Current status and trends of wheat genetic transformation studies in China

He, Yi; Wang, Qiong; Zeng, Jian; Sun, Tao; Yang, Guangxiao; He, Guangyuan

doi:10.1016/s2095-3119(14)60934-5

Cited by 11 publications

(6 citation statements)

References 108 publications

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“…There is general agreement that the efficiency of biolistic-mediated genetic transformation of cereals is influenced by several physical and biological factors [ 30 , 31 ]. In Triticeae , numerous systematic studies of the parameters affecting the efficiencies of gene transfer were undertaken in polyploid wheat [ 21 , 22 , 32 – 35 ], barley [ 36 , 37 ] and rye [ 38 ], although no studies have been published for diploid einkorn. Since the differences in cell size, cell division and morphogenesis patterns between einkorn and other cereals may affect the efficiency of particle penetration, three keys factors, i.e., helium pressure, particle size and explant type, were explored in this study.…”

Section: Discussionmentioning

confidence: 99%

Genetic transformation of einkorn (Triticum monococcum L. ssp. monococcum L.), a diploid cultivated wheat species

et al. 2018

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BackgroundDomesticated einkorn (Triticum monococcum L.) is one of the oldest cultivated cereal crops in the world. Its small genome size (~ 5.7 GB), low ploidy (2n = 2x = 14, AmAm) and high genetic polymorphism make this species very attractive for use as a diploid model for understanding the genomics and proteomics of Triticeae. Einkorn, however, is still a recalcitrant monocotyledonous species for the application of modern biotechnologies, including transgenesis. This paper reports the factors that may influence transgene delivery, integration, expression and inheritance in einkorn.ResultsIn this study, we report the successful genetic transformation of einkorn using biolistic-mediated DNA delivery. Immature embryo-derived tissues of spring einkorn were bombarded with a plasmid containing the reporter gene GFP (green fluorescent protein) driven by the rice actin promoter (act1) and the selectable bar gene (bialaphos resistance gene) driven by the maize ubiquitin promoter (ubi1). Adjustments to various parameters such as gas pressure, microcarrier size and developmental stage of target tissue were essential for successful transient and stable transformation. Bombarded einkorn tissues are recalcitrant to regenerating plants, but certain modifications of the culture medium have been shown to increase the production of transgenic events. In various experiments, independent transgenic plants were produced at frequencies ranging from 0.0 to 0.6%. Molecular analysis, marker gene expression and herbicide treatment demonstrated that gfp/bar genes were stably integrated into the einkorn genome and successfully inherited over several generations. The transgenes, as dominant loci, segregated in both Mendelian and non-Mendelian fashion due to multiple insertions. Fertile homozygous T1-T2 populations of transgenic einkorn that are resistant to herbicides were selected.ConclusionTo the best of our knowledge, this is the first report of the production of genetically modified einkorn plants. We believe that the results of our research could be a starting point for the application of the current biotechnological-based technologies, such as transgenesis and genome editing, to accelerate comparative functional genomics in einkorn and other cereals.Electronic supplementary materialThe online version of this article (10.1186/s12896-018-0477-3) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Genetic transformation of einkorn (Triticum monococcum L. ssp. monococcum L.), a diploid cultivated wheat species

et al. 2018

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“…Over the last two decades, the bombardment-mediated transformation has been widely used in gene functional studies and genetic improvement of wheat, and these achievements have been reviewed previously ( Jones et al., 2005 ; Li et al., 2012 ; He et al., 2015 ; Han et al., 2017 ; Shrawat and Armstrong, 2018 ; Hensel, 2020 ). While the importance of creating transgenic wheat, a limited number of labs are proficient in the transformation, which has been alleviated to some degree until the recent major improvements in Agrobacterium -mediated wheat transformation ( Richardson et al., 2014 ; Wang et al., 2016 , Wang et al., 2017 , Wang et al., 2022 ).…”

Section: Discussionmentioning

confidence: 99%

An established protocol for generating transgenic wheat for wheat functional genomics via particle bombardment

Wang

Zeng²,

Su³

et al. 2022

Front. Plant Sci.

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Wheat is one of the most important food crops in the world and is considered one of the top targets in crop biotechnology. With the high-quality reference genomes of wheat and its relative species and the recent burst of genomic resources in Triticeae, demands to perform gene functional studies in wheat and genetic improvement have been rapidly increasing, requiring that production of transgenic wheat should become a routine technique. While established for more than 20 years, the particle bombardment-mediated wheat transformation has not become routine yet, with only a handful of labs being proficient in this technique. This could be due to, at least partly, the low transformation efficiency and the technical difficulties. Here, we describe the current version of this method through adaptation and optimization. We report the detailed protocol of producing transgenic wheat by the particle gun, including several critical steps, from the selection of appropriate explants (i.e., immature scutella), the preparation of DNA-coated gold particles, and several established strategies of tissue culture. More importantly, with over 20 years of experience in wheat transformation in our lab, we share the many technical details and recommendations and emphasize that the particle bombardment-mediated approach has fewer limitations in genotype dependency and vector construction when compared with the Agrobacterium-mediated methods. The particle bombardment-mediated method has been successful for over 30 wheat genotypes, from the tetraploid durum wheat to the hexaploid common wheat, from modern elite varieties to landraces. In conclusion, the particle bombardment-mediated wheat transformation has demonstrated its potential and wide applications, and the full set of protocol, experience, and successful reports in many wheat genotypes described here will further its impacts, making it a routine and robust technique in crop research labs worldwide.

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“…For example, a mutant line shows strong resistance to powdery mildew that is generated by knocking out all six alleles encoding the mildew-resistance locus protein [103]. Additionally, the breakthrough of transformation technology bring the stable positive transformation efficiencies reaching to 13% in wheat, and the ratio of single transgene loci was more than 50% [104,105]. In a word, therefore, determining the functional role of these TF genes in wheat and tolerance to biotic stresses, and identification of target genes of TFs involved in pathogen responses are one of important current tasks, which will make great contribution to cut loss in wheat breeding because of the improvement of resistance in future.…”

Section: Conclusion and Future Prospection Of Tfs In Wheat Breedingmentioning

confidence: 99%

The role of transcription factor in wheat defense against pathogen and its prospect in breeding

Zhang¹,

Lv²,

Wang³

et al. 2018

J Plant Biol Crop Res

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The grain yield of a crop represents the expression of many thousands of genes integrated over the life of each of the component plants in the crop in response to environment, as well as severely affected by parasite, including pest and pathogen. Plant has no capacity to elude parasite other than by altering intrinsic gene expression or gene combinations to improve performance under pathogen. Transcriptional control is a crucial part of genes expression, especially in plant response to a range of stresses. Wheat (Triticum aestivum L.) is one of the most important cultivated crops, while its production isseverely affected by stripe rust and powdery mildew. Unfortunately, coupled with the loss of genetic diversity in wheat breeding programs, the disease resistance germplasms are more and more scarce due to the frequently variation of epidemic virulent race. Research carried out in the past few years has been productive in identifying TFs for regulating resistance to pathogen stress in wheat and other plant species. The increasing studies showed that Transcription Factors (TFs) are potent positive, negative, cis-and trans-regulators activating the functional gene. Herein, we highlighted the recent progress in elucidating the roles of TFs in wheat defense against pathogen, as well as the potential relationship between transcription factors family with regulating pathogen type, although there should be no expectation that everlasting favorable genes performance will be discovered for variable environments. Furthermore, we discussed new ways to improve varieties' resistance using biotechnology combining with empirical breeding program. This leads to new ideas to enhance wheat resistance or tolerance to disease in virtue of the progress of wheat genetic engineering. This maybe a new way to improve adaptive plasticity of wheat in highly variable environments as aresult of introducing greater diversity of resistance gene pool to cropping systems.

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Current status and trends of wheat genetic transformation studies in China

Cited by 11 publications

References 108 publications

Genetic transformation of einkorn (Triticum monococcum L. ssp. monococcum L.), a diploid cultivated wheat species

Genetic transformation of einkorn (Triticum monococcum L. ssp. monococcum L.), a diploid cultivated wheat species

An established protocol for generating transgenic wheat for wheat functional genomics via particle bombardment

The role of transcription factor in wheat defense against pathogen and its prospect in breeding

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