Primers for the Amplification of the Circular Chloroplast DNA from the A-genome Group of Cultivated Cotton

The availability of the plastid genome sequences is one of the bases for comparative,functional,and structural genomic studies of plastid-containing living organisms,in addition to the application

Visual Detection of Murray Valley Encephalitis Virus by Reverse Transcription Loop-Mediated Isothermal Amplification

A sensitive reverse transcription loop-mediated isothermal amplification （RT-LAMP） assay was developed for rapid visual detection of Murray valley encephalitis virus （MVEV） infection. The reaction was performed in one step in a single tube at 63 ~C for 60 min with the addition of the hydroxynaphthol blue （HNB） dye prior to amplification. The detection limit of the RT-LAMP assay was 100 copies per reaction based on 10-fold dilutions of in vitro transcribed RNA derived from a synthetic MVEV DNA template. No cross-reaction was observed with other encephalitis-associated viruses. The assay was further evaluated using spiked cerebrospinal fluid sample with pseudotype virus containing the NS5 gene of MVEV.

A study on the pathogen species and physiological races of tomato Fusarium wilt in Shanxi,China

In order to clarify the main pathogens of tomato Fusarium wilt in Shanxi Province, China, morphological identification, elongation factor 1 alpha （EF-1α） sequence analysis, specific primer amplification and pathogenicity tests were applied to study the isolates which were recovered from diseased plants collected from 17 different districts of Shanxi Province. The results were as follows： 1） Through morphological and molecular identification, the following 7 species of Fusarium were identified： F. oxysporum, F. solani, F. verticillioides, F. subglutinans, F. chlamydosporum, F. sporotrichioides, and F. semitectum; 2） 56 isolates of F. oxysporum were identified using specific primer amplification, among which, 29, 5 and 6 isolates were respectively identified as F. oxysporum f. sp. lycopersici physiological race 1, race 2, and race 3; 3） pathogenicity test indicated the significant pathogenicity of F. oxysporum, F. solani, F. verticillioides, and F. subglutinans to tomato plant. Therefore, among these 4 species confirmed as pathogenic to tomato in Shanxi, the highest isolation rate （53.3％） corresponded to F. oxysporum. Three physiological species, race 1, race 2, and race 3 of F. oxysporum f. sp. lycopersici are detected in Shanxi, among which race 1 is the most widespread pathogen and is also considered as the predominant race.

PCR-based Assay for the Detection of Xanthomonas campestris pv. mangiferaeindicae Causing Bacterial Black Spot in Mango

[Objective] This study aimed to develop a PCR assay for detecting Xanthomonas campestris pv. mangiferaeindicae(Xcm) in culture and in planta. [Method] Primers(Xcm HF and Xcm HR) were designed based on the partial sequence of hrp B gene from xanthomonads to develop a PCR assay for Xcm. Furthermore, specificity and sensitivity of the primer pairs were analyzed in detection of genomic DNA and cell from Xcm. [Result] Amplication was positive only with genomic DNA from positive control ATCC11637 and 12 Xcm strains; no PCR products were amplified with genomic DNA from ten other xanthomonads and seven other bacterial species. The sensitivity of detection was 2.4 pg/μl genomic DNA, and 1.8 × 104CFU/ml cells. The primers also worked well for pathogen detection in direct PCR assays of Xcm colonies grown on liquid medium and in PCR assays of total DNA from leaf, branch and fruit lesions. [Conclusion] A PCR assay was successfully established for rapid detection of Xcm in culture and in planta.