Analysis of gene functions by a syringe infiltration method of VIGS

Author： Wang X. Zhu H. Shao Y. Chen A. Ma Y. Luo Y. Zhu B.

ISSN： 1021-4437

Source： Russian Journal of Plant Physiology, Vol.57, Iss.4, 2010-07, pp. : 590-597

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Abstract

Virus-induced gene silencing (VIGS) is a powerful tool for characterization of functional genes in tomato. In this study, we improved the syringe infiltration method on detached tomato fruits. With PDS (phytoene desaturase gene) used as a reporter gene, harvested mature green tomato fruits were syringe-infiltrated with agrobacterium strain GV3101 containing pTRV1 and pTRV2-LePDS, which resulted in color changing on the fruit surface. The changes in pigments and transcript abundance of PDS were compared between silencing sectors and control. In addition, factors of environmental conditions, such as temperature and humidity, were optimized for gene silencing, and other factors, including volume of agrobacterial culture, were also analyzed. Furthermore, this silencing method was confirmed by silencing LeRIN and LeACS2, which influenced fruit ripening process as positively regulating genes, and LeCTR1, which took part in ethylene signal transduction as a negatively regulating gene. Transcripts of LeRIN, LeCTR1, and LeACS2 were measured in the following analysis of gene silencing. All results indicated that an infiltration-based method is efficient for gene characterization during development and ripening of tomato fruits.