Regeneration of istrian ecotypes of garlic through somatic embryogenesis

Abstract

Češnjak (Allium sativum L.) je poljoprivredna kultura koja se razmnožava vegetativno. To ga čini osjetljivim na razne bolesti, pogotovo na viruse koji se preko zaraženog materijala prenose u slijedeću generaciju i akumuliraju. U istraživanju regeneracije češnjaka somatskom embriogenezom korištena su dva istarska ekotipa (12 i 13) dobivena od Instituta za poljoprivredu i turizam u Poreču. Kao eksplantati su korištene baze češnja postavljene na dva hranidbena medija različitih sastava (MS i modificirani MS), a svaki od njih je imao dva različita sastava regulatora rasta. Kalogeneza je postignuta na oba ekotipa i svim tretmanima. Postotak embriogeneze je bio viši kod ekotipa 13, ali ne signifikantno različit od ekotipa 12. Broj regeneriranih biljaka se značajno razlikovao (P<0,05) između tretmana. Na tretmanu MS 2,4 D je bilo najviše regeneriranih biljaka po embriogenom kalusu u oba ekotipa. Uspješno je zakorijenjeno i posađeno u supstrat 239 regeneriranih biljaka od kojih je 70% preživjelo. Pomoću ELISA metode je provedena laboratorijska detekcija četiri virusa: virus žućenja i kržljavosti luka (OYDV), virus žute prugavosti poriluka (LYSV), obično latentni virus češnjaka (GCLV) i latentni virus kozjaka (SLV). Biljke regenerirane somatskom embriogenezom uspoređene su s matičnim biljkama. Od ukupno 82 ispitane regenerirane biljke, 16 ih je bilo potpuno oslobođeno od virusa, odnosno 19,5%. Rezultati ovog istraživanja ukazuju na to da je moguće pomoću somatske embriogeneze dobiti biljke oslobođene od virusa.Garlic (Allium sativum L.) is an agricultural crop which is propagated vegetatively. This makes garlic more susceptible to various diseases, especially viruses that are transmitted and accumulated through the infected material to the next generation. In the study of garlic regeneration by somatic embryogenesis two Istrian ecotypes (12 and 13) obtained from the Institute of Agriculture and Tourism in Poreč were used. Clove bases were used as explants and placed on two nutrient media of different compositions (MS and modified MS). Both media had also two different growth regulator compositions. Callogenesis was achieved on both ecotypes and all four treatments. The percentage of embryogenesis was higher for ecotype 13, but not significantly different than for ecotype 12. The number of regenerated plants differed significantly (P<0,05) between treatments. MS 2,4 D treatment had the highest number of regenerated plants per callus in both ecotypes. Successfully were planted and acclimatized 239 regenerated plants, of which 70% survived. Laboratory detection of four viruses was performed using the ELISA method, which included: onion yellow dwarf virus (OYDV), leek yellow stripe virus (LYSV), garlic common latent virus (GCLV) and shallot latent virus (SLV). Plants regenerated by somatic embryogenesis were compared with mother plants. Of the 82 regenerated plants tested, 16 were completely virus free (19,5%). The results of this study indicate that it is possible to obtain virus-free plants by somatic embryogenesis