Characterisation 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 re...

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Main Authors:	Wang, Duoduo, Samsulrizal, Nurul Hidayah, Cheng, Yan, Allcock, Natalie, Craigon, Jim, Blanco-Ulate, Barbara, Ortega-Salazar, Isabel, E. Marcus, Susan, Moeiniyan Bagheri, Hassan, Perez-Fons, Laura, D. Fraser, Paul, Foster, Timothy, Fray, Rupert, Knox, J. Paul, B. Seymour, Graham
Format:	Article
Language:	English English
Published:	American Society of Plant Biologists (ASPB) 2019
Subjects:	SB Plant culture
Online Access:	http://irep.iium.edu.my/70037/ http://irep.iium.edu.my/70037/ http://irep.iium.edu.my/70037/7/544.full.pdf http://irep.iium.edu.my/70037/8/Scopus%20-%20Document%20details.pdf

id	iium-70037
recordtype	eprints
spelling	iium-700372019-02-11T10:41:44Z http://irep.iium.edu.my/70037/ Characterisation of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato Wang, Duoduo Samsulrizal, Nurul Hidayah Cheng, Yan Allcock, Natalie Craigon, Jim Blanco-Ulate, Barbara Ortega-Salazar, Isabel E. Marcus, Susan Moeiniyan Bagheri, Hassan Perez-Fons, Laura D. Fraser, Paul Foster, Timothy Fray, Rupert Knox, J. Paul B. Seymour, Graham 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 remodelling 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 remodelling. 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 colour and weight. Pectin localisation, distribution and solubility in the pericarp cells of the CRISPR mutant fruits were investigated using the monoclonal antibody probes LM19 to de-esterified homogalacturonan (HG), INRA-RU1 to rhamnogalacturonan I, LM5 to β1-4-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. American Society of Plant Biologists (ASPB) 2019-02 Article PeerReviewed application/pdf en http://irep.iium.edu.my/70037/7/544.full.pdf application/pdf en http://irep.iium.edu.my/70037/8/Scopus%20-%20Document%20details.pdf Wang, Duoduo and Samsulrizal, Nurul Hidayah and Cheng, Yan and Allcock, Natalie and Craigon, Jim and Blanco-Ulate, Barbara and Ortega-Salazar, Isabel and E. Marcus, Susan and Moeiniyan Bagheri, Hassan and Perez-Fons, Laura and D. Fraser, Paul and Foster, Timothy and Fray, Rupert and Knox, J. Paul and B. Seymour, Graham (2019) Characterisation of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato. Plant Physiology, 179 (2). pp. 544-557. ISSN 1532-2548 http://www.plantphysiol.org
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English English
topic	SB Plant culture
spellingShingle	SB Plant culture Wang, Duoduo Samsulrizal, Nurul Hidayah Cheng, Yan Allcock, Natalie Craigon, Jim Blanco-Ulate, Barbara Ortega-Salazar, Isabel E. Marcus, Susan Moeiniyan Bagheri, Hassan Perez-Fons, Laura D. Fraser, Paul Foster, Timothy Fray, Rupert Knox, J. Paul B. Seymour, Graham Characterisation 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 remodelling 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 remodelling. 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 colour and weight. Pectin localisation, distribution and solubility in the pericarp cells of the CRISPR mutant fruits were investigated using the monoclonal antibody probes LM19 to de-esterified homogalacturonan (HG), INRA-RU1 to rhamnogalacturonan I, LM5 to β1-4-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.
format	Article
author	Wang, Duoduo Samsulrizal, Nurul Hidayah Cheng, Yan Allcock, Natalie Craigon, Jim Blanco-Ulate, Barbara Ortega-Salazar, Isabel E. Marcus, Susan Moeiniyan Bagheri, Hassan Perez-Fons, Laura D. Fraser, Paul Foster, Timothy Fray, Rupert Knox, J. Paul B. Seymour, Graham
author_facet	Wang, Duoduo Samsulrizal, Nurul Hidayah Cheng, Yan Allcock, Natalie Craigon, Jim Blanco-Ulate, Barbara Ortega-Salazar, Isabel E. Marcus, Susan Moeiniyan Bagheri, Hassan Perez-Fons, Laura D. Fraser, Paul Foster, Timothy Fray, Rupert Knox, J. Paul B. Seymour, Graham
author_sort	Wang, Duoduo
title	Characterisation of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato
title_short	Characterisation of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato
title_full	Characterisation of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato
title_fullStr	Characterisation of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato
title_full_unstemmed	Characterisation of CRISPR mutants targeting genes modulating pectin degradation in ripening tomato
title_sort	characterisation of crispr mutants targeting genes modulating pectin degradation in ripening tomato
publisher	American Society of Plant Biologists (ASPB)
publishDate	2019
url	http://irep.iium.edu.my/70037/ http://irep.iium.edu.my/70037/ http://irep.iium.edu.my/70037/7/544.full.pdf http://irep.iium.edu.my/70037/8/Scopus%20-%20Document%20details.pdf
first_indexed	2023-09-18T21:39:26Z
last_indexed	2023-09-18T21:39:26Z
_version_	1777413040245833728

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

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