Sprout vacuum-infiltration: a simple and efficient agroinoculation method for virus-induced gene silencing in diverse solanaceous species.
Virus-induced gene silencing (VIGS) is a robust technique for identifying the functions of plant genes. Tobacco rattle virus (TRV)-mediated VIGS has been commonly used in many plants. In order to overcome the limitations of existing agroinoculation methods, we report an easy and effective method of agroinoculation for virus-induced gene silencing-sprout vacuum-infiltration (SVI). Using sprout vacuum-infiltration, we have successfully silenced the expression of phytoene desaturase and Mg-protoporphyrin chelatase genes in four important solanaceous crops, including tomato, eggplant, pepper, and Nicotiana benthamiana. The gene-silenced phenotypes are conspicuous in 1-week-old plants. The method is simple, low cost and rapid compared to other techniques such as leaf infiltration or agrodrench. It may be more practical for studying gene function in the early stages of plant growth. An important aspect of SVI is that it will be used for high-throughput VIGS screens in the future. SVI will be an effective tool to overcome the limitations of current inoculation methods and to facilitate large-scale VIGS analysis of cDNA libraries.


CONCLUSIONS
SVI is a simple, low cost agroinoculation method for VIGS. It is practical for studying the function of genes expressed in early stages of plant growth and high-throughput VIGS screens.
Laboratory of Food Biotechnology, College of Food Science and Nutritional Engineering, China Agricultural University, No. 17 Qinghua Donglu Road, HaiDian District, Beijing, 100083, China.
Hua-xue
HX
Yan
Da-qi
DQ
Fu
Ben-zhong
BZ
Zhu
Hai-ping
HP
Liu
Xiao-ying
XY
Shen
Yun-bo
YB
Luo
D060054:Agrobacterium / Q000378:metabolism*; D002734:Chlorophyll / Q000378:metabolism; D035264:Flowers / Q000821:virology; D005638:Fruit / Q000821:virology; D020868:Gene Silencing; D005821:Genetic Techniques; D018525:Germination; D018551:Solanum lycopersicum / Q000821:virology; D010088:Oxidoreductases / Q000378:metabolism; D010641:Phenotype; D018515:Plant Leaves / Q000821:virology; D010942:Plant Viruses / Q000378:metabolism*; D011995:Recombination, Genetic / Q000235:genetics; D036226:Seedlings / Q000821:virology; D019657:Solanaceae / Q000254:growth & development / Q000821:virology*; D013045:Species Specificity; D014618:Vacuum
D002734:Chlorophyll; D010088:Oxidoreductases; C022398:phytoene dehydrogenase