Chloroplast Function Database II

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ID: An alphanumeric term identifying the code of each tagged line.

Disrupt locus: A genetic locus mapped on the basis of the flanking sequence of the tagged element.

Description: Descriptions are based on the definition accompanying TAIR gene annotation records.

Phenotype: The phenotypes of the 3-week-old seedlings of each line grown under a constant condition (approximately 75 µmol photon·m-2·s-1 at 22°C under a 16 h light/8 h dark cycle) on agar plates. We classified the mutants that exhibited phenotypes on the basis of their color or morphology into several phenotypic categories such as albino, pale green, variegated, and morphological abnormalities.

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Germination test

Condition

Generation: We examined the hereditary behavior of resistance to antibiotics of 3 generations (1 line of the 1st generation and 8 lines of the 2nd and 3rd generations). In this study, we defined the seeds of tagged lines, excluding RIKEN Ds-tagged lines, that could be obtained from several international seed stock centers as the F1 generation. In fact, the RIKEN Ds-tagged lines include seeds of the F3 generation.

Culture medium: The seeds were plated on Murashige & Skoog medium (MS) agar plates containing 1% sucrose with or without appropriate antibiotics. The Ds transposon of the RIKEN tagged lines contains the bacterial aph4 gene, which facilitates hygromycin (hyg) selection (Ito et al., 2002), whereas the Ds transposon of CSHL lines and the JIC GT lines contain the bacterial nptII gene, which allows for kanamycin (Km) selection (Sundaresan et al., 1995). In addition, the Spm transposon of the JIC SM lines contains the bacterial bar gene, which allows for selection by using the herbicide bialaphos (Tissier et al., 1999).

Results

Progeny: The offspring obtained from the parent line by self-fertilization. The seeds were harvested from 8 parent plants of transposon-tagged lines (A-H) or 20 parent plants of T-DNA-tagged lines (1-20) and observed their phenotypes. The data were excluded from the list if the seeds did not germinate or the plants did not grow on the soil.

Ratio: For the transposon-tagged lines, we obtained segregation data for each line using selection media, where (x:y:z) indicates (number of plants with visible phenotypes: number of plants showing resistance to antibiotics: number of plants showing sensitivity to antibiotics).

Test: For the T-DNA-tagged lines, we determined the genotype of each plant by performing PCR analysis with specific primers and T-DNA internal primers.

Abbreviations: HM, homozygote; HT, heterozygote; WT, wild-type

Trait: The phenotype classification of mutants.

Photos: We took 2 photographs (at different magnifications) of the mutants that exhibited visible phenotypes.

Remarks: Additional comments on the mutant stock.

RIKEN Plant Science Center