BnaHKL1基因过量表达与转基因油菜叶片中可溶性糖的变化
Overexpression of the BnaHKL1 Gene and Changes in Soluble Sugars in the Leaf Tissues of Transgenic Rapeseed
DOI: 10.12677/hjas.2024.1412165, PDF,    国家科技经费支持
作者: 刘佳桢, 钟 微, 杨 慧, 田闵玉, 阮 颖, 刘春林*:湖南农业大学,作物表观遗传调控与发育湖南省重点实验室,湖南 长沙;岳麓山实验室,湖南 长沙
关键词: HKL1过量表达转基因植株可溶性糖HKL1 Overexpression Transgenic Plants Soluble Sugars
摘要: 本研究构建了甘蓝型油菜BnaHKL1基因的过量表达载体pFGC5941-BnaHKL1,并采用根癌农杆菌介导的方法对甘蓝型油菜湘油15 (XY15)的下胚轴外植体进行转化。通过PCR鉴定,成功获得34株抗性植株,其中11株是具有基因过量表达的转基因植株。半定量RT-PCR分析结果表明,过表达转基因植株中BnaHKL1基因的表达水平显著高于XY15中的表达水平。与XY15相比,BnaHKL1过表达植株的淀粉含量显著降低,而蔗糖、可溶性糖和果聚糖的含量则显著升高。
Abstract: In this study, the overexpression vector pFGC5941-BnaHKL1 of the BnaHKL1 gene from Brassica napus was constructed, and the hypocotyl explants of Brassica napus Xiangyou 15 (XY15) were transformed by the Agrobacterium tumefaciens-mediated method. Through PCR identification, 34 resistant positive plants were successfully obtained, among which 11 were transgenic plants with overexpression cassette of the BnaHKL1 gene. The results of semi-quantitative RT-PCR analysis indicated that the expression level of the BnaHKL1 gene in the overexpression transgenic plants was significantly higher than that in XY15. Compared with XY15, the starch content in BnaHKL1 overexpression plants was significantly decreased, whereas the contents of sucrose, soluble sugars, and fructan were significantly increased.
文章引用:刘佳桢, 钟微, 杨慧, 田闵玉, 阮颖, 刘春林. BnaHKL1基因过量表达与转基因油菜叶片中可溶性糖的变化[J]. 农业科学, 2024, 14(12): 1307-1315. https://doi.org/10.12677/hjas.2024.1412165

参考文献

[1] 殷艳, 王汉中. 我国油菜生产现状及发展趋势[J]. 农业展望, 2011, 7(1): 43-45.
[2] 沈金雄, 傅廷栋. 我国油菜生产、改良与食用油供给安全[J]. 中国农业科技导报, 2011, 13(1): 1-8.
[3] 王汉中. 以新需求为导向的油菜产业发展战略[J]. 中国油料作物学报, 2018, 40(5): 613-617.
[4] 李积铭, 张耀文, 郭安强, 等. 油菜抗寒性和寒旱区油菜育种探析[J]. 华北农学报, 2023, 38(z1): 131-144.
[5] 李方一, 黄璜, 官梅, 等. 油菜理想株型研究进展[J]. 中国油料作物学报, 2023, 45(1): 4-16.
[6] 张哲, 殷艳, 刘芳, 等. 我国油菜多功能开发利用现状及发展对策[J]. 中国油料作物学报, 2018, 40(5): 618-623.
[7] Raza, A. (2020) Eco-Physiological and Biochemical Responses of Rapeseed (Brassica napus L.) to Abiotic Stresses: Consequences and Mitigation Strategies. Journal of Plant Growth Regulation, 40, 1368-1388. [Google Scholar] [CrossRef
[8] Liu, X., Wei, R., Tian, M., Liu, J., Ruan, Y., Sun, C., et al. (2022) Combined Transcriptome and Metabolome Profiling Provide Insights into Cold Responses in Rapeseed (Brassica napus L.) Genotypes with Contrasting Cold-Stress Sensitivity. International Journal of Molecular Sciences, 23, Article 13546. [Google Scholar] [CrossRef] [PubMed]
[9] 孙永梅, 刘丽杰, 冯明芳, 等. 植物在低温胁迫下的糖代谢研究进展[J]. 东北农业大学学报, 2015, 46(7): 95-102, 108.
[10] Ke, L., Lei, W., Yang, W., Wang, J., Gao, J., Cheng, J., et al. (2020) Genome-Wide Identification of Cold Responsive Transcription Factors in Brassica napus L. BMC Plant Biology, 20, Article No. 62. [Google Scholar] [CrossRef] [PubMed]
[11] Ma, L., Coulter, J.A., Liu, L., Zhao, Y., Chang, Y., Pu, Y., et al. (2019) Transcriptome Analysis Reveals Key Cold-Stress-Responsive Genes in Winter Rapeseed (Brassica rapa L.). International Journal of Molecular Sciences, 20, Article 1071. [Google Scholar] [CrossRef] [PubMed]
[12] 范军强, 路晓明, 王会文, 等. 低温胁迫下甘蓝型冬油菜抗寒性与叶片激素含量的关联性[J]. 江苏农业学报, 2023, 39(1): 15-21.
[13] 贾朋贺, 刘红, 杨定国, 等. 植物多糖的提取、修饰及其糖代谢作用[J]. 食品研究与开发, 2022, 43(5): 210-216.
[14] 叶香媛, 周文彬. 植物果糖激酶研究进展[J]. 科学通报, 2021, 66(22): 2820-2831.
[15] 俞锞, 李志邈, 万红建, 等. 高等植物蔗糖转化酶功能的研究进展[J]. 安徽农业科学, 2013, 41(33): 12815-12818, 12822.
[16] Moore, B., Zhou, L., Rolland, F., Hall, Q., Cheng, W., Liu, Y., et al. (2003) Role of the Arabidopsis Glucose Sensor HXK1 in Nutrient, Light, and Hormonal Signaling. Science, 300, 332-336. [Google Scholar] [CrossRef] [PubMed]
[17] 张超, 王彦杰, 付建新, 等. 高等植物己糖激酶基因研究进展[J]. 生物技术通报, 2012(4): 19-26.
[18] 张超, 付三雄, 周小婴, 等. 甘蓝型油菜GPDH基因克隆、表达分析及植物过表达载体构建[J]. 南方农业学报, 2019, 50(7): 1399-1407.
[19] 郑本川, 李浩杰, 张锦芳, 等. 甘蓝型油菜CONSTANS基因植物过表达载体构建及转基因植株的获得[J]. 分子植物育种, 2018, 16(1): 130-134.
[20] 陈吉杨, 阮颖, 刘博宇. 甘蓝型油菜含MATH结构域基因BnaM154过表达载体的构建与转化[J]. 分子植物育种, 2022, 20(13): 4377-4382.
[21] Zheng, B., Li, H., Zhang, J., Cui, C., Chai, L., Jiang, J., et al. (2018) Construction of Overexpression Vector of CONSTANS Gene Plant in Brassica napus and Production of Transgenic Plants. Molecular Plant Breeding, 9, 73-79. [Google Scholar] [CrossRef
[22] 马骊, 白静, 赵玉红, 等. 冷胁迫下甘蓝型冬油菜表达蛋白及BnGSTs基因家族的鉴定与分析[J]. 作物学报, 2023, 49(1): 153-169.
[23] 呼芳娣, 刘丽君, 马骊, 等. 低温胁迫下甘蓝型冬油菜BnYUCCA8基因差异表达及内源生长素含量变化[J]. 干旱地区农业研究, 2022, 40(3): 1-10.
[24] Janeczko, A., Gullner, G., Skoczowski, A., Dubert, F. and Barna, B. (2007) Effects of Brassinosteroid Infiltration Prior to Cold Treatment on Ion Leakage and Pigment Contents in Rape Leaves. Biologia Plantarum, 51, 355-358. [Google Scholar] [CrossRef
[25] Wilson, J.E. (2003) Isozymes of Mammalian Hexokinase: Structure, Subcellular Localization and Metabolic Function. Journal of Experimental Biology, 206, 2049-2057. [Google Scholar] [CrossRef] [PubMed]
[26] Sun, X., Zong, Y., Yang, S., Wang, L., Gao, J., Wang, Y., et al. (2020) A Fructan: The Fructan 1-Fructosyl-Transferase Gene from Helianthus tuberosus Increased the PEG-Simulated Drought Stress Tolerance of Tobacco. Hereditas, 157, Article No. 14. [Google Scholar] [CrossRef] [PubMed]
[27] Ozturk, M., Turkyilmaz Unal, B., García‐Caparrós, P., Khursheed, A., Gul, A. and Hasanuzzaman, M. (2020) Osmoregulation and Its Actions during the Drought Stress in Plants. Physiologia Plantarum, 172, 1321-1335. [Google Scholar] [CrossRef] [PubMed]