2.7 Arabidopsis plant material and growth condition
Arabidopsis thaliana (L.) Heynh ecotypes Col-0, homozygous mutant
and over expression (OE) lines used in this study. T-DNA insertion
mutant of rpoC1 (cs835205 ) was obtained from the
Arabidopsis Biological Resource Centre (ABRC, Ohio State University,
Columbus, OH, USA; https://abrc.osu.edu/). Seed sterilization and
plants growth were performed according to Tiwari et al. , (2021).
Homozygosity of rpoC1 mutant confirmed by polymerase chain
reaction (PCR) by using gene-specific right primer with a T-DNA left
border specific primer (pDAP110/ pCSA110) listed in Table S1 .
For over-expression of RpoC1 gene from two allele (B1K-09-07 and
B1K-50-04) in Col-0 background, coding sequence region (2046 bps without
stop codon) were synthesized from Genewiz (www.genewiz.com) in
pUC-GW-Amp vector. Further all the desired fragment were cloned in
binary vector (pICH86966 Addgene) using golden gate assembly (Plant
part: 1000000047 and toolkit: 1000000044, Addgene) (Figure S3 ).
RESULTS