Control of Pollen-Mediated Gene Flow in Transgenic Trees[W][OA]
Control of Pollen-Mediated Gene Flow in Transgenic Trees[W][OA]
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DOI：
10.1104/pp.112.197228
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发表时间：
2012-06
期刊：
影响因子：
7.4
通讯作者：
Chunsheng Zhang;Kim Norris-Caneda;W. Rottmann;J. Gulledge;Shujun Chang;Brian Kwan;Anita Thomas;Lydia C. Mandel;R. T. Kothera;A. D. Victor;L. Pearson;M. Hinchee
中科院分区：
文献类型：
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作者：
Chunsheng Zhang;Kim Norris-Caneda;W. Rottmann;J. Gulledge;Shujun Chang;Brian Kwan;Anita Thomas;Lydia C. Mandel;R. T. Kothera;A. D. Victor;L. Pearson;M. Hinchee
消除花粉提供了一种有效的遏制方法，以减少从转基因树到其野生近亲的直接基因流动。到目前为止，在控制树木花粉产生方面只取得了有限的成功。用辐射松雄球果特异启动子PrMC2驱动改良的barnase编码序列(barnaseH102E、barnaseK27A和barnaseE73G)，以确定它们在花粉消融中的有效性。表达盒PrMC2-barnaseH102E在烟草、松树和桉树中被发现能有效地去除花粉。大规模和多年的田间试验表明，95%以上的松树和桉树独立转化系完全防止了花粉产生。其中含有PrMC2-barnaseH102E表达盒。在转基因株系中获得了完整的花粉控制表型，并在多年、多个测试点以及PrMC2-barnaseH102E盒两侧有不同基因的情况下稳定表达。转PrMC2-barnaseH102E基因的松树和桉树(spp.)除了无花粉表型外，树的生长在所有观察到的属性上都与对照树相似。在田间生长的树木中能够实现完全控制花粉产生的能力可能是三个因素的独特组合的结果：PrMC2启动子的雄性球果/花药专一性，BarnaseH102E核糖核酸酶活性降低，以及与多倍体绒毡层相关的独特特征。PrMC2-barnaseH102E在有代表性的被子植物和裸子植物树上的田间表现表明，该基因可用于缓解大规模部署转基因树时的花粉介导的基因流。
Pollen elimination provides an effective containment method to reduce direct gene flow from transgenic trees to their wild relatives. Until now, only limited success has been achieved in controlling pollen production in trees. A pine (Pinus radiata) male cone-specific promoter, PrMC2, was used to drive modified barnase coding sequences (barnaseH102E, barnaseK27A, and barnaseE73G) in order to determine their effectiveness in pollen ablation. The expression cassette PrMC2-barnaseH102E was found to efficiently ablate pollen in tobacco (Nicotiana tabacum), pine, and Eucalyptus (spp.). Large-scale and multiple-year field tests demonstrated that complete prevention of pollen production was achieved in greater than 95% of independently transformed lines of pine and Eucalyptus (spp.) that contained the PrMC2-barnaseH102E expression cassette. A complete pollen control phenotype was achieved in transgenic lines and expressed stably over multiple years, multiple test locations, and when the PrMC2-barnaseH102E cassette was flanked by different genes. The PrMC2-barnaseH102E transgenic pine and Eucalyptus (spp.) trees grew similarly to control trees in all observed attributes except the pollenless phenotype. The ability to achieve the complete control of pollen production in field-grown trees is likely the result of a unique combination of three factors: the male cone/anther specificity of the PrMC2 promoter, the reduced RNase activity of barnaseH102E, and unique features associated with a polyploid tapetum. The field performance of the PrMC2-barnaseH102E in representative angiosperm and gymnosperm trees indicates that this gene can be used to mitigate pollen-mediated gene flow associated with large-scale deployment of transgenic trees.