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火炬松通过体细胞器官发生实现高频不定芽再生的基因型控制

Genotypic control of high-frequency adventitious shoot regeneration via somatic organogenesis in loblolly pine.

作者信息

Tang W, Whetten R, Sederoff R

机构信息

Forest Biotechnology Group, Department of Forestry, North Carolina State University, 27695-8008, Raleigh, NC, USA

出版信息

Plant Sci. 2001 Jul;161(2):267-272. doi: 10.1016/s0168-9452(01)00394-6.

Abstract

Mature zygotic embryos of 24 genotypes of loblolly pine (Pinus taeda L.) were used as explants to establish an adventitious shoot regeneration system through somatic organogenesis. Callus formation frequencies of 18.2 (genotype 11-1103) -77.7% (genotype 7-100) have been induced from mature zygotic embryos of all genotypes tested on callus induction medium (basal salts) containing 2,4-dichlorophenoxyacetic acid (2,4-D) or alpha-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and kinetin. Adventitious shoot regeneration via organogenesis with the frequency of 5.4 (genotype 11-1103 and 7-2) -77.2% (genotype 8-1082) was obtained from callus and tissue cultures derived from mature zygotic embryos of 24 genotypes of loblolly pine. The highest mean number of 18 adventitious buds per piece of callus 0.5x0.5 cm(2) in size was obtained from genotype 8-1082. Elongation of adventitious buds was achieved on TE medium supplemented with 0.5 mg/l indole-3-butyric acid (IBA) and 1 mg/l BA. After rooting, regenerated plantlets were established in soil. These results suggested that adventitious shoot regeneration via somatic organogenesis was influenced by the genotypes. The in vitro regeneration procedure established in this investigation could be used for clonal micropropagation of some genotypes of loblolly pine, as well as for establishing a transformation system in coniferous species.

摘要

以24种基因型的火炬松(Pinus taeda L.)成熟合子胚为外植体,通过体细胞器官发生建立不定芽再生体系。在含有2,4 - 二氯苯氧乙酸(2,4 - D)或α - 萘乙酸(NAA)、6 - 苄基腺嘌呤(BA)和激动素的愈伤组织诱导培养基(基础盐类)上,所有测试基因型的成熟合子胚均诱导出了愈伤组织形成频率为18.2%(基因型11 - 1103)至77.7%(基因型7 - 100)的愈伤组织。从24种基因型火炬松成熟合子胚产生的愈伤组织和组织培养物中,通过器官发生获得了不定芽再生,频率为5.4%(基因型11 - 1103和7 - 2)至77.2%(基因型8 - 1082)。从基因型8 - 1082中获得了每块大小为0.5×0.5 cm²愈伤组织平均最高18个不定芽的数量。不定芽在添加了0.5 mg/l吲哚 - 3 - 丁酸(IBA)和1 mg/l BA的TE培养基上实现伸长。生根后,再生植株在土壤中定植。这些结果表明,通过体细胞器官发生的不定芽再生受基因型影响。本研究建立的离体再生程序可用于某些基因型火炬松的克隆微繁殖,以及在针叶树种中建立转化系统。

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