Doubled haploid production via anther culture in annual, winter type of caraway (Carum carvi L.)

Abstract

Caraway (Carum carvi L.) is a traditional medicinal and spice cross-pollinated plant species. Although in vitro techniques are recently extensively applied in plant breeding programmes, these are not commonly utilized in caraway. Therefore, based on the protocol for anther culture in carrot (Daucus carota L., a closely related species of caraway in Daucaceae family), in vitro androgenesis in caraway has been studied with the aim to produce completely homozygous inbred lines. Various induction conditions, such as temperature pretreatments, carbon sources and combination of growth regulators in a culture medium as well as the effect of genotype on in vitro androgenesis were examined. Ten breeding lines of winter caraway representing third generation of forced (artificial) self-pollination were used as donor plant material. Cultured anthers produced embryogenic calli, and subsequently two types of regenerated plants were obtained, namely haploids with evident microspore origin, and diploids which may represent somatic (anther wall) regenerants or spontaneous doubled haploids. The ploidy status of regenerated plants was determined by flow cytometry. This is the first report on androgenic doubled haploid production in caraway.

Microspore Embryogenesis in the Apiaceae

“…Differences were observed between the growth conditions in terms of callus production, with greenhouse plants producing the most callus and fieldgrown plants producing the least; however, there was no difference in embryo production. Smykalova et al (2009) grew caraway donor plants for anther culture in the greenhouse, but provided different temperature regimes, including a cold treatment. The best response was from plants grown at 22°C for 9 days, followed by 19 days at 6°C.…”

“…In our experiments, several established basal media were compared in order to develop a DH protocol for fennel and caraway. We evaluated B5 medium, which is commonly used in many tissue culture protocols and has been used in Apiaceae anther culture and isolated microspore culture (Górecka et al 2005;Smykalova et al 2009;Dohya et al 1997). NLN is used for microspore culture in Brassica (Lichter 1982), N6 has been used in anther culture of cereals (Chu 1978), NN has been used in anther culture of dicots (Nitsch and Nitsch 1969), and K&M has been used with Vicia protoplasts (Kao and Michayluk 1975).…”

“…Limited response in terms of calli and/or embryoid production has been reported from anther cultures of celery, fennel (Foeniculum vulgare Mill. ), and mitsuba (Cryptotaenia japonica) (Dohya et al 1997), Bupleurum falcatum L. (Shon and Yoshida 1997), caraway (Smykalova et al 2009), and carrot (Adamus and Michalik 2003;Matsubara et al 1995;Górecka et al 2005Górecka et al , 2009). Plantlets were regenerated from celery, carrot, caraway, and B. falcatum anther culture; however, the efficiency of production was low.…”

Induction conditions for somatic and microspore-derived structures and detection of haploid status by isozyme analysis in anther culture of caraway (Carum carvi L.)

“…), parsley (Petroselinum crispum Miller), and carrot (Daucus carota L.), which have both nutritive and pharmaceutical properties, microspore-derived doubled haploid plants have been produced (Ferrie et al 2005Ferrie 2009). Production of haploids through anther culture has been reported for some varieties of caraway (Smýkalová et al 2009), where inductive conditions led to the production of true haploid plants, as well as plants of different ploidy levels. Analysis of the ploidy levels of various regenerated plants revealed haploid production in the range of 3-35% (Andersen et al 1990;Adamus and Michalik 2003;Gorećka et al 2005;Matsubara et al 2005;Smýkalová et al 2009).…”

Ploidy manipulation for genetic improvement in some Mediterranean fruit crops

“…In particular, for both registered parameters (AA: androgenic anthers and MC: morphogenic calli), the highest percentages (19.3 and 12.7 respectively) were detected in 'Sanfilippara', while the lowest percentages (1.94 and 0.97) were observed in 'Redonet'. In anthers culture, the effect of type and concentration of plant growth regulators has been widely studied (Oleszczuk et al 2004;Perera et al 2009;Smýkalová et al 2009;Germanà 2011a;Cardoso et al 2016). In woody species, different auxin types and concentrations, such as NAA, TDZ or 2.4-D, have been already employed to induce callus production in mandarin (Germanà and Chiancone 2003), papaya (Rimberia et al 2007) and also in loquat (Li et al 2008;Blasco et al 2015b).…”