Institute of Plant Biology, Biological Research Center, Hungarian Academy of Sciences, POB 521, H-6701, Szeged, Hungary.
Plant Cell Rep. 1995 Jan;14(4):232-5. doi: 10.1007/BF00233639.
Protoplasts of Helianthus giganteus and Helianthus annuus were fused using polyethylene glycol. Before fusion H.giganteus protoplasts were subjected to iodoacetic acid treatment to render them unable to divide. Fused protoplasts were cultured in V-KM medium containing benzylaminopurine and naphtaleneacetic acid. Hybrid calli were identified on the basis of their ability of embryogenic development contributed by the Helianthus giganteus parent. Fifty embryogenic calli were cultured on MS based medium without growth regulators to induce further development of somatic embryos. Elongated shoots were removed, rooted and transferred into growth chambers. Overall morphology of the plants was intermediate between the two parents. Their hybrid nature was confirmed by chromosome counting and by the analysis of esterase isozymes. The plants flowered within two to three months and later died. Thus the perennial nature of H.giganteus is a recessive trait in this interspecific hybrid. Seeds were obtained from two of the regenerated plants. From these seeds normal fertile F2 plants could be grown.
巨型向日葵和普通向日葵的原生质体通过聚乙二醇融合。融合前,用碘乙酸处理巨型向日葵原生质体,使其无法分裂。融合的原生质体在含有苄氨基嘌呤和萘乙酸的 V-KM 培养基中培养。杂种愈伤组织根据其巨型向日葵亲本的胚胎发生能力来鉴定。五十个胚性愈伤组织在没有生长调节剂的 MS 基础培养基上培养,以诱导体细胞胚的进一步发育。伸长的芽被去除,生根并转移到生长室中。这些植株的整体形态在双亲之间是中间型的。通过染色体计数和酯酶同工酶分析证实了它们的杂种性质。这些植株在两到三个月内开花,随后死亡。因此,巨型向日葵的多年生特性是这个种间杂种的隐性性状。从两个再生植株中获得了种子。从这些种子中可以培育出正常可育的 F2 植株。