Cultivar Chinese Spring (CS) was used for TaABCB1 gene cloning, Nullisomic-tetrasomic (NT) and Ditelosomic (DT) lines in CS background58 were used for mapping. Wheat cultivar Indian, lacking any known rht mutations was chosen for the Single-Strand Conformation Polymorphism (SSCP), real-time expression analysis, and Virus Induced Gene Silencing (VIGS). All plants were propagated in 6-inch pots using Sunshine#1 potting mixture (SunGro Horticulture, Bellevue, WA, USA) supplemented with 14 g Nutricote 14–14–14 fertilizer (Plantco Inc., Brampton, Ontario, Canada). Plants supplied with equal amount of soil and water, were grown in a Conviron PGR15 growth chamber equipped with high-intensity discharge lamps at 23 °C temperature and 16 hr light (500–700 μmol m−2 s−1).
Pgr15 growth chamber
Manufactured by Conviron
Sourced in Canada
The PGR15 growth chamber is a controlled-environment system designed for plant growth research. It provides a precisely regulated environment with adjustable temperature, lighting, and humidity settings to support the cultivation of various plant species. The PGR15 offers a consistent and reliable platform for controlled scientific experiments and plant growth studies.
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Lab products found in correlation
15 protocols using pgr15 growth chamber
1
Wheat Gene Cloning and Characterization
2
Optimized N. benthamiana Growth Protocol
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Experiments took place at Laval University in Québec City, QC, Canada (46°46′ N, 71°16′ W). N. benthamiana plants were grown from seeds kindly provided by Medicago Inc. (Québec City, QC). The seeds were soaked in water for 24 h at 20°C and then placed in peat moss substrate for germination in a PGR15 growth chamber (Conviron, Winnipeg MB, Canada). Seedlings were selected after 2 weeks based on uniformity, transplanted in 350-ml plastic pots filled with peat moss substrate, and let to grow in greenhouse for an additional 3 weeks at a culture density of 33 plants·m−2 under different cultural conditions before leaf agroinfiltration (see below). Day and night temperatures in the greenhouse were maintained at 29 and 27°C, respectively. Supplemental lighting was provided by 400-W high-pressure sodium lamps installed above the plant canopy (P.L. Light Systems, Beamsville ON, Canada). The plants were irrigated as needed with the Plant-Prod 12-2-14 Optimum complete nutrient solution supplemented with the Plant-Prod Chelated Micronutrient Mix (Plant Products, Laval QC, Canada). Electrical conductivity in the nutrient solution was maintained at 1.6 dS·m−1 for 1 week after transplantation, and then increased at 2.6 and 3.6 dS·m−1 for the second and third weeks, respectively.
3
Arabidopsis Seedling Growth and Tissue Dissection
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Arabidopsis seeds were surface sterilized in 12 % (w/v) bleach/0.1 % Tween 20 solution and washed extensively with sterile distilled water. Seeds were vernalized for 3 days at 4 °C in water, sown in two parallel rows on MS/agar plates (30 mM sucrose, 4.2 g Murashige and Skoog medium, and 0.8 % Phytagar, pH 5.8) covered with a 100 micron nylon membrane (Genesee Scientific). Seedlings were grown on vertical plates in the Conviron PGR15 growth chamber (12:12 h. light:dark, 21 °C, 50 % humidity, and 250 μmol/m2/s light intensity). Roots and leaves of 1 week old seedlings were dissected using surgical blade, flash-frozen in liquid nitrogen, and stored at −80 °C.
4
Abiotic Stress Responses in Brachypodium
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Brachypodium distachyon control plants were grown at 22°C with 16 hours light and 8 hours dark in a controlled environment growth room. Abiotic stress conditions included cold, heat, salt, and drought. All treatments were conducted with a light intensity of 200 µmol photons m−2s−1. For the heat experiments, Brachypodium plants were placed in a Conviron PGR 15 growth chamber at 42°C. Cold treatments were conducted in a walk-in cold room maintained at 4°C. Salt stress (soil saturation with 500 mM NaCl) and drought (simulated by removing plants from soil and placing them on paper towels to desiccate) treatments were conducted under the same light and temperature as the control samples. Three-week-old Brachypodium plants were placed under the respective conditions two hours after dawn (10 a.m.). Leaves and stems (total above ground tissues) from individual plants were collected at 1, 2, 5, 10, and 24 hours after exposure to the abiotic stress.
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Brachypodium distachyon control plants were grown at 22°C with 16 hours light and 8 hours dark in a controlled environment growth room. Abiotic stress conditions included cold, heat, salt, and drought. All treatments were conducted with a light intensity of 200 µmol photons m−2s−1. For the heat experiments, Brachypodium plants were placed in a Conviron PGR 15 growth chamber at 42°C. Cold treatments were conducted in a walk-in cold room maintained at 4°C. Salt stress (soil saturation with 500 mM NaCl) and drought (simulated by removing plants from soil and placing them on paper towels to desiccate) treatments were conducted under the same light and temperature as the control samples. Three-week-old Brachypodium plants were placed under the respective conditions two hours after dawn (10 a.m.). Leaves and stems (total above ground tissues) from individual plants were collected at 1, 2, 5, 10, and 24 hours after exposure to the abiotic stress.
5
Maize Seedling Etiolation Protocol
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Etiolated B73 maize (Zea mays) seedlings were prepared as follows. Kernels were imbibed for 24 h in room temperature water before being sown in moist ProMix (Sungro) soil. For 3 days, or until coleoptile emergence, plants were kept in a Conviron PGR15 growth chamber under a 12 h light (242 µmol)/12 h dark cycle, where temperatures and relative humidity were maintained at 26 °C/20 °C and 80%/60%, respectively. Plants were subsequently grown in complete darkness at 25 °C and 60% relative humidity for up to 2 weeks or until full expansion of the second true leaves.
6
Cultivation of Isodon and Nicotiana Plants
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Isodon rubescens plants were purchased from the Horizon Herbs nursery (Williams, OR), and cultivated in a greenhouse under ambient photoperiod and ~22/17°C day/night temperature. Nicotiana benthamiana plants were cultivated from seeds in a Conviron PGR15 growth chamber with 16 h light at 80 μmol m-2 sec-1 irradiance and a 24/17°C day/night temperature cycle.
7
Arabidopsis thaliana Seedling Growth Assay
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Arabidopsis thaliana ecotype Columbia 0 seeds were sterilized in a solution of 50% (v/v) bleach solution with 0.1% Tween 20 for 10 minutes, then rinsed extensively with sterile deionized water. Sterilized 100 micron nylon mesh was placed on top of solidified medium (30 mM sucrose, 4.2 g Murashige and Skoog medium (PhytoTech Labs), and 0.8 % Phytagar, pH adjusted to 5.8 with KOH) in large petri plates (Genesee Scientific). Following a 4 day vernalization period in water, sterilized seeds were suspended in a 0.75% agar solution and were transferred to each plate in two dense rows (~500 seeds per row) in a laminar flow hood under sterile conditions. Seedlings were grown vertically in a Conviron PGR15 growth chamber at 21C under a 12:12 hour light:dark cycle (50% humidity, and 250 mol/m2/s light intensity). At seven days, the seedlings were harvested and divided into three batches. For each batch, seedlings were dissected using a surgical blade and the root tissue was separated from the shoot tissue. For our purposes here, shoot tissues include the hypocotyl, cotelydons, and any stems and true leaves that had developed by the time of collection. Each batch of seedlings was handled identically, and harvested tissues were flash-frozen in liquid nitrogen, then stored at -80C until needed for the following protocols.
8
Etiolated Maize Seedling Cultivation
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Plant materials and growth conditions Etiolated B73 maize (Zea mays) seedlings were prepared as follows. Kernels were imbibed for 24 h in room temperature water before being sown in moist ProMix (Sungro) soil. For three days, or until coleoptile emergence, plants were kept in a Conviron PGR15 growth chamber under a 12 h light (242 µmol)/12 h dark cycle, where temperatures and relative humidity were maintained at 26°C/20°C and 80%/60%, respectively. Plants were subsequently grown in complete darkness at 25°C and 60% relative humidity for up to two weeks or until full expansion of the second true leaves.
9
Growth Conditions for Darnel Ryegrass
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Lolium temulentum L. (Lt, Darnel ryegrass) cv. Ceres seeds were planted, five seeds per pot, in TSD4 square pots (8.8 cm × 8.8 cm × 10 cm, 540 mL volume; McConkey Co., Sumner, WA) with Sun Gro Professional MM840 PC RSi (Sun Gro Horticulture, Hubbard, OR). Lolium plants were grown in Conviron PGR14 or PGR15 growth chambers (Conviron, Winnipeg, Canada), under 14 h photoperiods at 23 °C day and 18 °C night temperatures and fertilized with Technigro 20-18-20 all-purpose fertilizer (Sun Gro Horticulture, Hubbard, OR) weekly. All experiments were conducted using seeds from increases of Lolium temulentum cv. Ceres seeds originally provided by Dr. Lloyd T. Evans (CSIRO, Canberra Australia) in 2001.
10
Quantitative Expression Analysis of Flowering Genes
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Plants were grown for 3 weeks in a PGR15 growth chambers (Conviron) under long day conditions (16 h light/8 h dark) at a temperature of 20 °C during light period and 18 °C during dark. Samples were collected 4 h after the lights were turned on (Zeitgeber Time 4). RNA was extracted from leaves using the Spectrum Plant Total RNA Kit (Sigma-Aldrich). Quantitative PCR was performed using SYBR Green and a 7500 Fast Real-Time PCR system (Applied Biosystems). Primers for ACTIN are described in Distelfeld et al. (2009 (link)) and primers for ZCCT-B2 and ZCCT-D2 are described in Table S1. Transcript levels are expressed as linearized fold-ACTIN levels calculated by the formula 2(ACTIN CT – TARGET CT) ± SE of the mean. The resulting number indicates the ratio between the initial number of molecules of the target gene and the number of molecules of ACTIN.
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