Supplementary Figures Legend
Supplemental Figure S1 Secondary protein structure of ICPA 2039 and ICPB 2039 shows beta
sheets, helix and N-terminal loops. Comparative analysis of secondary protein structures of both
the lines showed significant differences in beta sheets and helix with a pre-dominant N-terminal
loop region. E denotes beta sheet and H denotes helix.
Supplemental Figure S2 Multiple sequence alignment and homology search of nad7a gene with
known genes available in NCBI database (a) Multiple sequence alignment of nad7a gene
amplified in ICPA 2039 of Cajanus cajan (1-162) and with other five species like Oryza
sativa (1-262), Arabidopsis
thaliana (1-143), Vigna
mungo (1-143),Glycince
max (1-143),
and Lotus japonicas (1-143), identified that the highly conserved nature of nad7a in legumes. (b)
Sequence homology search of 300bp upstream region uniquely present in pigonepea genome
showed maximum similarity to the Beta vulgaris mitochondrial orf124.
Supplemental Figure S3 CMS seed purity of ICPA 2039 seed lot using gene based
maker nad7a_del. Lane M: 100bp standard DNA ladder; Lane A: ICPA 2039 (male sterile line);
Lane B: ICPB 2039 (maintainer line); Lane 1-20 and Lane 21-40 (representative seed samples of
CMS line developed from ICPA 2039 × ICPB 2039). Fragments were resolved in 3.5%
Metaphore gel and the expected product sizes of CMS and maintainer lines are mentioned with
the arrow. The outliers are represented by arrow in the gel picture showing presence of ICPB
2039 sample among the seed lot of ICPA 2039.
Supplemental Figure S4 Validation of gene based maker nad7a_del in different set of
cytoplasm and their maintainer lines. Lane M: 100bp standard DNA ladder; Lane A4cytoplasm
A: CMS line ICPA 2039 and B: maintainer line ICPB 2039; Lane A1 cytoplasm A: CMS line
ICPA 2067 and B: maintainer line ICPB 2067; Lane A2 cytoplasm A: CMS line ICPA 2052 and
B: maintainer line ICPB 2052; Lane A6 cytoplasm A: CMS line ICPA 2209 and B: maintainer
line ICPB 2209; Lane A8 cytoplasm A: CMS line ICPW 74 and B: maintainer line Vyshali.
Fragments were resolved in 3.5% Metaphor agarose gel and the expected product sizes of CMS
and maintainer lines are mentioned with the arrow.