期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 20, 期 16, 页码 -出版社
MDPI
DOI: 10.3390/ijms20163976
关键词
agrobacterium; transient expression; virus-induced gene silencing (VIGS); tobacco rattle virus (TRV); cassava (Manihot esculenta)
资金
- National Key R&D Program of China [2018YFD1000500]
- National Natural Science Foundation of China [31760067]
- Scientific Research Foundation of Hainan University [kyqd1531]
- Crop Science Postgraduate Innovation Project of Hainan University Tropical Agriculture and Forestry College (College of Tropical Crops) [ZWCX2018005, ZWCX2018018]
Agrobacterium-mediated transient expression and virus-induced gene silencing (VIGS) are very useful in functional genomics in plants. However, whether these methods are effective in cassava (Manihot esculenta), one of the most important tropical crops, remains elusive. In this study, we used green fluorescent protein (GFP) and beta-glucuronidase (GUS) as reporter genes in a transient expression assay. GFP or GUS could be detected in the infiltrated leaves at 2 days postinfiltration (dpi) and were evidenced by visual GFP and GUS assays, reverse-transcription PCR, and Western blot. In addition, phytoene desaturase (PDS) was used to show the silencing effect in a VIGS system. Both Agrobacterium GV3101 and AGL-1 with tobacco rattle virus (TRV)-MePDS-infiltrated distal leaves showed an albino phenotype at 20 dpi; in particular, the AGL-1-infiltrated plants showed an obvious albino area in the most distal leaves. Moreover, the silencing effect was validated by molecular identification. Notably, compared with the obvious cassava mosaic disease symptom infiltrated by African-cassava-mosaic-virus-based VIGS systems in previous studies, TRV-based VIGS-system-infiltrated cassava plants did not show obvious virus-induced disease symptoms, suggesting a significant advantage. Taken together, these methods could promote functional genomics in cassava.
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