PtNF-Ys基因在油松花发育早期的表达与功能分析

doi:10.7525/j.issn.1673-5102.2026.02.001

摘要/Abstract

摘要：

为解析油松（Pinus tabuliformis）种子园童期长、产量低的成因并探索其调控机制，以山西吴城国家油松种子园成年油松为材料，对7—10月中旬的顶端分生组织进行转录组测序，研究花芽发育早期阶段开花相关基因的表达特征，解析核因子Y（NF-Y）家族成员的生物学功能。结果表明：PtNF-YB8等多个NF-Y家族基因在10月中旬表达量显著升高。启动子分析显示，PtNF-YB8基因含有赤霉素（GA）响应、光响应等顺式作用元件；对顶芽进行GA处理后PtNF-YB8基因表达量显著升高。在本氏烟草（Nicotiana benthamiana）细胞中，PtNF-YB8蛋白定位于细胞质和细胞核中，其转基因拟南芥（Arabidopsis thaliana）株系呈现晚花表型。酵母双杂交结果证明，PtNF-YB8蛋白能够与PtNF-YC9和PtNF-YC10蛋白互作。综上，PtNF-YB8蛋白可能与PtNF-YC9/10等NF-Y家族成员结合并进行信号传递，发挥延迟花期的作用。研究结果为进一步解析针叶树开花调控网络及研发种子园丰产稳产技术提供依据。

关键词: 油松, 花芽发育, 核因子家族, 赤霉素

Abstract:

To elucidate the causes of the long juvenile phase and low seed yield in Pinus tabuliformis seed orchards and explore the underlying regulatory mechanisms， this study focused on the flowering-related genes during early stages of flower bud differentiation. The aim was to reveal the gene expression characteristics during this period and clarify the biological functions of the nuclear factor Y（NF-Y） family members. Transcriptome sequencing（RNA-seq） was performed on apical meristems collected from adult trees in the Wucheng National Seed Orchard of P. tabuliformis， Shanxi， China， from mid-July to mid-October. The results revealed that the expression levels of PtNF-YB8 and many other NF-Y genes increased significantly in mid-October. Multiple cis-elements， including gibberellin（GA）-responsive and light-responsive elements， were found within the PtNF-YB8 promoter. Furthermore， exogenous GA treatment of apical buds significantly up-regulated the expression level of PtNF-YB8. Subcellular localization assays in tobacco cells showed that the PtNF-YB8 protein was localized in both the cytoplasm and the nucleus. Heterologous overexpression of PtNF-YB8 in Arabidopsis resulted in a distinct delayed flowering phenotype. Yeast two-hybrid assays confirmed that PtNF-YB8 interacts with PtNF-YC9 and PtNF-YC10. In conclusion， PtNF-YB8 may function by forming complexes with NF-Y members， such as PtNF-YC9/10， to facilitate signal transduction and negatively regulate flowering time. These findings provide a theoretical basis for elucidating the flowering regulatory network in conifers and developing techniques for high and stable yields in seed orchards.

Key words: Pinus tabuliformis, floral bud development, nuclear factor Y family, gibberellin

中图分类号:

Q945

丁文彬, 胡岳涵, 白寅甲, 杨宇寒, 薛华. PtNF-Ys基因在油松花发育早期的表达与功能分析[J]. 植物研究, 2026, 46(2): 195-208.

Wenbin DING, Yuehan HU, Yinjia BAI, Yuhan YANG, Hua XUE. Expression Characteristics and Functional Analysis of PtNF-Ys Gene during Early Flower Development in Pinus tabuliformis[J]. Bulletin of Botanical Research, 2026, 46(2): 195-208.

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基因名称 Gene name	正向引物序列（5′→3′） Forward primer sequence （5′→3′）	反向引物序列（5′→3′） Reverse primer sequence（5′→3′）
RqPtNF-YB8	ATGGCAGACTTAGCGAGCTCCG	GAGGACTTTCTTGGCTGGTGA
RqPtNF-YC1	ATGGACCACCACAACCACC	CCTGCTGCCCTCCACTCT
RqPtTFL2	ATGGCCAGGTTCAGAGAGCCT	ACATCGCCAATGACTCTTCCT
RqPtCOL1	ATGGTGAAGGAGGAGGATTGT	CCTTCACGATTCCTGCCTC
PtNF-YB8	ATGGCAGACTTAGCGAGCTCC	CCTAATATTTGGTAATGCTCCAAAA
PtNF-YB8-AD	CCATGGAGGCCAGTGAATTCATGGCAGACTTAGC GAGCTCC	CCATGGAGGCCAGTGAATTCCCTAATATTTGGTAATG CTCCAAAA
PtNF-YC4-BD	TGGCCATGGAGGCCGAATTCATGCATGATTTAGAA CAGACATCAGAC	CGCTGCAGGTCGACGGATCCCTAACTGCTAGAGGAT GGGCC
PtNF-YC10-BD	TGGCCATGGAGGCCGAATTCATGCATGATATAGAA CAGAC	CGCTGCAGGTCGACGGATCCCTATGCTGCTGGATGC
PtNF-YC9-BD	TGGCCATGGAGGCCGAATTCATGAGGAAGAAGCT CGATACCC	CGCTGCAGGTCGACGGATCCTATCCATCATCATTGTC ATAGTCATCCTC
PtNF-YA3-BD	TGGCCATGGAGGCCGAATTCATGACCGTTCGGGA ATTTTTGC	CGCTGCAGGTCGACGGATCCTCACTGGATCACAACA ACTCGT
PtNF-YA9-BD	TGGCCATGGAGGCCGAATTCATGGAGAACACGCC TACACATAC	CGCTGCAGGTCGACGGATCCCTAGGAATCCGTGTTC TGCTGT
PtNF-YB8-1300	AACACGGGGGACGAGCTCGGTACCATGGCAGAC TTAGCGAGCTCC	TTCTTCTCCTTTGCCCATGTCGACTCTAGACCTAATAT TTGGTAATGCTCCAAAA
PtActin	GGCTGACACCATCACCAGAATC	GTTGGTCGCCCTCGTCATACT
AtActin	CGTATGAGCAAAGAGATCAC	CACATCTGTTGGAAGGTGCT

序号 Number	序列 Sequence	数量 Quantity	位置 Location	顺式作用元件 Cis-acting regulatory element
1	TTACTTAA	1	342	Part of a light responsive element
2	GCCACT	2	420；529	Related to meristem expression
3	CCATCTTTTT	2	1 458；1 550	Involved in salicylic acid responsiveness
4	CACGTC	1	1 314	Involved in light responsiveness
5	CAAAGATATC	1	1 265	Involved in circadian control
6	TGAGTCA	2	752；943	Involved in endosperm expression
7	ACGTG	2	1 315；1 996	Involved in the abscisic acid responsiveness
8	ATGATAAGGTC	2	648；839	Part of a light responsive element
9	AAGATAAGGCT	1	1 217	Part of a light responsive element
10	TTCCAACAAACCCC	1	1 201	Gibberellin-responsive element and part of a light responsive element