Preliminary Construction of a Virus-induced Gene Silencing System in Asparagus officinalis

Purpose To establish a virus-induced gene silencing (VIGS) system in the perennial dioecious plant of Asparagus officinalis L., exploring the function of key genes.

Methods The green A. officinalis variety ‘Guelph Millennium’ was used as material, the phytoene dehydrogenase (AoPDS) and the magnesium chelatase subunit Ⅰ (AoCH42) were selected as report genes, and the apple latent spherical virus (ALSV) was used as silencing vector, the geminating A. officinalis seeds were infected by the resulted Agrobacterium tumefaciens with vacuum infiltration method. The VIGS system of A. officinalis was constructed by phenotype and qRT-PCR, and the function of AoNDUFS8 gene encoding A. officinalis NADH-dehydrogenase ferrithione subunit was further verified.

Results After 21 days of infection, both the AoPDS and AoCH42 silenced plants had an albino and etiolation phenotypes, while the AoNDUFS8 silenced plants grew thin and stunted. qRT-PCR results showed that: compared with the control group, the expression of the target gene decreased after VIGS, indicating that the silencing system was successfully established.

Conclusion The VIGS system can effectively silence the target gene(s) in A. officinalis, providing a practical basis for subsequent functional genomic analyses of A. officinalis.