Image_2_Optimizing RNAi-Target by Nicotiana benthamiana-Soybean Mosaic Virus System Drives Broad Resistance to Soybean Mosaic Virus in Soybean.TIF
Soybean mosaic virus (SMV) is a prevalent pathogen of soybean (Glycine max). Pyramiding multiple SMV-resistance genes into one individual is tedious and difficult, and even if successful, the obtained multiple resistance might be broken by pathogen mutation, while targeting viral genome via host-induced gene silencing (HIGS) has potential to explore broad-spectrum resistance (BSR) to SMV. We identified five conserved target fragments (CTFs) from S1 to S5 using multiple sequence alignment of 30 SMV genome sequences and assembled the corresponding target-inverted-repeat constructs (TIRs) from S1-TIR to S5-TIR. Since the inefficiency of soybean genetic transformation hinders the function verification of batch TIRs in SMV-resistance, the Nicotiana benthamiana-chimeric-SMV and N. benthamiana-pSMV-GUS pathosystems combined with Agrobacterium-mediated transient expression assays were invented and used to test the efficacy of these TIRs. From that, S1-TIR assembled from 462 bp CTF-S1 with 92% conservation rate performed its best on inhibiting SMV multiplication. Accordingly, S1-TIR was transformed into SMV-susceptible soybean NN1138-2, the resistant-healthy transgenic T1-plants were then picked out via detached-leaf inoculation assay with the stock-plants continued for progeny reproduction (T1 dual-utilization). All the four T3 transgenic progenies showed immunity to all the inoculated 11 SMV strains under individual or mixed inoculation, achieving a strong BSR. Thus, optimizing target for HIGS via transient N. benthamiana-chimeric-SMV and N. benthamiana-pSMV-GUS assays is crucial to drive robust resistance to SMV in soybean and the transgenic S1-TIR-lines will be a potential breeding source for SMV control in field.
History
References
- https://doi.org//10.1007/s11033-020-05583-4
- https://doi.org//10.1590/S0100-41582002000200005
- https://doi.org//10.1186/s13059-017-1381-1
- https://doi.org//10.1007/s10658-019-01870-4
- https://doi.org//10.1094/Phyto-69-467
- https://doi.org//10.1016/j.virusres.2005.03.020
- https://doi.org//10.3389/fpls.2020.00946
- https://doi.org//10.1038/cgt.2016.4
- https://doi.org//10.1016/j.virusres.2008.08.010
- https://doi.org//10.1007/s00122-015-2522-0
- https://doi.org//10.1094/PDIS-04-14-0405-RE
- https://doi.org//10.1094/MPMI-21-8-1015
- https://doi.org//10.1111/mpp.12644
- https://doi.org//10.1016/S1046-2023(03)00036-7
- https://doi.org//10.1016/B978-0-12-801246-8.00007-X
- https://doi.org//10.1007/BF02667740
- https://doi.org//10.1007/s00705-016-3123-1
- https://doi.org//10.1186/s13007-019-0442-8
- https://doi.org//10.1016/S1360-1385(02)02251-3
- https://doi.org//10.1007/s00438-017-1310-8
- https://doi.org//10.1007/s11816-016-0402-y
- https://doi.org//10.1007/s11816-013-0279-y
- https://doi.org//10.3390/microorganisms9040695
- https://doi.org//10.1094/PDIS-94-3-0351
- https://doi.org//10.3389/fpls.2017.00246
- https://doi.org//10.1371/journal.pone.0046451
- https://doi.org//10.1006/meth.2001.1262
- https://doi.org//10.3389/fpls.2019.01023
- https://doi.org//10.20546/ijcrbp.2016.307.004
- https://doi.org//10.3390/v5122931
- https://doi.org//10.1186/s13059-019-1678-3
- https://doi.org//10.1186/s13007-018-0343-2
- https://doi.org//10.1016/j.cub.2005.04.053
- https://doi.org//10.1093/nar/29.9.e45
- https://doi.org//10.1093/molbev/msx248
- https://doi.org//10.1016/j.virol.2009.07.007
- https://doi.org//10.5423/PPJ.2009.25.1.054
- https://doi.org//10.1371/journal.pone.0233366
- https://doi.org//10.1620/tjem.130.273
- https://doi.org//10.1016/S0042-6822(02)00018-1
- https://doi.org//10.1007/s00299-006-0186-6
- https://doi.org//10.3791/57258
- https://doi.org//10.1111/tra.12768
- https://doi.org//10.18388/abp.2006_3341
- https://doi.org//10.1016/j.coviro.2018.08.003
- https://doi.org//10.1111/pbi.13102
- https://doi.org//10.1016/j.virusres.2015.02.005
CC BY 4.0