Scientia Agricultura Sinica ›› 2026, Vol. 59 ›› Issue (19): 4208-4220.doi: 10.3864/j.issn.0578-1752.2026.19.002

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Identification of QTLs and Exploration of Candidate Genes Conferring Anaerobic Germination Tolerance in Rice

YANG HuiJuan1,2(), LIU YiFan2, YU XiaoMan2, HUANG DanLin2, CHEN DanDan2, SHU YaZhou2, WEI Hua2, HE HuiYing2, SHANG LianGuang2(), YUAN QiaoLing2()   

  1. 1 Qingdao Agricultural University, Qingdao 266000, Shandong
    2 Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences/Genome Analysis Laboratory of the Ministry of Agriculture and Rural Affairs/Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture/State Key Laboratory of Genome and Multi-omics Technologies, Shenzhen 518120, Guangdong
  • Received:2026-02-10 Accepted:2026-04-07 Online:2026-10-01 Published:2026-10-10
  • Contact: SHANG LianGuang, YUAN QiaoLing

Abstract:

【Objective】In direct-seeded rice production, low germination rates and uneven emergence under anaerobic conditions are major bottlenecks limiting yield stability and cultivation area. This study aims to elucidate the genetic regulation of anaerobic germination in rice and identify key genes controlling coleoptile elongation, thereby providing genetic resources for breeding submergence-tolerant direct-seeded rice varieties.【Method】Sixty-nine of 237 accessions were selected to measure anaerobic coleoptile length and seedling establishment rate, and the correlation between the two traits was evaluated. After anaerobic germination treatment, coleoptile length data was collected from all 237-rice pan-genome accessions. A genome-wide association study (GWAS) was performed using these phenotypic data together with high-density SNP markers to identify genetic loci underlying tolerance to anaerobic germination. Candidate genes were further screened through functional annotation, transcriptome analysis, and haplotype analysis.【Result】Coleoptile length was highly significantly correlated with seedling establishment rate (r2=0.4533), confirming that coleoptile elongation is a reliable indicator for anaerobic germination tolerance. GWAS identified two novel significant association loci on chromosomes 8 and 12, named qAG8 and qAG12, neither of which has been reported previously. Within these intervals, 46 and 48 genes were detected, respectively; after functional annotation, 29 genes from qAG8 and 14 from qAG12 were retained. Integration of transcriptome and haplotype analyses revealed four candidate genes potentially involved in anaerobic germination: LOC_Os08g39210 (encoding a cell wall-associated kinase OsWAK74, localized to the cell wall and negatively regulating cell proliferation), LOC_Os08g39290 (containing an EF-hand domain, possibly involved in calcium signaling), LOC_Os08g39550 (encoding a polygalacturonase inhibitor protein, localized to the cell wall), and LOC_Os12g39830 (a rice cyclin CycD5;2, involved in cell cycle regulation).【Conclusion】Two novel loci (qAG8 and qAG12) conferring tolerance to anaerobic germination were identified on chromosomes 8 and 12, and four candidate genes were uncovered through transcriptome and haplotype analyses. These findings provide new insights into the genetic basis of anaerobic germination and offer potential molecular targets for marker-assisted breeding of direct-seeded rice.

Key words: rice, anaerobic germination tolerance, coleoptile length, seedling establishment rate, genome-wide association study (GWAS)

Fig. 1

Population structure analysis of 237 Asian cultivated rice accessions A: Principal component analysis; B: Population structure distribution; C: Phylogenetic trees"

Fig. 2

Evaluation of anaerobic germination tolerance in rice based on coleoptile length A: Changes in coleoptile length under aerobic and anaerobic conditions; B: Histogram of the difference in coleoptile length between anaerobic and aerobic conditions"

Table 1

Information of ten submergence-tolerant germplasm resources"

品种
Accession
编号
No.
亚群
Subpopulation
有氧胚芽鞘长度
Aerobic coleoptile length (cm)
厌氧胚芽鞘长度
Anaerobic coleoptile length (cm)
9311 NH231 籼稻Indica 2.10 3.34
Taidongludao NH168 粳稻Japonica 2.12 3.46
NCS 458::IRGC 62290-1 TG52 籼稻Indica 3.18 3.48
Zinuo NH082 籼稻Indica 0.18 3.50
Nanoay P.A NH033 粳稻Japonica 1.78 3.56
L 301B NH133 籼稻Indica 1.90 3.58
2037(Rajahamsal) NH011 籼稻Indica 1.52 3.62
RY2 NH232 粳稻Japonica 2.88 3.78
J34 NH045 籼稻Indica 1.66 3.82
LJ TG7 粳稻Japonica 0.40 4.92

Fig. 3

Correlation analysis between coleoptile length and seedling establishment rate under direct-seeding flooding simulation A: Seedling establishment rate; B: Correlation between coleoptile length and seedling establishment rate"

Fig. 4

Genome-wide association study of anaerobic tolerance based on 237 Asian cultivated rice (Oryza sativa subsp japonica) accessions A: Manhattan plot; B: QQ plot"

Table 2

Candidate genes located in the qAG8 and qAG12 QTL region"

基因登录号
Accession
基因注释
Gene annotation
厌氧响应
Anaerobic response
LOC_Os08g39140 热激蛋白Heat shock protein —
LOC_Os08g39160 甲酰基转移酶Formyl transferase, putative, expressed —
LOC_Os08g39174 Ty1-copia反转录转座子蛋白Retrotransposon protein, putative, Ty1-copia subclass, expressed —
LOC_Os08g39180 OsWAK受体样蛋白激酶OsWAK73 OsWAK receptor-like protein kinase OsWAK73, expressed —
LOC_Os08g39210 壁相关激酶;类受体蛋白激酶
Wall-associated kinase 74; GRAIN WEIGHT AND NUMBER 1; receptor-like protein kinase
下调Down regulation
LOC_Os08g39220 OsWAK受体样蛋白激酶OsWAK75 OsWAK receptor-like protein kinase OsWAK75, expressed —
LOC_Os08g39240 OsWAK受体样细胞质激酶OsWAK76 OsWAK receptor-like cytoplasmic kinase OsWAK76, expressed —
LOC_Os08g39250 N6-腺嘌呤甲基转移酶样蛋白
ENHANCED DOWNY MILDEW 2-LIKE; N6-adenine methyltransferase-like (MTL) domain-containing protein
—
LOC_Os08g39270 细胞壁相关蛋白Fasciclin domain containing protein, expressed —
LOC_Os08g39290 EF手型结构域蛋白家族EF hand family protein, expressed 上调Up regulation
LOC_Os08g39300 丝氨酸:乙醛酸氨基转移酶Serine: glyoxylate aminotransferase —
LOC_Os08g39310 SWIB/MDM2结构域蛋白SWIB/MDM2 domain containing protein, expressed —
LOC_Os08g39330 皮肤分泌蛋xP2前体Skin secretory protein xP2 precursor, putative, expressed 上调Up regulation
LOC_Os08g39350 甘油磷酸二酯磷酸二酯酶家族蛋白
Glycerophosphoryl diester phosphodiesterase family protein, putative, expressed
—
LOC_Os08g39370 柠檬酸转运蛋白Citrate transporter, putative, expressed —
LOC_Os08g39380 β-1,2-木糖基转移酶β-1,2-xylT β-1,2-xylosyltransferase, β-1,2-xylT, reduced culm number11 —
LOC_Os08g39390 ZOS8-10:C2H2锌指蛋白ZOS8-10:C2H2 zinc finger protein, expressed —
LOC_Os08g39420 假尿苷-5′-磷酸糖苷酶
Pseudouridine-5′-phosphate glycosidase; Ψ-5′-phosphate glycosidase; homologous to both yeiN and PUMY
—
LOC_Os08g39430 叶绿体前体类囊体腔19 kDa蛋白Thylakoid lumenal 19 kDa protein, chloroplast precursor, putative, expressed —
LOC_Os08g39440 RNA结合蛋白RNA recognition motif containing protein, putative, expressed —
LOC_Os08g39450 A20/AN1型锌指蛋白A20/AN1 zinc-finger protein; Zinc-finger containing stress-associated protein —
LOC_Os08g39460 AGC激酶家族AGC_PVPK_like_CDK8.2-ACG kinases include homologs to PKA, PKG and PKC, expressed —
LOC_Os08g39500 60S核糖体蛋白L31 60S ribosomal protein L31, putative, expressed —
LOC_Os08g39540 锌指关节结构域蛋白Zinc knuckle domain containing protein, expressed —
LOC_Os08g39550 多聚半乳糖醛酸酶抑制蛋白Polygalacturonase-inhibiting protein 上调Up regulation
LOC_Os08g39560 半胱氨酸蛋白酶家族蛋白OTU-like cysteine protease family protein, putative, expressed —
LOC_Os08g39570 LRR结构域蛋白Leucine rich repeat family protein, expressed —
LOC_Os08g39580 锌指关节结构域蛋白Zinc knuckle family protein, expressed —
LOC_Os12g39750 保守假设蛋白Conserved hypothetical protein —
LOC_Os12g39830 细胞周期蛋白Rice cyclin gene 下调Down regulation
LOC_Os12g39850 bHLH转录因子Root Hair Defective-Six Like (RSL) Class Ⅱ family of transcription factor; bHLH transcription factor; basic helix-loop-helix (bHLH) transcription factor —
LOC_Os12g39860 磷酸核糖基转移酶Phosphoribosyl transferase, putative, expressed —
LOC_Os12g39980 含驱动蛋白马达结构域蛋白Kinesin motor domain containing protein, putative, expressed 下调Down regulation
LOC_Os12g39990 Dof转录因子;单锌指蛋白Dof transcription factor; DNA binding with one finger protein —
LOC_Os12g40070 B3 DNA结合结构域蛋白B3 DNA binding domain containing protein, expressed —
LOC_Os12g40080 B3 DNA结合结构域蛋白B3 DNA binding domain containing protein, expressed —
LOC_Os12g40090 B3 DNA结合结构域蛋白B3 DNA binding domain containing protein, expressed —
LOC_Os12g40120 B3 DNA结合结构域蛋白 B3 DNA binding domain containing protein, expressed —
LOC_Os12g40130 磷酸核糖基转移酶Phosphoribosyl transferase, putative, expressed —
LOC_Os12g40140 含F-box结构域蛋白OsFBX457 F-box domain containing protein OsFBX457, expressed —
LOC_Os12g40170 假设蛋白Hypothetical protein —
LOC_Os12g40190 超大GTP结合蛋白Extra-large GTP-binding protein —

Fig. 5

Heatmap of fold changes in expression of candidate genes at qAG8 and qAG12 loci under anaerobic and aerobic conditions a: Candidate genes associated with the qAG8 locus; b: Candidate genes associated with the qAG12 locus. A: Aerobic, N: Anaerobic"

Fig. 6

Haplotype analysis of candidate genes A and B: Haplotype analysis results of LOC_Os08g39210; C and D: Haplotype analysis results of LOC_Os08g39290; E and F: Haplotype analysis results of LOC_Os08g39550; G and H: Haplotype analysis results of LOC_Os12g39830; I and J: Haplotype combination analysis of OsLOX8 and CycD5. Different letters indicate significant differences among groups at P<0.05"

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