Abstract:F-box gene plays an important role in plant abiotic stress response. In order to study the function of F-box gene BoFBX117 (GenBank No. XM_013731217.1) in cabbage (Brassica oleracea var. capitata) response to low-temperature stress, in this study, the BoFBX117 over-expression vector was constructed, and the over-expression of BoFBX117 cabbage (OE-BoFBX117) plants were obtained by Agrobacterium tumefaciens mediated method. After 24 h of low-temperature stress at 4, 0, -2 and -4 ℃, the phenotype of the over-expression of BoFBX117 cabbage showed that the degree of leaf yellowing wilting was less than that of non-transgenic cabbage (WT). After 7 d of recovery, it was found that the over-expression of BoFBX117 cabbage had stronger ability to return to normal growth. The results of qPCR showed that the expression of BoFBX117 in transgenic plants was significantly increased under low-temperature stress (P<0.01), which was 1.87~4.80 times that of WT plants. Under low-temperature stress at 0 ℃, the activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) of transgenic plants were the highest, which were 80%, 16% and 99% higher than those of WT plants respectively. Under low-temperature stress, the content of malondialdehyde (MDA) continuously accumulated. At -4 ℃, the MDA content of transgenic plants was 0.62 μmol/g FW, 53.38% lower than WT. Under normal conditions and low-temperature stress, the proline and soluble sugar contents of transgenic plants were significantly higher than those of WT plants (P<0.05). The results showed that BoFBX117 might play a positive role in the response to low-temperature stress, and over-expression of BoFBX117 in cabbage could improve the low-temperature tolerance of cabbage. This study provides support for the breeding of low-temperature tolerance varieties of cabbage.
罗玉霞, 许涛, 戴倩, 陈瑶, 宋江华. 过表达F-box基因BoFBX117对甘蓝低温耐受性的影响[J]. 农业生物技术学报, 2024, 32(1): 70-79.
LUO Yu-Xia, XU Tao, DAI Qian, CHEN Yao, SONG Jiang-Hua. Effect of Overexpression of F-box Gene BoFBX117 on Low-temperature Tolerance of Cabbage (Brassica oleracea var. capitata). 农业生物技术学报, 2024, 32(1): 70-79.
[1] 陈秀秀, 张彤, 余倩文, 等. 2019. 紫花苜蓿F-box蛋白基因MsFTL的克隆及功能分析[J]. 植物遗传资源学报, 20(03), 750-759. (Chen X X, Zhang T, Yu Q W, et al.2019. Cloning and functional analysis of F-box protein gene MsFTL in Alfalfa (Medicago sativa L.)[J]. Journal of Plant Genetic Resources, 20(03), 750-759.) [2] 陈思琪, 孙敬爽, 麻文俊, 等. 2022. 植物低温胁迫调控机制研究进展[J].中国农学通报, 38(17):51-61. (Chen S Q, Sun J S, Ma W S, et al.2022. Regulation mechanism of low temperature stress on plants: Research progress[J]. Chinese Agricultural Science Bulletin, 38(17): 51-61.) [3] 段雪娇, 梁小红, 葛永强, 等. 2017. 大叶落地生根KdFBX基因的cDNA克隆与表达分析[J]. 农业生物技术学报, 25(12): 1961-1969. (Duan X J, Liang X H, Ge Y Q, et al.2019. cDNA cloning and expression analysis of KdFBX gene in Kalanchoe daigremontiana[J]. Journal of Agricultural Biotechnology, 25(12): 1961-1969.) [4] 高俊山, 蔡永萍. 2018. 植物生理学实验指导. 第2版[M]. 中国农业大学出版社, 北京. pp. 93-133. (Gao J S, Cai Y P.2018. Experimental Guidance on Plant Physiology. 2nd ed[M]. China Agricultural University Press, Beijing, China. pp. 93-133.) [5] 秦文斌, 山溪, 张振超, 等. 2018. 低温胁迫对甘蓝幼苗抗逆生理指标的影响[J]. 核农学报, 32(03): 576-581. (Qin W B, Shan X, Zhang Z C, et al.2018. Effect of low temperature stress on anti-stress physiological indexes of cabbage seedlings[J]. Journal of Nuclear Agricultural Sciences, 32(03): 576-581.) [6] 王芳, 王淇, 赵曦阳. 2019. 低温胁迫下植物的表型及生理响应机制研究进展[J]. 分子植物育种, 17(15): 5144-5153. (Wang F, Wang Q, Zhao X Y.2019. Research progress of phenotype and physiological response mechanism of plants under low temperature stress[J]. Molecular Plant Breeding, 17(15): 5144-5153.) [7] 王五宏, 汪精磊, 李必元, 等. 2020. 结球甘蓝抽薹性遗传规律和QTL定位分析[J]. 园艺学报, 47(05): 974-982. (Wang W H, Wang J L, Li B Yet al.2020. Genetic and QTL mapping analysis of bolting time in cabbage (Brassica oleracea)[J]. Acta Horticulturae Sinica, 47(05): 974-982.) [8] 许涛, 鲁正钊, 夏冬健, 等. 2021. 结球甘蓝BoFBX117基因的克隆与表达分析[J]. 核农学报, 35(05): 1060-1066. (Xu T, Lu Z Z, Xia D J, et al.2021. Cloning and expression analysis of BoFBX117 from Brassica oleracea[J]. Journal of Nuclear Agricultural Sciences, 35(05): 1060-1066.) [9] Bai C, Sen P, Hofmann K, et al.1996. SKP1 connects cell cycle regulators to the ubiquitin proteolysis machinery through a novel motif, the F-Box[J]. Cell, 86(2): 263-274. [10] Calderón-Villalobos L I A, Nill C, Marrocco K, et al.2007. The evolutionarily conserved Arabidopsis thaliana F-box protein AtFBP7 is required for efficient translation during temperature stress[J]. Gene, 392(1-2): 106-116. [11] Devireddy A R, Tschaplinski T J, Tuskan G A, et al.2021. Role of reactive oxygen species and hormones in plant responses to temperature changes[J]. International Journal of Molecular Sciences, 22(16): 8843. [12] Duplan V, Rivas S.2014. E3 ubiquitin-ligases and their target proteins during the regulation of plant innate immunity[J]. Frontiers in Plant Science, 5(3): 42-43. [13] Gao L T, Jia S Z, Cao L, et al.2022. An F-box protein from wheat, TaFBA-2A, negatively regulates JA biosynthesis and confers improved salt tolerance and increased JA responsiveness to transgenic rice plants[J]. Plant Physiology and Biochemistry, 182: 227-239. [14] Gu K Y, Hou S, Chen J F, et al.2021. The physiological response of different tobacco varieties to chilling stress during the vigorous growing period[J]. Scientific Reports, 11(1): 22136. [15] He R Q, Yu D S, Li X M, et al.2016. F-box gene FOA2 regulates GA- and ABA- mediated seed germination in Arabidopsis[J]. Science China (Life Sciences), 59(11): 1192-1194. [16] Horn-Ghetko D, Krist D T, Prabu J R, et al.2021. Ubiquitin ligation to F-box protein targets by SCF-RBR E3-E3 super-assembly[J]. Nature, 590(7847): 671-676. [17] Jain M, Nijhawan A, Arora R, et al.2007. F-Box proteins in Rice. Genome-wide analysis, classification, temporal and spatial gene expression during panicle and seed development, and regulation by light and abiotic stress[J]. Plant Physiology, 143(4): 1467-1483. [18] Kaul S, Koo H L, Jenkins J, et al.2000. Analysis of the genome sequence of the flowering plant Arabidopsis thaliana[J]. Nature, 408(6814): 796-815. [19] Khandra F, Kohei N, Fidela V.2015. Investigation on quantitative index of chilling injury in cucumber fruit based on the electrolyte leakage and malondialdehyde content[J]. International Journal on Advanced Science, Engineering and Information Technology, 5(3): 222-225. [20] Li Q X, Wang W Q, Wang W L, et al.2018. Wheat F-Box protein gene TaFBA1 is involved in plant tolerance to heat stress[J]. Frontiers in Plant Science, 9: 521. [21] Meitha K, Pramesti Y, Suhandono S.2020. Reactive oxygen species and antioxidants in postharvest vegetables and fruits[J]. International Journal of Food Science, 2020: 8817778. [22] Morel M, Shah K N, Long W W.2020. The F-box protein FBXL16 up-regulates the stability of C-MYC oncoprotein by antagonizing the activity of the F-box protein FBW7[J]. The Journal of Biological Chemistry, 295(23): 7970-7980. [23] Qu L N, Sun M S, Li X M, et al.2020. The Arabidopsis F-box protein FOF2 regulates ABA-mediated seed germination and drought tolerance[J]. Plant Science, 301: 110643. [24] Rameneni J J, Dhandapani V, Paul P, et al.2018. F-Box genes in Brassica rapa: Genome-wide identification, structural characterization, expressional validation, and comparative analysis[J]. Plant Molecular Biology Reporter, 36(3): 500-517. [25] Rao V, Virupapuram V.2021. Arabidopsis F-box protein At1g08710 interacts with transcriptional protein ADA2b and imparts drought stress tolerance by negatively regulating seedling growth[J]. Biochemical and Biophysical Research, 536: 45-51. [26] Šamec D, Piljac-Zagarac J, Bogovic M, et al.2011. Antioxidant potency of white (Brassica oleracea L. var. capitata) and Chinese (Brassica rapa L. var. pekinensis (Lour.) Brassica oleracea: The influence of development stage, cultivar choice and seed selection[J]. Scientia Horticulturae, 128(2): 78-83. [27] Venkatesh J, Kang M Y, Liu L, et al.2020. F-Box family genes, LTSF1 and LTSF2 regulate low-temperature stress tolerance in pepper (Capsicum chinense)[J]. Plants-Basel, 9(9): 1186. [28] Wei T L, Wang Z X, He Y F, et al.2022. Proline synthesis and catabolism-related genes synergistically regulate proline accumulation in response to abiotic stresses in grapevines[J]. Scientia Horticulturae, 305: 111373. [29] Xu G Y, Cui Y C, Wang M L, et al.2014. OsMsr9, a novel putative rice F-box containing protein, confers enhanced salt tolerance in transgenic rice and Arabidopsis[J]. Molecular Breeding, 34(3): 1055-1064.