tissue: 1.0-1.5 cm lateral root primordium zone (LRZ) Stage: V4 stage inbred line: Q319
Treatment protocol
The 1.0-1.5 cm lateral root primordium zone of maize were harvested at 2 days and 8 days, respectively, after transfer into the SP/LP nutrient solutions.Each biological repeat contains segments from 15~20 plants.
Growth protocol
Maize (Zea mays L.) inbred line Q319 was used in this study. Seeds of the maize inbred line Q319 were surface sterilized and held at 28°C in darkness. Seedlings (4 days old) were transferred to a sufficient phosphate (SP, 1,000 μM KH2PO4) solution (Ca(NO3)2.4H2O 2 mM, NH4NO3 1.25 mM, KCl 0.1 mM, K2SO4 0.65 mM, MgSO4 0.65 mM, H3BO3 10.0 mM, (NH4)6Mo7O24 0.5 mM, MnSO4 1.0 mM, CuSO4.5H2O 0.1 mM, ZnSO4.7H2O 1.0 mM, Fe-EDTA 0.1 mM), allowed to grow for 4 days (plants with 2–3 leaves), and endosperm was then removed carefully. After 2-3 days of re-culturing in SP nutrient solutions, half of the seedlings were transplanted into a low phosphate (LP, same composition as the SP solution, except that 5 μM KH2PO4 and 1 mM KH2PO4 were replaced with 1 mM KCl) nutrient solution. The plants were grown under a 32°C/25°C (day/night) temperature regime at a photon flux density (PFD) of 700 μmol m-2 s-1 with a 14 h/10 h light/dark cycle in a greenhouse with approximately 65% relative humidity.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted from the frozen samples using the method of McCarty (1986) and purified using Qiagen RNeasy MinElute columns and buffers (Qiagen Cat # 74204).
Label
Cy5
Label protocol
1.5 µg of the purified RNA was reverse transcribed, amplified and labeled using Cy5 or Cy3 (Cy3/5™ Mono-Reactive Dye Pack, Cat#PA23001) with the Aminoallyl Message Amp II kit (Ambion, Cat# 1753) according to the manufacturer’s instructions.Unlabelled primers and dyes were removed using Qiagen RNeasy MinElute columns and buffers (Qiagen Cat # 74204).
Channel 2
Source name
control maize Q319 roots clutrued in normal conditions
tissue: 1.0-1.5 cm lateral root primordium zone (LRZ) Stage: V4 stage inbred line: Q319
Treatment protocol
The 1.0-1.5 cm lateral root primordium zone of maize were harvested at 2 days and 8 days, respectively, after transfer into the SP/LP nutrient solutions.Each biological repeat contains segments from 15~20 plants.
Growth protocol
Maize (Zea mays L.) inbred line Q319 was used in this study. Seeds of the maize inbred line Q319 were surface sterilized and held at 28°C in darkness. Seedlings (4 days old) were transferred to a sufficient phosphate (SP, 1,000 μM KH2PO4) solution (Ca(NO3)2.4H2O 2 mM, NH4NO3 1.25 mM, KCl 0.1 mM, K2SO4 0.65 mM, MgSO4 0.65 mM, H3BO3 10.0 mM, (NH4)6Mo7O24 0.5 mM, MnSO4 1.0 mM, CuSO4.5H2O 0.1 mM, ZnSO4.7H2O 1.0 mM, Fe-EDTA 0.1 mM), allowed to grow for 4 days (plants with 2–3 leaves), and endosperm was then removed carefully. After 2-3 days of re-culturing in SP nutrient solutions, half of the seedlings were transplanted into a low phosphate (LP, same composition as the SP solution, except that 5 μM KH2PO4 and 1 mM KH2PO4 were replaced with 1 mM KCl) nutrient solution. The plants were grown under a 32°C/25°C (day/night) temperature regime at a photon flux density (PFD) of 700 μmol m-2 s-1 with a 14 h/10 h light/dark cycle in a greenhouse with approximately 65% relative humidity.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted from the frozen samples using the method of McCarty (1986) and purified using Qiagen RNeasy MinElute columns and buffers (Qiagen Cat # 74204).
Label
Cy3
Label protocol
1.5 µg of the purified RNA was reverse transcribed, amplified and labeled using Cy5 or Cy3 (Cy3/5™ Mono-Reactive Dye Pack, Cat#PA23001) with the Aminoallyl Message Amp II kit (Ambion, Cat# 1753) according to the manufacturer’s instructions.Unlabelled primers and dyes were removed using Qiagen RNeasy MinElute columns and buffers (Qiagen Cat # 74204).
Hybridization protocol
Hybridizations were performed according to the protocols at the website of the Maize Oligonucleotide Array Project.
Scan protocol
Scanned on an LuxScan-10KA (SN100-0094), Images were quantified using GENEPIX 6.0 software (version A.6.0).
Data processing
LOWESS normalized, background subtracted data. TM4 software was used.
