Abstract
Transgenic approaches to combating fungal pathogens, such as Fusarium graminearum, require the targeting of antifungal gene expression in tissues of developing seed spikes of cereal grains, especially lemmas and epicarps. The Lem2 gene of barley encodes a lectin-like protein that is strongly up-regulated by salicylic acid and is preferentially expressed in lemmas, paleas (lemma/palea) and coleoptiles. Transient expression studies have indicated that the proximal -75/+70 region (relative to the transcription start site) determines organ specificity. In the present study, Golden Promise barley stably transformed with Morex Lem2 promoter/gfp reporter constructs displayed cell- and development-specific expression of gfp (green fluorescent protein gene). This expression corresponded to the expression seen in Northern blots of Morex organs. Under the full-length promoter, strong GFP fluorescence was observed in the lemma/palea, glumes, coleoptile, auricle and ligule. Weak GFP fluorescence was also observed in the rachis, tips of primary leaves and the leaf sheath. Unexpectedly, strong expression occurred in the epicarp, even though Lem2 is not expressed in this organ in Morex. Studies showed that the Lem2 promoter is more highly methylated in the epicarp than in the lemma of Morex. In the lemma/palea, gfp underwent a temporal shift in expression from the mesophyll to specialized epidermal cork cells. Similar to the lemma/palea, expression in the leaf sheath was localized in the cork cells. Progressive 5' deletions of the promoter to nucleotide -75 gradually reduced the level of gfp expression, but tissue- and cell-specific expression was retained.
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Claims 10
Claim · model-written summary
Stable Lem2 promoter constructs produced cell- and development-specific gfp expression in barley.
- Taxon
- barley
.jpg?width=60)
Hordeum vulgaree0 - Material
- Golden Promisee0
- Gene
- gfpe0
- Outcome
- cell- and development-specific expression of gfpe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
.jpg?width=500)
Verbatim
e0“Golden Promise barley stably transformed with Morex Lem2 promoter/gfp reporter constructs displayed cell- and development-specific expression of gfp (green fluorescent protein gene)”
Claim · model-written summary
Reporter expression corresponded to expression in Northern blots of Morex organs.
- Taxon
- barleyHordeum vulgaret0
- Material
- Morexe0
- Outcome
- expressione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“This expression corresponded to the expression seen in Northern blots of Morex organs”
t0“barley”
Claim · model-written summary
The full-length promoter produced strong GFP fluorescence in the lemma/palea, glumes, coleoptile, auricle and ligule.
- Taxon
- barleyHordeum vulgaret0
- Organ / stage
- lemma/palea, glumes, coleoptile, auricle and ligulee0
- Treatment
- full-length promotere0
- Outcome
- strong GFP fluorescencee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Under the full-length promoter, strong GFP fluorescence was observed in the lemma/palea, glumes, coleoptile, auricle and ligule”
t0“barley”
Claim · model-written summary
The full-length promoter produced weak GFP fluorescence in the rachis, primary leaf tips and leaf sheath.
- Taxon
- barleyHordeum vulgaret0
- Organ / stage
- rachis, tips of primary leaves and the leaf sheathe0
- Outcome
- Weak GFP fluorescencee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Weak GFP fluorescence was also observed in the rachis, tips of primary leaves and the leaf sheath”
t0“barley”
Claim · model-written summary
Strong reporter expression occurred in the epicarp despite absent endogenous Lem2 expression there in Morex.
- Taxon
- barleyHordeum vulgaret0
- Material
- Morexe0
- Organ / stage
- epicarpe0
- Gene
- Lem2e0
- Outcome
- strong expressione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Unexpectedly, strong expression occurred in the epicarp, even though Lem2 is not expressed in this organ in Morex”
t0“barley”
Claim · model-written summary
The Lem2 promoter was more highly methylated in Morex epicarp than lemma.
- Taxon
- barleyHordeum vulgaret0
- Material
- Morexe0
- Organ / stage
- epicarpe0
- Gene
- Lem2e0
- Outcome
- Lem2 promoter is more highly methylatede0
- Direction
- more highlye0
- Comparator
- lemmae0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the Lem2 promoter is more highly methylated in the epicarp than in the lemma of Morex”
t0“barley”
Claim · model-written summary
gfp expression in lemma/palea shifted over time from mesophyll to epidermal cork cells.
- Taxon
- barleyHordeum vulgaret0
- Organ / stage
- lemma/paleae0
- Gene
- gfpe0
- Outcome
- temporal shift in expressione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“In the lemma/palea, gfp underwent a temporal shift in expression from the mesophyll to specialized epidermal cork cells”
t0“barley”
Claim · model-written summary
Leaf sheath expression was localized in cork cells.
- Taxon
- barleyHordeum vulgaret0
- Organ / stage
- leaf sheathe0
- Outcome
- localized in the cork cellse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“expression in the leaf sheath was localized in the cork cells”
t0“barley”
Claim · model-written summary
Progressive promoter deletions to -75 reduced gfp expression level.
- Taxon
- barleyHordeum vulgaret0
- Treatment
- Progressive 5' deletions of the promoter to nucleotide -75e0
- Outcome
- level of gfp expressione0
- Direction
- reducede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Progressive 5' deletions of the promoter to nucleotide -75 gradually reduced the level of gfp expression”
t0“barley”
Claim · model-written summary
Tissue- and cell-specific expression persisted after progressive promoter deletions.
- Taxon
- barleyHordeum vulgaret0
- Treatment
- Progressive 5' deletions of the promoter to nucleotide -75e0
- Outcome
- tissue- and cell-specific expression was retainede0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Progressive 5' deletions of the promoter to nucleotide -75 gradually reduced the level of gfp expression, but tissue- and cell-specific expression was retained”
t0“barley”
The same questions in other papers
loss of function · molecular: expression · Hordeum vulgare (barley)Positive and negative results
3 papers in the same direction as this paper · 9 in the opposite direction
- no effect
- negative
- negative
9 papers more in this group
See all 13 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.