The reproductive class profilins, PRF4 and PRF5, are mainly expressed in pollen. In this study, we examined the role of the first intron in the expression of the Arabidopsis profilin gene family using transgenic plants and a transient expression system. In transgenic plants, we examined PRF2 and PRF5, which represent vegetative and reproductive profilins. The expression of the PRF2 promoter fused with the b-glucuronidase (GUS) gene was observed in the vascular bundles, but transgenic plants carrying the PRF2 promoter-GUS with its first intron showed constitutive expression throughout the vegetative tissues. However, the first intron of PRF5 had little effect on the reporter gene expression pattern. Transgenic plants containing PRF5 promoter-GUS fusion with or without its first intron showed reproductive tissue-specific expression. To further investigate the different roles of the first two introns on gene expression, the first introns were exchanged between PRF2 and PRF5. The first intron of PRF5 had no apparent effect on the expression pattern of the PRF2 promoter. But, unlike the intron of PRF5, the first intron of PRF2 greatly affected the reproductive tissue-specific expression of the PRF5 promoter, confirming a different role for these introns. The results of a transient expression assay indicated that the first intron of PRF1 and PRF2 enhances gene expression, whereas PRF4 and PRF5 do not. These results suggest that the first introns of profilin genes are functionally distinctive and the first introns are required for the strong and constitutive gene expression of PRF1 and PRF2 in vegetative tissues.

HSP70, a heat shock protein, is a molecular chaperone responsive to various environmental stresses. Here, NtHSP70-1 was a drought-/ABA-inducible gene. We monitored the expression of CaERD15 (early responsive to dehydration) with exposing plants to progressive drought stress. Its activity was used as an indicator of water-deficit conditions. To analyze the protective role of HSP70, we obtained transgenic tobacco plants that constitutively expressed elevated levels of the tobacco HSP70, NtHSP70-1, as well as transgenic plants containing either the vector alone or else having NtHSP70-1 in the antisense orientation. Plants with enhanced levels of NtHSP70-1 in their transgenic sense lines exhibited tolerance to water stress. Under progressive drought, the amount of leaf NtHSP70-1 was correlated with maintenance of optimum water content, with contents being higher in the leaves of dehydrated transgenic sense plants than in those of either the control (vector-only) or the transgenic antisense plants. Moreover, the expression of CaERD15 was considerably reduced in tobacco plants that over-expressed NtHSP70-1. These results suggest that elevated levels of NtHSP70-1 can confer drought-stress tolerance.

Although heat shock response in plants has been extensively studied, there has been little investigation into the effect of heat shock on plant organ development. When heat shock was applied to Arabidopsis during flower development, floral organs such as sepals, petals, stamens, and carpels developed normally. However, the development of pollen inside the anther was disrupted in a stage-specific manner, with floral stage 9 primordia failing to produce any pollen grains. Morphological analyses suggested that heat shock causes a failure of separation of pollen mother cells followed by microspore differentiation and/or inhibition of male meiotic processes. Heat shock also caused sterility in floral stage 12 flowers but the sterility was due to the failure of pollen release from the pollen sacs.

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Nodules are formed on legume roots as a result of signaling between symbiotic partners and in response to the activities of numerous genes. We cloned fragments of differentially expressed genes in spot-inoculated soybean (Glycine max) roots. Many of the induced clones were similar to known genes related to oxidative stress, such as thioredoxin and b-carotene hydroxylase. The deduced amino acid sequences of full-length soybean cDNAs for thioredoxin and b-carotene hydroxylase were similar to those in other species. In situ RNA hybridization revealed that the thioredoxin gene is expressed on the pericycle of 2-d-old nodules and in the infected cells of mature nodules, suggesting that thioredoxin is involved in nodule development. The thioredoxin promoter was found to contain a sequence resembling an antioxidant responsive element. When a thioredoxin mutant of yeast was transformed with the soybean thioredoxin gene it became hydrogen peroxide tolerant. These observations prompted us to measure reactive oxygen species levels. These were decreased by 3-to 5-fold in 7-d-old and 27-d-old nodules, coincident with increases in the expression of thioredoxin and b-carotene hydroxylase genes. Hydrogen peroxide-producing regions identified with cerium chloride were found in uninoculated roots and 2-d-old nodules, but not in 7-d-old and 27-d-old nodules. RNA interference-mediated repression of the thioredoxin gene severely impaired nodule development. These data indicate that antioxidants such as thioredoxin are essential to lower reactive oxygen species levels during nodule development.

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