Molecular Mechanism of Cydonia oblonga Responding to NaHCO₃ Alkali Stress Based on Transcriptomic Analysis

doi:10.15933/j.cnki.1004-3268.2025.11.012

Journal of Henan Agricultural Sciences ›› 2025, Vol. 54 ›› Issue (11): 109-122.DOI: 10.15933/j.cnki.1004-3268.2025.11.012

• Horticulture • Previous Articles Next Articles

Molecular Mechanism of Cydonia oblonga Responding to NaHCO_₃ Alkali Stress Based on Transcriptomic Analysis

GUO Xianping^1,2,3，LÜ Zhenzhen^1,2,3，WANG Dongsheng^1,2,3，JIANG Hui¹，WU Zhongying¹，XU Lingfei⁴，HAN Yongping¹，GUO Peng¹，WANG Jiao¹

（1.Horticulture Research Institute，Henan Academy of Agricultural Sciences，Zhengzhou 450002，China；2.Henan Horticulture and Flower Engineering Research Center，Zhengzhou 450002，China；3.Henan Characteristic Melon and Fruit Engineering Technology Research Center，Zhengzhou 450002，China；4.College of Horticulture，Northwest A&F University，Yangling 712100，China）

Received:2025-05-04 Accepted:2025-06-26 Published:2025-11-15 Online:2025-11-24

基于转录组学分析榅桲响应NaHCO₃碱胁迫分子机制

郭献平^1,2,3，吕珍珍^1,2,3，王东升^1,2,3，蒋卉¹，吴中营¹，徐凌飞⁴，韩永平¹，郭鹏¹，王蛟¹

（1.河南省农业科学院园艺研究所，河南郑州 450002；2.河南省园艺与花卉工程研究中心，河南郑州 450002；3.河南省特色瓜果工程技术研究中心，河南郑州 450002；4.西北农林科技大学园艺学院，陕西杨凌 712100）

通讯作者: 王东升（1966-），男，河南武陟人，研究员，硕士，主要从事梨栽培生理研究。E-mail：wdse66@126.com
作者简介:郭献平（1982-），男，河南内黄人，副研究员，博士，主要从事梨栽培生理与分子生物学研究。E-mail：xianping2005@163.com
基金资助:
河南省科技攻关项目（242102110151，232102110196）；河南省农业科学院应用科技攻关专项（2024YY03）；河南省重大科技专项（221100110400）；河南省农业科学院科技创新团队项目（2024TD41）；国家梨产业技术体系项目（CARS-28）；河南省农业科学院自主创新项目（2025ZC37，2024ZC032）

Abstract

Abstract: To investigate the molecular mechanisms of pear dwarf rootstock quince（Cydonia oblonga）responding to NaHCO3‑induced alkaline stress，hydroponically cultivated quince seedlings were subjected to mild alkaline stress（T1）with 5 mmol/L NaHCO₃ and moderate alkaline stress（T2）with 10 mmol/L NaHCO₃，without NaHCO₃ as a control（CK）. After six weeks of treatment，chlorophyll content，net photosynthetic rate in leaves，and K⁺/Na⁺ ratios in leaves and roots were measured. Additionally，transcriptome sequencing and analysis were performed on the roots. The results showed that both chlorophyll content and net photosynthetic rate in quince leaves gradually decreased with alkaline stress increasing. Compared to CK，the chlorophyll a+b content in T1 and T2 decreased significantly by 15.71% and 45.38%，respectively，while the net photosynthetic rate decreased by 15.90% and 55.42%，respectively. The K⁺/Na⁺ ratios in leaves and roots of T1 and T2 were significantly reduced by 45.40% and 96.69%，and 92.04% and 96.32%，respectively，compared to CK.Transcriptome sequencing revealed a total of 4 974 differentially expressed genes（DEGs）between T1，T2，and CK，with T2 showing a significantly higher number of DEGs than T1.Gene ontology（GO）and Kyoto encyclopedia of genes and genomes（KEGG）pathway enrichment analyses indicated that the biological processes（BP）in T1 and T2 were mainly enriched in terms such as hydrogen peroxide catabolic process and response to wounding，while KEGG pathways were primarily enriched in phenylpropanoid biosynthesis. Gene set enrichment analysis（GSEA）showed that the phenylpropanoid biosynthesis pathway was more active under T1 and T2 compared to CK.Weighted gene co‑expression network analysis（WGCNA）revealed that the hub gene cydonia_oblonga_newGene_11700，annotated as a lignin metabolism transcription factor，was significantly induced by alkaline stress in T1.In T2，the hub genes cydonia_oblonga_newGene_11460 and cydonia_oblonga_newGene_14681，both annotated as carbonic anhydrases，were identified.The expression patterns of five selected DEGs were validated by qRT‑PCR，and the results were consistent with the RNA‑Seq analysis.In summary，quince mobilizes more genes in response to moderate alkaline stress compared to mild stress，and the lignin biosynthesis pathway and carbonic anhydrases play important roles in quince’s response to NaHCO₃‑induced alkaline stress.

Key words: Cydonia oblonga, Alkaline stress, Phenylpropanoid biosynthesis pathway, Carbonic anhydrase, Transcriptomic analysis

摘要： 为探究梨异属矮化砧木榅桲响应NaHCO₃碱胁迫的分子机制，对水培榅桲幼苗进行5（T1）、10 mmol/L（T2）NaHCO₃碱胁迫处理，以不加NaHCO3为对照（CK），处理6周后，测定叶片叶绿素含量、净光合速率、根系和叶片K⁺/Na⁺等指标，并对根系进行转录组测序和分析。结果表明，榅桲叶片叶绿素含量和净光合速率均随着碱胁迫程度的增加而逐渐降低，T1、T2处理叶绿素a+b含量比CK显著降低，降幅分别为15.71%、45.38%，叶片净光合速率比CK显著降低，降幅分别为15.90%、55.42%。T1、T2处理叶片和根系K+/Na+均比CK显著降低，降幅分别为45.40%和96.69%、92.04%和96.32%。转录组测序结果显示，与CK相比，T1、T2处理共筛选出4 974个差异表达基因，T2的差异表达基因数量显著高于T1。GO（Gene ontology）功能富集和KEGG（Kyoto encyclopedia of genes and genomes）通路富集分析显示，T1和T2处理在生物学过程（BP）主要富集到过氧化氢分解途径、对伤害的反应等条目，KEGG通路主要富集于苯丙烷生物合成等代谢通路。基因集富集分析（GSEA）结果显示，与CK相比，苯丙烷生物合成代谢通路在T1 和T2 碱胁迫处理下活性增强。加权基因共表达网络分析（WGCNA）显示，T1 处理枢纽基因cydonia_oblonga_newGene_11700表达受到碱胁迫的显著诱导，该基因被注释为木质素代谢转录因子。T2处理枢纽基因cydonia_oblonga_newGene_11460和cydonia_oblonga_newGene_14681均被注释为碳酸酐酶。选取5个差异表达基因进行qRT-PCR验证，其表达模式与RNA-Seq分析结果一致。综合分析，榅桲在10 mmol/L NaHCO₃条件下比5 mmol/L NaHCO₃下能调动更多的基因响应碱胁迫，且木质素合成代谢通路和碳酸酐酶在榅桲响应NaHCO₃碱胁迫时起到重要作用。

关键词: 榅桲, 碱胁迫, 苯丙烷生物合成代谢通路, 碳酸酐酶, 转录组分析

CLC Number:

S661.2

GUO Xianping, LÜ Zhenzhen, WANG Dongsheng, JIANG Hui, WU Zhongying, XU Lingfei, HAN Yongping, GUO Peng, WANG Jiao. Molecular Mechanism of Cydonia oblonga Responding to NaHCO_₃ Alkali Stress Based on Transcriptomic Analysis[J]. Journal of Henan Agricultural Sciences, 2025, 54(11): 109-122.

郭献平, 吕珍珍, 王东升, 蒋卉, 吴中营, 徐凌飞, 韩永平, 郭鹏, 王蛟. 基于转录组学分析榅桲响应NaHCO₃碱胁迫分子机制[J]. 河南农业科学, 2025, 54(11): 109-122.

Figures/Tables 11

References

［1］梁颖，吴雪梅，林梦桦，等. 我国梨全产业链标准体系现状分析与思考［J］. 农产品质量与安全，2024（6）：5‑11.
LIANG Y，WU X M，LIN M H，et al. Analysis and reflection on the current status of pear whole industry chain standard system in China［J］. Quality and Safety of Agro‑products，2024（6）：5‑11.
［2］国家统计局农村社会经济调查司. 2023中国农村统计年鉴［M］. 北京：中国统计出版社，2023：176‑179.
Rural Socioeconomic Survey Department，National Bureau of Statistics. China rural statistical yearbook 2023［M］. Beijing：China Statistics Press，2023：176‑179.

［3］WU J，WANG Z W，SHI Z B，et al. The genome of the pear（Pyrus bretschneideri Rehd.）［J］.Genome Research，2013，23（2）：396‑408.

［4］杨健，李坤，王龙，等. 国内西洋梨的生态适应性及栽培区域分析［J］. 中国农学通报，2021，37（16）：97‑101.
YANG J，LI K，WANG L，et al. Ecological adaptability and cultivation area of Pyrus communis in China［J］.Chinese Agricultural Science Bulletin，2021，37（16）：97‑101.
［5］谭旭，于云飞，王圣元，等. 梨种苗“砧木春早播和断根移栽”繁育技术研究［J］.中国果树，2022（2）：32‑37.
TAN X，YU Y F，WANG S Y，et al. Propagation of pear nursery stocks through seed sowing in early spring and transplanting with pruned roots［J］. China Fruits，2022（2）：32‑37.

［6］李莉，宣景宏，杜国栋，等. 梨矮化砧木选育及其应用研究进展［J］. 北方果树，2015（1）：1‑3.

LI L，XUAN J H，DU G D，et al. Progresses on the breeding and application of dwarfing rootstocks of pear［J］. Northern Fruits，2015（1）：1‑3.

［7］刘晨露. 富平楸子根系氨基酸代谢对碱胁迫的响应机制研究［D］. 杨凌：西北农林科技大学，2023.

LIU C L. Response mechanisms of amino acid metabolism in roots of Malus prunifolia under alkali stress［D］.Yangling：Northwest A&F University，2023.

［8］ZHANG Z Z，HE K N，ZHANG T，et al. Physiological responses of Goji berry（Lycium barbarum L.）to saline‑alkaline soil from Qinghai region，China［J］.Scientific Reports，2019，9（1）：12057.

［9］LI Q，LI M L，MA H Y，et al. Quantitative phosphoproteomic analysis provides insights into the sodium bicarbonate responsiveness of Glycine max［J］.Biomolecules，2023，13（10）：1520.

［10］VALIPOUR M，KHOSHGOFTARMANESH A H，BANINASAB B. Physiological responses of hawthorn（Crataegus persica Pojark.）and quince（Cydonia oblonga Mill. ）rootstocks to bicarbonate‑induced iron deficiency in nutrient solution［J］. Journal of Plant Nutrition and Soil Science，2018，181（6）：905‑913.

［11］FAN Y P，LU X K，CHEN X G，et al. Cotton transcriptome analysis reveals novel biological pathways that eliminate reactive oxygen species（ROS）under sodium bicarbonate（NaHCO₃） alkaline stress ［J］. Genomics，2021，113（3）：1157‑1169.

［12］魏天娇. 紫花苜蓿（Medicago sativa L.）品种耐盐碱性田间鉴定与抗逆生理机制的研究［D］. 北京：中国科学院大学，2021.

WEI T J. Field identification of saline‑alkaline tolerance of alfalfa（Medicago sativa L.）cultivars and its physiological mechanism of stress tolerance［D］.Beijing：University of Chinese Academy of Sciences，2021.

［13］PÉREZ‑MARTÍN L，ALMIRA M J，ESTRELA‑MURIEL L，et al. A role for root carbonic anhydrase βCA4 in the bicarbonate tolerance of Arabidopsis thaliana［J］.Physiologia Plantarum，2024，176（6）：e70026.

［14］PÉREZ‑MARTÍN L，BUSOMS S，TOLRÀ R，et al.Transcriptomics reveals fast changes in salicylate and jasmonate signaling pathways in shoots of carbonate‑tolerant Arabidopsis thaliana under bicarbonate exposure［J］.International journal of Molecular Sciences，2021，22（3）：1226.

［15］SONG J Q，WANG J C，QIN R，et al. RNA‑Seq‑based WGCNA reveals the physiological and molecular responses of poplar leaves to NaHCO3 stress［J］. Trees，2024，39（1）：3.

［16］XIANG G Q，MA W Y，GAO S W，et al. Transcriptomic and phosphoproteomic profiling and metabolite analyses reveal the mechanism of NaHCO₃‑induced organic acid secretion in grapevine roots［J］. BMC Plant Biology，2019，19（1）：383.

［17］KANG Y C，YANG X Y，LIU Y H，et al. Integration of mRNA and miRNA analysis reveals the molecular mechanism of potato（Solanum tuberosum L.）response to alkali stress［J］. International Journal of Biological Macromolecules，2021，182：938‑949.

［18］楚乐乐，罗成科，田蕾，等. 植物对碱胁迫适应机制的研究进展［J］.植物遗传资源学报，2019，20（4）：836‑844.
CHU L L，LUO C K，TIAN L，et al. Research advance in plants’adaptation to alkali stress［J］.Journal of Plant Genetic Resources，2019，20（4）：836‑844.

［19］郭献平，吴中营，王东升，等. 杜梨根系铁吸收关键基因生物信息学分析及缺铁胁迫对其表达的影响［J］.河南农业科学，2017，46（8）：96‑101.
GUO X P，WU Z Y，WANG D S，et al. Bioinformatics analysis of iron uptake key gene in the root of Pyrus betulifolia and the effect of iron deficiency on its expression［J］. Journal of Henan Agricultural Sciences，2017，46（8）：96‑101.

［20］LICHTENTHALER H K， WELLBURN A R.Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents［J］.Biochemical Society Transactions，1983，11（5）：591‑592.

［21］刘正祥，张华新，杨秀艳，等. NaCl胁迫下沙枣幼苗生长和阳离子吸收、运输与分配特性［J］. 生态学报，2014，34（2）：326‑336.
LIU Z X，ZHANG H X，YANG X Y，et al. Growth，and cationic absorption，transportation and allocation of Elaeagnus angustifolia seedlings under NaCl stress［J］.Acta Ecologica Sinica，2014，34（2）：326‑336.

［22］MORTAZAVI A，WILLIAMS B A，MCCUE K，et al.Mapping and quantifying mammalian transcriptomes by RNA‑Seq［J］. Nature Methods，2008，5（7）：621‑628.

［23］车玉红，杨波，郭春苗，等. 榅桲果实CAD基因的克隆、序列分析及表达［J］.新疆农业科学，2020，57（5）：814‑821.
CHE Y H，YANG B，GUO C M，et al. Cloning，sequence analysis and expression of CAD gene in quince（Cydonia oblonga Mill）fruit［J］. Xinjiang Agricultural Sciences，2020，57（5）：814‑821.
［24］董菁，张春宵，刘学岩，等. 碱胁迫条件下玉米苗期根部转录组表达谱分析［J］.分子植物育种，2022，23（19）：1‑17.
DONG J，ZHANG C X，LIU X Y，et al. Analysis of transcriptome expression profile of maize seedling roots under alkali stress［J］. Molecular Plant Breeding，2022，23（19）：1‑17.

［25］DONG N Q，LIN H X. Contribution of phenylpropanoid metabolism to plant development and plant –environment interactions［J］. Journal of Integrative Plant Biology，2021，63（1）：180‑209.

［26］MA Q，WANG H L，WU E G，et al. Comprehensive physiological，transcriptomic，and metabolomic analysis of the response of Panicum miliaceum L. roots to alkaline stress［J］. Land Degradation & Development，2023，34（10）：2912‑2930.

［27］ HU P，ZHANG K M，YANG C P. BpNAC012 positively regulates abiotic stress responses and secondary wall biosynthesis［J］. Plant Physiology，2019，179（2）：700‑717.

［28］崔亚宁，满奕，宋程威，等. 植物凯氏带化学成分、生理功能及相关调控机制的研究进展［J］.中国科学（生命科学），2020，50（2）：102‑110.
CUI Y N，MAN Y，SONG C W，et al. Chemical components，physiological functions and regulation mechanism of plant Casparian strips［J］.Scientia Sinica （Vitae），2020，50（2）：102‑110.

［29］UDDIN N，LI X，ULLAH M W，et al. Lignin developmental patterns and Casparian strip as apoplastic barriers：A review［J］. International Journal of Biological Macromolecules，2024，260：129595.

［30］赵继发，沙伟，马天意. 类黄酮物质在植物逆境胁迫中的研究进展［J］.高师理科学刊，2021，41（7）：53‑59.
ZHAO J F，SHA W，MA T Y. Research progress of flavonoids in plants stresses［J］. Journal of Science of Teachers’College and University，2021，41（7）：53‑59.

［31］POLISHCHUK O V. Stress‑related changes in the expression and activity of plant carbonic anhydrases［J］.Planta，2021，253（2）：58.

Molecular Mechanism of Cydonia oblonga Responding to NaHCO_₃ Alkali Stress Based on Transcriptomic Analysis

基于转录组学分析榅桲响应NaHCO₃碱胁迫分子机制

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 11

References

Related Articles 1

Recommended Articles

Metrics