| At present, the function researches about MATH-domain-containing genes were rarely reported in the field of plant. AtSb10 is a member of MATH gene family containing, with two consecutive MATH domains, but the other information about the gene, such as expression pattern, functions is unknown. In this paper, with the aid of bioinformatics, molecular biology, gene engineering technology, a preliminary study for AtSb10 gene was carried out and the main results were as follows:1. Bioinformatics analysis showed that AtSb10 gene was located on chromosome 3 of Arabidopsis thaliana and contained 1818bp with 6 exons and 5 introns, encoding 373 amino acids.The two MATH domains were located at 88 to 220,242 to 365 amino acids residues, respectively.2. Semi-quantitative RT-PCR was used to analyze the expressed pattern of AtSb10 gene in Arabidopsis. The result showed that AtSb10 gene was specifically expressed in root.3. AtSb10 gene promoter region had been cloned and pBI101-AtSb10p::GUS expression plasmid was constructed. The plasmid was transformed into Col-0 and seven transgenic plants were obtained. GUS histochemical staining demonstrated that AtSb10 gene was not only specifically expressed in root, but also in seedling bud site.4. T-DNA insertion homozygous mutant sb10 was screened and no expression of AtSb10 gene was detected by RT-PCR in sb10, indicating that AtSb10 gene was effectively silenced.5. Constructed pCAMBIA1300-35S::AtSb10 overexpression plasmid, transformed Col-0 and got 10 transgenic plants. RT-PCR assay showed AtSb10 gene were expressed in stems, leaves, flowers, siliques, root. Furthermore, the gene expression level in root of overexpressive plants was higher than the one in root of Col-0.6. Compared the root length, root morphology of T-DNA insertion mutants, overexpression transgenic plants and Col-0, the results showed that there were no obvious differences in phenotype.7. Under the condition of different concentration salt stress, seed germination of three materials were investigated. The results showed that compared with wild type, seeds germination rate of T-DNA insertion mutant was higher, whereas seeds germination rate of overexpression plant was lower.insertion mutant seeds germination rate were higher and overexpression transgenic plant were lower. |
