DNA methylation is a common and rapid evolving epigeneticmechanism among higher eukaryotic organisms including cytosine residues in CG and CNG sequences. NtROS2a gene isolated from tobacco plants (Nicotiana tabacum cv Xanthi nc) contains DNA glycosylase domain with significant similarities to Arabidopsis ROS1 and DME at the C-terminus. Purified NtROS2a protein gene is 5879 bp long and is designated as Nicotiana tabacum Repressor Of Silencing (NtROS: accession numbers; AB281588). RT-PCR analysis initially revealed the adaptive response of the lines overexpressing the gene in tobacco to various abiotic stresses. This was validated when phenotypic evaluation to cold, salt, simulated drought and osmotic stress tolerance was performed. Reduction in leaf wilting rate against high salinity treatment is remarkable in transgenic plants which showed at least 50% less wilting than the WT. Moreover, about 70% wilt rate difference was noted between the transgenic and WT in H₂O₂ treatment. This study showed that NtROS2a gene encoding DNA glycosylase which caused demethylation could regulate adaptive response of tobacco to various abiotic stress conditions.

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Plants have adapted the ability to respond to various abiotic stresses such as high salinity, osmotic stress, high and low temperatures, and drought in order to survive. Small heat shock proteins (sHsps) play important and extensive roles in plant defenses against abiotic stresses. Herein, we cloned an sHsp gene from the rice, which we named Oshsp16.9 based on the molecular weight of the protein. Real-time PCR analysis showed that expression of the Oshsp16.9 gene was rapidly and strongly induced by stresses including high-salinity (250 mM NaCl), osmotic stress (300 mM mannitol), 100 μM ABA, cold (4°C) and heat (45°C). Subcellular localization assay indicated that Oshsp16.9 was localized specifically in the cytoplasm. In addition, overexpression of Oshsp16.9 in rice conferred tolerance of transgenic plants to salt and drought stress. Taken together, these results suggest that the Oshsp16.9 gene is an important determinant of stress response in plants.

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C3HC4-type RING zinc finger proteins are known to be essential in the regulation of plant processes, including responses to abiotic stress. In order to explore the potential of the BrRZFP1 gene to enhance tolerance toward multiple stresses in different host plant genomes, we generated transgenic tomato (Solanum lycopersicum L. cv. Goldenbell) plants. The tomato plants overexpressing BrRZFP1 acquired a higher tolerance to drought stress. However, the transgenic plants did not appear to be more cold tolerant than the WT, in any tested condition. The data obtained indicate that the specificity and the degree of BrRZFP1 activity depend on the host genomic background. In physiological assessment of salt stress tolerance, transgenic plants showed more dry matter accumulation and maintained significantly higher levels of leaf chlorophyll content along with increasing levels of salt stress than the wild type plants. This study shows that BrRZFP1 is a candidate gene in the engineering of crops for enhanced drought and salt stress tolerance.

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Salt stress is by far the leading environmental stress limiting crop yields worldwide. Genetic engineering techniques hold great promise for developing crop cultivars with high tolerance to salt stress. In this study, the Brassica rapa var. SUN-3061 BrOAT1 gene was transferred into tomato through Agrobacterium-mediated leaf disc transformation. The transgenic status and transgene expression of the transgenic plants was confirmed by polymerase chain reaction (PCR) analysis and semi-quantitative one step RT-PCR analysis respectively. Subsequently, the growth status under salt stress, and physiological responses to salt stress of transgenic tomato were studied. The results showed that the transgenic plants exhibited better growth status under salt stress condition compared to the wild type plants. In physiological assessment of salt stress tolerance, transgenic plants showed more dry matter accumulation and maintained significantly higher levels of leaf chlorophyll content along with increasing levels of salt stress than the wild type plants. This study shows that BrOAT1 is a candidate gene in the engineering of crops for enhanced salt stress tolerance.

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• Overexpression of OsDUF868.12 enhances salt tolerance in rice
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In this study, we isolated cDNA with open reading frame encoding putative methyl-binding domain proteins from red pepper, which was designated as CaMBD1 (HQ171162). BLASTX search and phylogenetic analysis suggested that the CaMBD1 gene belonged to AtMBD10 group (subclass I) of MBD family. The expression profile of the CaMBD1 was studied via Q-RT-PCR and the results indicated that the CaMBD1 were differentially expressed in detected red pepper tissues. It was interesting to note that CaMBD1 was highly expressed in dry seeds and endosperms. Moreover, the differential expression pattern of CaMBD1 was observed in leaves and roots under water-stress. Also a GFP-CaMDB1 fusion construct introduced into the onion epidermal cells confirmed localization of CaMBD1 into the nuclei. To investigate the biological significance of CaMBD1 proteins, we transformed Arabidopsis using CaMBD1 gene. The resulting 35S::CaMBD1 plants showed a variety of phenotypic effects including aerial rosettes, serrated leaves, abnormal position of flowers, fertility problems and late flowering. Arabidopsis lines involved in chromatin remodeling show similar phenotypes. Our results suggest an important role of CaMBD1 biological in plant growth and development.

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The latest report on the draft genome of Brassica rapa sequence has been published. To elucidate the functions of these genes and to efficiently search for agriculturally useful genes, a Full-length cDNA Over-eXpressor (FOX) gene hunting system was used. The FOX library from Chinese cabbage was introduced into rice via Agrobacterium-mediated transformation. Approximately 1,150 FOX-rice lines were generated. Genomic PCR analysis indicated that the average length of FL-cDNAs introduced into individual lines was 900~1,200 bp. Basic Local Alignment System Tool (BLAST) analysis of the FL-cDNA genes revealed that 35.5% have unknown function. Most of the randomly selected transgenic rice lines showed overexpression (92%) of these genes relative to the wild-type Gopum. Moreover, 94% of the 850 transgenic rice lines were moderately tolerant (slightly yellow) to cold and 9 lines were tolerant (seedlings were light green). Morphological evaluation of the transgenic rice lines showed minimal phenotypic alteration (12%). Approximately 25.1% and 22% of the plants were significantly ahead in the days to heading and had elevated chlorophyll content, respectively. Other agronomic traits such as filled grains, number of tiller, panicle length, and culm and plant height were relatively less variable among the transgenic lines. These results provide a resource for defining genes that are associated with tolerance in transgenic rice lines.

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