Protocol detail

Genetic Analysis of tiller number in Arabidopsis

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A cross between the tin3 mutant and the parental accession TA4342-L96 was made. A total of 375 F₂ plants were grown and phenotyped for tiller number (Zadoks growth scale ~Z29). Leaf tissues were harvested and DNA was extracted using the BioSprint 96 DNA Plant Kit (Qiagen, 576) on a KingFisher Flex robot (Thermo Fisher Scientific, 5400610) according to the manufacturer’s instructions. Equimolar concentrations of 30 F₂ plants showing the tin3 mutant phenotype were pooled to create a mutant bulk. SEM samples were fixed in 3% glutaraldehyde, post-fixed in 1% osmium tetroxide and dehydrated in a graded acetone series, and these samples were processed and SEM images were captured as described previously^{114 (link),115 (link)}.

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Ahmed H.I., Heuberger M., Schoen A., Koo D.H., Quiroz-Chavez J., Adhikari L., Raupp J., Cauet S., Rodde N., Cravero C., Callot C., Lazo G.R., Kathiresan N., Sharma P.K., Moot I., Yadav I.S., Singh L., Saripalli G., Rawat N., Datla R., Athiyannan N., Ramirez-Gonzalez R.H., Uauy C., Wicker T., Tiwari V.K., Abrouk M., Poland J, & Krattinger S.G. (2023). Einkorn genomics sheds light on history of the oldest domesticated wheat. Nature, 620(7975), 830-838.

Publication 2023

BioSprint 96 DNA Plant Kit KingFisher Flex robot

Acetone Bulk Glutaraldehyde Leaf Osmium tetroxide Parental Phenotype Plants Tissues

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Variable analysis

independent variables

Cross between the tin3 mutant and the parental accession TA4342-L96

dependent variables

Tiller number (Zadoks growth scale ~Z29)

control variables

Leaf tissues were harvested and DNA was extracted using the BioSprint 96 DNA Plant Kit (Qiagen, 576) on a KingFisher Flex robot (Thermo Fisher Scientific, 5400610) according to the manufacturer's instructions.
Equimolar concentrations of 30 F₂ plants showing the tin3 mutant phenotype were pooled to create a mutant bulk.
SEM samples were fixed in 3% glutaraldehyde, post-fixed in 1% osmium tetroxide and dehydrated in a graded acetone series, and these samples were processed and SEM images were captured as described previously^{114 (link),115 (link)}.

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