PCR detection of ratoon stunting disease pathogen and natural resistance analysis in sugarcane core germplasms.

摘要

Exploring and utilizing resistant germplasm resources plays a pivotal role in breeding for disease resistance, while screening resistant germplasm is important for selecting and breeding varieties resistant to disease. In the present study, we used PCR to detect the ratoon stunting disease (RSD) bacterium Leifsonia xyli subsp. xyli (Lxx) in 137 sugarcane core germplasms from the National Nursery of Sugarcane Germplasm Resources (NNSGR, Kaiyuan, China) in 2009, 2010 and 2011. A total of 21 germplasms that tested negative for Lxx in 2009 and 2010 were selected for further Lxx detection after being subjected to artificial inoculation in 2011 and 2012. The 21 core germplasms that were negative for Lxx after natural infection and artificial inoculation can provide elite resistance source materials and reference frames for the effective breeding of RSD-resistant sugarcane varieties. Under natural conditions, 116 (84.67%) and 21 (15.33%) out of 137 germplasms were positive and negative for Lxx, respectively, as determined by PCR detection, which suggests that a relatively high ratio of sugarcane core germplasms was sensitive to RSD, while few were resistant to RSD. The sequencing and analysis of 30 randomly selected PCR products showed that all 30 sequences were identical or highly homologous to the corresponding Lxx genome region published in GenBank (99.54-100% similarity). Lxx can be detected effectively and precisely by PCR. We therefore recommend PCR as a rapid, low cost and simple procedure to score sugarcane core germplasms for RSD resistance.