Identification and Analysis of BrSAMS2 Protein in Winter Brassica rapa Under Low Temperature Stress
LI Bo-Wen1, TAO Xiao-Lei2, ZHANG Yuan-Yuan2, WU Jun-Yan2, WANG Yi-Fan2, SUN Hao2, LI Shi-Yi2, XU Zheng-Nan2, CAI Qian-Yi1, MA Li2,*
1 Gansu Provincial Agro-technology Extension Center, LanZhou 730020, China; 2 College of Agronomy/State Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Lanzhou 730070, China
Abstract:Winter Brassica rapa is an important oilseed crop in China, with strong adaptability, excellent agronomic traits and high production efficiency. Low-temperature adversity ranks among the most critical abiotic constraints throughout its developmental stages. To understand the molecular mechanism of cold resistance of winter B. rapa, in this study, two-dimensional electrophoresis was employed to identify differentially expressed proteins in the shoot apical meristem of winter B. rapa 'Longyou 7' exposed to low-temperature stress. Furthermore, bioinformatics approaches were utilized to perform functional annotation and pathway enrichment analysis on these differentially expressed proteins. The results showed that low-temperature stress induced 24 significantly differentially expressed proteins, among which 8 were up-regulated, 10 were down-regulated, and 6 were induced expression proteins. Go annotation results revealed that the majority of differentially expressed proteins participated in the response to low-temperature stress, antioxidant defense, energy metabolism, and cell structure maintenance processes. Analysis based on the KEGG database uncovered the pivotal roles played by differentially expressed proteins in carbon metabolism, photosynthetic carbon fixation, as well as multiple amino acid biosynthetic pathways. S-adenosylmethionine synthase 2 (SAMS2) gene was associated with low-temperature response and had the highest expression level at 12 h of low-temperature treatment. The encoded protein was a stable hydrophilic protein with a conserved domain, and its promoter region contained cis-acting elements such as low-temperature response. BrSAMS2 might participate in the low-temperature response by regulating methyl donors and ethylene synthesis. This study provides new candidate genes and protein targets for revealing the molecular mechanism of cold tolerance in winter rapeseed.
李博文, 陶肖蕾, 张媛媛, 武军艳, 王一帆, 孙浩, 李诗艺, 徐正南, 蔡芊依, 马骊. 低温胁迫下白菜型冬油菜BrSAMS2蛋白的鉴定与分析[J]. 农业生物技术学报, 2026, 34(7): 1383-1398.
LI Bo-Wen, TAO Xiao-Lei, ZHANG Yuan-Yuan, WU Jun-Yan, WANG Yi-Fan, SUN Hao, LI Shi-Yi, XU Zheng-Nan, CAI Qian-Yi, MA Li. Identification and Analysis of BrSAMS2 Protein in Winter Brassica rapa Under Low Temperature Stress. 农业生物技术学报, 2026, 34(7): 1383-1398.
[1] 陈姣荣, 孙万仓, 方彦, 等. 2012. 白菜型冬油菜在北方寒旱区的适应性分析[J]. 干旱地区农业研究, 30(06): 17-22+31. (Chen J R, Sun W C, Fang Y, et al.2012. Analysis of adaptility of Brassica rape winter cultivars in cold and dry regions of North China[J]. Agricultural Research in the Arid Areas, 30(06): 17-22+31.) [2] 刘自刚, 袁金海, 孙万仓, 等. 2016. 低温胁迫下白菜型冬油菜差异蛋白质组学及光合特性分析[J]. 作物学报, 42(10): 1541-50. (Liu Z G, Yuan J H, Sun W C, et al.2016. Differential proteomic analysis and photosynthetic characteristics of winter rapeseed under low temperature stress[J]. Acta Agronomica Sinica, 42(10): 1541-1550.) [3] 孙万仓, 马卫国, 雷建民, 等. 2007. 冬油菜在西北旱寒区的适应性和北移的可行性研究[J]. 中国农业科学, 40(12): 2716-2726. (Sun W C, Ma W G, Lei J M, et al.2007. Study on adaptation and introduction possibility of winter rapeseed to dry and cold areas in northwest China[J]. Scientia Agricultura Sinica, 40(12): 2716-2726.) [4] 孙万仓, 裴新梧, 马骊, 等. 2022. 我国北方冬季覆盖作物研究进展及发展前景[J]. 中国农业科技导报, 24(01): 128-36. (Sun W C, Pei X W, Ma L, et al.2022. Advances and outlook of winter cover crop development research in northern China[J]. Journal of Agricultural Science and Technology, 24(01): 128-136.) [5] 王毓洪, 孟秋峰, 郁勤飞, 等. 2021. 油菜冻害机理与抗寒机制研究[J]. 中国果菜, 41(11): 57-61. (Wang Y H, Meng Q F, Yu Q F, et al.2021. Reaserch on mechanism of freezing injury and low-temperature resistance of rapeseed[J]. China Fruit & Vegetable, 41(11): 57-61.) [6] 岳昌武, 何博文, 何明雄, 等. 2007. 甘薯S-腺苷甲硫氨酸合成酶基因克隆与表达[J]. 中国农学通报, 23(06): 121-125. (Yue C W, He B W, He M X, et al.2007. Cloning and expression of s-adenosylmethionine synthetase of sweetpotato[J]. Chinese Agricultural Science Bulletin, 23(06): 121-125.) [7] 邹娅, 米文博, 徐明霞, 等. 2021. 低温胁迫下北方强冬性区甘蓝型冬油菜的低温光合生理特征[J]. 甘肃农业大学学报, 56(02): 105-113. (Zou Y, Zhao Y H, Hou X F, et al.2021. Physiological and photosynthetic characteristics of winter rapeseeds under low temperature stress in northern strong winterness region[J]. Journal of Gansu Agricultural University, 56(02): 105-113.) [8] 周琦, 郑幸果, 何辉煌, 等. 2017. 植物S-腺苷甲硫氨酸合成酶的新功能展望[J]. 生命的化学, 37(04): 521-527. (Zhou Q, Zheng X G, He H H, et al.2017. Progresses in new functions of plant S-adenosylmethionine synthetase[J]. Chemistry of Life, 37(04): 521-527.) [9] 张寒冰, 张书发, 李毛, 等. 2021. 大麦 S-腺苷甲硫氨酸合成酶基因HvSAMS2对非生物胁迫响应的表达分析[J]. 农业生物技术学报, 29(1): 35-46. (Zhang H B, Zhang S F, Li M, et al.2021. Expression analysis of s-adenosylmethionine synthetase gene HvSAMS2 from Hordeum vulgare in response to abiotic stress[J]. Journal of Agricultural Biotechnology, 29(1): 35-46.) [10] 张红燕, 金参, 罗文. Kunitz型丝氨酸蛋白酶抑制剂的研究进展[J]. 浙江农业科学, 2025, 66(07): 1770-1778. (Zhang H Y, Jin C, Luo W.2025. Research progress of Kunitz-type serine protease inhibitors[J]. Journal of Zhejiang Agricultural Sciences, 66(07): 1770-1778.) [11] Blom N, Sicheritz-Pontén T, Gupta R, et al.2004. Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence[J]. Proteomics, 4(6): 1633-1649. [12] Conesa A, Götz S, García-Gómez J M, et al.2005. Blast2GO: A universal tool for annotation, visualization and analysis in functional genomics research[J]. Bioinformatics, 21(18): 3674-3676. [13] Fan J, Yang G, Wu J, et al.2025. Feasibility analysis of expanding winter rapeseed northwards in China[J]. Agricultural and Forest Meteorology, 361: 110297. [14] Frenette Charron J B, Ouellet F, Pelletier M, et al.2005. Identification, expression, and evolutionary analyses of plant lipocalins[J]. Plant Physiology, 139(4): 2017-2028. [15] Gao L, Pan L, Shi Y, et al.2024. Genetic variation in a heat shock transcription factor modulates cold tolerance in maize[J]. Molecular Plant, 17(9): 1423-1438. [16] He M W, Wang Y, Wu J Q, et al.2019. Isolation and characterization of S-Adenosylmethionine synthase gene from cucumber and responsive to abiotic stress[J]. Plant Physiology and Biochemistry, 141: 431-445. [17] Heidari P, Mazloomi F, Nussbaumer T, et al.2020. Insights into the SAM synthetase gene family and its roles in tomato seedlings under abiotic stresses and hormone treatments[J]. Plants, 9(5): 586. [18] Holaday A S, Martindale W, Alred R, et al.1992. Changes in activities of enzymes of carbon metabolism in leaves during exposure of plants to low temperature[J]. Plant Physiology, 98(3): 1105-1114. [19] Horton P, Park K-J, Obayashi T, et al.2007. WoLF PSORT: protein localization predictor[J]. Nucleic Acids Research, 35(suppl_2): W585-W587. [20] Hurry V M, Malmberg G, Gardestrom P, et al.1994. Effects of a short-term shift to low temperature and of long-term cold hardening on photosynthesis and ribulose-1, 5-bisphosphate carboxylase/oxygenase and sucrose phosphate synthase activity in leaves of winter rye (Secale cereale L.)[J]. Plant Physiology, 106(3): 983-990. [21] Letunic I, Khedkar S, Bork P.2021. SMART: Recent updates, new developments and status in 2020[J]. Nucleic Acids Research, 49(D1): D458-D460. [22] Li F, Liu B, Zhang H, et al.2024. Integrative multi-omics analysis of chilling stress in pumpkin (Cucurbita moschata)[J]. BMC Genomics, 25(1): 1042. [23] Li W, Zhang C, Lu Q, et al.2011. The combined effect of salt stress and heat shock on proteome profiling in Suaeda salsa[J]. Journal of Plant Physiology, 168(15): 1743-1752. [24] Li W, Han Y, Tao F, et al.2011. Knockdown of SAMS genes encoding S-adenosyl-l-methionine synthetases causes methylation alterations of DNAs and histones and leads to late flowering in rice[J]. Journal of Plant Physiology, 168(15): 1837-1843. [25] Liu X, Bulley S M, Varkonyi-Gasic E, et al.2023. Kiwifruit bZIP transcription factor AcePosF21 elicits ascorbic acid biosynthesis during cold stress[J]. Plant Physiology, 192(2): 982-999. [26] Liu T, Qu J, Fang Y,et al.2025. Polyamines: The valuable bio-stimulants and endogenous signaling molecules for plant development and stress response[J]. Journal of Integrative Plant Biology, 67(3): 582-595. [27] Mittler R, Zandalinas S I, Fichman Y, et al.2022. Reactive oxygen species signalling in plant stress responses[J]. Nature Reviews Molecular Cell Biology, 23(10): 663-679. [28] Nielsen H, Tsirigos K D, Brunak S, et al.2019. A brief history of protein sorting prediction[J]. The Protein Journal, 38(3): 200-216. [29] Peterson G L.1983. Determination of total protein[J]. Methods Enzymol, 91: 95-119. [30] Qiu H, Zhou M, Huang L, et al.2019. Overexpression of MfSAMS1 improves chilling tolerance in transgenic stylo[J]. Agronomy Journal, 111(5): 2287-2292. [31] Sahid S, Roy C, Paul S, et al.2020. Rice lectin protein r40c1 imparts drought tolerance by modulating S-adenosylmethionine synthase 2, stress-associated protein 8 and chromatin-associated proteins[J]. Journal of Experimental Botany, 71(22): 7331-7346. [32] Sheffield J, Taylor N, Fauquet C, et al.2006. The cassava (Manihot esculenta Crantz) root proteome: Protein identification and differential expression[J]. Proteomics, 6(5): 1588-1598. [33] Shi Y, Ding Y, Yang S.2018. Molecular regulation of CBF signaling in cold acclimation[J]. Trends in Plant Science, 23(7): 623-637. [34] Sicher R C, Hatch A L, Stumpf D K, et al.1981. Ribulose 1, 5-bisphosphate and activation of the carboxylase in the chloroplast[J]. Plant Physiology, 68(1): 252-255. [35] Stein O, Granot D.2018. Plant fructokinases: Evolutionary, developmental, and metabolic aspects in sink tissues[J]. Frontiers in Plant Science, 9: 339. [36] Wang X, Oh M, Komatsu S.2016. Characterization of S-adenosylmethionine synthetases in soybean under flooding and drought stresses[J]. Biologia Plantarum, 60(2): 269-278. [37] Ying Z, Fu S, Yang Y.2025. Signaling and Scavenging: Unraveling the complex network of antioxidant enzyme regulation in plant cold adaptation[J]. Plant Stress, 16: 100833. [38] Yin M, Huang Z, Aslam A, et al.2024. Genome-wide identification of SAMS gene family in Cucurbitaceae and the role of ClSAMS1 in watermelon tolerance to abiotic stress[J]. Plant Physiology and Biochemistry, 211: 108708. [39] Zhang B Q, Huang Y X, Zhou Z F, et al.2022. Cold-Induced physiological and biochemical alternations and proteomic insight into the response of Saccharum spontaneum to low temperature[J]. International Journal of Molecular Sciences, 23(22): 14244. [40] Zareei E, Karami F, Gholami M, et al.2021. Physiological and biochemical responses of strawberry crown and leaf tissues to freezing stress[J]. BMC Plant Biology, 21(1): 532. [41] Zhang Y, Xu J, Li R, et al.2023. Plants' response to abiotic stress: Mechanisms and strategies[J]. International Journal of Molecular Sciences, 24(13): 10915.