Utilization of wheat spike culture to assess Fusarium head blight disease progression and mycotoxin accumulation

Spike culture derived wheat (Triticum aestivum L.) variants exhibit improved resistance to multiple chemotypes of Fusarium graminearum

“…The immature spikes were excised from the plants and maintained in in vitro spike culture media (pH 6.2) containing sucrose (50 g/L), L-glutamine (0.4 g/L) and morpholinoethanesulfonic acid (0.5 g/L) for five days as previously described [37, 38]. Spikes were used for inoculation and disease severity assessment calculated as FHB severity (%) per spike = (number of symptomatic or infected spikelets × 100) / total number of spikelets as previously described [39].…”

“…The infected wheat spikes from each Fusarium strain were used to extract mycotoxin in an extraction buffer (acetonitrile:water, 84:16, v/v) using a recently optimized method [39]. The extracted mycotoxin was analyzed using a 4000 QTRAP LC-MS/MS system (AB SCIEX, Framingham, MA, USA) equipped with a high-performance liquid chromatography, an electrospray interface and a hybrid triple quadrupole ion trap mass spectrometer.…”

“…The extracted mycotoxin was analyzed using a 4000 QTRAP LC-MS/MS system (AB SCIEX, Framingham, MA, USA) equipped with a high-performance liquid chromatography, an electrospray interface and a hybrid triple quadrupole ion trap mass spectrometer. The chromatograms were evaluated using Analyst 1.6.2 software (AB SCIEX, Framingham, MA, USA) [39].…”

“…The selected SCDV lines at M 3 stage were grown in a greenhouse during May-July 2016 with 16/8 h of day/night cycle with average daily temperature of 27.3/20.6°C and relative humidity of 50.1/70.5%. The immature spikes in three replicates were collected and evaluated for FHB severity at 5, 7, 9, and 11 DAI, and calculated as described [39].…”

In Vitro Wheat Immature Spike Culture Screening Identified Fusarium Head Blight Resistance in Wheat Spike Cultured Derived Variants and in the Progeny of Their Crosses with an Elite Cultivar

“…The immature spike culture method was further adapted to evaluate FHB severity by point inoculation of the spikelets of the SCDV lines (Sharma et al, 2018). The in vitro spike culture method is simple, rapid, environment-independent, and is less resource demanding in terms of time and labour compared to field-based screening (Huang et al, 2020). To further support the validity of the immature spike culture screening method for FHB disease severity assessment and resistance identification, the objectives of this study were to (1) assess FHB severity in M 4 SCDV lines under field conditions in one location in Carman, Manitoba, Canada in 2016 and in parallel using immature spike cultures of the corresponding M 4 lines for comparison; (2) assess FHB severity in immature spike cultures of progeny derived from crosses of selected SCDV lines with elite cv.…”