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INSERTION OF AGROBACTERIUM RHIZOGENES ROLB GENE IN MANGO

机译:芒果农杆菌根瘤菌基因的插入

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摘要

Transgenic mango (Mangifera indica L.) plants were regenerated from somatic embryos inoculated with a wild Agrobacterium rhizogenes strain. The bacteria were grown in Luria Bertani medium supplemented or not with acetosyringone until an optical density of 1.0-1.5 was reached. Incubation time of somatic embryos from the Kent, Haden and Madame Francis varieties in bacterial suspension was 1h, 15 and 5min. Somatic embryos were transferred into Gamborg Miller Ojima semisolid medium for 48h, and finally placed on the same medium containing cefotaxi-melcarbencillin, in darkness at 27 ±1 ℃. In the first treatment, A. rhizogenes was eliminated after 20 washings with cefotaximelcar-bencillin, only in the Kent variety; while 1-5 washes were needed for the other treatments. There were no differences between pre-rnculture treatments of A. rhizogenes with or without acetosyringone. After infection embryos formed callus, then secondary embryos and, finally, plants were regenerated but hairy roots were not induced. The presence of rolB gene in leaf tissue was confirmed by PCR. An expected band of 720bp, corresponding to rol B gen amplification, was obtained only in the transformed plants. An efficient protocol has been developed for successful production of transgenic somatic embryos and plants of the Kent variety using A. rhizogenes. Twenty transgenic clones per 0.5g of inoculated tissues with 80% embryo survival were produced. The results constitute the first report of transgenic mango plants mediated by A. rhizogenes as an alternative tool for breeding improvement of this crop.
机译:从接种野生发根农杆菌菌株的体细胞胚再生转基因芒果(Mangifera indica L.)植物。使细菌在补充或不加入乙酰丁香酮的Luria Bertani培养基中生长,直到达到1.0-1.5的光密度。肯特,哈登和弗朗西斯女士的体细胞胚在细菌悬液中的孵育时间分别为1h,15和5min。将体细胞胚转移至Gamborg Miller Ojima半固体培养基中48h,最后在27±1℃的黑暗中置于含有头孢他昔-美卡培西林的相同培养基上。在第一个处理中,仅在肯特品种中,用头孢他西美卡苯-西林洗涤20次后就消除了发根农杆菌。其他处理则需要1-5次清洗。在有或没有乙酰丁香酮的发根农杆菌的培养前处理之间没有差异。感染后胚形成愈伤组织,然后形成次生胚,最后再生出植物,但未诱导毛状根。通过PCR确认了叶组织中rolB基因的存在。仅在转化植物中获得了预期的720bp条带,对应于rol B基因的扩增。已经开发出一种有效的方案,以使用发根农杆菌成功地生产肯特品种的转基因体细胞胚和植物。每0.5g接种的组织产生二十个转基因克隆,胚胎存活率为80%。该结果构成了由发根农杆菌介导的转基因芒果植物的首次报道,作为该作物育种改良的替代工具。

著录项

  • 来源
    《Interciencia》 |2010年第7期|p.521-525|共5页
  • 作者单位

    Agricultural Sciences, Universidad Central de Venezuela, (UCV). UCV, Venezuela;

    rnAgricultural Sciences, UCV, Venezuela. Researcher, Instituto Nacional de Investigaciones Agricolas (INIA), Venezuela Direction: Unidad de Biotecnologia Vegetal. Centro Nacional de Investigacio-nes Agropecuarias (CENIAP). Apartado 5653. Maracay 2105, Venezuela;

    rnAgricultural Sciences, UCV, Venezuela. Researcher, CENI-AP-INIA, Venezuela. 12/2009;

    rnAgricultural and Livestock Technician, Instituto Universitario de Tecnologia Alonso Gomero, Venezuela. CENIAP-INIA, Venezuela;

    rnApplied Multivariate Statistics, Universidad de Salamanca, EspaNa. Biostatitician, FundaciOn Instituto de Estudios Avanzados, Venezuela;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    genetic transformation; mangifera indica; oncogenes; somatic embryogenesis;

    机译:基因转化;印度芒果癌基因体细胞胚发生;
  • 入库时间 2022-08-17 13:08:14

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