J Integr Plant Biol. ›› 1997, Vol. 39 ›› Issue (7): -.

• Research Articles •    

RT-PCR Amplification and Analysis of Citrus Excortis Viroid

Hu Qin-xue, Lin Mu-lan, Zhang Chun-li, Wu De-xi and Chen Jie   

Abstract: Oligonucleotide primers Cl (–), C2 ( + ), C3 ( + ), T1 (–), T2 ( + ) were designed and synthesized according to the central and left-terminal regions of CEVd-A strain. Using these primers, reverse tnmscription-polymerase chain reaction (RT-PCR) was developed to detect and analyse citrus excortis vimid in citrus ( Citrus L. ) and etmg citron ( Citrus medica L. ) in China. RT-PCR amplified products were analysed by PAGE-silver staining method. Specific DNA bands about 210 bp and 370 bp were observed with primer pair C1/C3 and C1/C2, respectively, from the diseased citrus and etrog citron, while no products was obtained with T1/T2 pair. Under the same conditions, no amplified DNA was detected from nucleic acids of healthy samples with the above three primer pairs. Amplified products were further identified by dot blot hybridization with DIG-la-belled CEVd-cDNA probes. The results indicated that the Chinese isolate of CEVd is different in left-terminal region from the CEVd-A strain. With the established RT-PCR system, full-length CEVd-cDNA could be obtained from approximately 0.1 ng total nucleic acids of infected citrus tissues. This system is suitable for practical diagnosis.

Key words: Citrus excortis viroid, RT-PCR, DIG-labelled cDNA probe

Editorial Office, Journal of Integrative Plant Biology, Institute of Botany, CAS
No. 20 Nanxincun, Xiangshan, Beijing 100093, China
Tel: +86 10 6283 6133 Fax: +86 10 8259 2636 E-mail: jipb@ibcas.ac.cn
Copyright © 2021 by the Institute of Botany, the Chinese Academy of Sciences
Online ISSN: 1744-7909 Print ISSN: 1672-9072 CN: 11-5067/Q
备案号:京ICP备16067583号-22