Abstract
【Objective】A cold-regulated gene Cbcor15 a isolated from Chorispora bungeana was transferred into sugarcane embryogenic callus by Agrobacterium tumefaciens-mediated method to establish high-efficient genetic transformation system in order to provide basis on breeding sugarcane varieties with cold resistance. 【Method】Sugarcane variety ROC22 was used as tested material to optimize culture media for callus induction, bud differentiation and plantlet rooting, and screen suitable type and dosage of hormone, PPT and antibiotics dosage, which adapted for exogenous gene transferring to sugarcane. By using a bivalent plant expression vector named pCambia1300- Cbcor15a-bar,Cbcor15 a gene was transferred into callus of sugarcane using Agrobacterium tumefaciens-mediated method. 【Result】The results showed that OD6000.4 of Agrobacterium tumefaciens and 20 min of infecting time were beneficial to callus differentiation. The optimal dosage of PPT for callus induction and differentiation was 0.50 mg/L. After infected,adding 500.00 mg/L antibiotic Cef in co-culture could effectively inhabit growth of Agrobacterium tumefaciens, while adding 300.00 mg/L Cef could promote callus differentiation, and 200.00 mg/L Cef could promote plantlet rooting.Low dosage of NAA in MS medium was more beneficial to callus differentiation, and KT was superior to 6-BA in promoting cell division. By adding 5.00 mg/L NAA and 70.00 g/L sugar in MS medium could effectively promote plantlets rooting. After the preliminary selection using PPT, 286 anti- PPT regenerated plantlets transferred with Cbcor15 a gene were obtained. Eighty-three plantlets with well growing were selected to detect by PCR, out of which four anti-PPT regenerated plantlets were positive. 【Conclusion】The Agrobacterium tumefaciens-mediated genetic transformation system was established successfully for exogenous gene transferring to sugarcane, and Cbcor15 a gene had been integrated into genome of sugarcane.
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Claims 12
Claim · model-written summary
Agrobacterium OD6000.4 and 20 minutes of infection benefited sugarcane callus differentiation.
- Taxon
- sugarcanet0
- Treatment
- OD6000.4 of Agrobacterium tumefaciens and 20 min of infecting timee0
- Interactor
- Agrobacterium tumefaciens
Agrobacterium tumefaciense0other - Outcome
- callus differentiatione0
- Direction
- beneficiale0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“sugarcane”
e0“OD6000.4 of Agrobacterium tumefaciens and 20 min of infecting time were beneficial to callus differentiation”
Claim · model-written summary
The optimal PPT dose for sugarcane callus induction and differentiation was 0.50 mg/L.
Verbatim
t0“sugarcane”
e0“The optimal dosage of PPT for callus induction and differentiation was 0.50 mg/L.”
Claim · model-written summary
Cef at 500.00 mg/L inhibited Agrobacterium growth in sugarcane co-culture.
- Taxon
- sugarcanet0
- Treatment
- 500.00 mg/L antibiotic Cefe0
- Interactor
- Agrobacterium tumefaciensAgrobacterium tumefaciense0other
- Outcome
- growth of Agrobacterium tumefaciense0
- Direction
- inhabite0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“sugarcane”
e0“adding 500.00 mg/L antibiotic Cef in co-culture could effectively inhabit growth of Agrobacterium tumefaciens”
Claim · model-written summary
Cef at 300.00 mg/L promoted sugarcane callus differentiation.
Not reportedComparator
Verbatim
t0“sugarcane”
e0“adding 300.00 mg/L Cef could promote callus differentiation”
Claim · model-written summary
Cef at 200.00 mg/L promoted sugarcane plantlet rooting.
Not reportedComparator
Verbatim
t0“sugarcane”
e0“200.00 mg/L Cef could promote plantlet rooting”
Claim · model-written summary
Low dose NAA in MS medium benefited sugarcane callus differentiation.
- Taxon
- sugarcanet0
- Treatment
- Low dosage of NAA in MS mediume0
- Outcome
- callus differentiatione0
- Direction
- more beneficiale0
Not reportedComparator
Verbatim
t0“sugarcane”
e0“Low dosage of NAA in MS medium was more beneficial to callus differentiation”
Claim · model-written summary
KT was superior to 6-BA for promoting sugarcane cell division.
Verbatim
t0“sugarcane”
e0“KT was superior to 6-BA in promoting cell division”
Claim · model-written summary
Adding 5.00 mg/L NAA and 70.00 g/L sugar to MS medium promoted sugarcane plantlet rooting.
- Taxon
- sugarcanet0
- Treatment
- 5.00 mg/L NAA and 70.00 g/L sugar in MS mediume0
- Outcome
- plantlets rootinge0
- Direction
- promotee0
Not reportedComparator
Verbatim
t0“sugarcane”
e0“By adding 5.00 mg/L NAA and 70.00 g/L sugar in MS medium could effectively promote plantlets rooting.”
Claim · model-written summary
PPT selection yielded 286 anti-PPT regenerated sugarcane plantlets transferred with Cbcor15 a.
Verbatim
t0“sugarcane”
e0“After the preliminary selection using PPT, 286 anti- PPT regenerated plantlets transferred with Cbcor15 a gene were obtained.”
Claim · model-written summary
Four of 83 selected sugarcane plantlets tested positive by PCR.
Verbatim
t0“sugarcane”
e0“Eighty-three plantlets with well growing were selected to detect by PCR, out of which four anti-PPT regenerated plantlets were positive.”
Claim · model-written summary
The authors concluded that Cbcor15 a was integrated into the sugarcane genome.
Verbatim
t0“sugarcane”
e0“Cbcor15 a gene had been integrated into genome of sugarcane.”
Claim · model-written summary
The authors reported successful establishment of an Agrobacterium mediated sugarcane transformation system.
Not reportedComparatorValue
Verbatim
t0“sugarcane”
e0“The Agrobacterium tumefaciens-mediated genetic transformation system was established successfully for exogenous gene transferring to sugarcane”
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