An efficient in vitro-inoculation method for Tomato yellow leaf curl virus

Abstract

BackgroundTomato yellow leaf curl virus (TYLCV) is a member of the family Geminiviridae, genus Begomovirus. To test the infectivity of TYLCV in tomato plants, an improved protocol for inoculation of in vitro-cultured tomato plants was developed.ResultsA TYLCV isolate was cloned, sequenced and used to construct a 1.8-mer infectious clone. Three weeks old microshoots of TYLCV-susceptible tomato plants were inoculated with Agrobacterium tumefaciens harboring the infectious clone for the TYLCV isolate. After two weeks, the TYLCV symptoms started to appear on the in vitro-inoculated plants and the symptoms became more severe and pronounced eight weeks post-inoculation. The method was used efficiently to uncover the resistance mechanism against TYLCV in Solanum habrochaites accession LA 1777, a wild tomato known for its high resistance to whitefly and TYLCV.ConclusionsThe reported in vitro-inoculation method can be used to screen tomato genotypes for their responses to TYLCV under controlled conditions and it will be a useful tool for better understanding of the TYLCV biology in tomato plants.

The Contrasting Effects of Elevated CO2 on TYLCV Infection of Tomato Genotypes with and without the Resistance Gene, Mi-1.2

“…For agroinoculation of TYLCV, 25 8-week-old plants of each genotype in each OTC (25 plants × 4 OTC × 2 genotypes × 2 CO 2 levels and 400 plants in total) were selected and agroinoculated as described previously (Huang et al, 2012). The TYLCV infection of tomato plants was achieved using Agrobacterium tumefaciens -mediated infectious inoculation (Zhang et al, 2009; Al Abdallat et al, 2010), and the infectious 2 clone (pBINPLUS-SH2-1.4A) of TYLCV- Israel [China: Shangai2] was constructed into A. tumefaciens strain EHA105 as described previously (Zhang et al, 2009). The infectious clone of TYLCV was provided by Professor Xueping Zhou (State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, China).…”

Eugenol enhances the resistance of tomato against tomato yellow leaf curl virus

“…Agrobacterium tumefaciens strain EHA105 was inoculated in YEP medium supplemented with 50 mg L −1 kanamycin and 50 mg L −1 rifampicin, then cultured at 28 °C at 150 rpm until OD 600 was 0.6–0.8. Tomato stem phloem was inoculated with Agrobacterium tumefaciens strain EHA105 liquid using the puncture inoculation method …”

Lower incidence and severity of tomato virus in elevated CO2 is accompanied by modulated plant induced defence in tomato

“…The TYLCV infection of tomato plants was achieved using Agrobacterium tumefaciens ‐mediated infectious inoculation (Zhou et al. 2009; Al Abdallat et al. 2010), and the infectious clone (pBINPLUS‐SH2‐1.4A) of TYLCV‐ Israel [China: Shangai2] was constructed into A. tumefaciens strain EHA105 as described previously (Zhou et al.…”