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首页> 外文期刊>BMC Biotechnology >Regeneration of multiple shoots from transgenic potato events facilitates the recovery of phenotypically normal lines: assessing a cry9Aa2 gene conferring insect resistance
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Regeneration of multiple shoots from transgenic potato events facilitates the recovery of phenotypically normal lines: assessing a cry9Aa2 gene conferring insect resistance

机译：从转基因马铃薯事件中再生出许多芽，有助于表型正常系的恢复：评估赋予昆虫抗性的cry9Aa2基因

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Background The recovery of high performing transgenic lines in clonal crops is limited by the occurrence of somaclonal variation during the tissue culture phase of transformation. This is usually circumvented by developing large populations of transgenic lines, each derived from the first shoot to regenerate from each transformation event. This study investigates a new strategy of assessing multiple shoots independently regenerated from different transformed cell colonies of potato ( Solanum tuberosum L.). Results A modified cry 9Aa2 gene, under the transcriptional control of the CaMV 35S promoter, was transformed into four potato cultivars using Agrobacterium -mediated gene transfer using a npt II gene conferring kanamycin resistance as a selectable marker gene. Following gene transfer, 291 transgenic lines were grown in greenhouse experiments to assess somaclonal variation and resistance to potato tuber moth (PTM), Phthorimaea operculella (Zeller). Independently regenerated lines were recovered from many transformed cell colonies and Southern analysis confirmed whether they were derived from the same transformed cell. Multiple lines regenerated from the same transformed cell exhibited a similar response to PTM, but frequently exhibited a markedly different spectrum of somaclonal variation. Conclusions A new strategy for the genetic improvement of clonal crops involves the regeneration and evaluation of multiple shoots from each transformation event to facilitate the recovery of phenotypically normal transgenic lines. Most importantly, regenerated lines exhibiting the phenotypic appearance most similar to the parental cultivar are not necessarily derived from the first shoot regenerated from a transformed cell colony, but can frequently be a later regeneration event.
机译：背景技术克隆转化作物中高性能转基因品系的恢复受到转化组织培养阶段体细胞克隆变异的限制。通常通过发展大量的转基因品系来避免这种情况，每个转基因品系都来自第一个芽，并从每个转化事件中再生。这项研究调查了评估从马铃薯（Solanum tuberosum L.）的不同转化细胞菌落中独立再生的多个芽的新策略。结果在农杆菌介导的基因转移下，使用赋予卡那霉素抗性的npt II基因作为选择标记基因，在CaMV 35S启动子的转录控制下，将经修饰的cry 9Aa2基因转化为四个马铃薯品种。基因转移后，在温室实验中种植了291个转基因品系，以评估体细胞克隆变异和对马铃薯块茎蛾（PTM），小球藻（Zeller）的抗性。从许多转化的细胞集落中回收了独立再生的品系，Southern分析证实它们是否源自相同的转化细胞。从同一转化细胞再生的多个品系对PTM表现出相似的反应，但常常表现出体细胞克隆变异的显着不同。结论克隆作物遗传改良的新策略涉及从每个转化事件中再生和评估多个芽，以促进表型正常的转基因品系的恢复。最重要的是，表现出与亲本品种最相似的表型外观的再生品系不一定来自转化的细胞菌落再生的第一个芽，而通常是后来的再生事件。

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