This document provides information about micropropagation of blueberries. It discusses that blueberries can be propagated through generative or vegetative methods, but in vitro propagation is more efficient. Explants are taken from axillary buds and sterilized before culturing on AN or WPM medium supplemented with cytokinins like zeatin or TDZ for shoot initiation and multiplication. IBA is used for root formation in half-strength WPM at pH 4.5. The plantlets are hardened in a mist chamber and nursery before planting in soil.

1. Turkish Republic
Ministry of Higher Education and Scientific Research
University of Cukurova
Biotechnology and science
BLUEBERRIES MICROPROPAGATION
Awara M. HAMAKHAN
PhD. Student from plant tissue culture
Email: Awarahamakhan@yahoo.com
2021
Kurdistan Regional Government
Ministry of Higher Education and Scientific Research
Sulaimani Polytechnic University
Bakrajo Technical Agriculture Institute

2. General information of blueberry
 The genus Vaccinium, a member of the
family Ericaceae, has approximately 400
species distributed to all continents
 The blueberry bush is a flowering shrub 4
m for highbush
 Blueberries fruit are small 5–16
millimeters in diameter and feature a
flared crown at the end with purple color.

3. Fruit nutrient
 Blueberries are among the most nutrient are
low in calories but high in nutrients
From 1-cup (148-gram) serving of blueberries
contains :
 Fiber: 4 grams
 Vitamin C: 24%
 Vitamin K: 36%
 Manganese: 25%
 Water : 85%
 Calories :84
 Carbohydrates: 15 grams

4. Medicinal used
 Inside the fruit have anthocyanin flavonoid that
often powerful antioxidant effect they have been
shown to protect against heart disease and cancer
, and can also help maintain bone strength,
mental health, and healthful blood pressure.
 Plant foods may also promote hair and skin
health, increased energy, and overall lower
weight.

5. Plant propagation
Two different methods for propagation
 Generative (sexual) Propagation
 Vegetative (asexual) Propagation:
Softwood and hardwood cuttings
Traditional propagation methods are
also not particularly efficient as regards
the number
 in vitro propagation

6. Plant propagation
 in vitro propagation of blueberries take explant through
axillary leaf bud branching.

7. Plant sterilization
 Filed treatment: The best way for controlled
fungus from explant must be apply fungicide
before take the explant from the filed.
 For more controlled disease, better to apply
pesticide or fungicide by irrigation to mother
plant.

8. Plant surface sterilization
 Wash the brunch by tap water to remove the soil
 Cut them into smaller pieces 1-3 cm with single node
explants .
 Surface sterilization; with anti bacterial and fungicide
0.1 % for 10 minute.
 sodium hypochlorite 20 % and add Tween-20 (1%)
shake for 10 min.
 Wash the pieces several times with sterile water
 In some cases should be change sodium
hypochlorite concentration and time
 If the explants is not clean should be use 0.1%
solution of mercuric chloride for 2-5 min.

9. Medium used
 As a basal culture medium
Anderson’s Rhododendron
medium - AN (Anderson,1980)
is efficiently used for shoot
initiation
 Alternatively for shoot
induction and multiplication
modified WPM (McCown
Woody Plant Medium) (Lloyd
and McCown,1980)
 20 g/L sucrose, 6 g/L agar

10. Hormones
 Cytokinin group hormone commonly zeatin riboside
apply to Initiation and multiplication shoots in blueberry
 Zeatin 1.0 mg/L show high number shoot multiplication.
 2ip-isopen-tenyladenine 2.0 mg/l, is the alternative
hormone for shoot multiplication
 TDZ- Thidiazuron, 1.0 mg/L is second alternative
cytokinin for shoot multiplication.
 IBA- Indole-3-butyric acid 0.1- 1.0 mg/L, is auxins group
supported callus and root formation
 GA3- Gibberellic acid 0.1-0.5 mg/L, have supported plant
length

14. pH for the blueberry media
 In highbush blueberry
micropropagation, shoot growth or
plantlet development is also highly
sensitive to other environmental
conditions, including pH,
temperature, sucrose, humidity and
growing substrate
 pH is adjusted 5.1-5.5 for shoot
multiplication
 Root formation and shoot thickness
pH is 4.5-5.0.

15. Root formation
The best media for rooting :
 Anderson medium or WPM
supplemented with 0.8-1.0 mg/L IBA
and 1.0 g/L activated charcoal and the
pH adjusted 4.5
 Sucrose 15 mg/L
 Agar 6.5-7.0 g/L

16. Acclimatization process
 Primary hardening in mist chamber culture plants on peat substrate with green
leaves and roots were first kept in the mist chamber for 14 days.
 Graded plant were transferred nursery and spray NPK 19:19:19 5g/15 litter
water with fungicide and regular irrigation and gradually lowering the
humidity by ventilation for 2-4 weeks
 Secondary hardening plantlets in height 8-10 cm (after 2 months) were
replanted into bigger containers (12.0 cm in diameter) and transferred to open-
air conditions.

17. Tanks for listening