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兰考泡桐的直接体细胞胚胎发生与人工种子生产

Direct somatic embryogenesis and synthetic seed production from Paulownia elongata.

作者信息

Ipekci Z, Gozukirmizi N

机构信息

TUBITAK, Research Institute for Genetic Engineering and Biotechnology, P.O. Box 21, 41470 Gebze-Kocaeli, Turkey.

出版信息

Plant Cell Rep. 2003 Aug;22(1):16-24. doi: 10.1007/s00299-003-0650-5. Epub 2003 Jun 24.

Abstract

We have developed a reproducible system for efficient direct somatic embryogenesis from leaf and internodal explants of Paulownia elongata. The somatic embryos obtained were subsequently encapsulated as single embryos to produce synthetic seeds. Several plant growth regulators [6-benzylaminopurine, indole-3-acetic acid, alpha-naphthaleneacetic acid, kinetin and thidiazuron (TDZ)] alone or in combination were tested for their capacity to induce somatic embryogenesis. The highest induction frequencies of somatic embryos were obtained on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% Phytagel, 500 mg l(-1) casein hydrolysate and 10 mg l(-1) TDZ (medium MS10). Somatic embryos were induced from leaf (69.8%) and internode (58.5%) explants on MS10 medium after 7 days. Subsequent withdrawal of TDZ from the induction medium resulted in the maturation and growth of the embryos into plantlets on MS basal media. The maturation frequency of somatic embryos from leaf and internodal explants was 50.8% and 45.8%, respectively. Subculturing of mature embryos led to their germination on the same medium with a germination frequency of 50.1% and 29.8% from leaf and internode explants, respectively. Somatic embryos obtained directly on leaf explants were used for encapsulation in liquid MS medium containing different concentrations of sodium alginate with a 30-min exposure to 50 m M CaCl(2). A 3% sodium alginate concentration provided a uniform encapsulation of the embryos with survival and germination frequencies of 73.7% and 53.3%, respectively. Storage at 4 degrees C for 30 days or 60 days significantly reduced the survival and complete germination frequencies of both encapsulated and non-encapsulated embryos relative to those of non-stored somatic embryos. However, the survival and germination rates of encapsulated embryos increased following storage at 4 degrees C. After 30 days or 60 days of storage, the survival rates of encapsulated embryos were 67.8% and 53.5% and the germination frequencies were 43.2% and 32.4%, respectively. These systems could be useful for the rapid clonal propagation and dissemination of synthetic seed material of Paulownia elongata.

摘要

我们开发了一种可重复的系统,用于从兰考泡桐的叶片和节间外植体高效直接诱导体细胞胚胎发生。随后将获得的体细胞胚单个包埋以生产人工种子。单独或组合测试了几种植物生长调节剂[6-苄基腺嘌呤、吲哚-3-乙酸、α-萘乙酸、激动素和噻二唑素(TDZ)]诱导体细胞胚胎发生的能力。在添加了3%蔗糖、0.6%植物凝胶、500 mg l(-1)水解酪蛋白和10 mg l(-1) TDZ的Murashige和Skoog(MS)培养基(MS10培养基)上获得了最高的体细胞胚诱导频率。7天后,在MS10培养基上从叶片(69.8%)和节间(58.5%)外植体诱导出体细胞胚。随后从诱导培养基中去除TDZ,使胚在MS基本培养基上成熟并生长成小植株。来自叶片和节间外植体的体细胞胚的成熟频率分别为50.8%和45.8%。成熟胚的继代培养导致它们在同一培养基上萌发,来自叶片和节间外植体的萌发频率分别为50.1%和29.8%。直接在叶片外植体上获得的体细胞胚用于在含有不同浓度海藻酸钠的液体MS培养基中包埋,在50 mM CaCl(2)中暴露30分钟。3%的海藻酸钠浓度使胚得到均匀包埋,存活和萌发频率分别为73.7%和53.3%。相对于未储存的体细胞胚,在4℃下储存30天或60天显著降低了包埋和未包埋胚的存活和完全萌发频率。然而,包埋胚在4℃储存后的存活和萌发率有所提高。储存30天或60天后,包埋胚的存活率分别为67.8%和53.5%,萌发频率分别为43.2%和32.4%。这些系统可能有助于兰考泡桐人工种子材料的快速克隆繁殖和传播。

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