Objective: To identify the miRNAs that post-transcriptionally target the regulation of egfr in the antitumor mechanism of Cordyceps militaris. Methods: An aqueous extract of Cordyceps militaris was prepared using water-bath extraction combined with vacuum freeze-drying. Transcriptome sequencing was performed on A549 cells after 24 h treatment with 4 mg/mL Cordyceps militaris extract. Candidate miRNAs were selected on the basis of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Functional gain models were constructed by transient transfection of miRNA mimics, and the effects of miRNAs on cell viability were assessed using the MTT assay and Annexin V-FITC/PI double staining. Functional loss models were established by transient transfection of miRNA inhibitors, in combination with Cordyceps militaris extract treatment. The effect of Cordyceps-miRNA axis on cell viability was evaluated using the MTT assay, and its regulatory effects on EGFR transcription and protein levels in A549 cells were analyzed using qRT-PCR and immunocytochemical staining. Results: A total of 7211 DEGs were obtained and significantly enriched in the "MicroRNAs in cancer" signaling pathway (Sort 3). Eleven miRNAs were screened for their association with lung adenocarcinoma prognosis, miR-125a-3p, miR-125a-5p, miR-125b-1-3p, miR-125b-2-3p, miR-125b-5p and miR-145-5p were all associated with egfr in a target-regulatory manner, and patients with lung adenocarcinomas that showed low expression levels had a poorer prognosis (P<0.05). Cordyceps militaris treatment induced significant upregulation of miR-125b-1-3p, miR-125b-5p and miR-145-5p expression levels (P<0.01), and the seed sequences had potential binding sites with the egfr 3'-UTR region. miR-125b-1-3p, miR-125b-5p, and miR-145-5p exhibited tumor-specific pro-proliferative activities, and their high expression status induced a higher percentage of early apoptosis in A549 cells. Cordyceps militaris inhibited the proliferation of lung cancer A549 cells by promoting the expression of miR-125b-1-3p, miR-125b-5p, and miR-145-5p (P<0.05) and downregulating the expression of EGFR (P<0.05). Conclusion: Cordyceps militaris exerts anti-tumor activity by targeting and regulating the miRNAs (miR-125b-1-3p, miR-125b-5p and miR-145-5p)-EGFR axis, providing theoretical support for its development as a novel functional food for adjuvant tumor therapy.

Abstract Plant height is an important agronomic trait in rice and shows marked divergence between wild and cultivated germplasm. To identify loci associated with plant height variation from wild rice, we developed a BC 4 F 7 chromosome segment substitution line population of 308 lines derived from Yuanjiang common wild rice ( Oryza rufipogon ) and elite cultivar Youzhan 8 ( Oryza sativa ). Bulked segregant analysis coupled with sequencing identified a 13.6‐Mb quantitative trait loci region on chromosome 7 associated with plant height. Eight prioritized candidate genes were identified within this interval based on sequence polymorphism and functional annotation, including known regulators OsTCP21 and PROG1 . Quantitative RT‐PCR analysis of stem tissues at the jointing stage detected measurable expression for five of these genes. Four genes, OsTCP21 , OsFBL35 , OsFBA2 , and LOC_Os07g08190 , showed significant differential expression between tall and dwarf lines, whereas GID1L2 did not. The other three genes, including PROG1 , showed transcript levels too low for reliable detection. Haplotype analysis further identified coding and promoter variation in OsFBL35 , OsFBA2 , LOC_Os07g08190 , and GID1L2 , as well as promoter haplotypes in OsTCP21 . These results suggest that OsTCP21, OsFBL35 , OsFBA2 , LOC_Os07g08190 , and GID1L2 are potential candidate genes for plant height variation in this wild rice‐derived population. Functional annotation suggested that these candidates may be related to ubiquitination, hormone signaling, and growth regulation. These findings provide insight into the genetic basis of plant height variation derived from wild rice and offer candidate loci and molecular markers for rice improvement.

Rice (Oryza sativa L.) is a major staple crop that feeds nearly half of the global population. However, it is highly sensitive to low temperatures, which limits its cultivation and yield stability in high-latitude and high-altitude regions. Low-temperature stress suppresses growth, disrupts physiological processes, reduces yield, and in severe cases causes plant death, posing a serious threat to food security. With climate change, the frequency and intensity of low-temperature stress are expected to increase further. This review summarizes recent progress in understanding rice cold tolerance, emphasizing physiological responses, molecular and genetic regulatory mechanisms, and insights from omics studies. We also explored the natural variation in cold tolerance and its potential for breeding, and highlight current challenges and future directions in the context of climate change. This review aims to provide scientific insights aiming the breeding of cold-tolerant rice varieties and to enhance the protection of global food security.

Objective: From the perspective of the tumor microenvironment, this study explored the anti-tumor mechanisms of Ganoderma lucidum, Phellinus linteus, and Cordyceps militaris. Methods: Aqueous extracts (FAEs) of Ganoderma lucidum (RL), Phellinus linteus (BJ), and Cordyceps militaris (XG) were prepared via water bath extraction and vacuum freeze-drying. The effects of the FAEs and their conditioned media (CM) on the proliferative activity of A549 non-small cell lung cancer cells were determined using the MTT assay and Annexin V-FITC/PI double staining. Transcriptome sequencing was performed on cells treated with 4 mg/mL FAE, and KEGG pathway enrichment analysis was used to identify the response characteristics of the IL-17 signaling pathway to each extract. Additionally, immunocytochemical staining, antibody blocking assays, and ELISA were used to detect the effects of the FAEs on the expression levels of the mucins MUC5AC and MUC5B and the chemokines IL-6, CCL2, CCL20, CXCL8, and CXCL10. Results: The three FAEs and CM inhibited the proliferation of the A549 cells and induced their apoptosis. Transcriptome analysis results revealed that upregulated genes were predominant in the RL and BJ groups, whereas downregulated genes were predominant in the XG group. All groups showed significant enrichment in signaling pathways related to tumor microenvironment formation. Additionally, significant alterations were observed in the expression levels of key effector molecules downstream of the IL-17 signaling pathway (ko04657) in all groups. All three FAEs significantly inhibited MUC5B expression (P<0.01). However, BJ treatment did not significantly affect MUC5AC expression (P>0.05). MUC5AC antibody blocking weakened the inhibitory effects of RL and XG on cell proliferation (P<0.05), whereas MUC5B antibody blocking significantly reduced the anti-proliferative effects of RL and BJ on the cells (P<0.05). Furthermore, all three FAEs significantly inhibited the secretion of IL-6, CCL2, CCL20, and CXCL8 (P<0.01). Conclusion: In conclusion, Ganoderma lucidum, Phellinus linteus, and Cordyceps militaris can all remodel the tumor microenvironment to synergistically exert anti-tumor effects. This provides a new theoretical basis for their development as functional foods for adjuvant cancer therapy.

DNA demethylation is an essential epigenetic mechanism responsible for regulating the ripening and quality of fruit, yet its role in postharvest ethylene and volatile formation in oriental melon (Cucumis melo var. Makuwa) remains largely unexplored. This study combined multi-omics analysis with epigenetic intervention to systematically investigate how DNA methylation affects fruit ripening by regulating ethylene and volatile production. The promoter methylation levels of several core genes related to ethylene and aliphatic aromatic compound biosynthesis, including LOX (MELO3C011885 and MELO3C024348), ADH (MELO3C011043), AAT (MELO3C024771 and MELO3C024007), and ACO (MELO3C007425 and MELO3C026436), were negatively correlated with transcriptional activity. DNA methyltransferase-encoding gene (CMT, MET, and DRM) expression declined during postharvest ripening, whereas that of the gene encoding the demethylase (DML) was significantly upregulated on day 7. Treatment with DNA demethylation agent (5-azacytidine, 5-Aza) improved the ester content by 2.1-fold, enhanced lipoxygenase and alcohol dehydrogenase activities by 1.8- and 2.3-fold, and up-regulated ethylene production by up to 1.9-fold. In conclusion, DNA methylation dynamically regulates the expression of lipid metabolism and ethylene biosynthesis pathway genes, thereby significantly influencing the accumulation of aliphatic aromatic compounds and ethylene production in oriental melons. The findings provide insights into epigenetic mechanisms that can be targeted for fruit quality improvement.

Aluminum (Al) stress is an important factor that inhibits crop growth in acidic soils and poses a threat to pumpkin (Cucurbita moschata) production. In this study, we investigated the effect of endophyte (endophyte) strain J01 of blueberry (Vaccinium uliginosum) on the growth, development, and transcriptional regulatory mechanisms of pumpkin under aluminum stress. The results showed that the blueberry endophyte strain J01 significantly increased the root length of pumpkin under aluminum stress, promoted the growth of lateral roots, and increased root vigor; strain J01 reduced the content of MDA and the relative conductivity in the root system; strain J01 enhanced the activities of superoxide dismutase and catalase in the root system but inhibited ascorbate peroxidase activity. Transcriptome analysis further revealed that strain J01 significantly regulated the expression of key genes associated with aluminum tolerance, including the upregulation of transporter protein genes (aluminum-activated malate transporter and aquaporin), affecting the gene expression levels of genes encoding antioxidant enzymes (ascorbate peroxidase and glutathione S-transferase) and cell wall modification genes (xyloglucan endotransglucosylase/hydrolase and pectin methylesterase). This study provides a theoretical basis and practical guidance for using microbial resources to improve aluminum tolerance in cucurbit crops.

Methyl jasmonate (MeJA) has diverse effects on the preservation and quality of postharvest fruits. However, it remains unclear whether these effects are mediated by miRNAs and mRNAs. The present study aimed to provide insights into the effect of MeJA on peel carotenoid content in "Huaguniang" oriental melon fruit (Cucumis melo L.) by molecular analysis. Transcriptome and degradome analyses revealed 3571 mRNAs and 16 miRNAs of differentially expressed genes (DEGs) induced by MeJA. A weighted gene co-expression network analysis (WGCNA) divided the DEGs into 7 modules. WGCNA revealed that the "darkturquoise" module exhibited the strongest correlations with 4 major carotenoids. A transcriptional regulatory network was constructed using CmPYS1, a carotenoid-associated differentially expressed gene from this module. Analysis of the CmPYS1 promoter identified NAC-binding motifs, while yeast two-hybrid assays confirmed the interaction between CmPYS1 and CmNAC10 within the module. Dual-luciferase assays in tobacco further demonstrated that MeJA-suppressed miRNA164 interacts with CmNAC10. Collectively, our findings indicate that MeJA stimulates carotenoid accumulation in oriental melon through a regulatory cascade: suppression of miRNA164 expression elevates CmNAC10 levels, which subsequently enhances CmPYS1 expression. This study elucidates the molecular mechanisms through which MeJA regulates carotenoid biosynthesis.

N6-methyladenosine (m6A) methylation functions as a vital post-transcriptional and epigenetic modification in higher plants regulated by α-ketoglutarate-dependent dioxygenases (ALKBH). However, the role of ALKBH genes in oriental melon (Cucumis melo L.) fruit ripening has not been explored. Therefore, we treated oriental melon with an exogenous m6A demethylase inhibitor (mechlorfenamic acid) then analyzed endogenous ethylene production and ripening-related indicators to explore the effects of m6A methylation on ripening. Bioinformatics and real-time quantitative PCR analyses were used to determine the impact of ALKBH genes on key ethylene synthesis gene expression. Treatment effectively inhibited endogenous ethylene production, firmness changes, and soluble solid contents, thereby extending fruit ripening. Eight ALKBH gene family members belonging to five major groups were identified in the melon genome. All members were expressed in ripening fruits, with different expression patterns during ripening. CmALKBH6, CmALKBH7, and CmALKBH8 expression was inhibited by an ethylene inhibitor (1-methylcyclopropene). The transient overexpression (OE) of CmALKBH8 in oriental melon led to the increased expression of the ethylene synthesis genes CmACS1, CmACS2, and CmACO1. In summary, the ethylene-regulated gene CmALKBH8 may participate in oriental melon fruit ripening regulation by modulating the methylation levels of ethylene synthesis-related genes. These findings help us better understand how m6A methylation regulates melon ripening.

Dehydrins (DHNs; late embryogenesis-abundant D11 family) are a class of hydrophilic proteins involved in plant abiotic stress response. However, there is less information regarding DHN gene function in cucurbit crops. Herein, 34 DHN gene family members were identified and characterized in Cucumis sativus, Cucumis melo, Citrullus lanatus, Benincasa hispida, Lagenaria siceraria, and Cucurbita maxima. The DHN genes in the six cucurbit crops exhibited greater collinearity within subfamilies than between different subfamilies. Responses to stress (including low-temperature, salt, cadmium, and aluminum stress) varied among the DHN members, with a significant alteration in the expression of the acidic SnKn-type DHN gene CmDHN3 in response to aluminum stress. Subcellular localization analysis confirmed that CmDHN3 is expressed in the nucleus and cytoplasm. Virus-induced gene silencing (VIGS) revealed a remarkable decrease in CmDHN3 expression, which markedly increased malondialdehyde content, relative conductivity, and proline content in the roots and leaves of plants under aluminum stress. Transcriptome analysis showed that the decreased CmDHN3 expression reduced the expression of water channel protein-encoding genes. Interactions between CmDHN3 and CmAQP1 (MELO3C007188) and between CmDHN3 and CmAQP2 (MELO3C020774) were confirmed using yeast two-hybrid assays. These results clarify the pathway by which dehydrin genes are involved in the transcriptional-level response of melon to aluminum stress and provide a theoretical basis to comprehensively analyze the functions of this gene family in cucurbit crops.

Hybrid rice has made significant contributions to global food security. However, the efficient utilization of landraces to further enhance heterosis remains a key challenge in rice breeding. In this study, we collected a set of 1,392 landraces and 696 hybrid rice parental lines. A total of 770 hybrid combinations were constructed by crossing 517 accessions selected from 2088 rice accessions, and seven key yield-related traits were collected. Nearly 500,000 potential hybrid combinations were predicted, and comprehensive analysis revealed that landraces from South Asia played a significant role in improving multiple traits. Further investigation revealed substantial variation in the landraces' contribution to the optimal hybrid combinations for various characteristics, with landraces particularly contributing to improvements in grain width. We identified 171 quantitative trait loci (QTLs) for seven traits using three association analysis methods. Of these QTLs, 77 known genes were identified within 66 QTLs and 105 novel QTLs without any known genes near them. CRISPR/Cas9-mediated gene editing revealed that OsGRW5.1 plays a crucial role in regulating grain width and grain weight in rice. Furthermore, a strong correlation was observed between the accumulation of advantageous haplotypes and enhanced phenotypic performance. More than 45% of the advantageous haplotypes derived from landraces are likely to play a significant role in future hybrid rice improvement. A predictive platform was developed, which can output all seven phenotypes of potential hybrid combinations with a given genotype of both parents. Collectively, our study provides valuable data and practical insights for enhancing heterosis through the efficient utilization of landraces.

This study is to optimize microwave pretreatment of Acer truncatum seeds to enhance quality and production of Acer truncatum seed oil (ATSO). The optimum microwave power, microwave-treatment time, as well as microwave-treatment temperature were determined through single-factor and orthogonal experiments, and they were 600 W, 5 min, and 50 degrees C, respectively, resulting in an ATSO yield of 35.2 %. The lipid components of ATSO extracted from non-microwave-treated as well as microwave-treated A. truncatum seeds were qualitatively and quantitatively evaluated by lipidomic analysis. Through liquid chromatography-tandem mass spectrometry (LCMS/MS) analysis, 2005 lipid molecules belonging to 42 lipid subclasses were specified. The outcomes of lipidomic analysis demonstrated the microwave pretreatment of A. truncatum seeds not only increased levels of beneficial lipids in ATSO but also changed lipid composition of the oil, potentially enhancing the positive health effects of the oil. This research brings a scientific basis for improving the yield as well as quality of ATSO and helps in understanding lipid composition and health benefits of the oil, promoting the utilization of this valuable natural resource.

The membership of Camelina sativa in the Brassicaceae family has generated substantial interest in its potential as a high-value crop for food, feed, and fuel. Despite the critical regulatory roles played by Thaumatin-like proteins (TLPs) in plant growth and development, their presence in C. sativa has not been previously reported. Through a comprehensive analysis of the C. sativa genome, 34 CsTLP genes have been identified. The chromosomal localization and gene duplication of CsTLPs suggest that the expansion of the gene family was significantly influenced by segmental duplication, while the evolution of CsTLPs was primarily driven by purification selection. Synteny analysis indicates that C. sativa shares a greater number of homologous genes with Arabidopsis thaliana. Additionally, RNA-seq analysis suggests that numerous members of the CsTLPs gene family may potentially participate in abiotic stress. Significantly, the expression of the CsTLP29 gene exhibited a remarkable decrease in response to four distinct stress conditions. The comprehensive examination of the CsTLPs gene family suggests that these genes may have a crucial function in the adaptation of C. sativa to stress and establishes a foundation for further exploration of the TLPs gene family in C. sativa.

Alternative splicing (AS) is the direct cause of different transcripts in eukaryotes and plays a vital role in biological processes such as plant growth and signal transduction. Reproductive development is the key process for rice production. Although numerous studies on AS in plants have been identified, AS of reproductive development stage between japonica and indica rice has not been conducted. In this research, we identified 6994 genes (19.2

低温限制植物生长区域,决定种植范围;导致作物产量和品质下降,严重可能造成植株死亡.为了抵抗低温胁迫,植物进化出了复杂的防御机制.lncRNA是存在于细胞核和细胞质中、由体内基因组转录产生的一类转录本.近年来,已证实lncRNA可以通过多聚腺苷酸化、与某些蛋白酶协同作用、与miRNA竞争性结合等方式来响应植物低温胁迫.本文就lncRNA的定义、来源、分类以及低温条件下lncRNA在模式植物拟南芥和农经作物中的研究进展进行了总结,期望为植物耐低温机理研究及植物耐冷分子育种提供参考.

[目的]研究高粱生长素(IAA)与杂种优势之间的潜在关系,为高粱育种中杂种优势的利用提供重要理论依据.[方法]以11 个亲本及组配的11 个杂交种为材料,采用高效液相色谱技术测定杂交种及其亲本生长素在抽穗期和成熟期的含量,利用实时荧光定量PCR技术比较抽穗期、成熟期高粱不同品种的生长素基因表达的差异.[结果]11 个杂交种成熟期的生长素含量均高于抽穗期,成熟期杂种辽粘 3 号、辽杂 19、辽杂35、辽杂36、辽2297、辽2697、辽5397、辽糯7 号、辽糯11 号、辽夏梁1 号的生长素含量与其亲本相比,子代生长素含量更高,成熟期只有辽粘6 号的生长素含量比其亲本的低.以对应的母本或父本为对照,杂交种辽粘3 号和辽杂19 在抽穗期的生长素基因表达量低于其亲本,杂交种辽粘3 号、辽粘6 号、辽杂36 在成熟期的生长素基因表达量低于其亲本,其余杂交品种在抽穗期、成熟期生长素基因表达量均高于其亲本.[结论]内源生长素是调节高粱生长杂种优势的一个因素,但不是唯一因素.

实验教学体系是生命科学人才培养的重要组成部分，强化创新实践教育与实验教学的有效融合是高校实验教学体系 的重要人才培养任务。本文从高校实验教学中心的信息化建设和开放运行、校内实验教学培养体系的创新性与多元 化、实验教学资源的升级打造与深度共享、实验教学体系优化师资队伍结构优化和健全质量保障体系四个方面，为 高校生物类实验教学体系更好地实现创新应用型人才培养提供改革参考。

BACKGROUND:DNA mutations of diverse types provide the raw material required for phenotypic variation and evolution. In the case of crop species, previous research aimed to elucidate the changing patterns of repetitive sequences, single-nucleotide polymorphisms (SNPs), and small InDels during domestication to explain morphological evolution and adaptation to different environments. Additionally, structural variations (SVs) encompassing larger stretches of DNA are more likely to alter gene expression levels leading to phenotypic variation affecting plant phenotypes and stress resistance. Previous studies on SVs in rice were hampered by reliance on short-read sequencing limiting the quantity and quality of SV identification, while SV data are currently only available for cultivated rice, with wild rice largely uncharacterized. Here, we generated two genome assemblies for O. rufipogon using long-read sequencing and provide insights on the evolutionary pattern and effect of SVs on morphological traits during rice domestication.RESULTS:In this study, we identified 318,589 SVs in cultivated and wild rice populations through a comprehensive analysis of 13 high-quality rice genomes and found that wild rice genomes contain 49% of unique SVs and an average of 1.76% of genes were lost during rice domestication. These SVs were further genotyped for 649 rice accessions, their evolutionary pattern during rice domestication and potential association with the diversity of important agronomic traits were examined. Genome-wide association studies between these SVs and nine agronomic traits identified 413 candidate causal variants, which together affect 361 genes. An 824-bp deletion in japonica rice, which encodes a serine carboxypeptidase family protein, is shown to be associated with grain length.CONCLUSIONS:We provide relatively accurate and complete SV datasets for cultivated and wild rice accessions, especially in TE-rich regions, by comparing long-read sequencing data for 13 representative varieties. The integrated rice SV map and the identified candidate genes and variants represent valuable resources for future genomic research and breeding in rice.

为探讨不同高粱品种碳代谢相关酶的活性和SPS基因相对表达量的差异,以辽粘3号、辽杂11号、辽糯12号、辽杂53号4个高粱品种为试验材料,分别测定苗期、拔节期、抽穗期、成熟期高粱叶片和抽穗期、成熟期高粱籽粒碳代谢相关酶(SS、SPS、SSS、ADPG-ppase、UDPG-ppase)的活性,并比较分析4个高粱品种叶片及籽粒SPS基因表达的差异,探究它们之间的关系,从生理和基因角度选育出具有优良品质的高粱品种.分析结果发现,抽穗期、成熟期辽粘3号高粱叶片的SS、SPS、SSS、UDPG-ppase活性最高,显著高于其他3个高粱品种;成熟期辽粘3号高粱籽粒的SS、SPS、SSS、ADPG-ppase、UDPG-ppase活性也显著高于其他品种;对4个高粱品种的SPS基因表达量进行分析发现,苗期、抽穗期、成熟期辽粘3号高粱叶片的SPS基因表达量均为最高,抽穗期、成熟期高粱籽粒SPS基因表达量表现为辽糯12号>辽粘3号>辽杂53号>辽杂11号.综上所述,辽粘3号高粱品种叶片和籽粒在各个时期的碳代谢相关酶活性比较高,并且苗期、抽穗期、成熟期辽粘3号高粱叶片SPS基因相对表达量也相对较高,辽粘3号高粱籽粒SPS基因相对表达量仅次于辽糯12号,所以辽粘3号高粱品种可作为高粱生产中的首选品种,对筛选高质量的高粱品种具有重要的指导意义.