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

Abstract

Niazian M., Sadat Noori S.A., Galuszka P., Mortazavian S.M.M. (2017): Tissue culture-based Agrobacterium-mediated and in planta transformation methods. Czech J. Genet. Plant Breed., 53: 133−143.Gene transformation can be done in direct and indirect (Agrobacterium-mediated) ways. The most efficient method of gene transformation to date is the Agrobacterium-mediated method. The main problem of this 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. The development of a genotype-independent gene transformation method is of great interest in many plants. Some Agrobacteriummediated gene transformation methods independent of tissue culture 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 transformation methods based on tissue culture. Vacuum infiltration, injection of Agrobacterium culture into 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 faster than regular Agrobacterium-mediated gene transformation based on tissue culture and success can be achieved by non-experts. In the present review, we highlight all methods of in planta transformation comparing them with regular Agrobacterium-mediated transformation methods based on tissue culture. Finally, successful recent transformations using these methods are presented.

Perspectives of CRISPR/Cas-mediated cis-engineering in horticulture: unlocking the neglected potential for crop improvement

“…Therefore, tissue culture-free methods are highly desirable and necessary for transgenefree genome editing. In planta transformation takes advantage of natural biological processes, which makes it a valuable alternative to in vitro tissue culture and regeneration 132,133 . Various plant cells or tissues can be the ideal transformation targets such as germline or meristematic cells 116,134,135 and dormant buds 136 .…”

In Planta Genetic Transformation of Mungbean (Vigna radiata (L.) Wilczek) with Marker Gene

“…On the other hand, non-transformed tissues failed to develop blue colour in GUS histochemical assay. Such chimeric expression of gene is considered as a disadvantage in the in planta transformation method (Niazian et al 2017). In order to eliminate non-transformed and chimeric transformed plants in case of in planta transformation BASTA® selection is proven to be most effective (Kapildev et al 2016, Mayavan et al 2015, Mayavan et al 2013.…”

“…Considering such recalcitrant nature of the mungbean plants towards in vitro regeneration, a tissue culture-independent genetic transformation method called in planta genetic transformation has been conceptualized. Such a method is quicker, simpler, cheaper, more efficient and somaclonal variation free (Rao et al 2008, Niazian et al 2017. Hence, attempts have been made to develop an Agrobacterium-mediated in planta genetic transformation protocol in a locally grown Bangladeshi mungbean variety, namely BARI Mung-3 using a screenable marker gene called "gus"gene.…”

Genome editing for horticultural crop improvement

“…Although tissue culture-based transformation and regeneration is most widely used for genome editing, no well-established protocol for transformation and regeneration from tissue culture is available for many horticultural crops. In planta transformation, which is an alternative to in vitro tissue culture-based Agrobacterium transformation, refers to the infection of in vivo explants in which the targeted tissues are apical or auxiliary meristems, stigmas, pollens, or inflorescences 94 . This method has been successfully used to transform tomato 95 and Brassica species 96 and should be further explored for use in horticultural crops that are recalcitrant to traditional genetic transformation.…”