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Ugotavljanje citoplazmatske moške sterilnosti pri zelju (Brassica oleracea L. var. capitata f. alba)
Podržaj, Adriana (Author), Murovec, Jana (Mentor) More about this mentor... This link opens in a new window, Rudolf Pilih, Katarina (Co-mentor)

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Abstract
Žlahtnjenje hibridnih sort zelja je zapleten in dolgotrajen postopek. Pri križnicah se za pridobivanje hibridnega semena pogosto uporablja samoinkompatibilnost peloda, ki pa ne zagotovi v celoti hibridnega potomstva, saj samoopraševanja ne preprečuje popolnoma. Citoplazmatska moška sterilnost (CMS) je najbolj učinkovita in zanesljiva metoda pridobivanja hibridov. Zagotavlja nam popolnoma hibridno potomstvo, saj samooprašitev ni mogoča. CMS določajo geni na mitohondrijski DNA, ki se zato dedujejo po materi preko citoplazme. Poznanih je več virov CMS. Najpogostejši pri križnicah je tip Ogura CMS, ki je bil prvotno odkrit pri redkvici. V raziskavi smo želeli pri izbranih sortah zelja z molekulskimi in fenotipskimi markerji določiti prisotnost gena orf138, ki je odgovoren za pojav Ogura CMS. Za preverjanje prisotnosti Ogura CMS smo uporabili štiri že objavljene mikrosatelitne začetne oligonukleotide. Po pomnoževanju in sekvenciranju med sortami nismo opazili razlik. Oblikovali smo nove začetne oligonukleotide za pomnoževanje regije na kateri se nahaja gen orf138. Z njimi smo uspešno pomnožili želeno regijo. Dobljene sekvence so bile homologne objavljenim sekvencam za CMS pri redkvici in zelju. Gen za CMS je bil prisoten pri dveh hibridnih sortah zelja: 'Liberator F1' in 'Tolerator F1'. Opravili smo tudi izolacijo in gojenje mikrospor za preverjanje androgenetske odzivnosti sort in s tem fertilnosti mikrospor. Morfološki pregled cvetov nam je potrdil molekulske rezultate, saj so bili prašniki pri rastlinah s CMS zakrneli in niso tvorili peloda.

Language:Slovenian
Keywords:zelje, Brassicca oleracea var. capitata f. alba, citoplazmatska moška sterilnost, CMS, PCR, nukleotidno zaporedje
Work type:Master's thesis/paper (mb22)
Tipology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Year:2018
Publisher:[A. Podržaj]
UDC:635.342:601.4:577.212.3(043.2)
COBISS.SI-ID:9074297 This link opens in a new window
Views:759
Downloads:317
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Secondary language

Language:English
Title:Identification of cytoplasmic male sterility in white cabbage (Brassicca oleracea L. var. capitata f. alba)
Abstract:
Cabbage breeding is a very complicated and long-term procedure. In Brassicas the production of hybrid seeds generally relies on self-incompatibility system that does not provide fully hybrid progeny. CMS is an easier and faster method to achieve hybrid seeds. It ensures completely hybrid progeny, because self-pollination is not possible. CMS is inherited maternally through cytoplasm. There are few sources of CMS, but the most commonly used in Brassicca breeding is the Ogura type. It was introduced from radish CMS cytoplasm. In our research we wanted to confirm the presence of the gene orf138 in our genotypes with molecular and phenotypic markers. Isolated DNAs from 20 varieties of white cabbage were checked with published PCR based markers. The amplification and sequence analysis did not reveal any significant differences among the varieties. New primer pairs aimed at amplifying regions of the orf138 gene were therefore designed and tested. We successfully amplified the region including the orf138 gene. Sequence analysis showed homology with already known sequences for CMS in radish and cabbage. The orf138 gene, responsible for CMS, was confirmed in two studied hybrid varieties: 'Liberator F1' and 'Tolerator F1'. We also isolated microspores of our genotypes to check their responsiveness to microspore embryogenesis. With observation of flower morphology we confirmed molecular results, because there was no pollen formed at the stamens.

Keywords:cabbage, Brassicca oleracea var. capitata f. alba, cytoplasmic male sterility, CMS, PCR, nucleotide sequence

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