Abstract
Summary The stringency of spatial expression of phaseolin, the major storage protein of bean ( Phaseolus vulgaris ) seeds, has been rigorously evaluated using stable and transient transformation techniques. Transgenic tobacco plants known to be homozygous for the β‐glucuronidase ( gus ) reporter sequence under the regulation of various lengths of the β‐ phaseolin gene ( phas ) promoter were shown to express gus only in developing seed tissues. No expression was detected in calli initiated from stems, leaves and immature seeds, showing that expression was not leaky in undifferentiated tissues. Control plants and cultures containing gus fused to the CaMV 35S promoter actively expressed gus under identical conditions. It was not possible to induce expression in phas/gus calli with ABA, GA or jasmonic acid. Treatment of the cultures with 5‐azacytidine did not result in expression, excluding methyletlon as the major factor regulating the phas promoter. However, strong gus expression was detected in seed of plants regenerated from these callus cultures, confirming that neither gene rearrangements nor deletion were responsible for the lack of activity seen in tissues other than the developing seed. In contrast to the above observations, strong transient expression of gus was detected in tobacco, bean and soybean leaves following introduction of the phas/gus fusion constructs via biolistic approaches and in electroporated bean leaf and hypocotyl protoplasts. These experiments show unequi‐vocally that the phas promoter is under rigorous spatial control when integrated into the genome, but lacks spatial control when present as extrachromosomal naked DNA. A putative model explaining these differences is presented.
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Claims 11
Claim · model-written summary
Transgenic tobacco carrying phas/gus expressed gus only in developing seed tissues.
- Taxon
- tobacco
Nicotiana tabacume0 - Organ / stage
- developing seed tissuese1
- Gene
- guse1
- Outcome
- express guse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Transgenic tobacco plants”
e1“were shown to express gus only in developing seed tissues”
Claim · model-written summary
No gus expression was detected in calli initiated from stems, leaves or immature seeds of tobacco.
- Taxon
- tobaccoNicotiana tabacume0
- Organ / stage
- calli initiated from stems, leaves and immature seedse1
- Outcome
- expressione1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Transgenic tobacco plants”
e1“No expression was detected in calli initiated from stems, leaves and immature seeds”
Claim · model-written summary
Control plants and cultures with gus fused to the CaMV 35S promoter actively expressed gus under identical conditions.
- Taxon
- tobaccoNicotiana tabacume0
- Treatment
- gus fused to the CaMV 35S promotere1
- Outcome
- actively expressed guse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Transgenic tobacco plants”
e1“Control plants and cultures containing gus fused to the CaMV 35S promoter actively expressed gus under identical conditions”
Claim · model-written summary
ABA, GA or jasmonic acid did not induce expression in phas/gus calli.
- Taxon
- tobaccoNicotiana tabacume0
- Organ / stage
- phas/gus callie1
- Treatment
- ABA, GA or jasmonic acide1
- Outcome
- induce expressione1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Transgenic tobacco plants”
e1“It was not possible to induce expression in phas/gus calli with ABA, GA or jasmonic acid”
Claim · model-written summary
Treatment with 5-azacytidine did not result in expression from the phas promoter, which excludes methylation as the major regulating factor.
- Taxon
- tobaccoNicotiana tabacume0
- Treatment
- 5‐azacytidinee1
- Outcome
- expressione1
- Qualifier
- excluding methyletlon as the major factor regulating the phas promotere1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Transgenic tobacco plants”
e1“Treatment of the cultures with 5‐azacytidine did not result in expression, excluding methyletlon as the major factor regulating the phas promoter”
Claim · model-written summary
Strong gus expression was detected in seed of plants regenerated from the callus cultures, so gene rearrangement or deletion does not explain the lack of expression in other tissues.
- Taxon
- tobaccoNicotiana tabacume0
- Organ / stage
- seed of plants regenerated from these callus culturese1
- Outcome
- gus expressione1
- Qualifier
- confirming that neither gene rearrangements nor deletion were responsible for the lack of activity seen in tissues other than the developing seede2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Transgenic tobacco plants”
e1“strong gus expression was detected in seed of plants regenerated from these callus cultures”
e2“confirming that neither gene rearrangements nor deletion were responsible for the lack of activity seen in tissues other than the developing seed”
Claim · model-written summary
Strong transient gus expression was detected in tobacco leaves after biolistic introduction of phas/gus constructs.
- Taxon
- tobaccoNicotiana tabacume0
- Organ / stage
- leavese0
- Treatment
- biolistic approachese0
- Outcome
- transient expression of guse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“strong transient expression of gus was detected in tobacco, bean and soybean leaves following introduction of the phas/gus fusion constructs via biolistic approaches”
Claim · model-written summary
Strong transient gus expression was detected in bean leaves after biolistic introduction of phas/gus constructs.
Verbatim
e0“strong transient expression of gus was detected in tobacco, bean and soybean leaves following introduction of the phas/gus fusion constructs via biolistic approaches”
Claim · model-written summary
Strong transient gus expression was detected in soybean leaves after biolistic introduction of phas/gus constructs.
- Taxon
- soybean
Glycine maxe0 - Organ / stage
- leavese0
- Treatment
- biolistic approachese0
- Outcome
- transient expression of guse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“strong transient expression of gus was detected in tobacco, bean and soybean leaves following introduction of the phas/gus fusion constructs via biolistic approaches”
Claim · model-written summary
Strong transient gus expression was also detected in bean leaf and hypocotyl protoplasts after electroporation.
Verbatim
e0“strong transient expression of gus was detected in tobacco, bean and soybean leaves”
e1“in electroporated bean leaf and hypocotyl protoplasts”
Claim · model-written summary
The phas promoter is under rigorous spatial control when integrated into the genome but lacks spatial control as extrachromosomal naked DNA.
- Taxon
- tobaccoNicotiana tabacume0
- Treatment
- integrated into the genomee1
- Outcome
- spatial controle1
- Comparator
- extrachromosomal naked DNAe1
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Transgenic tobacco plants”
e1“is under rigorous spatial control when integrated into the genome, but lacks spatial control when present as extrachromosomal naked DNA”
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