Assessment of promoters and a selectable marker for development of strawberry intragenic vectors
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- 作者：Raquel F. Carvalho ; Kevin M. Folta
- 关键词：Strawberry ; Intragenic ; Cisgenic ; Constitutive promoter ; Transient expression ; Genetic engineering ; EPSPS gene
- 刊名：Plant Cell, Tissue and Organ Culture (PCTOC)
- 出版年：2017
- 出版时间：February 2017
- 年：2017
- 卷：128
- 期：2
- 页码：259-271
- 全文大小：
- 刊物类别：Biomedical and Life Sciences
- 刊物主题：Plant Sciences; Plant Physiology; Plant Genetics & Genomics; Plant Pathology;
- 出版者：Springer Netherlands
- ISSN：1573-5044
- 卷排序：128
文摘
Conventional breeding of the cultivated strawberry (Fragaria × ananassa) is hindered by a complicated octoploid genome and a paucity of molecular markers. Genetic improvement may be accelerated by direct gene-transfer techniques that could address single-gene deficiencies in otherwise commercially valuable selections. ‘Cisgenic’ or “intragenic” approaches face fewer regulatory hurdles, yet more characterized promoters and selectable markers are needed for the development of such technologies in strawberry. This report tests a number of putative promoters and a potential selectable marker for use in cisgenic vector development. Based on RNAseq data, the 5′-proximal noncoding regions from highly expressed transcripts were tested for activity. These putative promoter sequences were fused to eGFP and GUS reporter genes and transiently expressed in both Nicotiana benthamiana leaves and F. × ananassa strawberry fruits. The promoter sequences of FanAPA1-related and FanUBCE2 present constitutive promoter candidates that are active in all tissues tested. A potential selectable marker from strawberry sequence, the coding region of the strawberry 5-enolpyruvylshikimate 3 phosphatase synthase (EPSPS) gene, was cloned and mutagenized, and then stably transformed into Arabidopsis plants. Overexpression of the non-mutagenized form confers resistance to glyphosate, showing potential utility as a selectable marker. The results of these trials define new tools for the development of intragenic vectors in strawberry.