New method for arbuscular mycorrhizal fungus spore separation using a microfluidic device based on manual temporary flow
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SHORT NOTE
New method for arbuscular mycorrhizal fungus spore separation using a microfluidic device based on manual temporary flow diversion K. Srisom 1 & P. Tittabutr 2 & N. Teaumroong 2 & Y. Lapwong 3 & R. Phatthanakun 4 & S. Sirivisoot 1 & P. Kuntanawat 2 Received: 5 December 2019 / Accepted: 19 August 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Arbuscular mycorrhizal fungi are beneficial components often included in biofertilizers. Studies of the biology and utilization of these fungi are key to their successful use in the biofertilizer industry. The acquisition of isolated spores is a required step in these studies; however, spore quality control and spore separation are bottlenecks. Filtered and centrifuged spores have to be handpicked under a microscope. The conventional procedure is skill-demanding, labor-intensive, and time-consuming. Here, we developed a microfluidic device to aid manual separation of spores from a filtered and centrifuged suspension. The device is a single spore streamer equipped with a manual temporary flow diversion (MTFD) mechanism to select single spores. Users can press a switch to generate MTFD when the spore arrives at the selection site. The targeted spore flows in a stream to the collection chamber via temporary cross flow. Using the device, spore purity, the percentage of spore numbers against the total number of particles counted in the collecting chamber reached 96.62% (median, n = 10) which is greater than the spore purity obtained from the conventional method (88.89% (median, n = 10)). Keywords Arbuscular Mycorrhiza . Spore separation . Microfluidic device . Manual temporary flow diversion mechanism
Introduction Arbuscular mycorrhizal fungi (AMF) live symbiotically with plant roots and are known to benefit plant hosts in multiple ways (Amaya-Carpio et al. 2009; Nouri et al. 2014). (Göhre and Paszkowski 2006; Akhtar et al. 2011; Nouri et al. 2014; Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00572-020-00986-4) contains supplementary material, which is available to authorized users. * P. Kuntanawat [email protected] 1
Nanoscience and Nanotechnology Graduate Program, Faculty of Science, King Mongkut’s University of Technology Thonburi, 126 Pracha Uthit Rd, Bang Mod, Thung Khru, Bangkok 10140, Thailand
2
Institute of Agricultural Technology, Suranaree University of Technology, 111 University Avenue, Muang District, Nakhon Ratchasima 30000, Thailand
3
School of Life Science, University of Technology Sydney, PO Box 123, Broadway, Sydney, NSW 2007, Australia
4
Synchrotron Light Research Institute, 111 University Avenue, Muang District, Nakhon Ratchasima 30000, Thailand
Tamayo et al. 2014; Gehring et al. 2017). These capabilities of AMF are exploited in crop production. The spores of AMF are added as an active ingredient in biofertilizer. Biofertilizer is a major industry with great potential which had a market size in 2017 of around USD 202.1 million (Gosling et al. 2006; El Kinany e
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