An efficient Agrobacterium-mediated transformation method for the edible mushroom Hypsizygus marmoreus

The establishment of genetic transformation method is crucial for the functional genomics research in filamentous fungi. Although the transformation method has been developed in several types of fungi, a highly efficient and convenient transformation system is desperately needed in Lentinula edodes. Present work established the Agrobacterium-mediated transformation (ATMT) of basidiomycete L. edodes in both monokaryon and dikaryon mycelia by using constructed binary plasmid pCAMBIA-1300-GFP. Then, the transformation efficiency of ATMT was evaluated by using different mediums for recipient incubation and different varieties of L. edodes. The results showed that in dikaryon strain W1, the positive hygromycin-resistant transformants was observed in all medium with the positive frequency of selected transformants that ranged from 0 to 30%. While in the monokaryon strain W1-26, only the millet medium group obtained positive transformants with a positive frequency of 75.48%. Moreover, three dikaryotic wild strains (YS55, YS3334, and YS3357) and two dikaryotic cultivated strains (W1 and S606) showed the highest transformation efficiency, with 32.96% of the germination frequency, and 85.12% of positive frequency for hygromycin-resistant transformants. This work demonstrated that Agrobacterium-mediated transformation was successfully performed in L. edodes, and the genotype of recipients as well as the medium for mycelial incubation were suggested to play key roles in determining the transformation efficiency. These findings may provide new avenues for the genetic modification of edible mushroom and may extend the cognition of DNA-mediated transformation in filamentous fungi.

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Agrobacterium-mediated transformation of arthroconidia obtained from the edible mushroom Hypsizygus marmoreus

Journal of Microbiological Methods ◽

10.1016/j.mimet.2020.105878 ◽

2020 ◽

Vol 171 ◽

pp. 105878

Author(s):

Dapeng Bao ◽

Zhiheng Huang ◽

Yan Li ◽

Chenli Zhou ◽

Yan Li ◽

...

Keyword(s):

Edible Mushroom ◽

Hypsizygus Marmoreus ◽

Agrobacterium Mediated Transformation

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A PRELIMINARY STUDY ON GENOTYPE-INDEPENDENT AGROBACTERIUM-MEDIATED TRANSFORMATION METHOD TO OBTAIN GENETICALLY ENGINEERED APRICOT PLANTS

Acta Horticulturae ◽

10.17660/actahortic.2009.839.47 ◽

2009 ◽

pp. 369-374 ◽

Cited By ~ 1

Author(s):

H. Wang ◽

M.D. Nortes ◽

M. Faize ◽

L. Faize ◽

S. López-Noguera ◽

...

Keyword(s):

Transformation Method ◽

Genetically Engineered ◽

Agrobacterium Mediated Transformation ◽

Preliminary Study

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Tissue culture-based Agrobacterium-mediated and in planta transformation methods

Czech Journal of Genetics and Plant Breeding ◽

10.17221/177/2016-cjgpb ◽

2017 ◽

Vol 53 (No. 4) ◽

pp. 133-143 ◽

Cited By ~ 8

Author(s):

M. Niazian ◽

S.A. Sadat Noori ◽

P. Galuszka ◽

S.M.M. Mortazavian

Keyword(s):

Tissue Culture ◽

Transformation Method ◽

Gene Transformation ◽

In Planta Transformation ◽

In Planta ◽

Agrobacterium Mediated Transformation ◽

Advantages And Disadvantages ◽

Culture Independent ◽

Transformation Methods ◽

Mutant Lines

Gene transformation can be done in direct and indirect (Agrobacterium-mediated) ways. The most efficient method of gene transformation to date is Agrobacterium-mediated method. The main problem of Agrobacterium-method is that some plant species and mutant lines are recalcitrant to regeneration. Requirements for sterile conditions for plant regeneration are another problem of Agrobacterium-mediated transformation. Development of genotype-independent gene transformation method is of great interest in many plants. Some tissue culture-independent Agrobacterium-mediated gene transformation methods are reported in individual plants and crops. Generally, these methods are called in planta gene transformation. In planta transformation methods are free from somaclonal variation and easier, quicker, and simpler than tissue culture-based transformation methods. Vacuum infiltration, injection of Agrobacterium culture to plant tissues, pollen-tube pathway, floral dip and floral spray are the main methods of in planta transformation. Each of these methods has its own advantages and disadvantages. Simplicity and reliability are the primary reasons for the popularity of the in planta methods. These methods are much quicker than regular tissue culture-based Agrobacterium-mediated gene transformation and success can be achieved by non-experts. In the present review, we highlight all methods of in planta transformation comparing them with regular tissue culture-based Agrobacterium-mediated transformation methods and then recently successful transformations using these methods are presented.

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Genetic structure and evolutionary diversity of mating-type (MAT) loci in Hypsizygus marmoreus

IMA Fungus ◽

10.1186/s43008-021-00086-8 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Gang Wang ◽

Yuanyuan Wang ◽

Lianfu Chen ◽

Hongbo Wang ◽

Lin Guo ◽

...

Keyword(s):

Genetic Structure ◽

Mating Type ◽

Phylogenetic Analyses ◽

Schizophyllum Commune ◽

Edible Mushroom ◽

Hypsizygus Marmoreus ◽

Pheromone Receptor ◽

Recombination Hotspots ◽

Mating Compatibility ◽

Genetic Structure And Diversity

AbstractThe mating compatibility in fungi is generally governed by genes located within a single or two unlinked mating type (MAT) loci. Hypsizygus marmoreus is an edible mushroom in the order Agaricales with a tetrapolar system, which contains two unlinked MAT loci-homeodomain (HD) transcription factor genes and pheromone/pheromone receptor genes (P/R). In this study, we analyzed the genetic structure and diversity of MAT loci in tetrapolar system of H. marmoreus through sequencing of 54 heterokaryon and 8 homokaryon strains. Although within the HD loci, the gene order was conserved, the gene contents were variable, and the HD loci haplotypes were further classified into four types. By analyzing the structure, phylogeny, and the HD transmissibility based on the progeny of these four HD mating-type loci types, we found that they were heritable and tightly linked at the HD loci. The P/R loci genes were found to comprise three pheromone receptors, three pheromones, and two pheromone receptor-like genes. Intra- and inter-specific phylogenetic analyses of pheromone receptors revealed that the STE3 genes were divided into three groups, and we thus theorize that they diverged before speciation. Comparative analysis of the MAT regions among 73 Basidiomycete species indicated that the diversity of HD and P/R loci in Agaricales and Boletales may contribute to mating compatibility. The number of HD genes were not correlated with the tetrapolar or bipolar systems. In H. marmoreus, the expression levels of these genes at HD and P/R loci of compatible strains were found higher than in those of homonuclear/homokaryotic strains, indicating that these mating genes acted as switches for mating processes. Further collinear analysis of HD loci in interspecific species found that HD loci contains conserved recombination hotspots showing major rearrangements in Coprinopsis cinerea and Schizophyllum commune, suggesting different mechanisms for evolution of physically linked MAT loci in these groups. It seems likely that gene rearrangements are common in Agaricales fungi around HD loci. Together, our study provides insights into the genomic basis of mating compatibility in H. marmoreus.

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Agrobacterium-mediated in planta genetic transformation of winter wheat (Triticum aestivum L.)

Faktori eksperimental'noi evolucii organizmiv ◽

10.7124/feeo.v22.964 ◽

2018 ◽

Vol 22 ◽

pp. 293-298

Author(s):

S. I. Mykhalska ◽

A. G. Komisarenko ◽

V. M. Kurchii ◽

O. M. Tishchenko

Keyword(s):

Triticum Aestivum ◽

Transgenic Plants ◽

Genetic Transformation ◽

Winter Wheat ◽

Natural Frequency ◽

Triticum Aestivum L ◽

Transformation Method ◽

In Planta ◽

Agrobacterium Mediated Transformation ◽

Pollination Methods

Aim. To optimize the agrobacterium-mediated method of winter wheat transformation (Triticum aestivum L.); to select the conditions and period of inoculation to effectively transfer the genes during pollination. Methods. Agrobacterium-mediated in planta genetic transformation of winter wheat (Triticum aestivum L.) during pollination. Results. The conditions for agrobacterium-mediated transformation method of winter wheat during natural (frequency pollination was 1 %) and non-natural (frequency pollination was 4 %) pollination were defined. Conclusions. The possibility of integrating transgenes into the genome of winter wheat plants by the method of Agrobacterium-mediated transformation in planta in the process of forced and natural pollination is demonstrated. It is found that the transformation efficiency to a large extent depends on the plant genotype and the method of carrying out the transformation procedure. The selection of transgenic plants under water deficit conditions allowed to identify the plants with functional transgene. The signs of functioning transgene have been remaining in the next generation of genetically modified winter wheat. Keywords: Triticum aestivum L., Agrobacterium-mediated transformation in planta, transgenic plants, seeds.

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