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

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 sto...

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Autores principales: Rathnasamy, Sakthi Ambothi, Kambale, Rohit, Elangovan, Allimuthu, Mohanavel, Williams, Shanmugavel, Priyanka, Ramasamy, Gowtham, Alagarsamy, Senthil, Marimuthu, Rajavel, Rajagopalan, Veera Ranjani, Manickam, Sudha, Ramanathan, Valarmathi, Muthurajan, Raveendran, Vellingiri, Geethalakshmi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10217023/
https://www.ncbi.nlm.nih.gov/pubmed/37232714
http://dx.doi.org/10.3390/cimb45050245
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author Rathnasamy, Sakthi Ambothi
Kambale, Rohit
Elangovan, Allimuthu
Mohanavel, Williams
Shanmugavel, Priyanka
Ramasamy, Gowtham
Alagarsamy, Senthil
Marimuthu, Rajavel
Rajagopalan, Veera Ranjani
Manickam, Sudha
Ramanathan, Valarmathi
Muthurajan, Raveendran
Vellingiri, Geethalakshmi
author_facet Rathnasamy, Sakthi Ambothi
Kambale, Rohit
Elangovan, Allimuthu
Mohanavel, Williams
Shanmugavel, Priyanka
Ramasamy, Gowtham
Alagarsamy, Senthil
Marimuthu, Rajavel
Rajagopalan, Veera Ranjani
Manickam, Sudha
Ramanathan, Valarmathi
Muthurajan, Raveendran
Vellingiri, Geethalakshmi
author_sort Rathnasamy, Sakthi Ambothi
collection PubMed
description 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 T(0) progenies identified varying mutations (seven multiallelic, seven biallelic and three monoallelic mutations). T(0) mutant lines showed a 3.7–44.3% increase in the stomatal density, and all the mutations were successfully inherited into the T(1) generation. Evaluation of T(1) progenies through sequencing identified three homozygous mutants for one bp insertion. Overall, T(1) plants showed 54–95% increased stomatal density. The homozygous T(1) 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.
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spelling pubmed-102170232023-05-27 Altering Stomatal Density for Manipulating Transpiration and Photosynthetic Traits in Rice through CRISPR/Cas9 Mutagenesis Rathnasamy, Sakthi Ambothi Kambale, Rohit Elangovan, Allimuthu Mohanavel, Williams Shanmugavel, Priyanka Ramasamy, Gowtham Alagarsamy, Senthil Marimuthu, Rajavel Rajagopalan, Veera Ranjani Manickam, Sudha Ramanathan, Valarmathi Muthurajan, Raveendran Vellingiri, Geethalakshmi Curr Issues Mol Biol Article 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 T(0) progenies identified varying mutations (seven multiallelic, seven biallelic and three monoallelic mutations). T(0) mutant lines showed a 3.7–44.3% increase in the stomatal density, and all the mutations were successfully inherited into the T(1) generation. Evaluation of T(1) progenies through sequencing identified three homozygous mutants for one bp insertion. Overall, T(1) plants showed 54–95% increased stomatal density. The homozygous T(1) 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. MDPI 2023-04-30 /pmc/articles/PMC10217023/ /pubmed/37232714 http://dx.doi.org/10.3390/cimb45050245 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rathnasamy, Sakthi Ambothi
Kambale, Rohit
Elangovan, Allimuthu
Mohanavel, Williams
Shanmugavel, Priyanka
Ramasamy, Gowtham
Alagarsamy, Senthil
Marimuthu, Rajavel
Rajagopalan, Veera Ranjani
Manickam, Sudha
Ramanathan, Valarmathi
Muthurajan, Raveendran
Vellingiri, Geethalakshmi
Altering Stomatal Density for Manipulating Transpiration and Photosynthetic Traits in Rice through CRISPR/Cas9 Mutagenesis
title Altering Stomatal Density for Manipulating Transpiration and Photosynthetic Traits in Rice through CRISPR/Cas9 Mutagenesis
title_full Altering Stomatal Density for Manipulating Transpiration and Photosynthetic Traits in Rice through CRISPR/Cas9 Mutagenesis
title_fullStr Altering Stomatal Density for Manipulating Transpiration and Photosynthetic Traits in Rice through CRISPR/Cas9 Mutagenesis
title_full_unstemmed Altering Stomatal Density for Manipulating Transpiration and Photosynthetic Traits in Rice through CRISPR/Cas9 Mutagenesis
title_short Altering Stomatal Density for Manipulating Transpiration and Photosynthetic Traits in Rice through CRISPR/Cas9 Mutagenesis
title_sort altering stomatal density for manipulating transpiration and photosynthetic traits in rice through crispr/cas9 mutagenesis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10217023/
https://www.ncbi.nlm.nih.gov/pubmed/37232714
http://dx.doi.org/10.3390/cimb45050245
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