Session 7: Molecular basis of response to post-harvest treatment stresses

biosecurity built on science

PBCRC3149 Molecular basis of response to post-harvest

treatment stresses

Plant Biosecurity Cooperative Research Centre

Wei Xu, Ph.D.Murdoch University


Team Capability

David Eagling (Program Leader)

Murdoch University Dr. Wei Xu (Functional Genomics, Insect Molecular Biology)Prof. Yonglin Ren (Postharvest treatment)Dr. Kay Anantanawat (Molecular Entomology and Bioinformatics)Dr. Antonios Avgoustinos (Insect Genetics)

SARDIDr. Kelly Hill (Molecular Entomology)Dr. Peter Crisp (Fruit fly biology)

Hawkesburry Institute of Environment Dr. Alexie Papanicolaou (Genomics, Bioinformatics)

IAEAKostas Bourtzis (Fruit fly biology)

Industry Advisor Kim James


Objectives Identify the effects of post-harvest treatments

to fruit fly cells Build biomarkers for different treatments

Specific Aims1) Identify early response genes when a lethal stress is

applied2) Characterise their function in stress response, cell

death and resistance


Long Term goal

Towards more efficient bio-control methods

1. Understand how concurrent stresses can improve control efficiency.

2. Build methodology and knowledge for other species and treatment strategies.


Model system

Ceratitis capitata (Mediterranean fruit fly)

Bactocera tryoni (Queensland fruit fly)

Stages: Eggs and Larvae (L1 and L3)


Experiment Plan

T0 T1 T2

Stressors Recover

Next Generation Sequencing


Cold and Heat Treatment

Dr. Kay Anantanawat

CURATIONS_mRNA1521 NPC2 homologNCBI2_rna1712 PREDICTED: uncharacterized protein LOC101459691NCBI2_rna20008 transmembrane 147NCBI2_rna20059 heat shock 70NCBI2_rna20060 heat shock 70NCBI2_rna5626 FAM92A1 isoform X1NCBI2_rna820 nucleolar 16NCBI2_rna8180 heat shock 27NCBI2_rna8409 MAK16 homologNCBI2_rna8490 thiamin pyrophosphokinase 1 isoform X1NCBI2_rna8953 DNA-binding RFX2NCBI2_rna9453 leucine-rich repeat 1 (cell death)NCBI2_rna959 tigger transposable element-derived 1-likeNCBI2_rna11308 OCIA domain-containing 1NCBI2_rna14874 elongation of very long chain fatty acids AAEL008004NCBI2_rna16681 heat shock 68

Identified 16 Candidate Genes in Medfly Heat (T1 vs T0)

Dr. Amanda ChooCRISPR, gene mutate/knock out

Identified 77 Candidate Genes in Medfly Heat (T2 vs T0)


Irradiation

IAEA

gamma cell 2000 irradiator


Egg Radiation TreatmentMedfly

0Gy 10Gy 20Gy 30Gy 40Gy 50Gy 60Gy 70Gy 80Gy 90Gy 100Gy0

5

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45

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0Gy 0Gy 10Gy 20Gy 30Gy 40Gy 50Gy 60Gy 70Gy 80Gy 90Gy 100Gy0

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EggsEgg_hatchDead_L1PupationLive_PupaeAdults_emergedMalesFemalesAdults_DNE

Qfly


Summary of Irradiation Treatment

1. Eggs (0-200 Gy): Qfly more resistant, Low doses of up to 20 Gy are enough to prevent adult emergence. 2. L1: up to ~150 Gy do not seem to affect the pupation percentages severely.

3. L3: low doses ranging between 10-40Gy, lead to total prevention of adult emergence.

4. Collection of samples for RNA extraction to discover WHY


Cold Treatment on Medfly in Blue Berry

Mean number of pupae, adults, females, males and incompletely emerged flies from control and cold treated eggs at 0 in cold room 6. ℃


Genome Work

Funded by US governmentco-ordination from Al Handler (ARS-USDA)

Alexie Papanicolaou


Acknowledgements

PBCRC is established and supported under the Australian Government Cooperative Research Centres Programme

Please email [email protected]

Collaborations are welcome!

mailto:[email protected]

Science

Session 7: Molecular basis of response to post-harvest treatment stresses