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转基因梨砧木田间茎段扦插生根及有性和无性后代基因表达稳定性研究

The Rooting of Stem Cuttings and the Stability of Gene Expression in Generative and Vegetative Progeny of Transgenic Pear Rootstock in the Field.

作者信息

Lebedev Vadim

机构信息

Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Science avenue 6, Pushchino, Moscow Region 142290, Russia.

出版信息

Plants (Basel). 2019 Aug 19;8(8):291. doi: 10.3390/plants8080291.

Abstract

Adventitious rooting plays an important role in the commercial vegetative propagation of trees. Adventitious root formation is a complex biological process, but knowledge of the possible unintended effects induced by both the integration/expression of transgenes and in vitro conditions on the rooting is limited. The long-term stability of transgene expression is important both for original transformants of woody plants and its progeny. In this study, we used field-grown pear rootstock GP217 trees transformed with the reporter ß-glucuronidase () genes with and without intron and re-transformed with the herbicide resistance gene as model systems. We assessed the unintended effects on rooting of pear semi-hardwood cuttings and evaluated the stability of transgene expression in progeny produced by generative (seedlings) and vegetative (grafting, cutting) means up to four years. Our investigation revealed that: (1) The single and repeated transformations of clonal pear rootstocks did not result in unintended effects on adventitious root formation in cuttings; (2) stability of the transgene expression was confirmed on both generative and vegetative progeny, and no silenced transgenic plants were detected; (3) yearly variation in the gene expressions was observed and expression levels were decreased in extremely hot and dry summer; (4) the intron enhanced the expression of gene in pear plants approximately two-fold compared to gene without intron. The current study provides useful information on transgene expression in progeny of fruit trees under natural environmental conditions.

摘要

不定根形成在树木的商业营养繁殖中起着重要作用。不定根的形成是一个复杂的生物学过程,但关于转基因的整合/表达以及体外条件对生根可能产生的意外影响的了解有限。转基因表达的长期稳定性对于木本植物的原始转化体及其后代都很重要。在本研究中,我们使用了田间种植的梨砧木GP217树作为模型系统,这些树用带有和不带有内含子的报告基因β-葡萄糖醛酸酶(GUS)基因进行了转化,并再次用抗除草剂基因进行了转化。我们评估了对梨半硬枝插条生根的意外影响,并评估了通过有性(幼苗)和无性(嫁接、扦插)方式产生的后代中转基因表达的稳定性,长达四年。我们的研究表明:(1)克隆梨砧木的单次和重复转化对插条不定根的形成没有产生意外影响;(2)在有性和无性后代中都证实了转基因表达的稳定性,并且未检测到转基因沉默的植株;(3)观察到基因表达的年度变化,在极其炎热干燥的夏季表达水平下降;(4)与没有内含子的基因相比,内含子使梨植株中GUS基因的表达增强了约两倍。本研究为自然环境条件下果树后代中的转基因表达提供了有用信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fbfc/6724118/f0b7b9ec3779/plants-08-00291-g001.jpg

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