Optimization and standardization of transient expression assays for gene functional analyses in strawberry fruits

Strawberry is increasingly used as a model plant for research on fruit growth and development. The transient gene manipulation (TGM) technique is widely used to determine the function of plant genes, including those in strawberry fruits. However, its reliable application for the precise identification of gene function has been difficult owing to the lack of conditional optimization. In this study, we found that successful transient gene manipulation requires optimization, with the vector type, temperature, and fruit developmental stage being three major factors determining success. Notably, we found that transient gene manipulation was feasible only from the large green fruit stage onwards, making it especially suitable for identifying genes involved in strawberry fruit ripening. Furthermore, we established a method called percentage difference of phenotype (PDP), in which the functional effect of a gene could be precisely and efficiently identified in strawberry fruits. This method can be used to estimate the functional effect of a gene as a value from 0 to 100%, such that different genes can be quantitatively compared for their relative abilities to regulate fruit ripening. This study provides a useful tool for accelerating research on the molecular basis of strawberry fruit ripening.
Crop genetics: genetic tools to study strawberry ripening Chinese researchers have optimized transient gene manipulation (TGM) to probe the function of genes in strawberries. In TGM, the expression of a gene is temporarily altered and the effect on plant development measured. TGM experiments can be conducted in days rather than the months needed for experiments involving stable transformation. A team led by Jia Wensuo of China Agriculture University evaluated how the use of TGM in strawberry fruits was affected by various conditions, including different delivery vectors, temperatures, time periods, and fruit stages. They found that the correct delivery vector, temperature, and fruit stage were crucial for the success of TGM. The team also developed a TGM analysis method to quantify and compare the contribution of different genes. The techniques developed in this study provide valuable tools for the genetic analysis of strawberry ripening.