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