Virus-Induced Gene Silencing (VIGS) in Cassava Using Geminivirus Agroclones.

[en] Virus-induced gene silencing (VIGS) is an efficient, low-cost, and rapid functional validation tool for candidate genes in planta. The VIGS approach is particularly suitable to perform reverse genetics studies in crop species. Here we present a detailed method to perform VIGS in cassava, from target gene fragment to agroinoculation and VIGS quantitation.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Syed-Shan-e-Ali, Zaïdi ; Université de Liège - ULiège > Département GxABT > Plant Sciences

Vasudevan, Kumar ; Université de Liège - ULiège > Département GxABT > Plant Sciences

Lentz, Ezequiel Matias

Vanderschuren, Hervé ; Université de Liège - ULiège > Département GxABT > Plant Sciences

Language :

English

Title :

Virus-Induced Gene Silencing (VIGS) in Cassava Using Geminivirus Agroclones.

Publication date :

2020

Journal title :

Methods in molecular biology (Clifton, N.J.)

ISSN :

1064-3745

eISSN :

1940-6029

Volume :

2172

Pages :

51-64

Peer reviewed :

Peer reviewed

Available on ORBi :

since 09 July 2020

Statistics

Number of views

51 (8 by ULiège)

Number of downloads

4 (4 by ULiège)

More statistics

Scopus citations^®

Scopus citations^®
without self-citations

OpenCitations

Bibliography

Vanderschuren, H, Nyaboga, E, Poon, JS, Large-scale proteomics of the cassava storage root and identification of a target gene to reduce postharvest deterioration (2014) Plant Cell, 26, pp. 1913-1924
(2013) Save and grow: cassava. A guide to sustainable production intensification, , FAO ISBN: 978-92-5-107641-5, 140pp
Rey, C, Vanderschuren, H, Cassava mosaic and brown streak diseases: current perspectives and beyond (2017) Annu Rev Virol, 4, pp. 429-452
Vanderschuren, H, Strengthening African R&D through effective transfer of tropical crop biotech to African institutions (2012) Nat Biotechnol, 30, pp. 1170-1172
Mehta, D, Sturchler, A, Anjanappa, RB, Linking CRISPR-Cas9 interference in cassava to the evolution of editing-resistant geminiviruses (2019) Genome Biol, 20, p. 80
Beyene, G, Chauhan, RD, Wagaba, H, Loss of CMD2-mediated resistance to cassava mosaic disease in plants regenerated through somatic embryogenesis (2016) Mol Plant Pathol, 17, pp. 1095-1110
Bull, SE, Seung, D, Chanez, C, Accelerated ex situ breeding of GBSS-and PTST1-edited cassava for modified starch (2018) Sci Adv, 4, p. eaat6086
Rosas-Diaz, T, Zhang, D, Fan, P, A virus-targeted plant receptor-like kinase promotes cell-to-cell spread of RNAi (2018) Proc Natl Acad Sci U S A, 115, pp. 1388-1393
Bull, SE, Owiti, JA, Niklaus, M, Agrobacterium-mediated transformation of friable embryogenic calli and regeneration of transgenic cassava (2009) Nat Protoc, 4, pp. 1845-1854
Zainuddin, IM, Schlegel, K, Gruissem, W, Robust transformation procedure for the production of transgenic farmer-preferred cassava landraces (2012) Plant Methods, 8, p. 24
Niklaus, M, Gruissem, W, Vanderschuren, H, Efficient transformation and regeneration of transgenic cassava using the neomycin phosphotransferase gene as aminoglycoside resistance marker gene (2011) GM Crops, 2, pp. 193-200
Nyaboga, E, Njiru, J, Nguu, E, Unlocking the potential of tropical root crop biotechnology in east Africa by establishing a genetic transformation platform for local farmer-preferred cassava cultivars (2013) Front Plant Sci, 4, p. 526
Chetty, CC, Rossin, CB, Gruissem, W, Empowering biotechnology in southern Africa: establishment of a robust transformation platform for the production of transgenic industry-preferred cassava (2013) New Biotechnol, 30, pp. 136-143
Mehta, D, Hirsch-Hoffmann, M, Were, M, A new full-length circular DNA sequencing method for viral-sized genomes reveals that RNAi transgenic plants provoke a shift in geminivirus populations in the field (2018) Nucleic Acids Res, 47 (2), p. e9
Lentz, EM, Kuon, JE, Alder, A, Cassava geminivirus agroclones for virus-induced gene silencing in cassava leaves and roots (2018) Plant Methods, 14, p. 73
Fofana, IB, Sangare, A, Collier, R, A geminivirus-induced gene silencing system for gene function validation in cassava (2004) Plant Mol Biol, 56, pp. 613-624
Vanderschuren, H, Alder, A, Zhang, P, Dose-dependent RNAi-mediated geminivirus resistance in the tropical root crop cassava (2009) Plant Mol Biol, 70, pp. 265-272
Zhang, P, Vanderschuren, H, Futterer, J, Resistance to cassava mosaic disease in transgenic cassava expressing antisense RNAs targeting virus replication genes (2005) Plant Biotechnol J, 3, pp. 385-397
Fauquet, C, Fargette, D, African cassava mosaic-virus— etiology, epidemiology, and control (1990) Plant Dis, 74 (6), pp. 404-411
Soni, R, Murray, JA, Isolation of intact DNA and RNA from plant tissues (1994) Anal Biochem, 218, pp. 474-476
Moreno, I, Gruissem, W, Vanderschuren, H, Reference genes for reliable potyvirus quantitation in cassava and analysis of Cassava brown streak virus load in host varieties (2011) J Virol Methods, 177, pp. 49-54
Pfaffl, MW, A new mathematical model for relative quantification in real-time RT-PCR (2001) Nucleic Acids Res, 29, p. e45
Liu, J, Zheng, Q, Ma, Q, Cassava genetic transformation and its application in breeding (2011) J Integr Plant Biol, 53, pp. 552-569
Vanderschuren, H, Akbergenov, R, Pooggin, M, Transgenic cassava resistance to African cassava mosaic virus is enhanced by viral DNA-A bidirectional promoter-derived siRNAs (2007) Plant Mol Biol, 64, pp. 549-557
Bertani, G, Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli (1951) J Bacteriol, 62 (3), pp. 293-300