Repositorium

What is a repositorium?

The repositorium is a searchable database that provides data on relevant articles from journals, company web pages and web pages of governmental agencies about studies/applications of genome-editing in model plants and agricultural crops in the period January 1996 to May 2018. Search options are article type, technique, plant, traits or free text. The repositorium is based on the systematic map of Dominik Modrzejewski et al., published in the journal environmental evidence. (Download article PDF).

CRISPR/Cas9-mediated efficient editing in phytoene desaturase (PDS) demonstrates precise manipulation in banana cv. Rasthali genome


Typ / Jahr

Journal Article / 2018

Autoren

Kaur, Navneet; Alok, Anshu; Shivani; Kaur, Navjot; Pandey, Pankaj; Awasthi, Praveen; Tiwari, Siddharth

Abstract

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) has been reported for precise genome modification inmany plants. In the current study, we demonstrate a successful mutation in phytoene desaturase (RAS-PDS) ofbanana cv. Rasthali using the CRISPR/Cas9 system. Two PDS genes were isolated from Rasthali (RAS-PDS1 and RAS-PDS2), and their protein sequence analysis confirmed that both PDS comprises conserved motifs for enzyme activity. Phylogenetic analysis of RAS-PDS1 and RAS-PDS2 revealed a close evolutionary relationship with other monocot species. The tissue-specific expression profile of RAS-PDS1 and RAS-PDS2 in Rasthali suggested differential regulation of the genes. A single 19-bp guideRNA (gRNA) was designed to target the conserved region ofthese two RAS-PDSand transformedwith Cas9 in embryogenic cell suspension (ECS) cultures of cv. Rasthali. Complete albino and variegated phenotype were observed among regenerated plantlets. DNA sequencing of13 plants confirmed the indels with 59% mutation frequency in RAS-PDS, suggesting activation ofthe non-homologous end-joining (NHEJ) pathway. The majority ofmutations were either insertion (1–5) or deletion (1–4) ofnucleotides near to protospacer adjacent motif(PAM). These mutations have created stop codons in RAS-PDS sequences which suggest premature termination ofRAS-PDS protein synthesis. The decreased chlorophyll and total carotenoid contents were detected in mutant lines that revealed the functional disruption of both RAS-PDS genes. Our results demonstrate that genome editing through CRISPR/Cas9 can be applied as an efficient tool for banana genome modification.

Keywords
Banana transformation; carotenoid; Embryonic cell suspension; genome editing; Mutation
Periodical
Functional & integrative genomics
Periodical Number
1
Page range
89–99
Volume
18
DOI
10.1007/s10142-017-0577-5D

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
714 Tiwari, Siddharth
India
Banana cv. Rasthali CRISPR/Cas9
PDS
Albino phenotype
SDN1
Basic research
Basic research