Anther Culture Regeneration fromHelianthus Mollis(M-020) and its Hybrid Combinations

Abstract

Anthers from the He/ianthus mollis (M-020) species and from the hybrid combination H.mollis (Jvf-020) x H.annuus and H.annuus x H.mollis (lv/-020) were isolated. Different variants of nutrient media were tested. The most suitable medium for plant regeneration proved to be the B5 medium without hormones and with reduced sucrose content. Regeneration was obtainedji•om the H.mollis (M-020) as well asji•om the hybrid combination H.mollis (M-020) x H.annuus. On~y well formed embryos were obtained from the reciprocal crosses. Investigation on the ploidity level of the regenerants are still in progress.

The Production of Helianthus Haploids: A Review of Its Current Status and Future Prospects

“…The doubled haploids of sunflower can be produced in vitro from an anther culture by direct embryogenesis [ 9 , 17 , 38 , 39 ], indirect embryogenesis [ 13 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 ], or indirect shoot organogenesis [ 10 , 17 , 38 , 40 , 44 , 47 , 48 , 49 , 50 , 51 , 52 ] ( Figure 1 ). Several independent research groups have reported the formation of callus tissue from anther culture without any subsequent morphogenic response ( Table 1 and Table 2 ) [ 9 , 17 , 30 , 41 , 42 , 43 , 44 , 47 , 48 , 49 , 50 , 53 ].…”

“…The time interval from the beginning of anther culture to callus tissue formation and the subsequent organogenesis or embryogenesis varies considerably. In particular, callus is formed in vitro on days 5–10, depending on the genotype [ 40 , 42 , 51 ], on days 10–15 [ 45 , 54 ], on days 15–20 [ 14 , 52 ], and after 4 weeks of cultivation [ 41 , 44 ]. Indirect shoot organogenesis or embryogenesis is observable after 4 weeks of culture [ 10 ] and later, after 80–100 days [ 52 ] or 120 days [ 51 ].…”

“…In particular, callus is formed in vitro on days 5–10, depending on the genotype [ 40 , 42 , 51 ], on days 10–15 [ 45 , 54 ], on days 15–20 [ 14 , 52 ], and after 4 weeks of cultivation [ 41 , 44 ]. Indirect shoot organogenesis or embryogenesis is observable after 4 weeks of culture [ 10 ] and later, after 80–100 days [ 52 ] or 120 days [ 51 ]. Several researchers have noted that the complete cycle from the beginning of anther culture to the adaptation of in vitro regenerants to the soil conditions is rather long, lasting for up to 16–19 weeks [ 17 ].…”

“…The data listed in Table 1 and Table 2 demonstrate that the composition of the culture medium has a crucial effect on the morphogenesis efficiency of the anther cultures of different sunflower genotypes. Independent of the direct or indirect morphogenesis pathway in sunflower anther cultures, two culture media differing in their composition are most frequently used: callus induction medium (CIM), or embryo induction medium (EIM), and shoot induction medium (SIM) for callus tissue [ 13 , 17 , 38 , 39 , 40 , 41 , 42 , 44 , 45 , 47 , 48 , 49 , 51 , 52 ]. The use of anther culture in the same medium for callus induction, shoot regeneration, embryogenesis, and further development (EDM) is relatively rare [ 9 , 46 ].…”

“…A major part of both CIM and EIM is the basal components of Murashige and Skoog (MS) [ 55 ] culture medium [ 10 , 14 , 40 , 41 , 42 , 43 , 44 , 46 , 47 , 48 , 50 , 52 , 53 ]. As has been demonstrated, basal MS medium provides the best results for morphogenesis induction as compared with Gamborg (B 5 ) or Nitsch media [ 41 , 54 ].…”