Syeed Rukaya, Mujib A, Malik Moien Qadir, Gulzar Basit, Zafar Nadia, Mamgain Jyoti, Ejaz Bushra
Cellular Differentiation and Molecular Genetics Section, Department of Botany, Jamia Hamdard, New Delhi, India.
J Appl Genet. 2022 May;63(2):199-211. doi: 10.1007/s13353-021-00663-y. Epub 2021 Dec 3.
Caladium × hortulanum 'Fancy' is an important ornamental plant grown in pots and landscapes and known for its colorful leaves often used for interior decorations. In this work, we present a method of in vitro regeneration from three explants source through direct somatic embryogenesis (DSE) wherein the regenerated plants were screened for ploidy changes through flow cytometry analysis. Tuber, leaf and petiole explants were cultured on MS basal medium supplemented with 1-napthalene acetic acid (NAA), 6-benzyl amino purine (BAP) and N-phenyl-N'-1, 2,3-thiadiazol-5-ylurea (TDZ) concentrations. Tuber explants induced highest direct somatic embryos on NAA (1 mg L) + BAP (0.5 mg L) with 55.6 mean number of embryos per explant while as leaf and petiole explants amended with 1 mg L TDZ developed 18.7 and 12.27 mean number of embryos per explant respectively. The highest embryo conversion frequency was achieved on BAP (2 mg L) + NAA (0.2 mg L) with 44.2, 18.7 and 7.5 mean number of plantlets produced per tuber, leaf and petiole explant respectively after 4 weeks of culture. Plantlets were later rooted and maximum number of roots (6.33) per shoot was achieved on 2 mg L indolebutyric acid amended medium. Description of the process of DSE is presented through the histological and SEM evidences. The 2C DNA content of field grown plants and the DSE regenerants evaluated under flow cytometric analysis were 8.06 pg and 8.28 pg respectively showing no ploidy changes. Hence, a successful protocol of inducing direct somatic embryos from three explant types with efficient embryo conversion frequency was obtained with regenerants showing similar DNA ploidy as that of their parent plants.
花叶芋‘Fancy’是一种重要的观赏植物,常用于盆栽和园林景观,以其色彩斑斓的叶片而闻名,常被用于室内装饰。在本研究中,我们提出了一种通过直接体细胞胚胎发生(DSE)从三种外植体来源进行离体再生的方法,其中通过流式细胞术分析对再生植株进行倍性变化筛选。将块茎、叶片和叶柄外植体接种在添加了1-萘乙酸(NAA)、6-苄基氨基嘌呤(BAP)和N-苯基-N'-1,2,3-噻二唑-5-脲(TDZ)不同浓度的MS基本培养基上。块茎外植体在NAA(1 mg/L)+BAP(0.5 mg/L)培养基上诱导出的直接体细胞胚胎最多,每个外植体平均有55.6个胚胎,而用1 mg/L TDZ处理的叶片和叶柄外植体分别发育出每个外植体平均18.7个和12.27个胚胎。在BAP(2 mg/L)+NAA(0.2 mg/L)培养基上培养4周后,胚胎转化率最高,每个块茎、叶片和叶柄外植体分别平均产生44.2、18.7和7.5个小植株。随后小植株生根,在添加2 mg/L吲哚丁酸的培养基上每个芽的生根数最多(6.33条)。通过组织学和扫描电镜证据展示了DSE的过程。流式细胞术分析评估的田间生长植株和DSE再生植株的2C DNA含量分别为8.06 pg和8.28 pg,表明没有倍性变化。因此,我们获得了一种从三种外植体类型诱导直接体细胞胚胎并具有高效胚胎转化率的成功方案,再生植株的DNA倍性与其亲本植株相似。