Overexpression of Vitis GRF4-GIF1 improves regeneration efficiency in diploid Fragaria vesca Hawaii 4.

IF 4.7 2区生物学 Q1 BIOCHEMICAL RESEARCH METHODS Plant Methods Pub Date : 2024-10-18 DOI:10.1186/s13007-024-01270-8

Esther Rosales Sanchez, R Jordan Price, Federico Marangelli, Kirsty McLeary, Richard J Harrison, Anindya Kundu

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Abstract

Background: Plant breeding played a very important role in transforming strawberries from being a niche crop with a small geographical footprint into an economically important crop grown across the planet. But even modern marker assisted breeding takes a considerable amount of time, over multiple plant generations, to produce a plant with desirable traits. As a quicker alternative, plants with desirable traits can be raised through tissue culture by doing precise genetic manipulations. Overexpression of morphogenic regulators previously known for meristem development, the transcription factors Growth-Regulating Factors (GRFs) and the GRF-Interacting Factors (GIFs), provided an efficient strategy for easier regeneration and transformation in multiple crops.

Results: We present here a comprehensive protocol for the diploid strawberry Fragaria vesca Hawaii 4 (strawberry) regeneration and transformation under control condition as compared to ectopic expression of different GRF4-GIF1 chimeras from different plant species. We report that ectopic expression of Vitis vinifera VvGRF4-GIF1 provides significantly higher regeneration efficiency during re-transformation over wild-type plants. On the other hand, deregulated expression of miRNA resistant version of VvGRF4-GIF1 or Triticum aestivum (wheat) TaGRF4-GIF1 resulted in abnormalities. Transcriptomic analysis between the different chimeric GRF4-GIF1 lines indicate that differential expression of FvExpansin might be responsible for the observed pleiotropic effects. Similarly, cytokinin dehydrogenase/oxygenase and cytokinin responsive response regulators also showed differential expression indicating GRF4-GIF1 pathway playing important role in controlling cytokinin homeostasis.

Conclusion: Our data indicate that ectopic expression of Vitis vinifera VvGRF4-GIF1 chimera can provide significant advantage over wild-type plants during strawberry regeneration without producing any pleiotropic effects seen for the miRNA resistant VvGRF4-GIF1 or TaGRF4-GIF1.

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过表达葡萄 GRF4-GIF1 可提高二倍体 Fragaria vesca Hawaii 4 的再生效率。

背景：植物育种在将草莓从一种地域范围较小的小众作物转变为一种在全球种植的重要经济作物的过程中发挥了非常重要的作用。但是，即使是现代标记辅助育种也需要相当长的时间，经过多代植物才能培育出具有理想性状的植物。作为一种更快捷的替代方法，可以通过精确的基因操作，通过组织培养培育出具有理想性状的植物。过度表达以前已知的分生组织发育的形态发生调节因子--转录因子生长调节因子（GRFs）和 GRF 交互因子（GIFs）--提供了一种高效的策略，使多种作物的再生和转化更加容易：结果：我们在此介绍了在对照条件下二倍体草莓Fragaria vesca Hawaii 4（草莓）再生和转化的综合方案，并对不同植物物种异位表达不同的GRF4-GIF1嵌合体进行了比较。我们发现，异位表达葡萄 VvGRF4-GIF1 在再转化过程中的再生效率明显高于野生型植株。另一方面，抗 miRNA 版本的 VvGRF4-GIF1 或 Triticum aestivum（小麦）TaGRF4-GIF1 的表达失调会导致异常。不同嵌合 GRF4-GIF1 株系之间的转录组分析表明，FvExpansin 的不同表达可能是造成所观察到的多效应的原因。同样，细胞分裂素脱氢酶/加氧酶和细胞分裂素反应调节因子也出现了差异表达，这表明 GRF4-GIF1 通路在控制细胞分裂素平衡中发挥着重要作用：我们的数据表明，在草莓再生过程中，葡萄 VvGRF4-GIF1 嵌合体的异位表达比野生型植株具有显著优势，而不会产生任何 miRNA 抗性 VvGRF4-GIF1 或 TaGRF4-GIF1 的多生物效应。

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来源期刊

Plant Methods 生物-植物科学

CiteScore

9.20

自引率

3.90%

发文量

121

审稿时长

2 months

期刊介绍： Plant Methods is an open access, peer-reviewed, online journal for the plant research community that encompasses all aspects of technological innovation in the plant sciences. There is no doubt that we have entered an exciting new era in plant biology. The completion of the Arabidopsis genome sequence, and the rapid progress being made in other plant genomics projects are providing unparalleled opportunities for progress in all areas of plant science. Nevertheless, enormous challenges lie ahead if we are to understand the function of every gene in the genome, and how the individual parts work together to make the whole organism. Achieving these goals will require an unprecedented collaborative effort, combining high-throughput, system-wide technologies with more focused approaches that integrate traditional disciplines such as cell biology, biochemistry and molecular genetics. Technological innovation is probably the most important catalyst for progress in any scientific discipline. Plant Methods’ goal is to stimulate the development and adoption of new and improved techniques and research tools and, where appropriate, to promote consistency of methodologies for better integration of data from different laboratories.