玉米花粉中响应高温胁迫circRNA 的筛选及其功能初探

doi:10.15933/j.cnki.1004-3268.2026.01.003

摘要/Abstract

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

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

中图分类号:

S513

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.

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.

图/表 7

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