Articles
In vitro collection methods for Malus shoot cultures used for developing a cryogenic bank in Kazakhstan
Article number
1113_40
Pages
271 – 277
Language
English
Abstract
Initiation of shoot cultures from field-grown plants is often difficult due to the high level of bacterial and fungal organisms that populate trees.
This study compared shoot initiation techniques for apples.
Shoots were forced from dormant bud wood in the laboratory or directly taken from the field as new growth in the spring.
Fourteen promising apple cultivars, five clonal rootstocks (Malus × domestica Borkh.) and 10 wild Malus sieversii (Ledeb.
M. Roem.) were introduced into culture.
All shoots were treated with 0.1% HgCl2 for 10 min.
Cultures were initiated on liquid Murashige and Skoog medium containing 30 g L-1 of sucrose, 0.5 mg L-1 of 6-benzylaminopurine, 0.01 mg L-1 of indole-3-butyric acid, 1 mg L-1 of gibberellic acid and 1 mg L-1 of L-ascorbic acid, рН 5.7, for 2-4 weeks with daily transfers to fresh medium, and then moved to solid MS medium for propagation.
Cultures were indexed for bacteria and fungi using the 523 detection medium and any infected shoots were discarded.
Shoot initiation from forced dormant vegetative buds varied from 22.3 to 82.3% with a mean of 55.0% for the 35 genotypes.
Shoots, collected from new growth under field conditions, initiated clean shoots with a range from 5.1 to 44.8% and a mean of 18.4% for all genotypes.
Shoot cultures were established from all of the tested genotypes.
Many of these genotypes had never been cultured before, so this study is strategic for the security of important Malus germplasm.
These shoot cultures will be used for the creation of KazakhstanRSQUOs apple cryobank.
This study compared shoot initiation techniques for apples.
Shoots were forced from dormant bud wood in the laboratory or directly taken from the field as new growth in the spring.
Fourteen promising apple cultivars, five clonal rootstocks (Malus × domestica Borkh.) and 10 wild Malus sieversii (Ledeb.
M. Roem.) were introduced into culture.
All shoots were treated with 0.1% HgCl2 for 10 min.
Cultures were initiated on liquid Murashige and Skoog medium containing 30 g L-1 of sucrose, 0.5 mg L-1 of 6-benzylaminopurine, 0.01 mg L-1 of indole-3-butyric acid, 1 mg L-1 of gibberellic acid and 1 mg L-1 of L-ascorbic acid, рН 5.7, for 2-4 weeks with daily transfers to fresh medium, and then moved to solid MS medium for propagation.
Cultures were indexed for bacteria and fungi using the 523 detection medium and any infected shoots were discarded.
Shoot initiation from forced dormant vegetative buds varied from 22.3 to 82.3% with a mean of 55.0% for the 35 genotypes.
Shoots, collected from new growth under field conditions, initiated clean shoots with a range from 5.1 to 44.8% and a mean of 18.4% for all genotypes.
Shoot cultures were established from all of the tested genotypes.
Many of these genotypes had never been cultured before, so this study is strategic for the security of important Malus germplasm.
These shoot cultures will be used for the creation of KazakhstanRSQUOs apple cryobank.
Authors
N.V. Romadanova, S.A. Mishustina, G.N. Matakova, S.V. Kushnarenko, I.R. Rakhimbaev, B.M. Reed
Keywords
Malus × domestica, micropropagation, aseptic culture, cultivar, rootstock, wild form
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