Fabrication of plant array chip for high-throughput screening of arabidopsis thaliana

Abstract

Arabidopsis thaliana, which serves as a model for plant research, is used for various aspects of botanical studies. In particular, a more sophisticated and microscopic environment for the life of Arabidopsis is required due to the subdivision of the research field according to each growth stage. It is necessary to solve the spatial limitations of the methods of controlling and observing the existing germination and growth processes and to create a screening environment suitable for pursuing a more formalized method. In order to achieve this, an array type chip was developed using poly(dimethylsiloxane). While maintaining the existing solid batch type environment, 20 environmental conditions can be implemented on a single chip. This allows the germination and growth of the Arabidopsis seeds aligned in various environments to be tracked one by one over time. However, there is a limitation that only screening according to the initial medium environment can be performed. To improve this problem, we have developed a multi-well type plant array chip which has a solid medium form but can supply additional liquid medium as needed. In the new plant array chip, there are wells in which each Arabidopsis seeds are inserted independently, and holes are formed in the bottom of each well. This enables a substantial single seed study, and allows the addition of new materials, especially through holes in the bottom, to further diversify the media environment. Since the 384-well type plant array chip is a standard size, various modules can assemble with the chip. First, when the liquid medium is additionally supplied to the bottom hole, the plant hormone of different concentration is injected into each column or row of the array using a linear channel module to affect the germination of the Arabidopsis depending on the concentration of two hormones can be screened. In addition, a light gradient module, which can control the amount of light that affects germination, was able to create eight illuminations on one chip under the same light conditions. As a result, the germination rate could be screened according to the illuminance of 660 nm wavelength red light which affects germination.