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Effects of Seed Soaking with Brassinolide on Brassica napus Seedlings |
CUI Sai1, WANG Tong-Hua2,*, ZHANG Zhen-Qian1,* |
1 College of Agriculture, Hunan Agricultural University, Changsha 410128, China; 2 Hunan Crop Research Institute, Changsha 410128, China |
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Abstract Brassinolide has a promoting effect on crop growth. In order to clarify the effect of seed soaking on the seedlings of Brassica napus, the dry seeds were soaked in brassinolide, and the soaked in clear water were used as the control. The leaves at the 5~6 leaf stage were used for transcriptome sequencing analysis, and the leaves at young seedling stage, 5~6 leaf stage and budding stage were used for physiological and biochemical analysis. The results showed that transcriptome analysis found 4 066 up-regulated and 4 606 down-regulated differential genes. Three differential genes related to fatty acid synthesis were screened out: Fatty acyl carrier protein thioesterase (BnFAT), biological carboxyl carrier protein (BnBCCP), and stearoy acyl carrier protein desaturase (BnSAD); 3 differential genes related to photosynthesis were screened out: PhotoreactionⅠsystem centerⅤsubunit (BnPSAG), photosystemⅡreaction center protein PSB28 (BnPSB28) and chlorophyll ab-binding protein (BnCab).The qPCR method was used to verify the transcriptome results, and it was found that only BnSAD was inconsistent with the sequencing results, and the other 5 differential genes were consistent with the transcriptome sequencing results. Using the leaves at seedling stage, 5~6 leaf stage and budding stage as materials, the expression level of 6 differential genes was analyzed, and the soluble sugar, soluble protein content, superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and other physiological functions were analyzed, and the correlation between the differential gene expression in 3 periods and the physiological and biochemical indicators were studied. The results showed that the expression of BnFAT and BnBCCP genes were positively correlated with the content of soluble sugar (both correlation coefficients were 0.69), BnCab expression was positively correlated with the content of soluble protein and SOD activity (correlation coefficients was 0.89 and 0.62, respectively), BnPSAG gene expression was positively correlated with POD activity (correlation coefficient was 0.62). The results of this study indicate that the treatment of seeds with brassinolide could increase the soluble substance content and protective enzyme activity of B. napus, and there was a close correlation between the physiological and biochemical indexes and the differential gene expression. This research could provide theoretical and technical reference for high-yield cultivation of rape.
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Received: 26 April 2021
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Corresponding Authors:
*zzq770204@163.com; wangtonghua2004@163.com
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[1] 曹盈. 2013. 高通量测序技术在植物转录组研究中的应用[J]. 北京农业, (06): 6-7. (Cao Y. Application of high-throughput sequencing technology in plant transcriptome research[J]. Beijing Agriculture, (06): 6-7.) [2] 戴晓峰, 卢长明, 吴刚, 等. 2007. 甘蓝型油菜生物素羧基载体蛋白基因的克隆与结构分析[J]. 中国农业科学, (09): 1883-1889. (Dai X F, Lu C M, Wu G, et al. 2007. Cloning and structural analysis of the biotin carboxyl carrier protein gene in Brassica napus[J]. China Agricultural Sciences, (09): 1883-1889.) [3] 冯韬, 谭晖, 徐江林, 等. 2019. 油菜素内酯在不同生育期对两品系甘蓝型油菜的生长调控[J]. 中国油料作物学报, 41(06): 904-913. (Feng T, Tan H, Xu J L, et al.2019. The growth regulation of brassinolide in two lines of Brassica napus at different growth stages[J]. Chinese Journal of Oil Crops, 41(06): 904-913.) [4] 胡倩, 裴云霞, 倪天虹, 等. 2020. 长林4号油茶幼苗对低温胁迫的生理响应[J]. 湖北林业科技, 49(06): 5-12. (Hu Q, Pei Y X, Ni T H, et al.2020. Physiological response of Changlin No. 4 Camellia oleifera seedlings to low temperature stress[J]. Hubei Forestry Science and Technology, 49(06): 5-12.) [5] 黄克林. 2020. 油菜(Brassica napus)油脂相关的转录组和磷酸化蛋白组分析及脂肪酸运蛋白BnFAX6的功能研究[D]. 博士学位论文, 华中师范大学, 导师: 李学宝, pp. 1-3. (Huang K L.2020. Oilseed rape (Brassica napus) lipid-related transcriptome and phosphorylation proteome analysis and the functional study of fatty acid transport protein BnFAX6[D]. Thesis for Ph.D., Central China Normal University, Supervisor: Li X B, pp. 1-3.) [6] 黄一龙, 庞天德, 史静, 等. 2021. 2,4-表油菜素内酯对紫花苜蓿高温胁迫生理特性的影响[J]. 湖北农业科学, 60(03): 93-95, 99.(Huang Y L, Pang T D, Shi J, et al. 2021. Effects of 2,4-epibrassinolide on physiological characteristics of alfalfa under high temperature stress[J]. Hubei Agricultural Sciences, 60(03): 93-95, 99.) [7] 李兰. 2019. 油菜素内酯调控铜和铬胁迫下油菜生理生化与分子机制研究[D]. 博士学位论文, 浙江大学, 导师: 周伟军, pp. 20-21. (Li L.2019. Studies on brassinolide's regulation of physiological, biochemical and molecular mechanisms of Brassica napus under copper and chromium stress[D]. Thesis for Ph.D., Zhejiang University, Supervisor: Zhou W J, pp. 20-21.) [8] 马梅, 刘冉, 郑春芳, 等. 2015. 油菜素内酯对盐渍下油菜幼苗生长的调控效应及其生理机制[J]. 生态学报, 35(6): 1837. (Ma M, Liu R, Zheng C F, et al.2015. The regulatory effect of brassinolide on the growth of rape seedlings under salt and its physiological mechanism[J]. Acta Ecologica Sinica, 35(6): 1837.) [9] 孙玉珺. 2019. 玉米发芽期抗冷型筛选及低温胁迫下油菜素内酯对幼苗的调控效应研究[D]. 硕士学位论文, 东北农业大学, 导师: 杨德光, pp. 19-27 (Sun Y J. 2019. Screening of cold-tolerant types during germination of maize and the regulation effect of brassinolide on seedlings under low temperature stress[D]. Thesis for M.S, Northeast Agricultural University, Supervisor: Yang D G, pp. 19-27.) [10] 王伏林, 郎春秀, 吴关庭, 等. 2009. 大肠杆菌异质型ACCase亚基基因accB重组表达载体的构建和原核表达[J]. 中国农学通报, 25(23): 74-77. (Wang F L, Lang C X, Wu G T, et al.2009. Construction and prokaryotic expression of recombinant expression vector of E. coli heterogeneous ACCase subunit gene accB[J]. Chinese Agricultural Science Bulletin, 25(23): 74-77.) [11] 王晓丹, 肖钢, 张振乾, 等. 2019. 高油酸油菜脂肪酸代谢相关微效基因筛选及验证[J]. 农业生物技术学报, 27(07): 1171-1178. (Wang X D, Xiao G, Zhang Z Q, et al.2019. Screening and verification of minor genes related to fatty acid metabolism with high oleic acid[J]. Journal of Agricultural Biotechnology, 27(07): 1171-1178.) [12] 萧浪涛, 王三根. 2005. 植物生理学试验技术[M]. 北京: 中国农业出版社, pp. 211-215. (Xiao L T, Wang S G.2005. Experimental Technology of Plant Physiology[M]. China Agriculture Press, Beijing, China, pp. 211-215.) [13] 杨复康, 杨燕君, 宋永宏, 等. 2021. 不同杏品种抗寒性及生理指标[J]. 北方园艺, (03): 27-32. (Yang Y J, Yang F K, Song Y H, et al. 2021. Cold resistance and physiological indexes of different apricot varieties[J]. Northern Horticulture, (03): 27-32.) [14] 阳江华, 张希财, 邹智. 2019. 橡胶树捕光叶绿素a/b结合蛋白基因CAB2的克隆与分析[J]. 西南林业大学学报, 39(01): 88-94. (Yang J H, Zhang X C, Zhou Z.2019. Cloning and analysis of the light-harvesting chlorophyll a/b binding protein gene CAB2 of rubber tree[J].Journal of Southwest Forestry University, 39(01): 88-94.) [15] 张永霞, 赵锋, 张红玲. 2015. 中国油菜产业发展现状、问题及对策分析[J]. 世界农业,(04): 96-99, 203-204. (Zhang Y X, Zhao F, Zhang H L.2015. Analysis on the status, problems and countermeasures of China's rape industry development[J]. World Agriculture,(04): 96-99, 203-204.) [16] Agnieszka Z, Erik J P, Dario L, et al.2005. PhotosyemⅠlacking the PSI-G subunit has a higher affinity for plastocyanin and is sensitive to photodamage[J]. Biochimica et Biophysica Acta, 1708(2): 154-163. [17] Contreras C, Tambd D, Martin M, et al.2020. Characterization and validation of olive FAD and SAD gene families: Expression analysis in different tissues and during fruit development[J]. Molecular Biology Reports, 47(6): 4345-4355. [18] Grove M D, Spencer G F, Rohwedder W K, et al.1979. Brassinolide, a plant growth-promoting steroid isolated from Brassica napus pollen[J]. Nature, 281: 216-217. [19] Mutz K O, Heilkenbrinker A, Lnne M, et al.2013. Transcriptome analysis using next-generation sequencing[J]. Current Opinion in Biotechnology, 24(1): 22-30. [20] Salas J J, Ohlrogge J B.2002. Characterization of substrate specificity of plant FatA and FatB acyl-ACP thioesterases[J]. Archives of Biochemistry and Biophysics, 403(1): 25-34. [21] Sharma M, Mahajan P, Singh H P, et al.2019. 24-epibrassinolide pre-treatment reduces alkaline-induced oxidative stress in red rice seedlings[J]. Environmental Science and Pollution Research, 26(22): 23192-23197. [22] Wang S Q, Zhao H H, Zhao L M, et al.2020. Application of brassinolide alleviates cold stress at the booting stage of rice[J]. Journal of Integrative Agriculture, 19(4): 975-987. [23] Zhang W, Sheng J, Xu Y, et al.2019. Role of brassinosteroids in rice spikelet differentiation and degeneration under soil-drying during panicle development[J]. BMC Plant Biology, 19(1): 409. |
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