Screening and Preliminary Exploration of Function of circRNAs Responding to High Temperature Stress in Maize Pollen

doi:10.15933/j.cnki.1004-3268.2026.01.003

Abstract

Abstract: Zhengdan 958 with high tolerance to heat and Xianyu 335 with low tolerance to heat were used as research materials. The semi‐automatic retractable plastic greenhouse was used for high temperature（HT）stress treatment at flowering stage with normal growth condition as the control（CK）.The differentially expressed circRNAs in maize pollen were screened by high‐throughput sequencing of circRNA under high temperature stress. GO and KEGG enrichment analysis of the host genes of differentially expressed circRNAs were done. The differentially expressed circRNAs with miRNA binding sites were screened， and their downstream target genes were predicted.The potential circRNA‐miRNA‐mRNA co‐expression regulatory networks which were in response to high temperature stress in maize pollen were constructed. The molecular mechanism of regulating high temperature stress in maize pollen was analyzed from multiple aspects，so as to provide a theoretical basis for improving the heat tolerance of maize varieties. The results showed that a total of 1 843 different circRNAs were identified in different samples of Zhengdan 958 and Xianyu 335. The distribution of circRNAs in maize chromosomes was different. Each circRNA contained different number of exon. Most of them，624 circRNAs contained only one exon. A total of 1 563 circRNAs were identified from Zhengdan 958 pollen，305，213 and 356 circRNAs were identified from CK958‐1，CK958‐2 and CK958‐3，respectively，and 222，242 and 225 circRNAs were identified from HT958‐1，HT958‐2 and HT958‐3，respectively.A total of 1 423 circRNAs were identified from Xianyu 335 pollen，272，188 and 229 circRNAs were identified from CK335‐1，CK335‐2 and CK335‐3，respectively，and 259，237 and 238 circRNAs were identified from HT335‐1，HT335‐2 and HT335‐3，respectively. The highest proportions in different samples were all the exon‐circRNA. There was no one‐to‐one correspondence between circRNAs and their host genes，748 circRNA host genes generated one circRNA separately through back splicing mechanism，and 156 circRNA host genes generated two circRNAs separately through back splicing mechanism.Nine differentially expressed circRNAs were screened in the HT958 vs CK958 group，of which two circRNAs were up regulated. Their host genes were significantly enriched in pyrophosphatase activity，nucleoside phosphate metabolic process，glycosylphosphatidylinositol（GPI）anchor metabolic process and other 14 GO items，and two KEGG pathways，including GPI anchor biosynthesis and metabolic pathway.Only one differentially expressed circRNA was screened in the HT335 vs CK335 group，and no significant GO item and KEGG pathway were enriched. Seventeen differentially expressed circRNAs were screened in the HT958 vs HT335 group，of which six circRNAs were up regulated.Their host genes were significantly enriched in endomembrane system，golgi‐associated vesicle membrane，membrane protein proteolysis and other 13 GO items，but no significant KEGG pathway. Five circRNAs had miRNA binding sites，which could be used as miRNA sponge island to adsorb miRNAs and indirectly regulate the expression of downstream target genes. circRNA‐miRNA‐mRNA co‐expression regulatory networks，including five circRNAs，five miRNAs from different families and two mRNAs，were constructed. Fifty‐four circRNAs which contained the internal ribosomal entry sites（IRES）were screened，which could translate into peptides or proteins and directly act on their target genes.

Key words: Maize, Pollen, High temperature stress, circRNA, Host gene of circRNA, Screening, Preliminary exploration of function

摘要： 以高耐热性玉米品种郑单958、低耐热性玉米品种先玉335为试验材料，以正常生长条件为对照（CK），利用半自动伸缩高温棚进行花期高温胁迫（HT）处理，通过circRNA高通量测序筛选高温胁迫下不同玉米品种花粉中差异表达的环状RNA（circRNA），对其来源基因进行GO和KEGG富集分析，并筛选具有miRNA结合位点的差异表达circRNA，预测其下游目的基因，分析玉米花粉中响应高温胁迫的潜在circRNA-miRNA-mRNA共表达调控网络，从多层面解析玉米花粉中调控高温胁迫的分子作用机制，为提高玉米品种的耐热性提供理论依据。结果表明，在郑单958、先玉335不同样本中共鉴定出1 843个不同的circRNA，它们在玉米染色体中的分布不同。每个circRNA所包含的外显子数目也不相同，其中，大多数（624 个）circRNA 只含有1 个外显子。在郑单958 花粉中共鉴定出1 563 个circRNA，其中，CK958-1、CK958-2、CK958-3中分别鉴定出305、213、356个circRNA，HT958-1、HT958-2、HT958-3中分别鉴定出222、242、225个circRNA。在先玉335花粉中共鉴定出1 423个circRNA，其中，CK335-1、CK335-2、CK335-3中分别鉴定出272、188、229个circRNA，HT335-1、HT335-2、HT335-3中分别鉴定出259、237、238个circRNA。不同样本中占比最高的均为外显子circRNA。circRNA与其来源基因不是一一对应的关系，有748个circRNA来源基因通过反向剪接机制只形成1个circRNA，156个circRNA来源基因通过反向剪接机制各自形成2个circRNA。在郑单958高温胁迫花粉与对照花粉对比组（HT958 vsCK958）中共筛选到9个差异表达circRNA，其中2个circRNA呈上调表达，其来源基因显著富集到焦磷酸酶活性、核苷酸磷酸代谢过程、糖基磷脂酰肌醇（GPI）锚定代谢过程等17个GO条目，显著富集到GPI锚定生物合成、代谢途径等KEGG 通路。在先玉335 高温胁迫花粉与对照花粉对比组（HT335 vs CK335）中共筛选到1个差异表达circRNA，其来源基因没有显著富集到任何GO条目、KEGG通路。在郑单958 高温胁迫花粉与先玉335 高温胁迫花粉对比组（HT958 vs HT335）中共筛选到17 个差异表达circRNA，其中6个circRNA呈上调表达，其来源基因显著富集到内质网系统、高尔基相关囊泡膜、膜蛋白水解等16个GO条目中，没有显著富集到任何KEGG代谢通路。5个circRNA具有miRNA结合位点，可以作为海绵岛吸附miRNA间接调控下游靶标基因的表达，构建了包括5个circRNA、5个不同家族miRNA、2个mRNA在内的circRNA-miRNA-mRNA共表达调控网络。筛选到了54个circRNA包含内部核糖体进入位点（IRES），可以翻译表达多肽或者蛋白质直接作用于靶标基因。

关键词: 玉米, 花粉, 高温胁迫, 环状RNA, circRNA来源基因, 筛选, 功能初探

CLC Number:

S513

LI Chuan, ZHANG Panpan, ZHANG Meiwei, GUO Hanxiao, MU Weilin, NIU Jun, QIAO Jiangfang. Screening and Preliminary Exploration of Function of circRNAs Responding to High Temperature Stress in Maize Pollen[J]. Journal of Henan Agricultural Sciences, 2026, 55(1): 26-39.

李川, 张盼盼, 张美微, 郭涵潇, 穆蔚林, 牛军, 乔江方. 玉米花粉中响应高温胁迫circRNA 的筛选及其功能初探[J]. 河南农业科学, 2026, 55(1): 26-39.

Figures/Tables 7

References

［1］霍治国，张海燕，李春晖，等. 中国玉米高温热害研究进展［J］. 应用气象学报，2023，34（1）：1‐14.

HUO Z G，ZHANG H Y，LI C H，et al. Review on high temperature heat damage of maize in China［J］.Journal of Applied Meteorological Science，2023，34（1）：1‐14.

［2］宋英博. 气象灾害对我国玉米安全生产的影响及对策［J］.作物研究，2022，36（1）：80‐83.

SONG Y B.Influence of meteorological disasters on safe production of maize in China and its countermeasures［J］.Crop Research，2022，36（1）：80‐83.

［3］孙永江，王琪，邵琪雯，等. 高温胁迫对植物光合作用的影响研究进展［J］.植物学报，2023，58（3）：486‐498.

SUN Y J，WANG Q，SHAO Q W，et al. Research advances on the effect of high temperature stress on plant photosynthesis［J］. Chinese Bulletin of Botany，2023，58（3）：486‐498.

［4］李小凡，邵靖宜，于维帧，等. 高温干旱复合胁迫对夏玉米产量及光合特性的影响［J］.中国农业科学，2022，55（18）：3516‐3529.

LI X F，SHAO J Y，YU W Z，et al. Combied effects of high temperature and drougt on yield and photosynthetic characteristics of summer maize［J］. Scientia Agricultura Ainica，2022，55（18）：3516‐3529.

［5］穆心愿，马智艳，张兰薰，等. 不同耐/感玉米品种的叶片光合荧光特性、授粉结实和产量构成因素对花期高温的反应［J］. 中国生态农业学报（中英文），2022，30 （1）：57‐71.

MU X Y，MA Z Y，ZHANG L X，et al. Responses of photosynthetic fluorescence characteristics，pollination，and yield components of maize cultivars to high temperature during flowering［J］. Chinese Journal of Eco‐Agriculture，2022，30（1）：57‐71.

［6］孙武，韩淑华，刘建民，等. 不同玉米品种在小穗分化期和抽雄期对高温胁迫的响应差异［J］. 中国农学通报，2019，35（17）：12‐19.

SUN W，HAN S H，LIU J M，et al. Response of maize cultivars to high temperature stress at spikelet differentiation stage and tasseling stage［J］. Chinese Agricultural Science Bulletin，2019，35（17）：12‐19.

［7］吴伟华，柳家友，袁刘正，等. 花期高温对不同玉米品种主要农艺形状和产量的影响［J］. 安徽农业科学，2020，48（6）：33‐36.

WU W H，LIU J Y，YUAN L Z，et al. Effects of high temperature at florescence stage on the yield and major agronomic characters of different maize varieties［J］.Journal of Anhui Agricultural Sciences，2020，48（6）：33‐36.

［8］杨浩，刘晨，王志飞，等. 作物花粉高温应答机制研究进展［J］.植物学报，2019，54（2）：157‐167.

YANG H，LIU C，WANG Z F，et al. Advances in the regulatory mechanisms of pollen response to heat stress in crops［J］. Chinese Bulletin of Botany，2019，54（2）：157‐167.

［9］余梦奇，路梦莉，张雅婷，等. 灌浆期高温对玉米叶片光合特性及抗氧化酶活性的影响［J］. 中国农业气象，

2023，44（7）：599‐610.
YU M Q，LU M L，ZHANG Y T，et al. Effects of high temperature on photosynthetic characteristics and antioxidant enzyme activities of maize leaves during filling stage［J］. Chinese Journal of Agrometeorology，2023，44 （7）：599‐610.

［10］吴丽倩，王蕊，杨玉荣，等. 高温对玉米叶片衰老及产量的影响［J］.华北农学报，2022，37（S1）：110‐115.

WU L Q，WANG R，YANG Y R，et al. Effects of high temperature on leaf senescence and yield of maize［J］.Acta Agriculturae Boreali‐Sinica，2022，37（S1）：110‐115.

［11］王涛，冯敬磊，张翠. 高温胁迫影响玉米生长发育的分子机制研究进展［J］.植物学报，2024，59（6）：963‐977.

WANG T，FENG J L，ZHANG C. Research progress on molecular mechanisms of heat stress affecting the growth and development of maize［J］. Chinese Bulletin of Botany，2024，59（6）：963‐977.

［12］尹军良，马东方，刘乐承，等. 环状RNA的生物特征及其在植物中的研究进展［J］.西北植物学报，2017，37（12）：2510‐2518.

YIN J L，MA D F，LIU L C，et al. Biology features of circular RNAs and their research progress in plants ［J］. Acta Botanica Boreali‐Occidentalia Sinica，2017，37（12）：2510‐2518.

［13］周凤燕，杨青，朱熙春，等. 环状RNA的分子特征、作用机制及生物学功能［J］. 农业生物技术学报，2017，25 （3）：485‐501.

ZHOU F Y，YANG Q，ZHU X C，et al. Molecular feature，action mechanism and biology function of circular RNA ［J］. Journal of Agricultural Biotechnology，2017，25（3）：485‐501.

［14］PATOP I L，WÜST S，KADENER S. Past，present，and future of circRNAs［J］. The EMBO Journal，2019，38（16）：e100836.

［15］LI Z Y，HUANG C，BAO C，et al. Exon‐intron circular RNAs regulate transcription in the nucleus［J］. Nature Structural & Molecular Biology，2015，22（3）：256‐264.

［16］CHEN L L. The biogenesis and emerging roles of circular RNAs［J］.Nature Reviews Molecular Cell Biology，2016，17（4）：205‐211.

［17］ASHWAL‐FLUSS R，MEYER M，PAMUDURTI N R，et al. circRNA biogenesis competes with pre‐mRNA splicing［J］. Molecular Cell，2014，56（1）：55‐66.

［18］EBBESEN K K，KJEMS J，HANSEN T B. Circular RNAs：Identification，biogenesis and function［J］.Biochimica et Biophysica Acta （BBA） ‐ Gene Regulatory Mechanisms，2016，1859（1）：163‐168.

［19］ZUO J H，WANG Q，ZHU B Z，et al. Deciphering the roles of circRNAs on chilling injury in tomato［J］.Biochemical and Biophysical Research Communications，2016，479（2）：132‐138.

［20］GAO Z，LI J，LUO M，et al. Characterization and cloning of grape circular RNAs identified the cold resistance‐related Vv‑circATS1［J］. Plant Physiology，2019，180（2）：966‐985.

［21］CHENG J P，ZHANG Y，LI Z W，et al. A lariat‐derived circular RNA is required for plant development in Arabidopsis［J］. Science China Life Sciences，2018，61 （2）：204‐213.

［22］SONG Y P，BU C H，CHEN P F，et al. Miniature inverted repeat transposable elements cis‐regulate circular RNA expression and promote ethylene biosynthesis，reducing heat tolerance in Populus tomentosa［J］.Journal of Experimental Botany，2021，72 （5）：1978‐1994.

［23］CHEN L，ZHANG P，FAN Y，et al. Circular RNAs mediated by transposons are associated with transcriptomic and phenotypic variation in maize［J］.New Phytologist，2018，217（3）：1292‐1306.

［24］XU J，WANG Q，TANG X，et al. Drought‐induced circular RNAs in maize roots：Separating signal from noise［J］. Plant Physiology，2024，196（1）：352‐367.

［25］ZUO J H，WANG Y X，ZHU B Z，et al. Analysis of the coding and non‐coding RNA transcriptomes in response to bell pepper chilling［J］. International Journal of Molecular Sciences，2018，19（7）：2001.

［26］HUANG J，WANG Y L，YU J，et al. Evolutionary landscape of tea circular RNAs and its contribution to chilling tolerance of tea plant［J］. International Journal of Molecular Sciences，2023，24（2）：1478.

［27］ PAN T，SUN X Q，LIU Y X，et al. Heat stress alters genome‐wide profiles of circular RNAs in Arabidopsis［J］. Plant Molecular Biology，2018，96：217‐229.

［28］WANG X S，CHANG X C，JING Y，et al. Identification and functional prediction of soybean circRNAs involved in low‐temperature responses［J］. Journal of Plant Physiology，2020，250：153188.

［29］FANG F，YE S W，TANG J Y，et al. DWT1/DWL2 act together with OsPIP5K1 to regulate plant uniform growth in rice［J］. New Phytologist，2020，225（3）：1234‐1246.

［30］HAO C，YANG Y Z，DU J M，et al. The PCY‐SAG14 phytocyanin module regulated by PIFs and miR408 promotes dark‐induced leaf senescence in Arabidopsis［J］. Proceedings of the National Academy of Sciences of the United States of America，2022，119（3）：e2116623119.

［31］XU Z P， GAO Y H， GAO C X， et al.Glycosylphosphatidylinositol anchor lipid remodeling directs proteins to the plasma membrane and governs cell wall mechanics［J］. The Plant Cell，2022，34（12）：4778‐4794.

［32］张敏敏. 植物糖基化磷脂酰肌醇锚定蛋白协助受体激酶转运的机制研究［D］. 上海：华东师范大学，2019.
ZHANG M M. Study on the mechanism of plant glycosylated phosphatidylinositol ankyrin assisting receptor kinase transport［D］. Shanghai：East China Normal University，2019.

［33］杨春霞，胥猛，王明庥，等. 植物中miR160/miR167/miR390家族及其靶基因研究进展［J］.南京林业大学学报，2014，38（3）：155‐159.
YANG C X，XU M，WANG M X，et al. Advance on miR160/miR167/miR390 family and its target genes in plants［J］. Journal of Nanjing Forestry University，2014，38（3）：155‐159.

［34］赵思航，刘昊东，徐渴，等. 小麦中MIR160基因家族的生物信息学分析及靶基因鉴定［J］. 分子植物育种，2023，21（1）：27‐35.
ZHAO S H，LIU H D，XU K，et al. Bioinformatics analysis and target gene identification of MIR160 gene family in wheat［J］. Molecular Plant Breeding，2023，21 （1）：27‐35.

［35］ŞANL B A，ÖZTÜRK GÖKÇE Z N. Investigating effect of miR160 through overexpression in potato cultivars under single or combination of heat and drought stresses［J］. Plant Biotechnology Reports，2021，15（3）：335‐348.

［36］CHUCK G，MARK CIGAN A，SAETEURN K，et al.The heterochronic maize mutant Corngrass1 results from overexpression of a tandem microRNA［J］. Nature Genetics，2007，39（4）：544‐549.

［37］王多佳，王政委，任志鹏，等. 冬小麦miR172响应抗寒的特征分析［J］. 麦类作物学报，2023，43（10）：1304‐1310.
WANG D J，WANG Z W，REN Z P，et al. Analysis on the characteristics of miR172 responding to cold resistance in winter wheat［J］.Journal of Triticeae Crops，2023，43（10）：1304‐1310.

［38］ZHANG Y Q，WASEEM M，ZENG Z H，et al.microRNA482/2118，a miRNA superfamily essential for both disease resistance and plant development［J］.New Phytologist，2022，233（5）：2047‐2057.

［39］莫显兰，史列琴，陆秋利，等. Sl-miR482在番茄果实中的表达分析及STTM沉默载体的构建［J］.生物技术通报，2019，35（12）：50‐56.
MO X L，SHI L Q，LU Q L，et al. Expression analysis of Sl‑miR482 in tomato fruit and the construction of STTM silencing vector［J］. Biotechnology Bulletin，2019，35（12）：50‐56.

［40］姜孝成，田建红，黄科瑞，等. miR164家族及其调控种子发育与种子活力的研究进展［J］. 生命科学研究，2024，28（6）：471‐502.
JIANG X C，TIAN J H，HUANG K R，et al. Advances on the miR164 family and its roles in regulating seed development and vigor ［J］. Life Science Research，2024，28（6）：471‐502.

［41］牟桂萍，纪春艳，许东林，等. 植物miR164家族研究进展［J］.生命科学，2013，25（5）：525‐531.
MOU G P，JI C Y，XU D L，et al. Advances in plant miR164 family［J］. Chinese Bulletin of Life Sciences，2013，25（5）：525‐531.

［42］张好军. miR164调控玉米籽粒发育的信号通路解析［D］. 雅安：四川农业大学，2019.

ZHANG H J. Signal pathway analysis of miR164 regulating maize grain development［D］. Yaan：Sichuan Agricultural University，2019.

［43］LUAN M D，XU X Y，LU Y M，et al. Family‐wide survey of miR169s and NF‐YAs and Their expression profiles response to abiotic stress in maize roots［J］.PLoS One，2014，9（3）：e91369.

［44］RAGUPATHY R，RAVICHANDRAN S，MAHDI M S R， et al. Deep sequencing of wheat sRNA transcriptome reveals distinct temporal expression pattern of miRNAs in response to heat，light and UV［J］.Scientific Reports，2016，6：39373.

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