Journal: Current Protocols

Article Title: A Scalable Hydroponic System for Precise Nutrient and Stress Studies in Plants

doi: 10.1002/cpz1.70182

Figure Lengend Snippet: Overview of the hydroponic system workflow and associated experimental protocols. This schematic illustrates the sequential steps involved in establishing the modular hydroponic culture system for Arabidopsis thaliana , including preparation of agarose‐based planting units, seed sterilization and sowing, germination under controlled humidity, seedling acclimation to ambient conditions, and long‐term nutrient treatments.

Article Snippet: 1% agarose solution (see recipe) H 2 O, molecular‐grade (Milli‐Q water) Nutrient medium (see recipe) Surface‐sterilized Arabidopsis thaliana seeds (see recipe) 8‐strip PCR tubes (Genesee Scientific, cat. no. 27426) Beakers (for media preparation) Microwave (for agarose melting) Scissors (for trimming seedling roots) Pipette tip box and rack (Trupoint Pipet Tips, cat. no. 714715) Inoculating loops (Fisherbrand, cat. no. 22363598) Growth chamber, set to optimal Arabidopsis germination conditions (e.g., 22°C, 16 hr light/8 hr dark photoperiod) pH meter (Thermo Scientific Orion Star A2140, or equivalent)

Techniques:

Journal: Current Protocols

Article Title: A Scalable Hydroponic System for Precise Nutrient and Stress Studies in Plants

doi: 10.1002/cpz1.70182

Figure Lengend Snippet: Modular hydroponic culture system for Arabidopsis thaliana utilizing a 96‐well pipette tip box platform. ( A ) Preparation of the planting units: (1) remove the caps from 8‐strip PCR tubes; (2) fill each tube with 1% agarose solution; and (3) once solidified, trim the bottom ends of the tubes to allow root penetration into the liquid medium. ( B ) Assembled growth system: the modified PCR tubes are inserted into the wells of a standard 96‐well pipette tip box filled with nutrient solution. ( C ) Seed sowing: sterilized Arabidopsis seeds (1‐2 per tube) are sown directly onto the agarose surface. ( D ) Germination phase: the tip box is covered (e.g., with a transparent lid or Parafilm) to maintain high humidity until germination occurs. ( E ) Acclimation and early development: at 10‐15 days after sowing (DAS), the cover is removed to allow seedlings to adapt to ambient conditions. ( F ) Long‐term cultivation: Robust seedlings (8‐10 per experiment) are transferred to a fresh tip box containing new nutrient medium to support continued growth and experimental treatments. ( G ) Representative growth: phenotypic appearance of wild‐type Arabidopsis plants at 18 DAS under standard nutrient conditions. ( H ) Tissue accessibility: example of shoot and root tissues harvested from the same plant shown in ( G ), demonstrating the system's suitability for precise organ‐specific sampling and downstream analyses.

Article Snippet: 1% agarose solution (see recipe) H 2 O, molecular‐grade (Milli‐Q water) Nutrient medium (see recipe) Surface‐sterilized Arabidopsis thaliana seeds (see recipe) 8‐strip PCR tubes (Genesee Scientific, cat. no. 27426) Beakers (for media preparation) Microwave (for agarose melting) Scissors (for trimming seedling roots) Pipette tip box and rack (Trupoint Pipet Tips, cat. no. 714715) Inoculating loops (Fisherbrand, cat. no. 22363598) Growth chamber, set to optimal Arabidopsis germination conditions (e.g., 22°C, 16 hr light/8 hr dark photoperiod) pH meter (Thermo Scientific Orion Star A2140, or equivalent)

Techniques: Transferring, Stripping Membranes, Modification, Sampling

Journal: Current Protocols

Article Title: A Scalable Hydroponic System for Precise Nutrient and Stress Studies in Plants

doi: 10.1002/cpz1.70182

Figure Lengend Snippet: Split‐root hydroponic system using a 96‐well pipette tip box platform for differential nutrient treatment. ( A ) Schematic overview of the split‐root experimental procedure employing the modular hydroponic setup. ( B ) Twenty‐day‐old Arabidopsis thaliana plants (DAG 20) were carefully removed from the initial growth system, and their root systems were gently divided into two equal halves using sterile forceps. Each root half was then transferred into separate tip boxes containing either phosphate‐sufficient (+P) or phosphate‐deficient (–P) nutrient solutions. Aluminum foil was folded and shaped into a stable support stand, with custom perforations to securely hold the PCR tubes in an upright position, thereby maintaining root orientation and ensuring contact with the respective medium. This setup enables spatially distinct nutrient treatments while preserving a unified shoot, facilitating the investigation of local versus systemic signaling responses.

Article Snippet: 1% agarose solution (see recipe) H 2 O, molecular‐grade (Milli‐Q water) Nutrient medium (see recipe) Surface‐sterilized Arabidopsis thaliana seeds (see recipe) 8‐strip PCR tubes (Genesee Scientific, cat. no. 27426) Beakers (for media preparation) Microwave (for agarose melting) Scissors (for trimming seedling roots) Pipette tip box and rack (Trupoint Pipet Tips, cat. no. 714715) Inoculating loops (Fisherbrand, cat. no. 22363598) Growth chamber, set to optimal Arabidopsis germination conditions (e.g., 22°C, 16 hr light/8 hr dark photoperiod) pH meter (Thermo Scientific Orion Star A2140, or equivalent)

Techniques: Transferring, Sterility, Preserving

Journal: Current Protocols

Article Title: A Scalable Hydroponic System for Precise Nutrient and Stress Studies in Plants

doi: 10.1002/cpz1.70182

Figure Lengend Snippet: Phenotypic response of Arabidopsis thaliana to phosphorus deficiency under hydroponic conditions. Arabidopsis seedlings were initially grown for 12 days after sowing (DAS 12) under nutrient‐sufficient conditions. Following this period, plants were gently rinsed with distilled water to remove residual medium and subsequently transferred to either phosphorus‐sufficient (+P) or phosphorus‐deficient (–P) hydroponic media. After 8 days of treatment, marked phenotypic differences were observed. The plant on the left represents the control (+P), exhibiting normal vegetative growth, while the plant on the right was exposed to phosphorus‐limiting conditions, displaying growth retardation and anthocyanin accumulation. White scale bar = 1 cm.

Article Snippet: 1% agarose solution (see recipe) H 2 O, molecular‐grade (Milli‐Q water) Nutrient medium (see recipe) Surface‐sterilized Arabidopsis thaliana seeds (see recipe) 8‐strip PCR tubes (Genesee Scientific, cat. no. 27426) Beakers (for media preparation) Microwave (for agarose melting) Scissors (for trimming seedling roots) Pipette tip box and rack (Trupoint Pipet Tips, cat. no. 714715) Inoculating loops (Fisherbrand, cat. no. 22363598) Growth chamber, set to optimal Arabidopsis germination conditions (e.g., 22°C, 16 hr light/8 hr dark photoperiod) pH meter (Thermo Scientific Orion Star A2140, or equivalent)

Techniques: Control