Analiza potencialnih efektorjev fitoplazme 'Candidatus Phytoplasma solani' v transformiranih rastlinah vrste Nicotiana benthamiana

Strah, Rebeka (author), Pompe Novak, Maruša (mentor)

Fitoplazme so rastlinski intracelularni paraziti, ki živijo v floemu in jih prenašajo žuželke s sesanjem okuženega floemskega soka. V Sloveniji fitoplazma 'Candidatus. Phytoplasma solani' povzroča na vinski trti bolezen počrnelosti lesa. Bolezenska znamenja se pojavijo po vsej rastlini zaradi fitoplazemskih efektorskih proteinov. Da bi bolje preučili naravo efektorskih proteinov 'Ca. P. solani', smo z agrotransformacijo rastlin Nicotiana benthamiana prehodno izrazili šest potencialnih efektorjev spojenih z rumenim fluorescenčnim proteinom in jih šestnajst ur opazovali pod konfokalnim makroskopom. Z ustreznim redčenjem bakterijskih kultur smo transformirali večino celic na listni površini in posameznih celic, v katerih razdelkih smo videli fluorescenco. Pričakovanega prehajanja proteinov med rastlinskimi celicami nismo opazili ne pri potencialnih efektorskih proteinih, ne pri kontrolnem zelenem fluorescentnem proteinu. Domnevamo, da je nezmožnost difuzije proteinov med celicami v našem sistemu posledica imunskega odziva rastline na okužbo z Aagrobacterium tumefaciens, potrebno za transformacijo. Poznavanje mehanizmov navadne trsne rumenice je bistveno za njeno preprečevanje in zatiranje, zato moramo poglobiti raziskave potencialnih efektorskih proteinov, ki jo povzročajo.

• Language: Slovenian
• Keywords: fitoplazma, efektorski proteini, 'Candidatus Phytoplasma solani', Nicotiana benthamiana, prehodna transformacija
• Work type: Bachelor thesis/paper
• Typology: 2.11 - Undergraduate Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [R. Strah]
• Year: 2018
• PID: 20.500.12556/RUL-102947
• UDC: 602.6:582.930:632.3:577.21(043.2)
• COBISS.SI-ID: 9054585
• Publication date in RUL: 12.09.2018

Secondary language

• Language: English
• Title: Analysis of potential effectors of phytoplasma 'Candidatus Phytoplasma solani' in transformed Nicotiana bentamiana plants

Abstract

The phytoplasma are plant intracellular parasites that live in the phloem and are transmitted by insects ingesting infected floem sap. In Slovenia the phytoplasma 'Candidatus Phytoplasma solani' causes Bois noir disease in grape vine. The symptoms appear across the whole plant due to phytoplasma effector proteins. To better understand the nature of effector proteins, we transiently expressed six potential effectors fused with yellow fluorescent protein in Nicotiana benthamiana plants with agrotransformation and observed them for sixteen hours with a confocal macroscope. With appropriate dilutions of the bacterial cultures we successfully transformed most cells and single cells on the leaf surface and observed fluorescence in their compartments. We did not observe the expected movement of proteins between the plant cells for the potential effector proteins and the control green fluorescent protein. We assume that the inability of protein diffusion between cells in our system is a consequence of an immune response of the plants to infection with Aagrobacterium tumefaciens, needed for the transformation. Understanding the mechanisms of Bois noir disease is crucial for its prevention and suppression; it is therefore necessary to increase the research of the potential effector proteins that cause it.

• Keywords: phytoplasma, effector proteins, 'Candidatus Phytoplasma solani', Nicotiana benthamiana, transient transformation