Baoxing Du

Characterization of a DRE-binding transcription factor from a halophyte Atriplex hortensis

Environmental stresses, such as salinity, drought and cold, can induce the expression of a large amount of genes. Among these are many transcription factors that regulate the expression of downstream genes by specifically binding to cis-elements or forming transcriptional complexes with other proteins. In the present study, a DREB-like transcription factor gene, named AhDREB1, was isolated from a halophyte Atriplex hortensis. AhDREB1 encoded a protein containing a conserved EREBP/AP2 domain featuring the DREB family. In yeast one-hybrid analysis AhDREB1 protein was specifically bound to DRE elements and activated the expression of the reporter genes of HIS3 and LacZ. The AhDREB1 gene was expressed in roots, stems and leaves of A. hortensis. Salinity induced its expression in roots, but not in other organs. Overexpression of AhDREB1 in transgenic tobacco led to the accumulation of its putative downstream genes. The performance of the transgenic lines was also tested under stressed conditions and two lines were found to be stress-tolerant. These results suggest that the AhDREB1 protein functions as a DRE-binding transcription factor and play roles in the stress-tolerant response of A. hortensis.

OsGLU1, A Putative Membrane-bound Endo-1,4-ß-D-glucanase from Rice, Affects Plant Internode Elongation

A dwarf mutant glu was identified from screening of T-DNA tagged rice population. Genetic analysis of the T1 generation of glu revealed that a segregation ratio of wild-type:dwarf phenotype was 3:1, suggesting that the mutated phenotype was controlled by a single recessive nuclear locus. The mutated gene OsGLU1, identified by Tail-PCR, encodes a putative membrane-bound endo-1,4-β-D-glucanase, which is highly conserved between mono- and dicotyledonous plants. Mutation of OsGLU1 resulted in a reduction in cell elongation, and a decrease in cellulose content but an increase in pectin content, suggesting that OsGLU1 affects the internode elongation and cell wall components of rice plants. Transgenic glu mutants harboring the OsGLU1 gene complemented the mutation and displayed the wild-type phenotype. In addition, OsGLU1 RNAi plants showed similar phenotype as the glu mutant has. These results indicate that OsGLU1 plays important roles in plant cell growth. Gibberellins and brassinosteroids induced OsGLU1 expression. In rice genome, endo-1,4-β-D-glucanases form a multiple gene family with 15 members, and each may have a distinct expression pattern in different organs. These results indicate that endo-1, 4-β-D-glucanases may play diverse roles in growth and developmental process of rice plants.

Tobacco two-component geneNTHK2

By using a previously isolated tobacco twocomponent geneNTHK1 as a probe, we screened a cDNA library and obtained a homologous gene designatedNTHK2. Sequencing analysis revealed thatNTHK2 encoded a putative ethylene receptor homolog and contained a histidine kinase domain and a receiver domain. In the histidine kinase domain, the histidine at the phosphorylation site was replaced by an asparagine. Southern analysis indicated thatNTHK2 was present at low copies in tobacco genome. The expression ofNTHK2 was studied using a competitive RT-PCR method. It was found that, in young flower buds,NTHK2 was expressed abundantly, while in other organs or tissues, it was expressed in a low level. When leaf was subjected to wounding (cutting) treatment,NTHK2 expression was increased. When tobacco seedlings were stressed with PEG and heat shock,NTHK2 transcription was also enhanced. Other treatments showed little effects. These results indicated that NTHK2 might be involved in the developmental processes and in plant responses to some environmental stresses.

Isolation and characterization of a new defense gene from soybean.

A cDNA clone of a single-copy gene designatedSbPRP was isolated and characterized from 2-week-old soybean seedlings. It putatively encodes a bimodular protein similar to developmentally regulated proteins in other plant species. The deduced amino acid sequence consists of 126 amino acids with a distinct proline-rich domain (17 amino acids) and a long hydrophobically cysteine-rich domain (84 amino acids), plus a signal peptide of 25 amino acids in N terminal.SbPRP mRNA transcripts accumulated in an organ specific manner. It can be detected in leaves and epicotyls of soybean seedlings, whereas virtually expression signal ofSbPRP was not detected in cotyledons, hypocotyls and roots. Further Northern hybridization suggested thatSbPRP steady-state mRNA level accumulated differentially not only in response to salicylic acid, but to the inoculation of soybean mosaic virus Sa strain. Also it was responsive to drought treatment and salt (NaCl) stress. Therefore it is likely thatSbPRP functions as a defense gene in soybean.