CRISPR-Cas 9 mediated genome editing of selected early flowering genes in Wheat

Abstract

The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat) Cas9 method is the current favorite among the available gene editing methods, as it has shown to function as editing method in many species, while following a set of simple target specific design rules. However, it may not ultimately result in the introduction of foreign DNA at the target location. Despite being robust, developed and most successful genome editing tool, it has been frequently called into question, yet it is used as most accurate available genome editing tool. It maintains the great potential of improving crop plant performance by enhancing yield, nutritional value in crop and capability to cope with biotic as well as abiotic stresses. Different technique or guidelines have been developed specifically for different plant species. Since the domestication of wheat, flowering period has been intensively modified to get high yield in varying condition and environment and also, it provided a great variety of adaptation mechanism for study and research. Hexaploid – a type of wheat, ancestrally a long day plant - may not be compatible with many environments and such environments may require the types or varieties of wheat having PI (Photoperiod Insensitivity) so that it can flower in short days. CRISPR / Cas9 mediated protocol was used for modifying genome in wheat, along with detailed record on design of RNA, construction of vector, and transformation process in plants. Mutations on A B or D genome of Ppd-1 gene causes PI and also, flowering is affected through an alternative route, most likely, by up regulating FT as a result of mutation in Ppd-1. That suggests that a strong direct quantitative relationship exists between Ppd-1 mutation and FT expression and flowering