Effect of different sterilisation methods on establishing apricot (prunus armeniaca l.) tissue cultures

V letu 2017 smo na Fakulteti za kmetijstvo in biosistemske vede v Mariboru opravili poskus, v katerem smo proučevali vpliv različnih načinov sterilizacije, vpliv tipa brstov ter vpliv dodanega antibiotika v gojišče na vzpostavitev tkivne kulture marelice. Uporabili smo 6 različnih načinov sterilizacije brstov, katere smo predhodno tretirali 60 sekund v 70 % etanolu: 1. sedem minut DICA, 2. deset minut DICA, 3. petnajst minut DICA, 4. petnajst minut 0,75 % NaClO, 5. deset minut 5 % NaClO in 6. dvajset minut 10 % Ca(ClO)2. Uporabili smo dva tipa brstov, in sicer zaprte ter odprte. Razkužene brste smo prenesli na dve različni gojišči, eno z dodanim antibiotikom (Cefotaxime sodium), drugo brez. Kot najboljša metoda sterilizacije se je izkazala sterilizacija z deset minutnim tretiranjem z DICA. Pri tem načinu sterilizacije je ostalo vitalnih 15 % brstov. Največ brstov se je okužilo (74 %) pri petnajst minutni sterilizaciji z 0,75 % NaClO. Pri šestem načinu sterilizacije, kjer smo brste 20 minut mešali v Ca(ClO)2, je večina brstov propadla (79 %). Preživel ni noben brst. Statistično značilne razlike so se pokazale tudi med tipi brstov. Pri odprtih brstih smo opazili večjo stopnjo okužbe. Zaprtih brstov pa je več propadlo, kot se jih je okužilo. Gojišče z dodanim antibiotikom se ni statistično značilno razlikovalo od gojišča brez antibiotika.In this experiment, that was carried out in year 2017 at the Faculty of Agriculture and Life Sciences of Maribor, the effect of different sterilisation methods, the effect of the type of the buds and the effect of added antibiotic in culture medium on establishing apricot tissue cultures were investigated. We used 6 different ways of sterilisation. At first, we sterilized buds in 70 % ethanol for 60 seconds. At 1st sterilisation buds were treated with DICA for seven minutes, 2nd with DICA for ten minutes, 3rd with DICA for fifteen minutes, 4th with 0,75 % NaClO for fifteen minutes, 5th with 5 % NaClO for ten minutes and at 6th with 10 % Ca(ClO)2 for twenty minutes. We used two types of buds: opened and closed. Sterilized buds were transferred on two different medias. In one media we added antibiotic (Cefotaxime sodium), in the other one there was none. A ten minute treatment with DICA turned out as the best method of sterilisation. At this treatment 15 % of buds remained vital. At fifteen minute treatment with 0,75 % NaClO most buds were contaminated (74 %). At twenty minute treatment with Ca(ClO)2 most of the buds deceased (79 %), none of the buds survived. Statistically significant differences were also found among the types of buds. With open buds, the higher rate of contamination was observed, on the other hand the higher number of buds that did not survive the sterilisation were closed. Medium with antibiotic was not statistically different than medium without antibiotic