Open AccessTemporal and spatial Bean pod mottle virus ‐induced gene silencing in soybean
Author(s) -
Juvale Parijat S.,
Hewezi Tarek,
Zhang Chunquan,
Kandoth Pramod Kaitheri,
Mitchum Melissa G.,
Hill John H.,
Whitham Steven A.,
Baum Thomas J.
Publication year - 2012
Publication title -
molecular plant pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.945
H-Index - 103
eISSN - 1364-3703
pISSN - 1464-6722
DOI - 10.1111/j.1364-3703.2012.00808.x
Subject(s) - biology , mottle , point of delivery , gene silencing , virology , plant virus , gene , virus , genetics , botany
Summary Virus‐induced gene silencing ( VIGS ) is a powerful reverse genetics tool in plant science. In this study, we investigated the temporal and spatial silencing patterns achieved by B ean pod mottle virus ( BPMV )‐based VIGS in soybean using virus constructs targeting green fluorescence protein ( GFP ). Silencing GFP enabled an in‐depth analysis of silencing in soybean tissues over time in a transgenic line constitutively expressing GFP . We discovered evidence for variable GFP silencing based on insert orientation and targeted region in the coding sequence. A 3′ sequence in reverse orientation produced the strongest silencing phenotypes. Furthermore, we documented that BPMV VIGS can achieve widespread silencing in a broad range of tissues, including leaves, stems, flowers and roots. Near‐complete silencing was attained in leaves and flowers. Although weaker than in shoots, the observed gene silencing in soybean roots will also allow reverse genetics studies in this tissue. When GFP fluorescence was assayed in cross‐sections of stems and leaf petioles, near‐complete and uniform silencing was observed in all cell types. Silencing was observed from as early as 2 weeks post‐virus inoculation in leaves to 7 weeks post‐virus inoculation in flowers, suggesting that this system can induce and maintain silencing for significant durations.