Trypsin Inhibitor Assessment with Biochemical and Molecular Markers in a Soybean Germplasm Collection and Hybrid Populations for Seed Quality Improvement

Abstract

A soybean germplasm collection was studied for the identification of accessions with low trypsin inhibitor content in seeds. Twenty-nine accessions, parental plants, and two hybrid populations were selected and analyzed using genetic markers for alleles of the Ti3 locus, encoding Kunitz trypsin inhibitor (KTI). Most of the accessions had high or very high KTI (49.22–73.53 Trypsin units inhibited (TUI/mg seeds), while the two local Kazakh cultivars, Lastochka and Ivushka, were found to have a moderately high content of KTI, at 54.16–54.87 TUI/mg. In contrast, two soybean cultivars from Italy, Hilario and Ascasubi, showed the lowest levels of trypsin units inhibited, at 25.47–27.87 TUI/mg. Electrophoresis of seed proteins in a non-denaturing system showed a simple discrimination pattern and very clear presence/absence of bands corresponding to KTI. The SSR marker Satt228 was the most effective diagnostic marker among the three examined, and it confirmed the presence of the homozygous null-allele ti3/ti3 in cultivars Ascasubi and Hilario, which were used for hybridization with the local cv. Lastochka. Heterozygote F1 hybrid plants and homozygous ti3/ti3 lines in F2 segregating populations were successfully identified using Satt228. Finally, through marker-assisted selection with Satt228, prospective homozygous ti3/ti3 lines were produced for further application in the breeding program aimed at improving soybean seed quality in Kazakhstan.

Research Article Genetic removal of trypsin inhibitor and lipoxygenase isozymes form soybean seeds (<i>Glycine</i> <i>max</i>) by simple sequence repeat marker assisted selection

“…These observations fit a 3:1 ratio proving that the character is controlled by a single gene. Molecular analysis of Kti locus encoding KTI was carried out with four SSR markers located on linkage group A2 of the USDA/Iowa State University soybean molecular linkage map (Cregan et al 1999), the markers were selected on base published data (Kim et al 2006, Moraes et al 2006and Bulatova et al 2019. Kim et al (2006) reported Satt228 and Satt409 to be tightly linked in the mapping populations Jinpumkomg2 x C242 (source of null allele of kunitz trypsin inhibitor protein) and Clark x C242.…”

“…Kim et al (2006) reported Satt228 and Satt409 to be tightly linked in the mapping populations Jinpumkomg2 x C242 (source of null allele of kunitz trypsin inhibitor protein) and Clark x C242. Bulatova et al (2019) assessed the populations derived from two soybean cultivars Lastochka x Ascasubi and Lastochka x Hilario for kunitz trypsin inhibitor protein with biochemical and molecular markers (Satt 228, Satt409 and KTi/kti gene specific). They found that the two soybean cultivars from Italy Hilario and Ascasubi, showed the lowest levels of trypsin units inhibited, and F 2:3 segregating populations were successfully identified using Satt228, and it confirmed the presence of the homozygous null-allele kti/kti in cultivars Ascasubi and Hilario.…”

“…The organization and regulation of the genes encoding KTI have been elucidated (Orf & Hymowitz, 1979;Walling et al, 1986;Jofuku et al, 1989;Krishnan, 2001;Moraes et al, 2006;Kumar et al, 2015;Bulatova et al, 2019;Choi et al, 2020). Although soybean, during the plant life cycle, expresses at least 10 distinct genes for KTI, Kti 3 is the mutant gene that encodes the predominant trypsin inhibitor in soybean seeds (Jofuku et al, 1989).…”

Global Status of Vegetable Soybean

“…KTI free grain-type soybean varieties have been developed in several countries [ 102 , 103 , 104 ]. For this purpose, the plant breeders deployed KTI null allele specific marker [ 105 ] and Ti linked SSR markers [ 106 , 107 ].…”

Monitoring Quality and Yield Capacity of Soybean Varieties during the Creation of Various Ecotypes in Kazakhstan

“…In Kazakhstan, KazNIIZiR Limited Liability Partnership organized a study of the genetic diversity of soybean varieties of various maturity groups in terms of productivity and quality (Abugalieva & Didorenko, 2016;Novikova et al, 2018), the composition of fatty acids (Lee, Spankulova, Orazbayeva, Didorenko, & Atabayeva, 2016a;2016b), and the content of anti-nutrients (Bulatova et al, 2019). The work aims to monitor the quality indicators and yield of new highly productive varieties of soybeans were adaptive to various ecological and geographical zones of Kazakhstan, with a high content of protein and oil in seeds.…”