analysis of differential gene expression in blueberries · blue results expression pattern of chs...
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Analysis of differential gene expression in Blueberries
Sandra Martins, Manuela Matos, Valdemar Carnide, Berta Gonçalves
Linha ISAC – P2
Lowbush
Rabbiteye
Highbush - (Vaccinium corymbosum L.)
Vaccinium corymbosum L.
Northern Highbush Blueberry - 700 to 1000 chill hours
Southern Highbush Blueberry - 150 a 600 chill hours
Subgenus Vaccinium
Reduce DNA damage, which
may help protect against ageing
and cancerProtect
cholesterol in the blood from becoming damaged
May lower blood pressure
May help prevent heart disease
Can help maintain brain function and
improve memory
Can have anti-diabetic effects
May help fight urinary tract
infections
Chemical composition
Vitamins
Minerals
Anthocyanins
Polyphenols
Pectin
Tannin
Citric acid, Malic acid and Tartaric acid
Blueberries are the King of Antioxidant
Consumption
fresh
bakery
Jam
Ice cream
deserts
Duke Goldtraub
OzarkblueBluecrop
Green
Green/Pink
Blue
Aims
Four varieties
Three maturation phases
Evaluation of differential gene expression:
1st and 2nd year samples collection
RNA Extraction
Qualitative evaluation
cDNA
Semi-quantitative analysis Quantitative analysis – Real Time PCR
Material and Methods
Quantitative evaluation
Genes:
CHS
F3’5’H
ANS
ANR
UFGT Housekeeping genes:
Actina
EF-1α
GAPDH
Results
Green BlueGreen/Pink
Qualitative evaluation of RNA
2nd year - 2017
Pattern of RNA in the three phases of fruits maturation in Goldtraub variety
Expression pattern of ANS gene in the three phases of fruits maturation in each variety
Goldtraub Ozarkblue
139pb
Green Green/Pink Blue Green Green/Pink Blue
Results
Semi-quantitative analysis
2nd year - 2017
115pb
Goldtraub Ozarkblue
Green Green/Pink Blue Green Green/Pink Blue
Results
Expression pattern of CHS gene in the three phases of fruits maturation in each variety
Semi-quantitative analysis
2nd year - 2017
Results
Quantitative analysis – Real Time PCR
For the six genes the conditions of:
annealing temperature
number of cycles
dissociation curves
calibration lines
1st year - 2016
Standard curve of Actin geneAmplification curve of CHS gene
Dissociation curve of CHS gene
Results Quantitative analysis – Real Time PCR
1st year - 2016
cDNA
Complete the semi-quantitative analysis
Begin the quantitative analysis - Real Time PCR for the six genes
Future Work :
1st year - 2016
Complete the quantitative analysis - Real Time PCR, for four genes
2nd year - 2017
RNA extraction of two varieties
cDNA
Thank you