Humidity assay for studying plant-pathogen interactions in miniature controlled discrete humidity environments with good throughput

Xu, Zhen and Jiang, Huawei and Sahu, Binod Bihari and Kambakam, Sekhar and Singh, Prashant and Wang, Xinran and Wang, Qiugu and Bhattacharyya, Madan K. and Dong, Liang (2016) Humidity assay for studying plant-pathogen interactions in miniature controlled discrete humidity environments with good throughput. Biomicrofluidics, 10 (3). ISSN 1932-1058

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Abstract

This paper reports a highly economical and accessible approach to generate different discrete relative humidity conditions in spatially separated wells of a modified multi-well plate for humidity assay of plant-pathogen interactions with good throughput. We demonstrated that a discrete humidity gradient could be formed within a few minutes and maintained over a period of a few days inside the device. The device consisted of a freeway channel in the top layer, multiple compartmented wells in the bottom layer, a water source, and a drying agent source. The combinational effects of evaporation, diffusion, and convection were synergized to establish the stable discrete humidity gradient. The device was employed to study visible and molecular disease phenotypes of soybean in responses to infection by Phytophthora sojae, an oomycete pathogen, under a set of humidity conditions, with two near-isogenic soybean lines, Williams and Williams 82, that differ for a Phytophthora resistance gene (Rps1-k). Our result showed that at 63% relative humidity, the transcript level of the defense gene GmPR1 was at minimum in the susceptible soybean line Williams and at maximal level in the resistant line Williams 82 following P. sojae CC5C infection. In addition, we investigated the effects of environmental temperature, dimensional and geometrical parameters, and other configurational factors on the ability of the device to generate miniature humidity environments. This work represents an exploratory effort to economically and efficiently manipulate humidity environments in a space-limited device and shows a great potential to facilitate humidity assay of plant seed germination and development, pathogen growth, and plant-pathogen interactions. Since the proposed device can be easily made, modified, and operated, it is believed that this present humidity manipulation technology will benefit many laboratories in the area of seed science, plant pathology, and plant-microbe biology, where humidity is an important factor that influences plant disease infection, establishment, and development.

Item Type:
Journal Article
Journal or Publication Title:
Biomicrofluidics
Additional Information:
Copyright 2016 American Institute of Physics. The following article appeared in Biomicrofluidics, 10, (3) 2016 and may be found at http://dx.doi.org/10.1063/1.4950998 This article may be downloaded for personal use only. Any other use requires prior permission of the author and the American Institute of Physics.
Uncontrolled Keywords:
/dk/atira/pure/subjectarea/asjc/3100/3104
Subjects:
ID Code:
79983
Deposited By:
Deposited On:
03 Oct 2016 13:30
Refereed?:
Yes
Published?:
Published
Last Modified:
07 Apr 2020 03:56