magistrsko delo
Miha Slapnik (Author), Jana Murovec (Mentor), Marko Flajšman (Co-mentor)

Abstract

Molekulsko določanje spola z DNA markerji je določanje spola na genetskem nivoju. S pomočjo markerjev je mogoče spol določiti že pri zelo mladih rastlinah ali celo semenih. Določanje spola rastline pred njegovim fenotipskim izražanjem je koristno v žlahtniteljskih procesih. Indukcija moških cvetov na dvodomnih ženskih rastlinah je postopek, pri katerem z aplikacijo rastnih regulatorjev vplivamo na izražanje spola rastlin. Praktičen namen metode indukcije moškihcvetov na dvodomnih ženskih rastlinah je omogočiti samooplodnjo ali križanje rastlin, ki so genetsko ženskega spola. Na ta način lahko pridemo do čistih linij, pridobimo pa tudi femenizirana semena. Cilji magistrske naloge so bili preizkusiti markerje SCAR119, SCAR323 in MADC2 in optimizacija PCR mešanice ter temperatur prileganja začetnih oligonukleotidov za določanje spola pri navadni konoplji ter preveriti vpliv rastnih regulatorjev (GA3, STS) in različnih tretiranj na izražanje spola dvodomnih ženskih rastlin medicinske konoplje. Za najučinkovitejše tretiranje se je izkazalo tretiranje z neredčenim STS, pri katerem se je tvorilo največ moških socvetij in cvetov. Kot najučinkovitejši marker za določanje spola se je izkazal marker SCAR119. Za najbolj natančno tehniko brez odstopanja od fenotipskih ocen se je izkazala tehnika PCR s predhodno izolacijo DNA po CTAB metodi, pri kateri smo uporabili marker SCAR119. Optimizirali smo tudi protokol a določanje spola s t.i. direktnim PCR, kjer predhodna izolacija DNA ni potrebna, in ga preizkusili na 200 rastlinah navadne konoplje. Uspešnost določitve spola je bila 75 %.

Keywords

Cannabis sativa;industrijska konoplja;medicinska konoplja;indukcija spola;določanje spola;SCAR markerji;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [M. Slapnik]
UDC: 633.522:601.4:577.21(043.2)
COBISS: 9040249 Link will open in a new window
Views: 1287
Downloads: 493
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Other data

Secondary language: English
Secondary title: Sex determination in cannabis (Cannabis sativa L.) with use of DNA markers and influences on sex expression
Secondary abstract: Molecular sex determination with DNA markers is a gender determination on genetic level. With their help gender can be determined in very young plants or even seeds. Sex determination of plants before its phenotipic expression is useful in plant breeding processes. Induction of male flowers on dioecious female plants is a process in which with the help of growth regulators we influence the expression of sex. The practical purpose of the method of male-flower induction on dioecious female plants is to enable self-polination or crossing plants that are genetically female. In this way we can obtain pure lines and we also obtain feminized seeds. The goals of the master 's thesis were to test the markers for cannabis sex determination SCAR119, SCAR323 and MADC2, to optimize the PCR mixture and the temperatures of primers annealing for sex determination in cannabis and to test the effect of growth regulators (GA3, STS), and their various treatments on the expression of sex on dioecious female plants of medical cannabis. The most efficient treatment was the use of undiluted STS after which we obtained the highest number of male inflorescences and flowers. The marker SCAR119 proved to be the most effective marker for gender determination. The best technique for obtaining results in accordance with phenotypic observations was the technique with the use of isolated DNA and its amplification with the SCAR119 marker. We also optimized a sex determination protocol for i.e. direct PCR, where preliminary DNA isolation is not required, and we tested it on 200 hemp plants. The success for gender determination was 75%.
Secondary keywords: industrial hemp;medical cannabis;gender induction;sex determination;SCAR markers;
Type (COBISS): Master's thesis/paper
Study programme: 0
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Oddelek za agronomijo
Pages: XIII, 57 f., [15] f. pril.
ID: 10961060