Abstract
Cottonseed is the most important byproduct of cotton production. However, high free gossypol contents limit the application of cottonseed in the food or feed industry. In this study, CRISPR/Cas9 technology was used to knock out the (+)-δ-cadinene synthase gene (GhCAD) to decrease the gossypol content. Gossypol levels decreased approximately 64% in cottonseeds and leaves following the targeted mutation of GhCAD. If only GhCAD1-A was edited, the seed gossypol content decreased by approximately 46%, but there were no major changes in the leaf gossypol content. In addition, the protein and fatty acid (C16:0, C18:1, and C18:2) profiles of the transgenic cotton seeds were similar to those of the control cotton seeds. Furthermore, transcriptome analysis revealed that the jasmonic acid signal transduction pathway was significantly enriched among the DEGs, and GhMYC2-D09 expression was down-regulated. Silencing of GhMYC2-D09 via virus-induced gene silencing decreased the expression of gossypol biosynthesis-related genes, ultimately restricting the accumulation of gossypol in cotton leaves. In contrast, the overexpression of GhMYC2-D09 in hairy roots had the opposite effect. Dual-luciferase assays revealed that GhMYC2-D09 can activate the expression of GhCAD1-A and GhCAD1-C, but Y1H assays revealed that GhMYC2-D09 cannot bind directly to GhCAD promoters. In conclusion, we used CRISPR/Cas9 technology to silence GhCAD expression and developed new genetic resources for generating low-gossypol cotton materials. Furthermore, we characterized GhMYC2-D09 as a transcription factor that increases gossypol biosynthesis. These findings may provide new insights to further elucidate the regulatory network of gossypol biosynthesis.
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Claims 15
Claim · model-written summary
Targeted GhCAD mutation reduced gossypol levels in cottonseed and leaves by about 64%.
- Taxon
- cotton
Gossypium hirsutumt0 - Organ / stage
- cottonseeds and leavese0
- Treatment
- targeted mutation of GhCADe0
- Gene
- GhCADe0
- Outcome
- Gossypol levelse0
- Direction
- decreasede0
- Value
- approximately 64%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Gossypol levels decreased approximately 64% in cottonseeds and leaves following the targeted mutation of GhCAD”
t0“cottonseed”
Claim · model-written summary
Editing only GhCAD1-A reduced seed gossypol content by about 46%.
- Taxon
- cottonGossypium hirsutumt0
- Organ / stage
- seede0
- Treatment
- only GhCAD1-A was editede0
- Gene
- GhCAD1-Ae0
- Outcome
- seed gossypol contente0
- Direction
- decreasede0
- Value
- approximately 46%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“If only GhCAD1-A was edited, the seed gossypol content decreased by approximately 46%”
t0“cottonseed”
Claim · model-written summary
Editing only GhCAD1-A caused no major change in leaf gossypol content.
- Taxon
- cottonGossypium hirsutumt0
- Organ / stage
- leafe0
- Treatment
- only GhCAD1-A was editede0
- Gene
- GhCAD1-Ae0
- Outcome
- leaf gossypol contente0
- Direction
- no major changese0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“If only GhCAD1-A was edited, the seed gossypol content decreased by approximately 46%, but there were no major changes in the leaf gossypol content”
t0“cottonseed”
Claim · model-written summary
Transgenic and control cotton seeds had similar protein profiles.
- Taxon
- cottonGossypium hirsutume0
- Treatment
- transgenic cotton seedse0
- Outcome
- proteine0
- Direction
- similare0
- Comparator
- control cotton seedse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the protein and fatty acid (C16:0, C18:1, and C18:2) profiles of the transgenic cotton seeds were similar to those of the control cotton seeds”
Claim · model-written summary
Transgenic and control cotton seeds had similar fatty acid profiles.
- Taxon
- cottonGossypium hirsutume0
- Treatment
- transgenic cotton seedse0
- Outcome
- fatty acid (C16:0, C18:1, and C18:2) profilese0
- Direction
- similare0
- Comparator
- control cotton seedse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the protein and fatty acid (C16:0, C18:1, and C18:2) profiles of the transgenic cotton seeds were similar to those of the control cotton seeds”
Claim · model-written summary
Jasmonic acid signal transduction was significantly enriched among the differentially expressed genes.
- Taxon
- cottonGossypium hirsutumt0
- Outcome
- jasmonic acid signal transduction pathwaye0
- Direction
- enrichede0
- Qualifier
- among the DEGse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the jasmonic acid signal transduction pathway was significantly enriched among the DEGs”
t0“cottonseed”
Claim · model-written summary
GhMYC2-D09 expression was down-regulated.
- Taxon
- cottonGossypium hirsutumt0
- Gene
- GhMYC2-D09e0
- Outcome
- GhMYC2-D09 expressione0
- Direction
- down-regulatede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GhMYC2-D09 expression was down-regulated”
t0“cottonseed”
Claim · model-written summary
Silencing GhMYC2-D09 decreased expression of gossypol biosynthesis genes.
- Taxon
- cottonGossypium hirsutumt0
- Treatment
- Silencing of GhMYC2-D09 via virus-induced gene silencinge0
- Gene
- GhMYC2-D09e0
- Outcome
- expression of gossypol biosynthesis-related genese0
- Direction
- decreasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Silencing of GhMYC2-D09 via virus-induced gene silencing decreased the expression of gossypol biosynthesis-related genes”
t0“cottonseed”
Claim · model-written summary
Silencing GhMYC2-D09 restricted gossypol accumulation in cotton leaves.
- Taxon
- cottonGossypium hirsutume0
- Organ / stage
- leavese0
- Treatment
- Silencing of GhMYC2-D09 via virus-induced gene silencinge0
- Gene
- GhMYC2-D09e0
- Outcome
- accumulation of gossypol in cotton leavese0
- Direction
- restrictinge0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Silencing of GhMYC2-D09 via virus-induced gene silencing decreased the expression of gossypol biosynthesis-related genes, ultimately restricting the accumulation of gossypol in cotton leaves”
Claim · model-written summary
Overexpressing GhMYC2-D09 in hairy roots had the opposite effect on gossypol accumulation.
- Taxon
- cottonGossypium hirsutumt0
- Organ / stage
- hairy rootse0
- Treatment
- overexpression of GhMYC2-D09e0
- Gene
- GhMYC2-D09e0
- Outcome
- opposite effecte0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the overexpression of GhMYC2-D09 in hairy roots had the opposite effect”
t0“cottonseed”
Claim · model-written summary
GhMYC2-D09 activated GhCAD1-A expression in dual-luciferase assays.
- Taxon
- cottonGossypium hirsutumt0
- Gene
- GhMYC2-D09e0
- Outcome
- expression of GhCAD1-Ae0
- Qualifier
- activatee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GhMYC2-D09 can activate the expression of GhCAD1-A and GhCAD1-C”
t0“cottonseed”
Claim · model-written summary
GhMYC2-D09 activated GhCAD1-C expression in dual-luciferase assays.
- Taxon
- cottonGossypium hirsutumt0
- Gene
- GhMYC2-D09e0
- Outcome
- GhCAD1-Ce0
- Qualifier
- activatee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GhMYC2-D09 can activate the expression of GhCAD1-A and GhCAD1-C”
t0“cottonseed”
Claim · model-written summary
GhMYC2-D09 did not bind directly to GhCAD promoters in Y1H assays.
- Taxon
- cottonGossypium hirsutumt0
- Gene
- GhMYC2-D09e0
- Outcome
- cannot bind directly to GhCAD promoterse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“GhMYC2-D09 cannot bind directly to GhCAD promoters”
t0“cottonseed”
Claim · model-written summary
The study developed genetic resources for low-gossypol cotton materials.
- Taxon
- cottonGossypium hirsutume0
- Outcome
- new genetic resources for generating low-gossypol cotton materialse0
Not reportedComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“developed new genetic resources for generating low-gossypol cotton materials”
Claim · model-written summary
GhMYC2-D09 was characterized as a transcription factor that increases gossypol biosynthesis.
- Taxon
- cottonGossypium hirsutumt0
- Gene
- GhMYC2-D09e0
- Outcome
- gossypol biosynthesise0
- Qualifier
- transcription factore0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“we characterized GhMYC2-D09 as a transcription factor that increases gossypol biosynthesis”
t0“cottonseed”
The same questions in other papers
loss of function · molecular: metabolites · Gossypium hirsutum (cotton)Positive and negative results
7 papers in the same direction as this paper · 32 in the opposite direction
- positive
- negative
- negative
36 papers more in this group
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