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Altering Stomatal Density for Manipulating Transpiration and Photosynthetic Traits in Rice through CRISPR/Cas9 Mutagenesis

Authors :
Sakthi Ambothi Rathnasamy
Rohit Kambale
Allimuthu Elangovan
Williams Mohanavel
Priyanka Shanmugavel
Gowtham Ramasamy
Senthil Alagarsamy
Rajavel Marimuthu
Veera Ranjani Rajagopalan
Sudha Manickam
Valarmathi Ramanathan
Raveendran Muthurajan
Geethalakshmi Vellingiri
Source :
Current Issues in Molecular Biology, Vol 45, Iss 5, Pp 3801-3814 (2023)
Publication Year :
2023
Publisher :
MDPI AG, 2023.

Abstract

Stomata regulates conductance, transpiration and photosynthetic traits in plants. Increased stomatal density may contribute to enhanced water loss and thereby help improve the transpirational cooling process and mitigate the high temperature-induced yield losses. However, genetic manipulation of stomatal traits through conventional breeding still remains a challenge due to problems involved in phenotyping and the lack of suitable genetic materials. Recent advances in functional genomics in rice identified major effect genes determining stomatal traits, including its number and size. Widespread applications of CRISPR/Cas9 in creating targeted mutations paved the way for fine tuning the stomatal traits for enhancing climate resilience in crops. In the current study, attempts were made to create novel alleles of OsEPF1 (Epidermal Patterning Factor), a negative regulator of stomatal frequency/density in a popular rice variety, ASD 16, using the CRISPR/Cas9 approach. Evaluation of 17 T0 progenies identified varying mutations (seven multiallelic, seven biallelic and three monoallelic mutations). T0 mutant lines showed a 3.7–44.3% increase in the stomatal density, and all the mutations were successfully inherited into the T1 generation. Evaluation of T1 progenies through sequencing identified three homozygous mutants for one bp insertion. Overall, T1 plants showed 54–95% increased stomatal density. The homozygous T1 lines (# E1-1-4, # E1-1-9 and # E1-1-11) showed significant increase in the stomatal conductance (60–65%), photosynthetic rate (14–31%) and the transpiration rate (58–62%) compared to the nontransgenic ASD 16. Results demonstrated that the genetic alterations in OsEPF1 altered the stomatal density, stomatal conductance and photosynthetic efficiency in rice. Further experiments are needed to associate this technology with canopy cooling and high temperature tolerance.

Details

Language :
English
ISSN :
14673045 and 14673037
Volume :
45
Issue :
5
Database :
Directory of Open Access Journals
Journal :
Current Issues in Molecular Biology
Publication Type :
Academic Journal
Accession number :
edsdoj.baec2927c8f49b19d5eb15e2c7fdde3
Document Type :
article
Full Text :
https://doi.org/10.3390/cimb45050245