The procedure providing enhanced Agrobacterium-mediated transformation of wheat
2014
Тип документа:
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт:
The examinations of conditions for establishing a variety independent
Agrobacterium-mediated transformation procedure for wheat are preferable
since many of cultivars and breeding lines remain recalcitrant for
biotechnological manipulation, mainly due to low efficiency of plant
regeneration in vitro, which is highly genotype specific. This paper
describes and discusses an improved protocol for enhanced and
low-genotype dependent Agrobacterium-mediated transformation using a
super-binary vector LBA4404/pTOK233 carrying reporter gus-intron gene
and hygromycin (hpt) and kanamicyn (nptII) selectable marker genes. The
protocol was optimized on highly responsive common wheat cv. Vesna.
Transient expression monitored by the gus-intron on explants after 3, 6
and 25 days of co-cultivation, followed by GUS expression and hygromycin
resistance in whole plants indicated the protocol including a
co-cultivation of freshly isolated immature embryos in the presence of
ascorbic acid, and acetosyringone added only in the bacteria-containing
infection medium combined with a delayed and stepwise increasing
hygromycin B selection procedure significantly enhanced the
transformation efficiency in cv. Vesna that exceed 7\% of treated
explants from previously 0.41\%. Explant pre-cultivation did not
additionally improve transformation efficiency. The optimized protocol
was successful in evoking satisfactory transformation efficiencies from
3.6\% to 10.8\% in 5 less-responsive wheat genotypes. All 57 T0
hygromycin-resistant and GUS-positive lines were phenotypically normal
and fertile. Therefore, the conditions employed in this study may serve
as a base to facilitate the transformation in other, particularly
recalcitrant wheat cultivars.
Кључне речи:
Agrobacterium; immature embryos; transformation frequency; regeneration; wheat genotypes.Извор:
Biologia, 2014, 69, 12, 1668-1677Финансирање / пројекти:
- Биотехнологија ин витро - гајене, лековите и угрожене биљне врсте (RS-MESTD-Basic Research (BR or ON)-173015)
DOI: 10.2478/s11756-014-0477-2
ISSN: 1336-9563