Marker-assisted stacking of null Kunitz trypsin inhibitor and off-flavour generating lipoxygenase-2 in soybean

2021 ◽  
pp. 1-9
Author(s):  
V. Kumar ◽  
A. Rani ◽  
A. K. Anshu ◽  
T. Tayalkar
Keyword(s):  
Ssr Markers ◽  
Trypsin Inhibitor ◽  
Null Allele ◽  
Protein Digestibility ◽  
Null Alleles ◽  
Human Consumption ◽  
Recurrent Parent ◽  
Kunitz Trypsin Inhibitor ◽  
Soybean Genotypes ◽  
Parental Combination

Abstract Soybean varieties genetically free from Kunitz trypsin inhibitor (KTI) and lipoxygenase-2 (Lox2) are desirable to increase human consumption, as the former is an antinutritional factor that affects protein digestibility while the latter is a principal contributor to off-flavour. In the present investigation, soybean genotypes free from both these undesirable components were developed by introgression of null allele of Lox2 from NRC109 (lox2lox2) into two KTI-free soybean genotypes derived from genotypes JS97-52 and NRC7. Foreground selection of plants in F1, F2, BC1F1, BC1F2, BC2F1 and BC2F2 generations developed from two cross combinations i.e. NRC7-derived KTI-free genotype (N7KTIF)×NRC109 (parental combination 1) and JS97-52 derived KTI free genotype (JKTIF)×NRC109 (parental combination 2) was performed using null allele specific markers and tightly linked simple sequence repeat (SSR) markers for both KTI and Lox2 genes for the identification of homozygous recessive (titilox2lox2) plants. Background selection was performed using 239 and 241 polymorphic SSR markers across the genome. This resulted in the development of 9 and 13 soybean lines stacked for null alleles of both KTI and Lox2 (titilox2lox2) exhibiting recurrent parent genome content more than 97 and 96%, respectively. Days-to-flowering, days-to-maturity, 100-seed weight and yield per plant of the stacked lines developed from both the parental combinations were at par with the respective recurrent parents.

scholarly journals Bowman-Birk inhibitor in soybean: Genetic variability in relation to total trypsin inhibitor activity and elimination of Kunitz trypsin inhibitor

2021 ◽  
Vol 13 (1) ◽  
pp. 10836
Author(s):  
Priyanka MITTAL ◽  
Vineet KUMAR ◽  
Anita RANI ◽  
S. M. GOKHALE
Keyword(s):  
Trypsin Inhibitor ◽  
Genetic Background ◽  
Soy Flour ◽  
Trypsin Inhibitor Activity ◽  
Recurrent Parent ◽  
Kunitz Trypsin Inhibitor ◽  
Inhibitor Activity ◽  
Commercial Preparation ◽  
Defatted Soy Flour ◽  
Soybean Genotypes

Bowman-Birk inhibitor (BBI) is a soybean seed serine protease inhibitor whose nutraceutical properties far exceed its anti-nutritional property arising from its trypsin and chymotrypsin inhibitor activity. High BBI soybean genotypes are sought for the commercial preparation of this nutraceutical biomolecule while low BBI content in Kunitz trypsin inhibitor (KTI)-free genetic background is desirable in soymeal manufacturing. In the present investigation, monoclonal antibody assay of 95 soybean genotypes revealed 11-fold genetic variation for BBI concentration. The study led to the identification of 3 very high (>20 mg/g defatted soy flour) and 5 very low BBI (< 4 mg/g defatted soy flour) soybean genotypes. Contribution of BBI to total trypsin inhibition activity ranged from 2.2 to 53.5% with average value of 11.6%. Genotypes with varying level of BBI raised consecutively for two years exhibited non-significant (p>0.05) effect of growing year on the trait. BBI content in BC3F2 derived KTI-free lines was at par with the recurrent parent. Low and high BBI content genotypes with diverse genetic background identified in the study may be exploited to develop mapping population to identify genomic regions underlying BBI in soybean.

scholarly journals Length polymorphisms of simple sequence repeat (SSR) markers in assisted selection for lipoxygenase-free in soybean

2021 ◽  
Vol 3 (1) ◽  
Author(s):  
VALERIA CARPENTIERI-PIPOLO ◽  
Thales Pereira Barreto ◽  
Daiana Alves Silva ◽  
Ricardo Vilela Abdelnoor ◽  
Silvana Rockenbach Marin ◽  
...  
Keyword(s):  
Ssr Markers ◽  
Null Allele ◽  
Seed Quality ◽  
Soybean Seed ◽  
Hybrid Population ◽  
Human Consumption ◽  
F1 Hybrid ◽  
Hybrid Plants ◽  
Length Polymorphisms ◽  
Glycine Max L

Beany flavor of soybean (Glycine max (L.) Merr.) is caused by oxidation of polyunsaturated fatty acids by the action of three lipoxygenases (LOX1, LOX2 and LOX3) present in mature seeds. The unpleasant flavor restricts human consumption of soybean products. This problem could be solved through genetic elimination of alleles that code these enzymes. Parental cultivars and two hybrid population were selected and analyzed using genetic markers for alleles locus, encoding Lox1, Lox2 and Lox3 free. The SSR marker Satt212 confirmed the presence of the homozygous null-allele Lx3 in the cultivar BRS 213, which were used for hybridization with BR 36. Heterozygote F1 hybrid plants and homozygous Lx3 lines in F2 segregating populations were successfully identified.  The SSR markers Sat090 and Sat417 was the most effective diagnostic marker among the all SSR markers tested. Satt090 and Satt417 confirmed the presence of the homozygous Lx2 null-allele in the parental cultivar BRS 213 by flanking Lx2 loci at 3,00 and 2,77 cM, respectively. The presence of Lx2 null allele in The F2 segregating populations between BRS 213 and BRS 155 were successfully identified with a selection efficiency of 98% and have great potential for further application in the Brazilian breeding program aimed at improving soybean seed quality.

scholarly journals Development of Breeder-Friendly KASP Markers for Low Concentration of Kunitz Trypsin Inhibitor in Soybean Seeds

2021 ◽  
Vol 22 (5) ◽  
pp. 2675
Author(s):  
M. Luciana Rosso ◽  
Chao Shang ◽  
Qijian Song ◽  
Diana Escamilla ◽  
Jay Gillenwater ◽  
...  
Keyword(s):  
Trypsin Inhibitor ◽  
Protein Digestibility ◽  
Population Based ◽  
Hplc Method ◽  
Snp Markers ◽  
Soybean Seeds ◽  
Kunitz Trypsin Inhibitor ◽  
Breeding Lines ◽  
Specific Pcr ◽  
Kasp Assay

Trypsin inhibitors (TI), a common anti-nutritional factor in soybean, prevent animals’ protein digestibility reducing animal growth performance. No commercial soybean cultivars with low or null concentration of TI are available. The availability of a high throughput genotyping assay will be beneficial to incorporate the low TI trait into elite breeding lines. The aim of this study is to develop and validate a breeder friendly Kompetitive Allele Specific PCR (KASP) assay linked to low Kunitz trypsin inhibitor (KTI) in soybean seeds. A total of 200 F3:5 lines derived from PI 547656 (low KTI) X Glenn (normal KTI) were genotyped using the BARCSoySNP6K_v2 Beadchip. F3:4 and F3:5 lines were grown in Blacksburg and Orange, Virginia in three years, respectively, and were measured for KTI content using a quantitative HPLC method. We identified three SNP markers tightly linked to the major QTL associated to low KTI in the mapping population. Based on these SNPs, we developed and validated the KASP assays in a set of 93 diverse germplasm accessions. The marker Gm08_44814503 has 86% selection efficiency for the accessions with low KTI and could be used in marker assisted breeding to facilitate the incorporation of low KTI content in soybean seeds.

scholarly journals The influence of soybean genotypes and HTC processing method on trypsin inhibitor activity of soymilk

2016 ◽  
Vol 61 (3) ◽  
pp. 271-279 ◽  
Author(s):  
Sladjana Stanojevic ◽  
Miroljub Barac ◽  
Mirjana Pesic ◽  
Biljana Vucelic-Radovic
Keyword(s):  
Trypsin Inhibitor ◽  
Strong Dependence ◽  
Total Content ◽  
Human Consumption ◽  
Trypsin Inhibitor Activity ◽  
Inhibitor Activity ◽  
Soybean Genotype ◽  
Kunitz Inhibitor ◽  
Soybean Genotypes ◽  
The Impact

Kunitz inhibitor (KTI) and Bowman-Birk trypsin inhibitor (BBI) are inhibitors of digestive enzymes in raw soybeans. Due to their antinutritive properties in the active state, their inactivation by heat treatment is commonly used. Soymilk is a turbid and stable colloidal solution, obtained by thermal treatment of soybean. In this study soymilk was made on a pilot-plant scale from six soybean cultivars using hydrothermal cooking (HTC) as the production method. This procedure is significantly different from the traditional one. The aim of this investigation was to evaluate the impact of the HTC processing for soymilk production and different soybean genotypes on trypsin inhibitor content and activity. Obtained soymilk contained BBI in trace amounts, in the BBI-polymeric forms. The BBI monomeric forms were not detected. The soymilk of the investigated soybean genotypes had very similar KTI levels (2.34-2.99%). Results have suggested that the soybean genotype does not have substantial effects on the levels of KTI, as well as on the value of residual trypsin inhibitor activity (rTIA). The total content of TI and rTIA showed a strong dependence (r=0.91; p<0.05). HTC-soymilk rTIA was <20% (7.15-19.89%). These results have indicated that HTC processed soymilk is applicable for human consumption.

scholarly journals Marker assisted accelerated introgression of null allele of kunitz trypsin inhibitor in soybean

2015 ◽  
Vol 65 (5) ◽  
pp. 447-452 ◽  
Author(s):  
Vineet Kumar ◽  
Anita Rani ◽  
Reena Rawal ◽  
Vaishali Mourya
Keyword(s):  
Trypsin Inhibitor ◽  
Null Allele ◽  
Kunitz Trypsin Inhibitor

scholarly journals Agronomic characteristics, isoflavone content and Kunitz trypsin inhibitor of vegetable soybean genotypes

2011 ◽  
Vol 29 (2) ◽  
pp. 222-227 ◽  
Author(s):  
Renata Castoldi ◽  
Hamilton César de O Charlo ◽  
Pablo F Vargas ◽  
Leila T Braz ◽  
Mercedes C Carrão-Panizzi
Keyword(s):  
Trypsin Inhibitor ◽  
Agronomic Traits ◽  
Block Design ◽  
Total Yield ◽  
Vegetable Soybean ◽  
Kunitz Trypsin Inhibitor ◽  
Fresh Weight ◽  
Agronomic Characteristics ◽  
Immature Seeds ◽  
Soybean Genotypes

Five vegetable soybean genotypes were evaluated for agronomic characteristics, functional and antinutritional properties. The experiments were conducted in the field, in the School of Agricultural and Veterinary Sciences, in Jaboticabal, São Paulo State, Brazil. The experimental design was a randomized block design with five treatments (genotypes) and four replications. The genotypes JLM003, JLM004, JLM010, JLM024 and JLM030 were evaluated in the following characteristics: number of pods per plant, number of seeds per pod, fresh weight of pods per plant (g), fresh weight of 100 seeds (g), estimated total yield of immature seeds (kg ha-1), moisture content, proteins, lipids, ash, carbohydrates, isoflavones and Kunitz trypsin inhibitor. In the assessment of the major agronomic traits, in the conditions of this study, the genotype JLM010 was the most recommended for the region of Jaboticabal. This genotype showed better agronomic characteristics of production as a vegetable, producing 136.04 g of fresh weight of pods per plant, 92.52 g of fresh weight/100 seeds and 11.12 t/ha of total immature seeds, and better functional properties.

scholarly journals Genetic diversity analysis of soybean genotypes using SSR markers for salinity tolerance

2019 ◽  
Vol 30 (1) ◽  
pp. 1-9
Author(s):  
M Moniruzzaman ◽  
RM Saiem ◽  
RM Emon ◽  
MS Haque ◽  
NR Saha ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Genetic Distance ◽  
Ssr Markers ◽  
Gene Diversity ◽  
Dry Weight ◽  
Null Alleles ◽  
Root Weight ◽  
Root And Shoot Length ◽  
Soybean Genotypes ◽  
Ssr Loci

Soil salinity is a major constraint to soybean production. Five soybean genotypes were grown in pots with hydroponic culture under control and different salt stressed conditions to observe salt tolerance capacity on the basis of phenotypic screening and measure genetic diversity and relatedness among the genotypes. Minimum effects of salinity on root and shoot length was observed in Binasoyben-3, GC840 and Binasoyben-5at different salt stresses. Root dry weight and shoot dry weight of different soybean genotypes under different salt stresses were depicted. The highest reduction in root weight was noted in Binasoybean-1. The same genotypes were used to assess genetic diversity among them with simple sequence repeat (SSR) markers. A total of 33 alleles were detected among 5 soybean genotypes by using 10SSR markers. The number of alleles per locus ranged from 2 to 5, with an average of 3.33 alleles across the 10 loci. Rare alleles were observed at 10 SSR loci with an average of 2.8 alleles per locus. In this experiment, two SSR loci were found to be null alleles. The average values of null allele were 0.2. PIC values ranged from 0.27 in Satt184 to 0.77 in Satt339 with the average value of 0.56.The major allele frequency of the most common allele at each locus ranged from 0.80 in Satt184 to 0.20 in Satt339with a mean frequency of 0.48. The size of the different major alleles at different loci ranged from 173 bp for Satt509 to 407 bp for Satt339. The highest gene diversity (0.80) was observed in loci Satt339 and the lowest gene diversity (0.32) was observed in loci Satt184 with the mean diversity of 0.61. The lowest genetic distance (0.60) was observed in Asset vs Binasoybean-3 and Binasoybean-5 vs Binasoybean-3. The highest genetic distance (1.0) was observed between a numbers of genotype pairs with GC840 vs Asset. The UPGMA cluster analysis led to the grouping of the 5 genotypes into two major clusters. GC840, an advanced line identified to be salt tolerant, together with Binasoybean-5 and Binasoybean-3 clustered in the same sub group. The results from morphological and molecular study suggested that GC840 and Binasoybean-3 are moderately tolerant to salt stress. Progressive Agriculture 30 (1): 1-9, 2019

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