Extraction for plant hormones has been conducted utilizing a previously established protocol with minor modifications [6]. Briefly, leaves, roots, and stems were individually homogenized in liquid nitrogen using a TissueLyser II (Thermo Fisher Scientific). In total, 50 mg of the material was extracted with 1 ml cold 50% acetonitrile. The extract was then purified on a non-selective reversed-phase solid-phase extraction (RP-SPE) using an Oasis HLB cartridge (Waters). The column was activated with 100% methanol and ultrapure water, followed by equilibration with 50% acetonitrile. The sample was loaded onto the cartridge and flow-through collected. The residues of the target hormones were then eluted with 1 ml of 30% acetonitrile. The flow-through and eluted fractions were mixed and were evaporated to dryness in vacuum concentrator for 3 hours. Dried residuals were dissolved in 100 μl of 30% acetonitrile. The protocol was deposited in protocols.io (http://dx.doi.org/10.17504/protocols.io.bqy6mxze).
Copyright and License information: ©2021 Hashiguchi et al ©2021 Raffaella BalestriniThis is an open access article distributed under the terms of the , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ©2021 Raffaella BalestriniThis is an open access article distributed under the terms of the , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ©2021 Raffaella BalestriniThis is an open access article distributed under the terms of the , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.