A Rapid, Equipment-Free Method for Detecting Avirulence Genes of Pyricularia oryzae Using a Lateral Flow Strip-Based RPA Assay

Rice blast, caused by Pyricularia oryzae, is one of the most destructive rice diseases worldwide. Using resistant rice varieties is the most cost-effective way to control rice blast. Consequently, it is critical to monitor the distribution frequency of avirulence (Avr) genes in rice planting fields...

Ausführliche Beschreibung

Bibliographische Detailangaben
Veröffentlicht in:Plant disease. - 1997. - 108(2024), 8 vom: 01. Aug., Seite 2283-2290
1. Verfasser: Qi, Zhongqiang (VerfasserIn)
Weitere Verfasser: Ju, Fangyi, Guo, Yunxia, Du, Yan, Yu, Junjie, Zhang, Rongsheng, Yu, Mina, Cao, Huijuan, Song, Tianqiao, Pan, Xiayan, Dai, Tingting, Liu, Yongfeng
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2024
Zugriff auf das übergeordnete Werk:Plant disease
Schlagworte:Journal Article Pyricularia oryzae RPA-LFD rapid detection
LEADER 01000caa a22002652 4500
001 NLM370772148
003 DE-627
005 20240823232231.0
007 cr uuu---uuuuu
008 240408s2024 xx |||||o 00| ||eng c
024 7 |a 10.1094/PDIS-10-23-2098-SR  |2 doi 
028 5 2 |a pubmed24n1510.xml 
035 |a (DE-627)NLM370772148 
035 |a (NLM)38587798 
040 |a DE-627  |b ger  |c DE-627  |e rakwb 
041 |a eng 
100 1 |a Qi, Zhongqiang  |e verfasserin  |4 aut 
245 1 2 |a A Rapid, Equipment-Free Method for Detecting Avirulence Genes of Pyricularia oryzae Using a Lateral Flow Strip-Based RPA Assay 
264 1 |c 2024 
336 |a Text  |b txt  |2 rdacontent 
337 |a ƒaComputermedien  |b c  |2 rdamedia 
338 |a ƒa Online-Ressource  |b cr  |2 rdacarrier 
500 |a Date Completed 22.08.2024 
500 |a Date Revised 22.08.2024 
500 |a published: Print-Electronic 
500 |a Citation Status MEDLINE 
520 |a Rice blast, caused by Pyricularia oryzae, is one of the most destructive rice diseases worldwide. Using resistant rice varieties is the most cost-effective way to control rice blast. Consequently, it is critical to monitor the distribution frequency of avirulence (Avr) genes in rice planting fields to facilitate the breeding of resistant rice varieties. In this study, we established a rapid recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) detection system for the identification of AvrPik, Avr-Piz-t, and Avr-Pi9. The optimized reaction temperature and duration were 37°C and 20 min, indicating that the reaction system could be initiated by body temperature without relying on any precision instruments. Specificity analysis showed that the primer and probe combinations targeting the three Avr genes exhibited a remarkable specificity at genus-level detection. Under the optimized condition, the lower detected thresholds of AvrPik, Avr-Piz-t, and Avr-Pi9 were 10 fg/μl, 100 fg/μl, and 10 pg/μl, respectively. Notably, the detection sensitivity of the three Avr genes was much higher than that of PCR. In addition, we also successfully detected the presence of AvrPik, Avr-Piz-t, and Avr-Pi9 in the leaf and panicle blast lesions with the RPA-LFD detection system. In particular, the genomic DNA was extracted using the simpler PEG-NaOH rapid extraction method. In summary, we developed an RPA detection system for AvrPik, Avr-Pi9, and Avr-Piz-t, combined with the PEG-NaOH rapid DNA extraction method. The innovative approach achieved rapid, real-time, and accurate detection of the three Avr genes in the field, which is helpful to understand the distribution frequency of the three Avr genes in the field and provide theoretical reference for the scientific layout of resistant rice varieties 
650 4 |a Journal Article 
650 4 |a Pyricularia oryzae 
650 4 |a RPA-LFD 
650 4 |a rapid detection 
700 1 |a Ju, Fangyi  |e verfasserin  |4 aut 
700 1 |a Guo, Yunxia  |e verfasserin  |4 aut 
700 1 |a Du, Yan  |e verfasserin  |4 aut 
700 1 |a Yu, Junjie  |e verfasserin  |4 aut 
700 1 |a Zhang, Rongsheng  |e verfasserin  |4 aut 
700 1 |a Yu, Mina  |e verfasserin  |4 aut 
700 1 |a Cao, Huijuan  |e verfasserin  |4 aut 
700 1 |a Song, Tianqiao  |e verfasserin  |4 aut 
700 1 |a Pan, Xiayan  |e verfasserin  |4 aut 
700 1 |a Dai, Tingting  |e verfasserin  |4 aut 
700 1 |a Liu, Yongfeng  |e verfasserin  |4 aut 
773 0 8 |i Enthalten in  |t Plant disease  |d 1997  |g 108(2024), 8 vom: 01. Aug., Seite 2283-2290  |w (DE-627)NLM098181742  |x 0191-2917  |7 nnns 
773 1 8 |g volume:108  |g year:2024  |g number:8  |g day:01  |g month:08  |g pages:2283-2290 
856 4 0 |u http://dx.doi.org/10.1094/PDIS-10-23-2098-SR  |3 Volltext 
912 |a GBV_USEFLAG_A 
912 |a SYSFLAG_A 
912 |a GBV_NLM 
912 |a GBV_ILN_350 
951 |a AR 
952 |d 108  |j 2024  |e 8  |b 01  |c 08  |h 2283-2290