Study of chicory germacrene-A synthase function in guaianolide biosynthesis through amiRNA-induced gene silencing
2015
Аутори:
Dragićević, MilanBogdanović, Milica
Cankar, Katarina
Beekwilder, Jules
Bouwmeester, Harro
Todorović, Slađana
Simonović, Ana
Остала ауторства
Uzelac, BrankaТип документа:
Конференцијски прилог (Објављена верзија)
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© 2015 the Serbian Biological Society
Метаподаци
Приказ свих података о документуАпстракт:
Germacrene A synthase (GAS) catalyzes an important step in guaianolide biosynthesis converting farnesyl pyrophosphate to germacrene A – universal sesquiterpene precursor in chicory. The function of two genes coding for GAS – CiGASlo and CiGASsh, was studied by gene silencing. Chicory was transformed with amiRNA constructs designed to specifically silence either of the two genes, and regenerated transgenic plants were tested for changes in gene expression and sesquiterpene lactones content. Silencing was successful in most of the clones, which had reduced gene expression. However, the amiRNA constructs were not specific for CiGASlo or CiGASsh, so that both genes were silenced in most transgenic plants. To verify that the gene silencing had impact on sesquiterpene lactone levels, three representative guaianolide oxalates (lactucin-15-oxalate, deoxilactucin-15-oxalate and lactucopicrin-15-oxalate) were identified and quantified by UPLC-MS/MS in roots and shoots of transgenic and control plants. Data analyzes showed unequal segregation of targeted compounds in the plant tissues, and implied that the synthesis of these compounds highly depends on the expression of GAS genes in the shoot and that these metabolites are pooled in the shoot, while the root guaianolide oxalates content is regulated on a level different from GAS expression. Nevertheless gene silencing reduced the targeted metabolites levels significantly. It was concluded that both genes are involved in guaianolide biosynthesis, and that their complex pattern of expression in different parts of the plant determines the amount of guaianolides and their derivatives.
Кључне речи:
germacrene A synthase; silencing; amiRNA; chicory; guaianolidesФинансирање / 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)
- Plant Terpenoids for Human Health: a chemical and genomic approach to identify and produce bioactive compounds (EU-FP7-227448)
У:
- Uzelac B, editor. Book of Abstracts: 2nd International Conference on Plant Biology, 21th Symposium of the Serbian Plant Physiology Society, and CОST Action FA1106 Quality Fruit Workshop; 2015 Jun 17-20; Petnica, Serbia. Belgrade: Serbian Plant Physiology Society: Institute for Biological Research “Siniša Stanković”; 2015. p. 20.