耐弱光马铃薯品种筛选及转录因子编码基因StPIF3的克隆与功能分析

doi:10.3724/SP.J.1006.2026.54135

作物学报 ›› 2026, Vol. 52 ›› Issue (6): 1631-1645.doi: 10.3724/SP.J.1006.2026.54135

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

耐弱光马铃薯品种筛选及转录因子编码基因StPIF3的克隆与功能分析

王文辕¹(), 燕雪嘉¹, 刘玉霖¹, 孙晓彤¹, 李亚楠¹, 唐鑫华¹^,^2,*(), 石瑛¹^,^2,*()

¹ 东北农业大学农学院, 黑龙江哈尔滨 150030
² 寒地粮食作物种质创新与生理生态教育部重点实验室, 黑龙江哈尔滨 150030

收稿日期:2025-10-28 接受日期:2026-03-16 出版日期:2026-06-12 网络出版日期:2026-03-27
通讯作者: * 石瑛, E-mail: shiyingneau@sina.com;唐鑫华, E-mail: tangxinhua821@sina.com
作者简介:王文辕, E-mail: 2465378925@qq.com
基金资助:
财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-09)

Screening of low-light-tolerant potato varieties and cloning and functional analysis of the transcription factor gene StPIF3

Wang Wen-Yuan¹(), Yan Xue-Jia¹, Liu Yu-Lin¹, Sun Xiao-Tong¹, Li Ya-Nan¹, Tang Xin-Hua¹^,^2,*(), Shi Ying¹^,^2,*()

¹ College of Agriculture, Northeast Agricultural University, Harbin 150030, Heilongjiang, China
² Key Laboratory of Germplasm Innovation and Physio-Ecology of Food Crops in Cold Regions, Ministry of Education, Harbin 150030, Heilongjiang, China

Received:2025-10-28 Accepted:2026-03-16 Published:2026-06-12 Published online:2026-03-27
Contact: * Shi Ying, E-mail: shiyingneau@sina.com; Tang Xin-Hua, E-mail: tangxinhua821@sina.com
Supported by:
China Agriculture Research System of MOF and MARA(CARS-09)

摘要/Abstract

摘要：

根据9个马铃薯品种弱光处理下生长指标及荧光参数指标的测定比较, 经主成分分析(principle component analysis, PCA)筛选出马铃薯品种“大西洋”为耐弱光品种。为研究马铃薯耐弱光品种“大西洋”在弱光逆境下响应的相关基因, 对10 μmol m^-2 s^-1光强下处理48 h的“大西洋”进行转录组学分析, 通过GO富集、KEGG通路分析及转录因子预测, 筛选出马铃薯光响应相关转录因子编码基因StPIF3, 并进行全长克隆。使用农杆菌介导法对烟草进行遗传转化, 自交获得T₁代转基因烟草株系。采用生物信息学、RT-qPCR、荧光参数测定对其功能及性质进行分析验证。生物信息学分析表明, StPIF3蛋白由716个氨基酸组成, 分子质量、等电点分别为76.77 kD、7.26, 为亲水性不稳定蛋白。StPIF3蛋白具有典型HLH结构域, 属于bHLH家族成员, 进化树显示StPIF3与烟草的亲缘关系最近。在50 μmol m^-2 s^-1光强下, 转基因烟草的F_v/F_m、q_P、ETR、ETRmax、SPAD均显著高于野生型烟草, F₀低于野生型烟草。综上所述, StPIF3过量表达会显著提高电子传递效率和光能利用效率, 进而提高植物光合性能与弱光耐性。

关键词: 马铃薯, 弱光耐性, StPIF3转录因子, bHLH, 过表达

Abstract:

Based on measurements and comparisons of growth traits, chlorophyll fluorescence parameters, and other related indices in nine potato varieties under low-light treatment, the cultivar “Atlantic” was identified as low-light tolerant by principal component analysis. To investigate genes involved in the low-light response of “Atlantic”, we performed transcriptome analysis after treatment with a light intensity of 10 μmol m^-2 s^-1 for 48 h. Using GO enrichment, KEGG pathway analysis, and transcription factor prediction, we identified the light-responsive transcription factor gene StPIF3 and cloned its full-length cDNA. StPIF3 was introduced into tobacco via agrobacterium-mediated transformation, and T₁ transgenic lines were obtained by selfing. Bioinformatics analysis, RT-qPCR, and chlorophyll fluorescence measurements were used to characterize and validate its function. StPIF3 encodes a 716-amino-acid protein with a predicted molecular weight of 76.77 kD and an isoelectric point of 7.26, and it is predicted to be hydrophilic and unstable. The StPIF3 protein contains a typical HLH domain and belongs to the bHLH family; phylogenetic analysis indicated that StPIF3 is most closely related to the tobacco homolog. Under 50 μmol m^-2 s^-1 light intensity, transgenic tobacco plants showed significantly higher F_v/F_m, q_P, ETR, ETRmax, and SPAD values than wild-type plants, whereas F₀ was lower. Overall, overexpression of StPIF3 markedly improves electron transport and light energy use efficiency, thereby enhancing photosynthetic performance and low-light tolerance.

Key words: Solanum tuberosum, weak light tolerance, transcription factors StPIF3, bHLH, overexpression

王文辕, 燕雪嘉, 刘玉霖, 孙晓彤, 李亚楠, 唐鑫华, 石瑛. 耐弱光马铃薯品种筛选及转录因子编码基因StPIF3的克隆与功能分析[J]. 作物学报, 2026, 52(6): 1631-1645.

Wang Wen-Yuan, Yan Xue-Jia, Liu Yu-Lin, Sun Xiao-Tong, Li Ya-Nan, Tang Xin-Hua, Shi Ying. Screening of low-light-tolerant potato varieties and cloning and functional analysis of the transcription factor gene StPIF3[J]. Acta Agronomica Sinica, 2026, 52(6): 1631-1645.

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链接本文: https://zwxb.chinacrops.org/CN/10.3724/SP.J.1006.2026.54135

https://zwxb.chinacrops.org/CN/Y2026/V52/I6/1631

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名称 Name	正向引物序列 Forward primer sequence (5′-3′)	反向引物序列 Reverse primer sequence (5′-3′)	用途 Purpose
StPIF3T	gagaacacgggggactctagaATGCCTCTCTCTGAGTTCTTGAAAA	catggatcctctgtttctagaTCAGTTGGTCAACCTATTCG	载体连接 Vector ligation
StPIF3q	AAAGGAGGAGAAAAAGTGGC	GAACATACAGATGAGCAAGC	荧光定量PCR RT-qPCR
MYC4	ATAAGAACAAGCGTAGAGCGTCGTC	TTCTCTGTTCCAGTAACCCAAATCG
TIFY/10A	AGCAAGGGAAACAACACACATAAAC	AGTTTTCACTGCAAATTGACCACCC
TIFY/10B	AATTTGTGTGACGGCCGCAGG	TGTTACTTGTCAAAGAAGCAGGGGC
BER12	GGATCGTTGAAGATGATGGCACCAC	CTAGGGAATGACGATGATGTTGGGC
Aux-RF4	AAGACTGTTTCCCCCC	CTCCACCTTCTCCCCT
scarecrow-like 8	CGATGAGTGGTCGGAAAC	CTTGAAACACGGGGATAT
Aux/IAA16-like	GTTTGGTAAATGGAGGGC	AGCAGAAGCAACGGTGAG
Actin	GATGTTGTGCCAAAGGATGT	AACTTGGTCAATGCGAGA

培养时期 Culture period	培养基主要成分 Main component of medium	培养条件 Culture condition
菌体活化 Bacterial activation	LB+50 mg L^-1 Kan+50 mg L^-1 Rif	28℃, 2 days
菌体重悬 Bacterial resuspension	4.4 g L^-1 MS+30 g L^-1蔗糖(pH = 6.0) 4.4 g L^-1 MS+30 g L^-1 sucrose (pH = 6.0)	在超净台内 Inside the laminar flow hood
共培养 Co-cultivation	4.4 g L^-1 MS+30 g L^-1蔗糖+7 g L^-1琼脂+1 mg L^-1 6-BA+0.1 mg L^-1 NAA (pH = 6.0) 4.4 g L^-1 MS+30 g L^-1 sucrose+7 g L^-1 agar+1 mg L^-1 6-BA+0.1 mg L^-1 NAA (pH = 6.0)	23℃, 暗培养, 2 d 23℃, dark culture, 2 days
选择培养 Selective cultivation	4.4 g L^-1 MS+30 g L^-1蔗糖+7 g L^-1琼脂+1 mg L^-1 6-BA+0.1 mg L^-1 NAA+300 mg L^-1 cef+5 mg L^-1 PPT (pH = 6.0) 4.4 g L^-1 MS+30 g L^-1 sucrose+7 g L^-1 agar+1 mg L^-1 6-BA+0.1 mg L^-1 NAA+300 mg L^-1 cef+5 mg L^-1 PPT (pH = 6.0)	23℃, 16 h光照/8 h黑暗, 4周 23℃, 16 h light/8 h dark, 4 weeks
生根培养 Rooting cultivation	4.4 g L^-1 MS+30 g L^-1蔗糖+7 g L^-1琼脂+300 mg L^-1 cef (pH = 6.0) 4.4 g L^-1 MS+30 g L^-1 sucrose+7 g L^-1 agar+300 mg L^-1 cef (pH = 6.0)	至出苗到高3 cm左右 Until the seedlings grow to approximately 3 cm in height

品种名称 Varieties name	基于形态指标 Based on morphological indicators		得分 Score	基于荧光参数 Based on fluorescence parameters		得分 Score
品种名称 Varieties name	主成分1 Principle component 1	主成分2 Principle component 2	得分 Score	主成分1 Principle component 1	主成分2 Principle component 2	得分 Score
东农310 Dongnong 310	-1.12	-0.23	-0.92	-0.01	2.04	0.35
大西洋 Atlantic	1.52	-0.54	1.13	1.54	0.59	1.21
兴佳2 Xingjia 2	-0.92	0.60	-0.64	-1.42	-0.31	-1.08
克新13 Kexin 13	-0.55	-0.29	-0.48	-0.26	-1.32	-0.42
克新19 Kexin 19	-0.74	-0.74	-0.69	-0.05	-0.58	-0.13
东农311 Dongnong 311	1.46	-0.20	1.12	0.73	-0.89	0.37
东农303 Dongnong 303	0.22	-0.90	0.05	0.89	-0.39	0.58
延薯4 Yanshu 4	0.36	2.41	0.63	-1.48	0.50	-0.98
尤金 Youjin	-0.35	-0.12	-0.29	0.05	0.36	0.10
特征值Eigenvalue	3.17	0.58		2.86	0.68
累计贡献率 Cumulative contribution rate (%)	79.34	93.70		71.90	88.91

样本 Sample	高质量读段 Clean reads	高质量碱基 Clean bases	GC含量 GC content (%)	Q30以上碱基比率 ≥ Q30 (%)
CK1	24,085,504	7,204,888,144	43.02	94.43
CK2	25,404,108	7,597,106,052	42.94	94.45
CK3	24,370,569	7,292,240,504	42.68	94.47
L241	24,360,702	7,282,788,098	42.88	94.57
L242	23,705,884	7,094,796,702	43.15	94.48
L243	25,114,630	7,513,499,908	42.83	94.59
L481	22,923,926	6,856,284,542	42.58	94.59
L482	23,873,572	7,137,387,592	42.92	95.05
L483	24,583,906	7,348,163,688	42.80	94.86

基因ID Gene ID	趋势 Trend	表达倍数 log₂(FC)
LOC102577631	下降Down	-0.755650532
LOC102604891	上升Up	1.360512855
LOC102582060	下降Down	-0.750062461
LOC102601108	下降Down	-0.674340874
LOC102592798	下降Down	-1.048333044
LOC102593308	下降Down	-0.953968101
LOC102601000	下降Down	-0.682094614
LOC102590668	下降Down	-0.644804900
LOC102592910	下降Down	-0.838504449
LOC102601372	下降Down	-1.392340176
LOC102597649	下降Down	-1.414059764
LOC102599201	下降Down	-0.685843864
LOC102592864	下降Down	-0.410146215
LOC102602099	下降Down	-0.884948358

耐弱光马铃薯品种筛选及转录因子编码基因StPIF3的克隆与功能分析

Screening of low-light-tolerant potato varieties and cloning and functional analysis of the transcription factor gene StPIF3

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基因ID Gene ID	家族 Family	类别 Type	表达倍数 log₂(FC)	趋势 Trend
LOC102577631	AUX/IAA	TR	-0.755650532	下降Down
LOC102604891	bHLH	TF	1.360512855	上升Up
LOC102582060	BES1	TF	-0.750062461	下降Down
LOC102592798	BES1	TF	-1.048333044	下降Down
LOC102601000	GARS	TF	-0.682094614	下降Down
LOC102590668	AUX/IAA	TR	-0.644804900	下降Down
LOC102592910	Tify	TF	-0.838504449	下降Down
LOC102601372	bHLH	TF	-1.392340176	下降Down
LOC102577631	AUX/IAA	TR	-0.755650532	下降Down

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