Transgenic hairy root in vitro culture system of apple as a tool for allelopathic studies
2018
Authors:
Stanišić, MarianaĆosić, Tatjana
Savić, Jelena
Krstić Milošević, Dijana
Mišić, Danijela
Smigocki, Ann
Ninković, Slavica
Banjac, Nevena
Contributors
Uzelac, BrankaDocument Type:
Conference object (Published version)
,
© 2018 by the Serbian Plant Physiology Society
Metadata
Show full item recordAbstract:
Since the investigation of plant allelopathic interactions in the rhizosphere is difficult to perform
because of its high complexity, microbial and environmental factors interactions, the generation
of autonomous root culture could be appropriate system for the studies of allelopathic potential of apple (Malus x domestica Borkh.). Thus, we have focused our efforts on the generation
of transformed hairy-roots in apple cv. Golden Delicious using the 15834 strain of Agrobacterium
rhizogenes. The agrobacteria inoculation by dipping the basal cut off end of the stem of in vitro
grown apple shoots into bacterial suspension proved to be the best option for transformation,
providing efficiency of 39.3%. Molecular characterization of hairy roots by RT-PCR analyses confirmed
their transgenic nature. In vitro conditions for apple hairy root growth and maintenance
were optimized using different mineral solutions and auxins. The liquid half-strength MS medium
with the addition of IBA at 0.025 mg L-1 was chosen for the initial stimulation of growth. For their
prolonged cultivation and maintenance, the alternation of IBA-containing and IBA-free media is
recommendable. Fast-growing lines continuously maintained in IBA-free medium were used for
further research. Potential allelochemicals including phenolic acids (chlorogenic, caffeic, syringic,
p-coumaric and ferulic), glycosilated flavonoids (phloridzin, rutin, quercitrin, isoquercitrin, kaempferol-
3-glucoside) and flavonoid aglycons (floretin, quercetin and naringenin) have been detected
by UHPLC/DAD/(+/-)HESI-MS/MS analyses in the growth medium, in which these transformed
roots were maintained for 4 weeks, as well as in the root tissue itself. The obtained hairy-root
cultures offer a complementary system for studying and validating the allelopathic potential of
apple roots.
Keywords:
Agrobacterium rhizogenes,; allelopathy; allelochemicals; growth conditions; hairy rootsFunding / projects:
- Biotechnology in vitro - crop, medicinal and endangered plant species (RS-MESTD-Basic Research (BR or ON)-173015)
In:
- Uzelac B, editor. Book of abstracts. 3rd International Conference on Plant Biology (22nd SPPS Meeting); 2018 Jun 9-12; Belgrade, Serbia. Belgrade: Serbian Plant Physiology Society; Institute for Biological Research "Siniša Stanković"; Faculty of Biology; 2018. p. 55-6.