Characterization of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato

Tomato (Solanum lycopersicum) is a globally important crop with an economic value in the tens of billions of dollars, and a significant supplier of essential vitamins, minerals, and phytochemicals in the human diet. Shelf life is a key quality trait related to alterations in cuticle properties and r...

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Main Authors:	Wang, Duoduo, Samsulrizal, Nurul Hidayah, Yan, Cheng, Allcock, Natalie, Craigon, Jim, Blanco-Ulate, Barbara, Ortega-Salazar, Isabel, E. Marcus, Susan, Bagheri, Hassan Moeiniyan, Perez, Laura, Fraser, Paul D, Foster, Timothy, Fray, Rupert, Knox, J. Paul, Seymour, Graham B
Format:	Article
Language:	English English English
Published:	NLM (Medline) 2019
Subjects:	QK Botany SB Plant culture
Online Access:	http://irep.iium.edu.my/62266/ http://irep.iium.edu.my/62266/ http://irep.iium.edu.my/62266/1/62266_Characterization%20of%20CRISPR%20Mutants.pdf http://irep.iium.edu.my/62266/2/62266_Characterization%20of%20CRISPR%20Mutants_SCOPUS.pdf http://irep.iium.edu.my/62266/3/62266_Characterization%20of%20CRISPR%20Mutants_WOS.pdf

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recordtype	eprints
spelling	iium-622662019-07-12T07:44:29Z http://irep.iium.edu.my/62266/ Characterization of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato Wang, Duoduo Samsulrizal, Nurul Hidayah Yan, Cheng Allcock, Natalie Craigon, Jim Blanco-Ulate, Barbara Ortega-Salazar, Isabel E. Marcus, Susan Bagheri, Hassan Moeiniyan Perez, Laura Fraser, Paul D Foster, Timothy Fray, Rupert Knox, J. Paul Seymour, Graham B QK Botany SB Plant culture Tomato (Solanum lycopersicum) is a globally important crop with an economic value in the tens of billions of dollars, and a significant supplier of essential vitamins, minerals, and phytochemicals in the human diet. Shelf life is a key quality trait related to alterations in cuticle properties and remodeling of the fruit cell walls. Studies with transgenic tomato plants undertaken over the last 20 years have indicated that a range of pectin-degrading enzymes are involved in cell wall remodeling. These studies usually involved silencing of only a single gene and it has proved difficult to compare the effects of silencing these genes across the different experimental systems. Here we report the generation of CRISPR-based mutants in the ripening-related genes encoding the pectin-degrading enzymes pectate lyase (PL), polygalacturonase 2a (PG2a), and β-galactanase (TBG4). Comparison of the physiochemical properties of the fruits from a range of PL, PG2a, and TBG4 CRISPR lines demonstrated that only mutations in PL resulted in firmer fruits, although mutations in PG2a and TBG4 influenced fruit color and weight. Pectin localization, distribution, and solubility in the pericarp cells of the CRISPR mutant fruits were investigated using the monoclonal antibody probes LM19 to deesterified homogalacturonan, INRA-RU1 to rhamnogalacturonan I, LM5 to β-1,4-galactan, and LM6 to arabinan epitopes, respectively. The data indicate that PL, PG2a, and TBG4 act on separate cell wall domains and the importance of cellulose microfibril-associated pectin is reflected in its increased occurrence in the different mutant lines. © 2019 American Society of Plant Biologists. All Rights Reserved. NLM (Medline) 2019-02-01 Article PeerReviewed application/pdf en http://irep.iium.edu.my/62266/1/62266_Characterization%20of%20CRISPR%20Mutants.pdf application/pdf en http://irep.iium.edu.my/62266/2/62266_Characterization%20of%20CRISPR%20Mutants_SCOPUS.pdf application/pdf en http://irep.iium.edu.my/62266/3/62266_Characterization%20of%20CRISPR%20Mutants_WOS.pdf Wang, Duoduo and Samsulrizal, Nurul Hidayah and Yan, Cheng and Allcock, Natalie and Craigon, Jim and Blanco-Ulate, Barbara and Ortega-Salazar, Isabel and E. Marcus, Susan and Bagheri, Hassan Moeiniyan and Perez, Laura and Fraser, Paul D and Foster, Timothy and Fray, Rupert and Knox, J. Paul and Seymour, Graham B (2019) Characterization of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato. Plant Physiology, 179 (2). pp. 544-557. ISSN 0032-0889 E-ISSN 1532-2548 http://www.plantphysiol.org/content/plantphysiol/179/2/544.full.pdf
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English English English
topic	QK Botany SB Plant culture
spellingShingle	QK Botany SB Plant culture Wang, Duoduo Samsulrizal, Nurul Hidayah Yan, Cheng Allcock, Natalie Craigon, Jim Blanco-Ulate, Barbara Ortega-Salazar, Isabel E. Marcus, Susan Bagheri, Hassan Moeiniyan Perez, Laura Fraser, Paul D Foster, Timothy Fray, Rupert Knox, J. Paul Seymour, Graham B Characterization of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato
description	Tomato (Solanum lycopersicum) is a globally important crop with an economic value in the tens of billions of dollars, and a significant supplier of essential vitamins, minerals, and phytochemicals in the human diet. Shelf life is a key quality trait related to alterations in cuticle properties and remodeling of the fruit cell walls. Studies with transgenic tomato plants undertaken over the last 20 years have indicated that a range of pectin-degrading enzymes are involved in cell wall remodeling. These studies usually involved silencing of only a single gene and it has proved difficult to compare the effects of silencing these genes across the different experimental systems. Here we report the generation of CRISPR-based mutants in the ripening-related genes encoding the pectin-degrading enzymes pectate lyase (PL), polygalacturonase 2a (PG2a), and β-galactanase (TBG4). Comparison of the physiochemical properties of the fruits from a range of PL, PG2a, and TBG4 CRISPR lines demonstrated that only mutations in PL resulted in firmer fruits, although mutations in PG2a and TBG4 influenced fruit color and weight. Pectin localization, distribution, and solubility in the pericarp cells of the CRISPR mutant fruits were investigated using the monoclonal antibody probes LM19 to deesterified homogalacturonan, INRA-RU1 to rhamnogalacturonan I, LM5 to β-1,4-galactan, and LM6 to arabinan epitopes, respectively. The data indicate that PL, PG2a, and TBG4 act on separate cell wall domains and the importance of cellulose microfibril-associated pectin is reflected in its increased occurrence in the different mutant lines. © 2019 American Society of Plant Biologists. All Rights Reserved.
format	Article
author	Wang, Duoduo Samsulrizal, Nurul Hidayah Yan, Cheng Allcock, Natalie Craigon, Jim Blanco-Ulate, Barbara Ortega-Salazar, Isabel E. Marcus, Susan Bagheri, Hassan Moeiniyan Perez, Laura Fraser, Paul D Foster, Timothy Fray, Rupert Knox, J. Paul Seymour, Graham B
author_facet	Wang, Duoduo Samsulrizal, Nurul Hidayah Yan, Cheng Allcock, Natalie Craigon, Jim Blanco-Ulate, Barbara Ortega-Salazar, Isabel E. Marcus, Susan Bagheri, Hassan Moeiniyan Perez, Laura Fraser, Paul D Foster, Timothy Fray, Rupert Knox, J. Paul Seymour, Graham B
author_sort	Wang, Duoduo
title	Characterization of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato
title_short	Characterization of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato
title_full	Characterization of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato
title_fullStr	Characterization of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato
title_full_unstemmed	Characterization of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato
title_sort	characterization of crispr mutants targeting genes modulating pectin degradation in ripening tomato
publisher	NLM (Medline)
publishDate	2019
url	http://irep.iium.edu.my/62266/ http://irep.iium.edu.my/62266/ http://irep.iium.edu.my/62266/1/62266_Characterization%20of%20CRISPR%20Mutants.pdf http://irep.iium.edu.my/62266/2/62266_Characterization%20of%20CRISPR%20Mutants_SCOPUS.pdf http://irep.iium.edu.my/62266/3/62266_Characterization%20of%20CRISPR%20Mutants_WOS.pdf
first_indexed	2023-09-18T21:28:16Z
last_indexed	2023-09-18T21:28:16Z
_version_	1777412338614272000

Characterization of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato

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