Noga Angelika, Skrzypek Edyta, Warchoł Marzena, Czyczyło-Mysza Ilona, Dziurka Kinga, Marcińska Izabela, Juzoń Katarzyna, Warzecha Tomasz, Sutkowska Agnieszka, Nita Zygmunt, Werwińska Krystyna
Department of Biotechnology, Polish Academy of Sciences, The Franciszek Górski Institute of Plant Physiology, Niezapominajek 21, 30-239 Kraków, Poland.
Department of Plant Breeding and Seed Science, University of Agriculture, Łobzowska 24, 31-140 Kraków, Poland.
In Vitro Cell Dev Biol Plant. 2016;52(6):590-597. doi: 10.1007/s11627-016-9788-z. Epub 2016 Nov 4.
Obtaining oat DH lines is only effective via wide crossing with maize. Seven hundred haploid embryos from 21 single F progeny obtained from wide crosses with maize were isolated, divided into four groups according to their size (<0.5 mm, 0.5-0.9 mm, 1.0-1.4 mm, and ≥1.5 mm), and transferred into 190-2 regeneration medium with different growth regulators: 0.5 mg L kinetin (KIN) and 0.5 mg L 1-naphthaleneacetic acid (NAA); 1 mg L zeatin (ZEA) and 0.5 mg L NAA; or 1 mg L dicamba (DIC), 1 mg L picloram (PIC), and 0.5 mg L kinetin (KIN). Among all isolated embryos, approximately 46.1% were between 1.0-1.4 mm, while the smallest group of embryos (7.1%) were those <0.5 mm. The ability of haploid embryos to germinate varied depending on oat genotypes and the size of embryos. Haploid embryos <0.5 mm were globular and did not germinate, whereas embryos ≥1.5 mm had clearly visible coleoptiles, radicles, and scutella, and were able to germinate. Germination of oat haploid embryos varied depending on growth regulators in the regeneration medium. Most haploid embryos germinated on medium with 0.5 mg L NAA and 0.5 mg L KIN, while the fewest germinated on medium with 1 mg L DIC, 1 mg L PIC, and 0.5 mg L KIN. One hundred thirty germinated haploid embryos converted into haploid plants. Fifty oat DH lines were obtained after colchicine treatment.
只有通过与玉米进行远缘杂交才能有效地获得燕麦双单倍体系。从与玉米远缘杂交得到的21个单F子代中分离出700个单倍体胚胎,根据其大小(<0.5毫米、0.5 - 0.9毫米、1.0 - 1.4毫米和≥1.5毫米)分为四组,并转移到含有不同生长调节剂的190 - 2再生培养基中:0.5毫克/升激动素(KIN)和0.5毫克/升1 - 萘乙酸(NAA);1毫克/升玉米素(ZEA)和0.5毫克/升NAA;或1毫克/升麦草畏(DIC)、1毫克/升毒莠定(PIC)和0.5毫克/升激动素(KIN)。在所有分离出的胚胎中,约46.1%的胚胎大小在1.0 - 1.4毫米之间,而最小的一组胚胎(7.1%)是<0.5毫米的胚胎。单倍体胚胎的萌发能力因燕麦基因型和胚胎大小而异。<0.5毫米的单倍体胚胎呈球形,不能萌发,而≥1.5毫米的胚胎有明显可见的胚芽鞘、胚根和盾片,能够萌发。燕麦单倍体胚胎的萌发因再生培养基中的生长调节剂而异。大多数单倍体胚胎在含有0.5毫克/升NAA和0.5毫克/升KIN的培养基上萌发,而在含有1毫克/升DIC、1毫克/升PIC和0.5毫克/升KIN的培养基上萌发的胚胎最少。130个萌发的单倍体胚胎转化为单倍体植株。秋水仙素处理后获得了50个燕麦双单倍体系。