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Določanje spola pri navadni konoplji (Canabis sativa L.) s pomočjo DNA markerjev in vplivanje na njegovo izražanje
ID Slapnik, Miha (Author), ID Murovec, Jana (Mentor) More about this mentor... This link opens in a new window, ID Flajšman, Marko (Co-mentor)

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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 %.

Language:Slovenian
Keywords:Cannabis sativa, industrijska konoplja, medicinska konoplja, indukcija spola, določanje spola, SCAR markerji
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Publisher:[M. Slapnik]
Year:2018
PID:20.500.12556/RUL-103521 This link opens in a new window
UDC:633.522:601.4:577.21(043.2)
COBISS.SI-ID:9040249 This link opens in a new window
Publication date in RUL:19.09.2018
Views:2352
Downloads:555
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Secondary language

Language:English
Title:Sex determination of hemp and medical cannabis (Cannabis Sativa L.) with DNA markers and influence on sex expression
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%.

Keywords:Cannabis sativa, industrial hemp, medical cannabis, gender induction, sex determination, SCAR markers

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