siRNAs Trigger Efficient Silencing of a Parasitism Gene in Plant Parasitic Root-Knot Nematodes
Abstract
:1. Introduction
2. Results and Discussion
2.1. Localization of siRNAs within Soaked Nematodes
2.2. siRNAs Trigger RNAi in Infective Juveniles
siRNA | Sequence 5'→3' | Target Matching Region | |
---|---|---|---|
Size (bp) | GC (%) | ||
CRT-577 | F: AAGGCAGAAAGTGGAGAGCTTCCTGTCTC | 21 | 47.61 |
R: AAAAGCTCTCCACTTTCTGCCCCTGTCTC | |||
CRT-761 | F: AAGACTGGGATGATGATATGGCCTGTCTC | 21 | 42.85 |
R: AACCATATCATCATCCCAGTCCCTGTCTC | |||
CRT-1039 | F: AAGGAAACATTTGAGCCATTGCCTGTCTC | 21 | 38.09 |
R: AACAATGGCTCAAATGTTTCCCCTGTCTC | |||
CRT-1102 | F: AACGCAAGAAGTTTGACGAGGCCTGTCTC | 21 | 47.61 |
R: AACCTCGTCAAACTTCTTGCGCCTGTCTC | |||
Control siRNA | F: AAGTACATTTAACGCCAGCCTCCTGTCTC | 21 | 42.85 |
R: AAAGGCTGGCGTTAAATGTACCCTGTCTC |
2.3. Analysis of Gene-Silencing in Parasitic Stages
2.4. Nematode Viability after siRNA-Mediated RNAi
Gene | name | primer sequence (5'→ 3') | size | T° |
---|---|---|---|---|
Target gene: | ||||
Mi-CRT | CRT9 | GCCTGAGGATTGGGATGAG | 19 | 60 |
CRT10 | TTTGGTGGCTCCCATTCTC | 19 | 60 | |
Reference genes: | ||||
ACT | ACT23 | AAGACGAAGCAGCTGTAGCC | 20 | 60 |
ACT24 | GGTGTTACGCACACAGTTCC | 20 | 60 | |
GAPDH | GAP17 | GGTTATCTCAGCTCCGTCTGC | 21 | 60 |
GAP18 | TAACCTTCGCAAGAGGAGCA | 20 | 60 | |
Mi-eri-1: | ||||
Qp2F | CTGAATATCCGGCGTTCATT | 20 | 58 | |
Qp2R | AGCGTCATCCATTCCACAAT | 20 | 58 | |
Metabolism genes: | ||||
ACDH | ACD19 | GCTCTGATGTTTCCGGACTT | 20 | 60 |
ACD20 | AGCATCACCATCCACAACAA | 20 | 60 | |
GK | GK30 | AGCATGGAATTGAGCCTGAG | 20 | 60 |
GK31 | ACGTTATCCAGAAGCCAACG | 20 | 60 | |
PK | PK38 | GCTGAGGGAAGTGATGTTGC | 20 | 60 |
PK39 | ACACGAAATAGCCGCAGAAG | 20 | 60 | |
ACO-C | ACO44 | GGAGAGGGCTCTTCTCGTG | 19 | 60 |
ACO45 | CTGGAGCAAGAGTGTTGAGC | 20 | 60 | |
CTS | CTS21 | TCGTGAAGGAACATCTGTGG | 20 | 60 |
CTS22 | ATTGCCGCAGAGAAAGAGAG | 20 | 60 | |
GIP7 | GIP46 | TGGTACAGGCTGTGTTCCAT | 20 | 60 |
GIP47 | TACAACAGATCCGGCACGTA | 20 | 60 |
3. Experimental Section
4. Conclusions
Acknowledgments
References
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Arguel, M.-J.; Jaouannet, M.; Magliano, M.; Abad, P.; Rosso, M.-N. siRNAs Trigger Efficient Silencing of a Parasitism Gene in Plant Parasitic Root-Knot Nematodes. Genes 2012, 3, 391-408. https://doi.org/10.3390/genes3030391
Arguel M-J, Jaouannet M, Magliano M, Abad P, Rosso M-N. siRNAs Trigger Efficient Silencing of a Parasitism Gene in Plant Parasitic Root-Knot Nematodes. Genes. 2012; 3(3):391-408. https://doi.org/10.3390/genes3030391
Chicago/Turabian StyleArguel, Marie-Jeanne, Maëlle Jaouannet, Marc Magliano, Pierre Abad, and Marie-Noëlle Rosso. 2012. "siRNAs Trigger Efficient Silencing of a Parasitism Gene in Plant Parasitic Root-Knot Nematodes" Genes 3, no. 3: 391-408. https://doi.org/10.3390/genes3030391