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Fig. 1 | BMC Plant Biology

Fig. 1

From: An efficient and improved method for virus-induced gene silencing in sorghum

Fig. 1

Schematic representation of BMV based VIGS in sorghum. For VIGS, the BMV genome is cloned into two Agrobacterium strains (see Zhu et al. 2014). One Agrobacterium strain has RNA1 and RNA2 of BMV, and another strain has modified RNA3 with restriction sites to insert plant gene fragments for silencing. The two Agrobacterium strains are multiplied to 1.5 OD600 and mixed in equal amounts to infiltrate N. benthamiana to reconstitute and multiply the virus. The virus is extracted from N. benthamiana leaves. RT-PCR is performed using viral RNA as template and primers flanking the insert to confirm the presence of gene fragment. The sap of infected N. benthamiana is used to inoculate the plants with BMV. The inoculated Chenopodium and barley plants can be kept at 22 °C, whereas inoculated sorghum plants should be maintained at 18 °C

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