Asparagine Synthetase Gene <i>OsASN2</i> Is Crucial for Rice Seed Development and Germination

Seed development plays a critical role in determining both crop yield and grain quality in rice. As a key nutrient storage organ, the rice endosperm development not only contributes to grain filling but also plays an essential role during the early stages of seed germination. Amino acid metabolism i...

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Main Authors: Rui Hu, Kaiming Liang, Xiangyu Hu, Meijuan Li, Qunhuan Ye, Yuanhong Yin, Cai Tang, Xinyu Wang, Youqiang Fu, Junfeng Pan, Mingyong Zhang, Xuhua Zhong
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Language:English
Published: MDPI AG 2025-06-01
Series:Plants
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Online Access:https://www.mdpi.com/2223-7747/14/13/1999
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author Rui Hu
Kaiming Liang
Xiangyu Hu
Meijuan Li
Qunhuan Ye
Yuanhong Yin
Cai Tang
Xinyu Wang
Youqiang Fu
Junfeng Pan
Mingyong Zhang
Xuhua Zhong
author_facet Rui Hu
Kaiming Liang
Xiangyu Hu
Meijuan Li
Qunhuan Ye
Yuanhong Yin
Cai Tang
Xinyu Wang
Youqiang Fu
Junfeng Pan
Mingyong Zhang
Xuhua Zhong
author_sort Rui Hu
collection DOAJ
description Seed development plays a critical role in determining both crop yield and grain quality in rice. As a key nutrient storage organ, the rice endosperm development not only contributes to grain filling but also plays an essential role during the early stages of seed germination. Amino acid metabolism is active during the process of seed development and seed germination. Asparagine is a primary amino acid responsible for long-distance organic nitrogen transport in plants. Asparagine synthetase catalyzes the synthesis of asparagine from aspartate and glutamine. In this study, CRISPR/Cas9-mediated knockout mutants of the <i>OsASN2</i> gene of rice were generated. Homozygous mutants exhibited complete failure of seed germination, and heterozygotes could not produce homozygous offspring. Endosperm development of homozygous mutant seeds showed severe defects. Additionally, interacting protein screening combined with pull-down and co-immunoprecipitation (Co-IP) assays confirmed that OsASN2 physically interacted with pyruvate phosphate dikinase OsPPDKB, the mutants of which showed impaired endosperm development. These findings collectively indicate that <i>OsASN2</i> plays a critical role in seed development and germination in rice.
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spelling doaj-art-36aed8cd8a66429fb79743f86467f67c2025-08-20T03:16:50ZengMDPI AGPlants2223-77472025-06-011413199910.3390/plants14131999Asparagine Synthetase Gene <i>OsASN2</i> Is Crucial for Rice Seed Development and GerminationRui Hu0Kaiming Liang1Xiangyu Hu2Meijuan Li3Qunhuan Ye4Yuanhong Yin5Cai Tang6Xinyu Wang7Youqiang Fu8Junfeng Pan9Mingyong Zhang10Xuhua Zhong11Rice Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Rice Science and Technology/Guangdong Rice Engineering Laboratory, Guangzhou 510640, ChinaRice Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Rice Science and Technology/Guangdong Rice Engineering Laboratory, Guangzhou 510640, ChinaRice Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Rice Science and Technology/Guangdong Rice Engineering Laboratory, Guangzhou 510640, ChinaRice Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Rice Science and Technology/Guangdong Rice Engineering Laboratory, Guangzhou 510640, ChinaRice Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Rice Science and Technology/Guangdong Rice Engineering Laboratory, Guangzhou 510640, ChinaRice Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Rice Science and Technology/Guangdong Rice Engineering Laboratory, Guangzhou 510640, ChinaRice Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Rice Science and Technology/Guangdong Rice Engineering Laboratory, Guangzhou 510640, ChinaRice Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Rice Science and Technology/Guangdong Rice Engineering Laboratory, Guangzhou 510640, ChinaRice Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Rice Science and Technology/Guangdong Rice Engineering Laboratory, Guangzhou 510640, ChinaRice Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Rice Science and Technology/Guangdong Rice Engineering Laboratory, Guangzhou 510640, ChinaGuangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, ChinaRice Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Genetics and Breeding of High Quality Rice in Southern China (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Rice Science and Technology/Guangdong Rice Engineering Laboratory, Guangzhou 510640, ChinaSeed development plays a critical role in determining both crop yield and grain quality in rice. As a key nutrient storage organ, the rice endosperm development not only contributes to grain filling but also plays an essential role during the early stages of seed germination. Amino acid metabolism is active during the process of seed development and seed germination. Asparagine is a primary amino acid responsible for long-distance organic nitrogen transport in plants. Asparagine synthetase catalyzes the synthesis of asparagine from aspartate and glutamine. In this study, CRISPR/Cas9-mediated knockout mutants of the <i>OsASN2</i> gene of rice were generated. Homozygous mutants exhibited complete failure of seed germination, and heterozygotes could not produce homozygous offspring. Endosperm development of homozygous mutant seeds showed severe defects. Additionally, interacting protein screening combined with pull-down and co-immunoprecipitation (Co-IP) assays confirmed that OsASN2 physically interacted with pyruvate phosphate dikinase OsPPDKB, the mutants of which showed impaired endosperm development. These findings collectively indicate that <i>OsASN2</i> plays a critical role in seed development and germination in rice.https://www.mdpi.com/2223-7747/14/13/1999asparagineamino acid metabolismendosperm developmentrice
spellingShingle Rui Hu
Kaiming Liang
Xiangyu Hu
Meijuan Li
Qunhuan Ye
Yuanhong Yin
Cai Tang
Xinyu Wang
Youqiang Fu
Junfeng Pan
Mingyong Zhang
Xuhua Zhong
Asparagine Synthetase Gene <i>OsASN2</i> Is Crucial for Rice Seed Development and Germination
Plants
asparagine
amino acid metabolism
endosperm development
rice
title Asparagine Synthetase Gene <i>OsASN2</i> Is Crucial for Rice Seed Development and Germination
title_full Asparagine Synthetase Gene <i>OsASN2</i> Is Crucial for Rice Seed Development and Germination
title_fullStr Asparagine Synthetase Gene <i>OsASN2</i> Is Crucial for Rice Seed Development and Germination
title_full_unstemmed Asparagine Synthetase Gene <i>OsASN2</i> Is Crucial for Rice Seed Development and Germination
title_short Asparagine Synthetase Gene <i>OsASN2</i> Is Crucial for Rice Seed Development and Germination
title_sort asparagine synthetase gene i osasn2 i is crucial for rice seed development and germination
topic asparagine
amino acid metabolism
endosperm development
rice
url https://www.mdpi.com/2223-7747/14/13/1999
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