Virus-induced gene silencing in Medicago truncatula and Lathyrus odorata
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Virus-induced gene silencing in Medicago truncatula and Lathyrus odorata. / Grønlund, Mette; Kjær, Gabriela Didina Constantin; Piednoir, Elodie; Kovacev, Jordan; Johansen, Elisabeth; Lund, Ole Søgaard.
In: Virus Research, Vol. 135, No. 2, 2008, p. 345-349.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Virus-induced gene silencing in Medicago truncatula and Lathyrus odorata
AU - Grønlund, Mette
AU - Kjær, Gabriela Didina Constantin
AU - Piednoir, Elodie
AU - Kovacev, Jordan
AU - Johansen, Elisabeth
AU - Lund, Ole Søgaard
PY - 2008
Y1 - 2008
N2 - Virus-induced gene silencing (VIGS) has become an important reverse genetics tool for functional genomics. VIGS vectors based on Pea early browning virus (PEBV, genus Tobravirus) and Bean pod mottle virus (genus Comovirus) are available for the legume species Pisum sativum and Glycine max, respectively. With the aim of extending the application of the PEBV VIGS vector to other legumes, we examined susceptibility of 99 accessions representing 24 legume species including 21 accessions of Medicago truncatula and 38 accessions Lotus japonicus. Infectivity of PEBV was tested by agro-inoculation with a vector carrying the complete ß-glucuronidase (GUS) coding sequence. In situ histochemical staining analysis indicated that 4 of 21 M. truncatula and three of three Lathyrus odorata accessions were infected systemically by GUS tagged PEBV, while none of 38 L. japonicus accessions displayed GUS staining of either inoculated or uninoculated leaves. Agro-inoculation of plants representing PEBV-GUS susceptible M. truncatula and L. odorata accessions with PEBV carrying a fragment of Phytoene desaturase (PDS) resulted in development of a bleaching phenotype suggesting a down-regulation of PDS expression. In M. truncatula this was supported by quantification of PDS mRNA levels by real-time PCR
AB - Virus-induced gene silencing (VIGS) has become an important reverse genetics tool for functional genomics. VIGS vectors based on Pea early browning virus (PEBV, genus Tobravirus) and Bean pod mottle virus (genus Comovirus) are available for the legume species Pisum sativum and Glycine max, respectively. With the aim of extending the application of the PEBV VIGS vector to other legumes, we examined susceptibility of 99 accessions representing 24 legume species including 21 accessions of Medicago truncatula and 38 accessions Lotus japonicus. Infectivity of PEBV was tested by agro-inoculation with a vector carrying the complete ß-glucuronidase (GUS) coding sequence. In situ histochemical staining analysis indicated that 4 of 21 M. truncatula and three of three Lathyrus odorata accessions were infected systemically by GUS tagged PEBV, while none of 38 L. japonicus accessions displayed GUS staining of either inoculated or uninoculated leaves. Agro-inoculation of plants representing PEBV-GUS susceptible M. truncatula and L. odorata accessions with PEBV carrying a fragment of Phytoene desaturase (PDS) resulted in development of a bleaching phenotype suggesting a down-regulation of PDS expression. In M. truncatula this was supported by quantification of PDS mRNA levels by real-time PCR
KW - Former LIFE faculty
KW - Reverse genetics
KW - Leguminosae
U2 - 10.1016/j.virusres.2008.04.005
DO - 10.1016/j.virusres.2008.04.005
M3 - Journal article
C2 - 18495283
VL - 135
SP - 345
EP - 349
JO - Virus Research
JF - Virus Research
SN - 0168-1702
IS - 2
ER -
ID: 8114450