TABLE 1.

Advantages and limitations of transient expression in rice seedlings.

Advantages
Transient expression in rice seedling represents the in-vivo conditions of intact rice cells.
It does not require expensive enzymes or specialized equipment or large amount of vector DNA like the methods involving protoplast or particle bombardment do.
The problem of protoplast bursting because of changes in osmoticum does not exist in transient expression of rice seedlings.
Localization of proteins in cells of different tissues of rice plant like root, coleoptile, epidermal cells and mesophyll cells of leaf can be studied which is not possible by methods involving protoplasts or particle bombardment.
Changes in protein localization on application of different abiotic stress or hormonal treatment can be studied by this method which is not possible by the method involving protoplast.
The first leaf of rice is thinner as compared to mature leaves and since they are grown at 20°C, the problem of light scattering by thick waxy cuticle which impedes high resolution fluorescent protein visualization is largely absent.
This method can be used to test the efficiency of gRNAs before raising CRISPR/CAS9 stable transgenics.
Disadvantages
The first leaf is not flat like a mature leaf and is convex in shape as it encircles the leaf sheath. As a result, problem of focal drift may arise if the tissue is not properly mounted on the slide.
If the seeds are not sterilized properly, infection can be a problem and can hamper healthy growth of rice seedlings.
Since 20°C is not the natural growth temperature, the growth of seedlings is sensitive to the water content in the phyta-jar. Germination rate is also altered with the age of the seeds with fresh seeds germinating faster than old seeds at 20°C.
Slight modifications of co-cultivation stage may be required if using different variety of seeds.