Abstract
Agrobacterium-mediated plant transformation is typically conducted by inoculating plant tissues with an Agrobacterium suspension containing approximately 108–109 bacteria mL−1, followed by a 2–3-d co-culture period. Use of longer co-culture periods could potentially increase transformation efficiencies by allowing more time for Agrobacterium to interact with plant cells, but bacterial overgrowth is likely to occur, leading to severe tissue browning and reduced transformation and regeneration. Low bacterial inoculum levels were therefore evaluated as a means to reduce the negative outcomes associated with long co-culture. The use of low inoculum bacterial suspensions (approximately 6 × 102 bacteria mL−1) followed by long co-culture (15 d) led to the production of an average of three transformed sunflower shoots per explant while the use of high inoculum (approximately 6 × 108 bacteria mL−1) followed by short co-culture (3 d) led to no transformed shoots. Low inoculum and long co-culture acted synergistically, and both were required for the improvement of sunflower transformation. Gene expression analysis via qRT-PCR showed that genes related to plant defense response were generally expressed at lower levels in the explants treated with low inoculum than those treated with high inoculum during 15 d of co-culture, suggesting that low inoculum reduced the induction of plant defense responses. The use of low inoculum with long co-culture (LI/LC) led to large increases in sunflower transformation efficiency. This method has great potential for improving transformation efficiencies and expanding the types of target tissues amenable for transformation of different plant species.
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Claims 3
Claim · model-written summary
Low inoculum and long co-culture yielded three transformed sunflower shoots per explant, versus none under high inoculum and short co-culture.
- Taxon
- sunflower
Helianthus annuuse0 - Treatment
- low inoculum bacterial suspensions (approximately 6 × 102 bacteria mL−1) followed by long co-culture (15 d)e0
- Outcome
- transformed sunflower shoots per explante0
- Direction
- no transformed shootse0
- Value
- threee0
- Comparator
- high inoculum (approximately 6 × 108 bacteria mL−1) followed by short co-culture (3 d)e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“low inoculum bacterial suspensions (approximately 6 × 102 bacteria mL−1) followed by long co-culture (15 d) led to the production of an average of three transformed sunflower shoots per explant while the use of high inoculum (approximately 6 × 108 bacteria mL−1) followed by short co-culture (3 d) led to no transformed shoots”
Claim · model-written summary
Low inoculum and long co-culture acted synergistically and both were needed to improve transformation.
- Taxon
- sunflowerHelianthus annuuse0
- Outcome
- improvement of sunflower transformatione0
- Qualifier
- both were requirede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Low inoculum and long co-culture acted synergistically, and both were required for the improvement of sunflower transformation.”
Claim · model-written summary
Defense-related genes were generally expressed less under low versus high inoculum during 15 days of co-culture.
- Taxon
- sunflowerHelianthus annuust0
- Treatment
- low inoculume0
- Interactor
- Agrobacterium tumefaciens
Agrobacterium tumefacienst0other - Outcome
- genes related to plant defense responsee0
- Direction
- lowere0
- Qualifier
- during 15 d of co-culturee0
- Comparator
- high inoculume0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“genes related to plant defense response were generally expressed at lower levels in the explants treated with low inoculum than those treated with high inoculum during 15 d of co-culture”
t0“Agrobacterium tumefaciens inoculum levels and a long co-culture period lead to reduced plant defense responses and increase transgenic shoot production of sunflower (Helianthus annuus L.)”
The same questions in other papers
inoculation · Agrobacterium tumefaciens · Helianthus annuus (common sunflower)Positive and negative results
1 in the opposite direction
- negative
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.