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6 September 2019 Evaluation of virus-induced gene silencing methods for forage legumes including alfalfa, sainfoin, and fenugreek
Champa Wijekoon, Stacy D. Singer, Randall J. Weselake, Udaya Subedi, Surya N. Acharya
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Abstract

Virus-induced gene silencing (VIGS) is a rapid reverse genetics tool that has been developed in a wide variety of plant species for assessing gene functions. However, while VIGS has been utilized successfully in the diploid model leguminous species Medicago truncatula (Gaertn.) (barrel medic), such a platform has yet to be established in forage legume crop species. Therefore, we evaluated the effectiveness of this method in forage legumes using a previously developed PEBV (pea early browning virus) system whereby a fragment of the pea (Pisum sativum L.) PHYTOENE DESATURASE (PDS) gene was transferred into a range of alfalfa (Medicago sativa L.), sainfoin (Onobrychis viciifolia Scop.), and fenugreek (Trigonella foenum-graecum L.) cultivars using leaf infiltration and apical meristem injection. Barrel medic was used as a positive control. Gene silencing was observed after 10–15 d through the presence of a leaf bleaching phenotype, and was confirmed using quantitative real-time RT-PCR. Silencing of PDS was achieved in a selection of cultivars in all species assessed, with the highest silencing efficiency apparent in fenugreek. The introduction of a highly homologous gene fragment from a heterologous plant species to target endogenous genes for transient VIGS-based silencing in a range of species of interest represents a potentially useful strategy for the rapid functional characterization of candidate genes in forages.

© Her Majesty the Queen in Right of Canada 2019. Permission for reuse (free in most cases) can be obtained from Rightslink.
Champa Wijekoon, Stacy D. Singer, Randall J. Weselake, Udaya Subedi, and Surya N. Acharya "Evaluation of virus-induced gene silencing methods for forage legumes including alfalfa, sainfoin, and fenugreek," Canadian Journal of Plant Science 99(6), 917-926, (6 September 2019). https://doi.org/10.1139/cjps-2018-0329
Received: 21 December 2018; Accepted: 30 July 2019; Published: 6 September 2019
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KEYWORDS
alfalfa
barrel medic
Fenugreek
phytoene desaturase
sainfoin
VIGS
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