Transformation of tomato cultivar Moneymaker with Agrobacterium tumefaciens
2018
Аутори:
Simonović, AnaBogdanović, Milica
Bergau, Nick
Tissier, Alain
Todorović, Slađana
Остала ауторства
Uzelac, BrankaТип документа:
Конференцијски прилог (Објављена верзија)
,
© 2018 by the Serbian Plant Physiology Society
Метаподаци
Приказ свих података о документуАпстракт:
Tomato (Solanum lycopersicum) is not only one of the most important vegetable crops, but
a widely used model for genetic improvement of dicotyledonous crops. Even though the first
report on tomato transformation using Agrobacterium tumefaciens has been published over 30
years ago, numerous protocols published since then demonstrate that tomato transformation is
neither standardized nor routine. Tomato cultivars and genotypes greatly differ in their susceptibility
to transformation and response to growth regulator and antibiotics combinations used during
regeneration and selection. Hereby we report a successful protocol for “Moneymaker” transformation
with A. tumefaciens GV3101:pSM90 bearing pAGT174 with Kanamycin resistance cassette.
Whole larger leaves (from ~10 cm high plantlets), rather than leaf segments were selected for
inoculation, as the explant size significantly affected regeneration rate. Combination of zeatin (1
mg L-1) and IAA (0.1 mg L-1) was used for preculture and throughout the selection process, with
addition of Timentin (200 mg L-1) and increasing concentrations of Kanamycin (from 20 to 50 mg
L 1) during the selection and regeneration. The regeneration on Kan occurred almost exclusively
in transformed explants and proceeded via organogenesis with the callusing stage. Using Timentin
instead of commonly used Cefotaxime to kill the bacteria proved beneficial for Moneymaker
regeneration. In this system the regeneration frequency was 7%. Plantlets that survived Kan selection
were PCR-tested, and 89.6% of them were transformed. The protocol is convenient and robust
in terms of very low false-positive rate (10.3%).
Кључне речи:
Moneymaker; transformation; Timentin; ZeatinФинансирање / пројекти:
- Физиолошка, хемијска и молекуларна анализа диверзитета одабраних ретких и угрожених биљних врста у циљу еx ситу заштите и продукције биолошки активних једињења (RS-MESTD-Basic Research (BR or ON)-173024)
У:
- 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. 21.