From bf6ad06d38a8a2cdb9afef38b6aebeeca4390bdc Mon Sep 17 00:00:00 2001 From: Gesa Helmsorig <gesa.helmsorig@hhu.de> Date: Wed, 13 Nov 2024 12:28:16 +0000 Subject: [PATCH] Update 2 files - /studies/generating_lwd1_mutants/protocols/plant_material_growth_conditions.md - /studies/generating_lwd1_mutants/README.md --- studies/generating_lwd1_mutants/README.md | 10 ++++++---- .../plant_material_growth_conditions.md | 20 ++++++------------- 2 files changed, 12 insertions(+), 18 deletions(-) diff --git a/studies/generating_lwd1_mutants/README.md b/studies/generating_lwd1_mutants/README.md index 5b27823..b1c8549 100644 --- a/studies/generating_lwd1_mutants/README.md +++ b/studies/generating_lwd1_mutants/README.md @@ -1,10 +1,10 @@ -**Generating *lwd1* mutants** +## Generating *lwd1* mutants Using CRISPR-Cas9 to generate *lwd1* mutants to confirm *LWD1* as the gene underlying the *eam7* locus. GP-fast embryos were transformed with vectors carrying sgRNAs that target the start or the end of the *LWD1* coding sequence. The resulting T0 plants were genotyped to screen for different induced mutations. Promising T1 plants were sown, and genotyped again. From this, three different homozygous mutants were selected for gene expression analysis and phenotyping: *lwd1-26*, *lwd1-390* and *lwd1-402*. -**study overview** +**Study overview:** - WT transformation plants (GP-fast) + input: WT transformation plants (GP-fast) -> assay: embryo transformation output: T0 mutant plants @@ -19,4 +19,6 @@ Using CRISPR-Cas9 to generate *lwd1* mutants to confirm *LWD1* as the gene under output: T1 mutant genotype info -> assay: plant propagation - output: T2 mutant plants (used in studies gene_expression_lwd1_mutants" and "phenotyping_lwd1_mutants) \ No newline at end of file + output: T2 mutant plants (used in studies + gene_expression_lwd1_mutants and + phenotyping_lwd1_mutants) \ No newline at end of file diff --git a/studies/generating_lwd1_mutants/protocols/plant_material_growth_conditions.md b/studies/generating_lwd1_mutants/protocols/plant_material_growth_conditions.md index a74a7bc..119026c 100644 --- a/studies/generating_lwd1_mutants/protocols/plant_material_growth_conditions.md +++ b/studies/generating_lwd1_mutants/protocols/plant_material_growth_conditions.md @@ -1,28 +1,20 @@ -**Plant material and growth conditions** +## Plant material and growth conditions -**plant material** +**Plant material:** -species: Hordeum vulgare +Species: *Hordeum vulgare* +Genotypes: - GP-fast: Ppd-H1 introgression line in Golden Promise background, lwd1 mutant background, Gol et al. (2021) -**growth condtions** +**Growth condtions:** -parents: - long day: (LD) 16h light / 8h dark, 20/16 °C, 60% humidity, ~ 250 PAR) - 1.5 L pots (HerkuPlast Kubern GmbH, pot volume 75 cm^3) - Einheitserde ED73 (Einheitserde Werkverband e.V.) with 7% sand and 4 g/L Osmocote Exact Hi.End 3-4M, 4th generation (ICL Group Ltd.) -- F1 crosses: -- short day (SD): 8h light / 16h dark, 20/16 °C, 60% humidity, ~ 250 PAR) -- 7x7x7.5 cm pots (pot volume: 367.5 cm^3) -- Einheitserde ED73 (Einheitserde Werkverband e.V.) with 7% sand and 4 g/L Osmocote Exact Hi.End 3-4M, 4th generation (ICL Group Ltd.) - - -**References** -Druka A, Franckowiak J, Lundqvist U, Bonar N, Alexander J, Houston K, Radovic S, Shahinnia F, Vendramin V, Morgante M, et al. Genetic dissection of barley morphology and development. Plant Physiol. 2011:155(2):617–627. https://doi.org/10.1104/pp.110. -166249 +**References:** Gol L, Haraldsson EB, von Korff M. Ppd-H1 integrates drought stress signals to control spike development and flowering time in barley. J Exp Bot. 2021:72(1):122–136. https://doi.org/10.1093/jxb/eraa261 -- GitLab