Abstract
Raffinose family oligosaccharides (RFOs) are major soluble carbohydrates in soybean seeds that cannot be digested by human and other monogastric animals. Hence, a major goal is to reduce RFO levels to improve the nutritional quality of soybean. In this study, we utilized a dual gRNAs CRISPR/Cas9 system to induce knockouts in two soybean galactinol synthase (GOLS) genes, GmGOLS1A and its homeolog GmGOLS1B . Genotyping of T0 plants showed that the construct design was efficient in inducing various deletions in the target sites or sequences spanning the two target sites of both GmGOLS1A and GmGOLS1B genes. A subset of induced alleles was successfully transferred to progeny and, at the T2 generation, we identified null segregants of single and double mutant genotypes without off-target induced mutations. The seed carbohydrate analysis of double mutant lines showed a reduction in the total RFO content of soybean seed from 64.7 mg/g dry weight to 41.95 mg/g dry weight, a 35.2% decrease. On average, the stachyose content, the most predominant RFO in soybean seeds, decreased by 35.4% in double mutant soybean, while the raffinose content increased by 41.7%. A slight decrease in verbascose content was also observed in mutant lines. Aside from changes in soluble carbohydrate content, some mutant lines also exhibited increased protein and fat contents. Otherwise, no difference in seed weight, seed germination, plant development and morphology was observed in the mutants. Our findings indicate that GmGOLS1A and GmGOLS1B contribute to the soybean oligosaccharide profile through RFO biosynthesis pathways, and are promising targets for future investigation, as well as crop improvement efforts. Our results also demonstrate the potential in using elite soybean cultivars for transformation and targeted genome editing.
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Claims 9
Claim · model-written summary
The construct induced deletions at both GOLS targets.
- Taxon
- soybean
Glycine maxt0 - Gene
- GmGOLS1Ae0
- Outcome
- various deletionse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the construct design was efficient in inducing various deletions in the target sites or sequences spanning the two target sites of both GmGOLS1A and GmGOLS1B genes”
t0“Soybean Seeds”
Claim · model-written summary
Single and double mutant null segregants were identified without off-target induced mutations.
- Taxon
- soybeanGlycine maxt0
- Outcome
- null segregants of single and double mutant genotypes without off-target induced mutationse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we identified null segregants of single and double mutant genotypes without off-target induced mutations”
t0“Soybean Seeds”
Claim · model-written summary
Double mutants had 35.2% lower total seed RFO content.
- Taxon
- soybeanGlycine maxt0
- Outcome
- total RFO contente0
- Direction
- decreasee0
- Value
- 35.2%e0
- Qualifier
- 64.7 mg/g dry weight to 41.95 mg/g dry weighte0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“reduction in the total RFO content of soybean seed from 64.7 mg/g dry weight to 41.95 mg/g dry weight, a 35.2% decrease”
t0“Soybean Seeds”
Claim · model-written summary
Stachyose content decreased 35.4% in double mutants.
- Taxon
- soybeanGlycine maxt0
- Material
- double mutant soybeane0
- Outcome
- stachyose contente0
- Direction
- decreasede0
- Value
- 35.4%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“stachyose content, the most predominant RFO in soybean seeds, decreased by 35.4% in double mutant soybean”
t0“Soybean Seeds”
Claim · model-written summary
Raffinose content increased 41.7% in double mutants.
- Taxon
- soybeanGlycine maxt0
- Outcome
- raffinose contente0
- Direction
- increasede0
- Value
- 41.7%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“raffinose content increased by 41.7%”
t0“Soybean Seeds”
Claim · model-written summary
Verbascose content decreased slightly in mutant lines.
- Taxon
- soybeanGlycine maxt0
- Material
- mutant linese0
- Outcome
- verbascose contente0
- Direction
- decreasee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A slight decrease in verbascose content was also observed in mutant lines”
t0“Soybean Seeds”
Claim · model-written summary
Some mutant lines had increased protein and fat contents.
- Taxon
- soybeanGlycine maxt0
- Material
- mutant linese0
- Outcome
- protein and fat contentse0
- Direction
- increasede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“some mutant lines also exhibited increased protein and fat contents”
t0“Soybean Seeds”
Claim · model-written summary
Mutants showed no difference in seed weight, germination, development, or morphology.
- Taxon
- soybeanGlycine maxt0
- Material
- mutantse0
- Outcome
- seed weight, seed germination, plant development and morphologye0
- Direction
- no differencee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“no difference in seed weight, seed germination, plant development and morphology was observed in the mutants”
t0“Soybean Seeds”
Claim · model-written summary
GmGOLS1A and GmGOLS1B contribute to the soybean oligosaccharide profile.
- Taxon
- soybeanGlycine maxt0
- Gene
- GmGOLS1A and GmGOLS1Be0
- Outcome
- soybean oligosaccharide profilee0
- Qualifier
- RFO biosynthesis pathwayse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GmGOLS1A and GmGOLS1B contribute to the soybean oligosaccharide profile through RFO biosynthesis pathways”
t0“Soybean Seeds”