Clonal fidelity of Iris sibirica plants regenerated by somatic embryogenesis and organogenesis in leaf-base culture - RAPD and flow cytometer analyses
2015
Аутори:
Stanišić, MarianaRaspor, Martin
Ninković, Slavica
Milošević, Snežana
Ćalić, Dušica
Bohanec, Borut
Trifunović-Momčilov, Milana
Petrić, Marija
Subotić, Angelina
Jevremović, Slađana
Тип документа:
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт:
Efficient protocols, safe from somaclonal variation, were developed for
regeneration of Iris sibirica plants via organogenesis and somatic
embryogenesis from leaf-base explants cultivated on Murashige and Skoog
media supplemented with thidiazuron (TDZ, 1.0 mg/l) or
2,4-dichlorophenoxyacetic acid (2,4-D, 1.0 mg/l). The morphogenic
response and callus formation efficiency differed significantly between
2,4-D (80.9\%) and TDZ (67\%) morphogenesis induction treatments. TDZ
induced only organogenic calli, while calli obtained with 2,4-D were
composed of three types differing in color and consistency: white,
friable - embryogenic calli (4.5\%, 3.8 mg/explant), green, compact -
organogenic calli (12.4\%, 48.4 mg/explants) and yellow -
non-regenerative calli (773\%, 254.4 mg/explant). The cultivation of
embryogenic calli on medium with 2,4-0 and Kinetin resulted in further
development of somatic embryos (54 embryos/g of calli) which germinated
with a frequency of 62\% after being transferred to a medium without
plant growth regulators. Stable shoot cultures were established by
transferring organogenic calli with shoot primordia to media with 0.1
mg/l alpha-naphthaleneacetic acid (NAA) and 1.0 mg/l 6-benzylaminopurine
(BA), while further cultivation on media of the same composition (TDZ or
2,4-D) resulted in the reduced growth and rhizogenesis, respectively.
The TDZ induction treatment resulted in higher number of shoots per
explant (7.9) than the 2,4-0 treatment (43). After successful rooting
and ex vitro acclimatization, plants were grown in the field and
flowered to seed production. Flow cytometry, chromosome counting and
random amplification of polymorphic DNA (RAPD) analysis indicated no
evidence of genetic variation in plants regenerated via somatic
embryogenesis or organogenesis. The results suggest that established
protocols are safe for use in genetic transformation procedures or
large-scale production of true-to-type I. sibirica plants. (C) 2014
SAAB. Published by Elsevier B.V. All rights reserved.
Кључне речи:
Chromosome counting; Flow cytometry; Somaclonal variation; Tissue cultureИзвор:
South African Journal of Botany, 2015, 96, 42-52Финансирање / пројекти:
- Биотехнологија ин витро - гајене, лековите и угрожене биљне врсте (RS-MESTD-Basic Research (BR or ON)-173015)
- Развој и примена биотехнолошких поступака у добијању здравог садног материјала украсних биљака (RS-MESTD-Technological Development (TD or TR)-31019)
DOI: 10.1016/j.sajb.2014.10.014
ISSN: 1727-9321