Escobedo-Gracia-Medrano Rosa Maria, Cruz-Cárdenas Carlos Iván, Sánchez-Cach Lucila Aurelia, Ku-Cauich José Roberto, González-Kantún Wilma Aracely
Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán A.C., Mérida, Yucatán, Mexico.
Centro Nacional de Recursos Genéticos, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (INIFAP), Tepatitlán de Morelos, Jalisco, Mexico.
Methods Mol Biol. 2018;1815:215-226. doi: 10.1007/978-1-4939-8594-4_14.
Most cultivated bananas (Musa spp.) are polyploids, and their fruits are seedless and propagated exclusively vegetatively; however, they can also be cloned by micropropagation techniques, viz., direct organogenesis (DO) or somatic embryogenesis (SE). Banana indirect SE (ISE), with an embryogenic callus phase, is possible using young male or female flowers as direct explant depending on the genotype or shoot tips (scalps). For the False Horn Plantain, cv. Curraré (AAB, plantain subgroup), which has a degenerating male bud, female flowers are used to regenerate plants through ISE. Here, a protocol for increasing the number of initial explant material from a single mother plant and its embryogenic response is described. For those purposes, hands with young female buds are in vitro proliferated in the presence of 1 μM indole-3-acetic acid and 2.5 μM thidiazuron. Friable embryogenic cultures, here called ISE-2, obtained from the new proliferative secondary female bud clusters are initiated on medium containing auxins. Embryogenic suspensions are then established from the ISE-2 cultures. Regeneration of plants is achieved from embryogenic suspensions after plating on semisolid medium free of plant growth regulators; greenhouse acclimatized plantlets are ready for banana farming. This study demonstrates that proliferative female buds are a proper choice for ISE.
大多数栽培香蕉(芭蕉属)是多倍体,其果实无籽,仅通过营养繁殖;然而,它们也可以通过微繁殖技术进行克隆,即直接器官发生(DO)或体细胞胚胎发生(SE)。香蕉间接体细胞胚胎发生(ISE),具有胚性愈伤组织阶段,根据基因型或茎尖(芽尖),可以使用幼嫩雄花或雌花作为直接外植体。对于退化雄芽的假角大蕉品种Curraré(AAB,大蕉亚组),雌花用于通过ISE再生植株。在此,描述了一种增加来自单个母株的初始外植体材料数量及其胚性反应的方案。为此,带有幼嫩雌芽的果穗在1 μM吲哚-3-乙酸和2.5 μM噻苯隆存在下进行离体增殖。从新的增殖性次生雌芽簇获得的易碎胚性培养物,在此称为ISE-2,在含有生长素的培养基上起始。然后从ISE-2培养物建立胚性悬浮液。将胚性悬浮液接种在不含植物生长调节剂的半固体培养基上后可实现植株再生;温室驯化的幼苗可用于香蕉种植。本研究表明,增殖性雌芽是ISE的合适选择。