title: MR219_Control
organism: Oryza sativa Indica Group
description: Protocols: Homogeneus seeds of all eleven varieties were sowed in sterilized sand, kept at 25-30°C for three weeks to germinate and grown. Similarly and healthy grown 21 days old seeds were selected and moved to different plastics pots containing soil medium under well-watered (control) and drought stress condition in the glasshouse. Seedlings of both experiments including well-watered (control) and drought stress were arranged in a complete randomized design in a green house using pots 1 m in length and 40 cm in diameter. The mercury-type tensiometer at 15 cm and 30 cm of soil depths was performed to monitor soil water potential and determine time of re-watering. Moisture content of soil was monitored every 3 days to differentiate drought stress quantitatively. Then growth response of plants were recorded by measuring height, total leaf area per plant, and weekly recording electrophysiology the potential differences of leaves. Three seedlings of each variety were harvested from respective drought stress treatments with three untreated rice seedlings as controls. The whole leaves were collected for analysis. The treatment of drought stress was imposed from vegetative stage and continued to panicle initiation Homogeneus seeds of all eleven varieties were sowed in sterilized sand, kept at 25-30°C for three weeks to germinate and grown The harvested leaves were frozen immediately in liquid nitrogen and kept at -80°C for downstream processing. Total RNA was extracted from the both treated and control plant leaves using the Trizol reagent (FavorPrep™ Tri-RNA Reagent, Taiwan) following manufacturer instructions and treated with DNase I (New England Biolabs, Hitchin, UK).. The mRNA from both control and treated samples were purified using PolyATtract® mRNA Isolation Systems III (Promega, USA) according to the manufacturer instructions. Isolated mRNA from both treated and control samples were used to synthesis cDNA library for each variety. The First Strand Super Script II (Invitrogen, USA) was performed to generate first-strand cDNAs from both samples for two rice varieties, separately. The Low Input Library Prep Kit (Clontech Laboratories, Inc., USA) was performed to construct cDNA library for Illumina sequencing according to manufacturer instruction. Then the PCR products of two varieties were analysed using 2% agarose gel and TAE buffer. Verification of cDNA libraries were carried out through determining the average molecule length using the Agilent 2100 bioanalyzer (Agilent DNA 1000 Reagents), then quantification of cDNA libraries were done performing real-time PCR (TaqMan Probe) in Beijing Genomics Institute, Shenzhen, China (BGI).
attributes.tag: ENA first public
attributes.value: 2017-07-27
attributes.tag: ENA last update
attributes.value: 2017-07-03
attributes.tag: External Id
attributes.value: SAMEA104150501
attributes.tag: INSDC center alias
attributes.value: University Putra Malaysia
attributes.tag: INSDC center name
attributes.value: University Putra Malaysia
attributes.tag: INSDC first public
attributes.value: 2017-07-27T17:40:14Z
attributes.tag: INSDC last update
attributes.value: 2017-07-03T10:20:35Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-5402:MR219_Control
attributes.tag: age
attributes.value: 4
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: long-grained rice
attributes.tag: cultivar
attributes.value: MR219
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: growth condition
attributes.value: Glasshouse, using pots 1 m in length and 40 cm in diameter
attributes.tag: organism part
attributes.value: leaf
attributes.tag: sample name
attributes.value: E-MTAB-5402:MR219_Control
attributes.tag: sampling site
attributes.value: Glasshouse (25933.2N 1014318.3E) in the Universiti Putra Malaysia