Transgenic Banana Resistant to Banana Bunchy Top Virus
John Hu, et al.Plant & Environmental Protection Sciences
Transgenic Banana Project at UHM
This work is a result of contributions made by many individuals: Dr. Eden PerezDr. John Hu
Dr. Kheng Cheah Dr. Diane SetherDr. Said Khalil
Mr. Jason Lutz Dr. Wayne BorthMr. Doug Gaskill
Ms. Bethany Kimokeo (UH) Mr. Scott Henderson (UH) Dr. Douglas Becker (Aus)
Previous Work• ‘Dwarf Brazilian’ meristems were transformed• About 300 transformed plants were produced• These were screened for BBTV resistance in
bioassays using aphid inoculations• 100% of non-transformed control plants developed
symptoms within 3-4 weeks• 7 transformed plants did not develop BBTV
symptoms for one year after inoculation– BUT…all 7 eventually developed symptoms
Initiation of Cell Suspension Cultures
Bracts and ‘hands’Male inflorescence
Surface sterilization
Apical ~3cm
Production of EmbryogenicCell Suspensions
Male Inflorescence Male Flowers EmbryogenicCallus
2-4 weeks
EmbryogenicCell Suspension
Homogeneous Cell Suspension
1-6 months
Transformation of EmbryogenicCell Suspensions
Agrobacterium tumefaciens mediated
BBTV constructs:Coat Protein (CP)Coat Protein Mutation (M1)Replication Associated Protein (Rep)
What is BBTV?
100 nm ORF 1: Replication-associated protein (Rep)
ORF 3: Coat protein (CP)
Genome Component Functions
Screening for BBTV Resistance
Adults collected 5-10 adult aphids allowed 10 day
inoculation access
Aphids raised on BBTV-infected banana
Current Status of Transgenic Banana Lines
• # inoculated / # of lines– M1: 40 / 60– AS: 59 / 90– Rep∆2/5: 3 / 113– ∆2/5: 0 / 59– Control: 23 / 49
• Total # lines = 322• Total # inoculated = 102
Future Work
• Continue screening transgenic banana lines for BBTV resistance
• Further characterize any BBTV-resistant transgenic banana lines
• Establish and maintain new ‘Dwarf Brazilian’ embryogenic cell cultures
• Transform these embryogenic cell cultures with new constructs
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