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利用营养组织作为外植体进行体细胞胚胎培养 值得学习和尝试

已有 2121 次阅读 2017-10-28 11:57 |个人分类:自然与社会|系统分类:科研笔记

目前比较常用的外植体是未成熟的合子胚。这对于保存母体的优势是不利的,而采用营养组织作为外植体进行体细胞胚胎培养 就可以实现。


Propagation of mature Quercus ilex L. (holm oak) trees by somatic embryogenesis

作者:Martinez, MT (Martinez, M. T.)[ 1 ] ; San Jose, MC (San Jose, M. C.)[ 1 ] ; Vieitez, AM (Vieitez, A. M.)[ 1 ] ; Cernadas, MJ (Cernadas, M. J.)[ 1 ] ; Ballester, A (Ballester, A.)[ 1 ] ; Corredoira, E (Corredoira, E.)[ 1 ]


PLANT CELL TISSUE AND ORGAN CULTURE


卷: 131

期: 2

页: 321-333

DOI: 10.1007/s11240-017-1286-4

出版年: NOV 2017

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摘要

Somatic embryogenesis from in vitro leaf and shoot apex explants excised from axillary shoot cultures established from two mature Quercus ilex trees has been developed. Somatic embryos (SE) were obtained from both explant types and genotypes evaluated, although embryogenic frequencies were influenced by the genotype, auxin concentration, and explant type. The explants were cultured on Murashige and Skoog salts and vitamins, supplemented with 500 mg L- casein hydrolysate (CH) and different concentrations of indole-3-acetic acid or a-naphthalene acetic acid (NAA) in combination with 2.22 mu M 6-benzylaminopurine (BA). In both genotypes, shoot apex explants were more responsive than leaf explants. The best results were obtained with apex explants of clone Q3 (11%) cultured on medium with 21.48 mu M NAA plus 2.22 mu M BA. This combination was also effective for initiating SE from leaf explants, although the induction rates were lower (1-3%). Embryogenic lines were maintained by repetitive embryogenesis following culture of nodular embryogenic structures on Schenk and Hildebrand medium without plant growth regulators. Low embryo multiplication rates were obtained when torpedo or early cotyledonary SE were used as initial explant for embryo proliferation, or when glutamine or CH (500 mg L-1) was added to proliferation medium. For germination, cotyledonary-stage SE were isolated and stored at 4 degrees C for 2 months. After cold storage, SE were cultured on germination medium consisting of Gresshoff and Doy medium, supplemented with 0.44 mu M BA and 20 mu M silver thiosulphate. Under these conditions, plantlets were regenerated from 21 to 66.7% of the SE generated for both genotypes.




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