Deciphering Regulatory Networks Governing Seedling Emergence in Deep-Sown Direct-Seeded Rice Cultivation

Abstract The rise of direct-seeded rice cultivation as a suitable alternative to transplanted puddled rice depends on developing genotypes with high seedling emergence under deep sown conditions. Two rice genotypes (IRGC 128442 and PR126) were screened for contrasting seedling emergence and subjecte...

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Main Authors: Jasneet Singh, Nitika Sandhu, Aman Kumar, Om Prakash Raigar, Sutej Bains, Gaurav Augustine, Muskan Gupta, Ekta Kharche, Anu Kalia, Navtej Singh Bains, Arvind Kumar
Format: Article
Language:English
Published: SpringerOpen 2025-02-01
Series:Rice
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Online Access:https://doi.org/10.1186/s12284-025-00760-0
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author Jasneet Singh
Nitika Sandhu
Aman Kumar
Om Prakash Raigar
Sutej Bains
Gaurav Augustine
Muskan Gupta
Ekta Kharche
Anu Kalia
Navtej Singh Bains
Arvind Kumar
author_facet Jasneet Singh
Nitika Sandhu
Aman Kumar
Om Prakash Raigar
Sutej Bains
Gaurav Augustine
Muskan Gupta
Ekta Kharche
Anu Kalia
Navtej Singh Bains
Arvind Kumar
author_sort Jasneet Singh
collection DOAJ
description Abstract The rise of direct-seeded rice cultivation as a suitable alternative to transplanted puddled rice depends on developing genotypes with high seedling emergence under deep sown conditions. Two rice genotypes (IRGC 128442 and PR126) were screened for contrasting seedling emergence and subjected to high-throughput RNA sequencing under varying sowing depths (4 cm and 10 cm) and time intervals (5, 10, and 15 days after sowing). On average, a total of 2702 differentially expressed genes were identified across twelve inter- and intra-genotypic pairwise differential expression analyses, with a false discovery rate ≤ 0.05 and log2 fold change ≥ ± 2. The DEGs specifically showing differential expression under deep-sowing stress were prioritized and further refined based on their corresponding gene ontology terms, gene set enrichment analysis and KEGG and plant reactome pathway. From this pool of DEGs, 24 genes were validated using qRT-PCR. Among these, two genes (LOC_Os04g51460 and LOC_Os02g45450) contribute to cell wall remodelling and membrane stability, while three genes (LOC_Os04g48484, LOC_Os06g04399, and LOC_Os07g15440) play key roles in mitigating abiotic stress. Transcriptional regulators (LOC_Os06g33940 and LOC_Os01g45730) drive stress responses and growth. Notably, high fold changes in LOC_Os03g22720 and LOC_Os07g01960 underscore their importance in early stress responses and metabolic adjustments. The transcriptome analysis also highlighted the role of 29 heat shock proteins in response to deep sowing stress. Differential expression of key components in the abscisic acid (ABA)-mediated signalling pathway such as OsABI5 (LOC_Os01g64000), phosphatase 2C-like (PP2C) (LOC_Os09g15670) and OsPYL (LOC_Os06g36670) indicated downregulation of ABA signalling in the genotype IRGC 128442. Additionally, a role for miRNA-mediated regulation of auxin response factors was hypothesized in seedling emergence regulation. The study brings us closer to understanding the genetic control of seedling emergence under deep sown conditions. Functional validation of the key candidate genes and pathways could provide new targets for genetic improvement, potentially contributing to the development of rice cultivars optimized for direct-seeded rice cultivation.
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spelling doaj-art-7fc63faa2a0a4165b52f3fb96fb373ed2025-02-09T12:54:26ZengSpringerOpenRice1939-84251939-84332025-02-0118111710.1186/s12284-025-00760-0Deciphering Regulatory Networks Governing Seedling Emergence in Deep-Sown Direct-Seeded Rice CultivationJasneet Singh0Nitika Sandhu1Aman Kumar2Om Prakash Raigar3Sutej Bains4Gaurav Augustine5Muskan Gupta6Ekta Kharche7Anu Kalia8Navtej Singh Bains9Arvind Kumar10Punjab Agricultural UniversityPunjab Agricultural UniversityPunjab Agricultural UniversityPunjab Agricultural UniversityPunjab Agricultural UniversityPunjab Agricultural UniversityPunjab Agricultural UniversityPunjab Agricultural UniversityPunjab Agricultural UniversityPunjab Agricultural UniversityDelta AgrigeneticsAbstract The rise of direct-seeded rice cultivation as a suitable alternative to transplanted puddled rice depends on developing genotypes with high seedling emergence under deep sown conditions. Two rice genotypes (IRGC 128442 and PR126) were screened for contrasting seedling emergence and subjected to high-throughput RNA sequencing under varying sowing depths (4 cm and 10 cm) and time intervals (5, 10, and 15 days after sowing). On average, a total of 2702 differentially expressed genes were identified across twelve inter- and intra-genotypic pairwise differential expression analyses, with a false discovery rate ≤ 0.05 and log2 fold change ≥ ± 2. The DEGs specifically showing differential expression under deep-sowing stress were prioritized and further refined based on their corresponding gene ontology terms, gene set enrichment analysis and KEGG and plant reactome pathway. From this pool of DEGs, 24 genes were validated using qRT-PCR. Among these, two genes (LOC_Os04g51460 and LOC_Os02g45450) contribute to cell wall remodelling and membrane stability, while three genes (LOC_Os04g48484, LOC_Os06g04399, and LOC_Os07g15440) play key roles in mitigating abiotic stress. Transcriptional regulators (LOC_Os06g33940 and LOC_Os01g45730) drive stress responses and growth. Notably, high fold changes in LOC_Os03g22720 and LOC_Os07g01960 underscore their importance in early stress responses and metabolic adjustments. The transcriptome analysis also highlighted the role of 29 heat shock proteins in response to deep sowing stress. Differential expression of key components in the abscisic acid (ABA)-mediated signalling pathway such as OsABI5 (LOC_Os01g64000), phosphatase 2C-like (PP2C) (LOC_Os09g15670) and OsPYL (LOC_Os06g36670) indicated downregulation of ABA signalling in the genotype IRGC 128442. Additionally, a role for miRNA-mediated regulation of auxin response factors was hypothesized in seedling emergence regulation. The study brings us closer to understanding the genetic control of seedling emergence under deep sown conditions. Functional validation of the key candidate genes and pathways could provide new targets for genetic improvement, potentially contributing to the development of rice cultivars optimized for direct-seeded rice cultivation.https://doi.org/10.1186/s12284-025-00760-0ABADeep sown stressDSRHSPmiRNA riceRNA sequencing
spellingShingle Jasneet Singh
Nitika Sandhu
Aman Kumar
Om Prakash Raigar
Sutej Bains
Gaurav Augustine
Muskan Gupta
Ekta Kharche
Anu Kalia
Navtej Singh Bains
Arvind Kumar
Deciphering Regulatory Networks Governing Seedling Emergence in Deep-Sown Direct-Seeded Rice Cultivation
Rice
ABA
Deep sown stress
DSR
HSP
miRNA rice
RNA sequencing
title Deciphering Regulatory Networks Governing Seedling Emergence in Deep-Sown Direct-Seeded Rice Cultivation
title_full Deciphering Regulatory Networks Governing Seedling Emergence in Deep-Sown Direct-Seeded Rice Cultivation
title_fullStr Deciphering Regulatory Networks Governing Seedling Emergence in Deep-Sown Direct-Seeded Rice Cultivation
title_full_unstemmed Deciphering Regulatory Networks Governing Seedling Emergence in Deep-Sown Direct-Seeded Rice Cultivation
title_short Deciphering Regulatory Networks Governing Seedling Emergence in Deep-Sown Direct-Seeded Rice Cultivation
title_sort deciphering regulatory networks governing seedling emergence in deep sown direct seeded rice cultivation
topic ABA
Deep sown stress
DSR
HSP
miRNA rice
RNA sequencing
url https://doi.org/10.1186/s12284-025-00760-0
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