Transcriptomic Analysis of Peanut Pods in Response to Drought and Waterlogging Stresses

doi:10.7668/hbnxb.20195949

Abstract

Abstract:

To elucidate the molecular mechanisms underlying peanut pod responses to water stress,this study employed pot experiments combined with transcriptomic analysis.Using well-watered conditions as the control, we systematically investigated the effects of periodic drought and waterlogging stress during the flowering-pegging stage on yield,quality,and gene expression in peanut pods. Results demonstrated that both drought and waterlogging stresses significantly reduced peanut pod yield(by 26.43% and 77.69%,respectively)and crude fat content in kernels (by 9.46, 6.71 percentage points,respectively).Transcriptomic analysis further revealed 1 525 and 1 382 differentially expressed genes(DEGs)in the drought-stress and waterlogging-stress groups compared to the control, respectively, with down-regulated expression being predominant in both sets of DEGs. Specifically, drought stress suppressed six key metabolic processes related to lipid and fatty acid metabolism in peanut pods,with 88.38% of associated genes showing downregulated expression,indicating that lipid metabolic disruption may be the primary cause of yield and quality reduction under drought. Waterlogging stress predominantly interfered with pod metabolism and defense functions by downregulating genes associated with catalytic activity,transmembrane transport,redox reactions,and biosynthesis of secondary metabolites.Moreover, KEGG enrichment analysis indicated that metabolic pathways and biosynthesis of secondary metabolites were significantly affected under both water stress conditions. Key gene validation via qRT-PCR corroborated the RNA-seq data, confirming the reliability of the transcriptomic findings. In summary, this research elucidates the molecular basis of peanut pod response to water stress at the transcriptome level, demonstrating that lipid metabolic disruption is the primary factor underlying yield reduction and quality deterioration under drought stress, whereas peanut pods mitigate the adverse effects of waterlogging mainly by modulating the expression of genes associated with redox homeostasis and metabolic pathways.

Key words: Peanut pod, Drought stress, Waterlogging stress, Transcriptome, Kernel quality, Yield

CLC Number:

S565.2

YU Tianyi, FAN Zhaobo, ZHANG Jialei, LU Ya, WU Juxiang, YANG Jishun, LI Shangxia, WU Zhengfeng, WAN Shubo. Transcriptomic Analysis of Peanut Pods in Response to Drought and Waterlogging Stresses[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(6): 31-39. doi: 10.7668/hbnxb.20195949.

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Figures/Tables 8

Tab.1 Primers sequence information

基因名称 Gene name	基因注释 Gene annotation	上下调情况 Up-down situation	引物序列(5'—3') Primer sequence (5'—3')
LOC112697400	硬脂酰-[酰基-载体蛋白]9-去饱和酶6%2C叶绿体	D vs CK:不显著 W vs CK:上调	F:GGATCAGGAGGCTGCAAGAG R:TCTTGTGATGCAAGCAAAGCC
LOC112710465	亚油酸 9S-脂氧合酶	D vs CK:上调 W vs CK:下调	F:AGCACAGATGCCGTACACTC R:CAATACTGAAGAACCATAACTCCCT
LOC112727170	乙醇脱氢酶三类	D vs CK:不显著 W vs CK:下调	F:AACAGCTTTCGGAGGCTTCA R:AGAACACAACGGAGACACCC
LOC112738386	蔗糖合成酶	D vs CK:不显著 W vs CK:不显著	F:TGAACCTTGTCGTGGTAGCC R:GTGTCGCAGATCACACGGTA
LOC112764235	葡萄糖-1-磷酸腺苷基转移酶小亚基2%2C叶绿体	D vs CK:不显著 W vs CK:不显著	F:CACTCACACAACTCTCGCAAT R:CGGAAAGTTGTGAGGAGGAGA
LOC112766166	1% 2c4 -葡聚糖分支酶1%2C叶绿体/淀粉体	D vs CK:上调 W vs CK:不显著	F:CAGCGGGTGGTGTGTGTA R:AGTTCTCCCTCAAGCTACTGG
LOC112776097	亚油酸 9S-脂氧合酶	D vs CK:上调 W vs CK:下调	F:ATGCAAACTCGTGCGGAG R:CGGCATGAATTGCCTGCTAA
LOC112802430	β-淀粉酶1%2C叶绿体	D vs CK:不显著 W vs CK:不显著	F:CAGCAGCCTATCTCTCTGGC R:CGGCATTACGCTACATGAATGG
LOC112802492	乙醛脱氢酶家族2成员B7%2C线粒体异构体X3\|\|乙醛脱氢酶家族2成员B7%2C线粒体%2C转录变体	D vs CK:不显著 W vs CK:上调	F:GGAAGTGGTACAGAGGGCAA R:TCCCTTGTCCACTCATCTTGT

Tab.2 Effects of drought and waterlogging stress on peanut yield,yield components and dry matter weight

处理 Treatments	产量/(g/株) Yield	生物产量/(g/株) Biomass	收获指数/% Harvest index	单株果数 Pod number per plant	千克果数 Pod number per kg
CK	29.32±2.74a	54.80±3.89a	53.53±3.98a	25.42±3.26a	875.39±161.60b
D	21.57±0.78b	43.64±4.49b	49.66±3.58a	21.42±3.39a	991.15±130.26b
W	6.54±1.73c	37.41±2.47b	17.33±3.64b	8.17±1.18b	1 286.36±235.88a

Fig.1 Effects of drought and waterlogging stress on kernel quality of peanut Different lowercase letters on the column indicate significant difference of 0.05 among different treatments.

Tab.3 Summary of the RNA-Seq results

样本名称 Sample name	过滤后读数 Clean reads	Q20/%	Q30/%	GC/%
CK1	46 203 906	98.31	94.64	46.47
CK2	51 365 320	98.29	94.57	46.28
CK3	45 946 732	98.22	94.43	45.57
D1	46 391 496	98.16	94.28	46.58
D2	50 736 972	98.30	94.67	47.55
D3	45 234 596	97.96	93.81	45.43
W1	54 049 742	98.25	94.50	46.50
W2	52 552 574	98.02	93.96	45.41
W3	49 924 880	97.89	93.60	45.52

Fig.2 Analysis of differential expression gene in peanut pods under drought and waterlogging stress A.The number of differentially expressed genes;B.Venn diagram of differentially expressed genes.

Fig.3 GO enrichment of differentially expressed genes A. Drought and control group;B. Waterlogging and control group;C. Drought and waterlogging group.GO:0003824. Catalytic activity;GO:0003854. 3-beta-hydroxy-delta5-steroid dehydrogenase activity;GO:0004312. Fatty acid synthase activity;GO:0004315. 3-oxoacyl-[acyl-carrier-protein]synthase activity;GO:0004553. Hydrolase activity,hydrolyzing O-glycosyl compounds;GO:0004601. Peroxidase activity;GO:0004866. Endopeptidase inhibitor activity;GO:0005506. Iron ion binding;GO:0005975. Carbohydrate metabolic process;GO:0006629. Lipid metabolic process;GO:0006631. Fatty acid metabolic process;GO:0006633. Fatty acid biosynthetic process;GO:0008171. O-methyltransferase activity;GO:0008610. Lipid biosynthetic process;GO:0016053. Organic acid biosynthetic process;GO:0016229. Steroid dehydrogenase activity;GO:0016491. Oxidoreductase activity;GO:0016684. Oxidoreductase activity,acting on peroxide as acceptor;GO:0016705. Oxidoreductase activity,acting on paired donors,with incorporation or reduction of molecular oxygen;GO:0016746. Transferase activity,transferring acyl groups;GO:0016747. Transferase activity,transferring acyl groups other than amino-acyl groups;GO:0016758. Transferase activity,transferring hexosyl groups;GO:0016798. Hydrolase activity,acting on glycosyl bonds;GO:0020037. Heme binding;GO:0030001. Metal ion transport;GO:0030414. Peptidase inhibitor activity;GO:0032787. Monocarboxylic acid metabolic process;GO:0033764. Steroid dehydrogenase activity,acting on the CH-OH group of donors,NAD or NADP as acceptor;GO:0044255. Cellular lipid metabolic process;GO:0044281. Small molecule metabolic process;GO:0044283. Small molecule biosynthetic process;GO:0046394. Carboxylic acid biosynthetic process;GO:0046906. Tetrapyrrole binding;GO:0055085. Transmembrane transport;GO:0055114. Oxidation-reduction process;GO:0060962. Regulation of ribosomal protein gene transcription by RNA polymerase Ⅱ;GO:0061135. Endopeptidase regulator activity;GO:0072330. Monocarboxylic acid biosynthetic process.

Fig.4 Enrichment map of Top 20 KEGG of differentially expressed genes A.Drought and control group;B.Waterlogging and control group;C.Drought and waterlogging group.adu00010:Glycolysis/Gluconeogenesis;adu00040.Pentose and glucuronate interconversions;adu00052.Galactose metabolism;adu00061.Fatty acid biosynthesis;adu00071.Fatty acid degradation;adu00232.Caffeine metabolism;adu00330.Arginine and proline metabolism;adu00350.Tyrosine metabolism;adu00430.Taurine and hypotaurine metabolism;adu00460.Cyanoamino acid metabolism;adu00500.Starch and sucrose metabolism;adu00520.Amino sugar and nucleotide sugar metabolism;adu00561.Glycerolipid metabolism;adu00591.Linoleic acid metabolism;adu00592.alpha-Linolenic acid metabolism;adu00710.Carbon fixation in photosynthetic organisms;adu00905.Brassinosteroid biosynthesis;adu00909.Sesquiterpenoid and triterpenoid biosynthesis;adu00910.Nitrogen metabolism;adu00940.Phenylpropanoid biosynthesis;adu00941.Flavonoid biosynthesis;adu00943.Isoflavonoid biosynthesis;adu00944.Flavone and flavonol biosynthesis;adu00945.Stilbenoid,diarylheptanoid and gingerol biosynthesis;adu00950.Isoquinoline alkaloid biosynthesis;adu00965.Betalain biosynthesis;adu01100.Metabolic pathways;adu01110.Biosynthesis of secondary metabolites;adu04016.MAPK signaling pathway-plant;adu04075.Plant hormone signal transduction;adu04130.SNARE interactions in vesicular transport;adu04712.Circadian rhythm-plant.

Fig.5 The qRT-PCR detection of key differentially expressed genes

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