1.OsCOI1, a Putative COI1 in Rice, Show MeJA and ABA Dependent Expression
Tingzhang HU ; Weiping WANG ; Kaiming CAO ; Xiping WANG
Progress in Biochemistry and Biophysics 2006;33(4):388-393
A novel gene, which was a homologue of Arabidopsis COI1 was isolated from rice (Oryza sativa L.) by RT-PCR and designated as OsCOI1. It encoded a protein of 595 amino acids. The similar F-box motif and 16 leucine-rich repeats were found in the deduced protein OsCOI1. OsCOI1 and COI1 showed high homology (74%) at amino acid level. Semi-quantitative RT-PCR and Northern blot analysis demonstrated that the expression of OsCOI1 in rice varied obviously after treatment with MeJA and ABA but was not affected by SA and ET, suggesting that the specific function of OsCOI1 in JA signal pathway and related ABA pathway.
2.Comparison of Three Detection Methods for Burkholderia Cepacia Complex
Jue LI ; Yinhuan WANG ; Tingzhang WANG ; Linshuang ZHANG ; Huan CHEN ; Jun LI ; Xiaoling ZHENG ; Zhijian WANG
Chinese Journal of Modern Applied Pharmacy 2024;41(8):1091-1098
OBJECTIVE
To establish a rapid and accurate PCR method for detecting 24 strains of Burkholderia cepacia complex(Bcc) by comparing three detection methods of loop-mediated isothermal amplification(LAMP), SYTO 9 dye method based on polymerase chain reaction(PCR) and TaqMan probe real-time fluorescent quantitative PCR method( TaqMan probe method).
METHODS
According to the molecular biological information of 24 strains of Bcc in the NCBI database, multiple candidate sequence fragments unique to Bcc were screened out, and specific primer and probe that could simultaneously detect 24 strains of Bcc were designed. At the same time, the detection methods of LAMP, SYTO 9 dye method based on PCR and Taqman probe were explored, and the optimal annealing temperature was optimized and screened. The 39 experimental strains were used to verify the Bcc detection method.
RESULTS
LAMP method could not effectively detect Bcc, SYTO 9 dye method and TaqMan probe method could effectively detect more than 20 strains of Bcc, while TaqMan probe method had higher amplification effect, better detection sensitivity, repeatability and stability, which could meet the requirements of this study.
CONCLUSION
In this study, a TaqMan probe method for rapid detection of Bcc was established. Compared with LAMP method and SYTO 9 dye method, this method has the advantages of fast, simple and high sensitivity, and provides technical support for the rapid detectionof Bcc.