Construction of a high-density mutant library in soybean and development of a mutant retrieval method using amplicon sequencing

Table 3 Mutations in Glyma20g25000 detected by high resolution melting analysis

Line name	Target region	Amplicon size (bp)*	Base change	Mode of mutation	Chromosome	Position (bp)	Amino acid substitution
EnT-0541	Ln ex1	332	G > A	hetero	Gm20	34688627	Met1Ile
EnT-0685	Ln ex1	332	C > T	hetero	Gm20	34688652	Leu10Phe
EnT-1168	Ln ex1	332	G > A	homo^**	Gm20	34688672	Syn
EnT-0112	Ln ex1	332	G > A	homo^**	Gm20	34688682^a	Asp20Asn
EnT-0044	Ln ex1	332	G > A	hetero	Gm20	34688682^a	Asp20Asn
EnT-1589	Ln ex1	332	G > A	hetero	Gm20	34688682^a	Asp20Asn
EnT-1376	Ln ex1	332	G > A	hetero	Gm20	34688686^b	Gly21Asp
EnT-0621	Ln ex1	332	G > A	hetero	Gm20	34688686^b	Gly21Asp
EnT-1048	Ln ex1	332	C > T	homo^**	Gm20	34688696^c	Syn
EnT-1306	Ln ex1	332	C > T	homo^**	Gm20	34688696^c	Syn
EnT-0987	Ln ex1	332	C > T	homo^**	Gm20	34688713	Ser30Phe
EnT-0160	Ln ex1	332	C > T	hetero	Gm20	34688719	Ser32Phe
EnT-0634	Ln ex2	231	G > A	homo^**	Gm20	34689275^d	Gly40Ser
EnT-0749	Ln ex2	231	G > A	homo^**	Gm20	34689275^d	Gly40Ser
EnT-0439	Ln ex2	231	C > T	hetero	Gm20	34689313	Syn
EnT-1084	Ln ex2	231	G > A	hetero	Gm20	34689360	Gly68Glu
EnT-1281	Ln ex3	114	C > T	hetero	Gm20	34689704	Ala102Val
EnT-1312	Ln ex3	114	A > T	hetero	Gm20	34689710	His104Leu
EnT-0601	Ln ex4	467	G > A	homo^**	Gm20	34690049	Syn
EnT-0155	Ln ex4	467	G > A	hetero	Gm20	34690058	Syn
EnT-0687	Ln ex4	467	C > T	homo^**	Gm20	34690175^e	Syn
EnT-0862	Ln ex4	467	C > T	homo**	Gm20	34690175^e	Syn
EnT-1619	Ln ex4	467	G > A	hetero	Gm20	34690246	Gly213Asp
EnT-1265	Ln ex4	467	C > T	homo^**	Gm20	34690247^f	Syn
EnT-0510	Ln ex4	467	C > T	hetero^***	Gm20	34690247^f	Syn
EnT-0383	Ln ex4	467	C > T	hetero	Gm20	34690247^f	Syn

*The amplicon size does not include primer sequences
**Base changes in a homozygous state probably occurred in M1 plants
***Base changes in a heterozygous state are probably derived from the same M1 plants as those labeled with **
Syn indicates synonymous site at which a base substitution does not cause an amino acid substitution
Superscripts a to f indicate that the mutation was duplicated in plants labeled with the same letter

ISSN: 1471-2164