施硫对不结球白菜硝酸盐累积及氮硫同化关键基因表达的影响

doi:10.3864/j.issn.0578-1752.2016.11.018

摘要/Abstract

摘要： 【目的】确定降低不结球白菜硝酸盐累积效果最佳的硫素形态，从转录水平筛选影响不结球白菜硝酸盐累积的关键基因，为完善不结球白菜科学施硫技术及进一步揭示硝酸盐累积分子调控机制、指导分子育种奠定基础。【方法】选取4种硫素形态及3个施用浓度处理不结球白菜，测定其对植株叶片及叶柄中硝酸盐含量的影响；利用半定量PCR技术从转录水平分析施硫对氮及硫代谢同化网络中30个基因表达的影响。【结果】不同形态硫处理均显著增加了不结球白菜的地上生物量，其中30 mg·kg^-1 Na₂SO₄处理的增幅最大，比对照增加49.76%，以Na₂SO₄处理的增幅最大。在降低小白菜硝酸盐含量中，以Na₂SO₄、Na₂S₂O₃处理效果相对显著，其中Na₂SO₄降低叶片中硝酸盐12.23%—23.55%，叶柄中33.08%—41.98%，降幅与浓度呈正相关，30 mg·kg^-1 Na₂SO₄处理的降幅最大；Na₂S₂O₃处理降低植株叶片中硝酸盐15.34%—33.08%，叶柄中11.95%—19.68%。硫处理在一定程度上促进氮同化，对照叶片中NR-1、NADH-GOGAT-1、NADH-GOGAT-2、Cytoplasm-GS-4、Cytoplasm-GS-5、GDH-3表达量低于其他处理，对照叶柄中NR-1、NADH-GOGAT-2、Cytoplasm-GS-1、GDH-2表达量低于其他处理，其中，各处理NADH-GOGAT-2表达量与叶片及叶柄中硝酸盐含量变化呈现一定规律性。硫处理对植株硫同化基因也产生一定影响，对照叶片中ATPS-2、ATPS-3、ATPS-4、APSR-3、SIR、SAT1.1、SAT2.1表达量较低，而对照叶柄中仅SIR、OASTL-A表达量明显低于其他处理。【结论】Na₂SO₄是降低不结球白菜硝酸盐效果较为显著的硫素，且能够显著提高产量，30 mg·kg^-1的Na₂SO₄为较优处理。NADH-GOGAT-2表达量与不结球白菜内硝酸盐含量呈负相关，推测其可能是影响氮同化的关键基因。

关键词: 不结球白菜, 硫, 硝酸盐, 氮硫同化, 基因表达

Abstract: 【Objective】The objective of this paper is to identify the best sulfate to reduce nitrate accumulation in non-heading Chinese cabbage, and analyze the key genes involved in nitrogen and sulphur assimilation. The results of this study will provide new insights into sulphur fertilization and investigation of the molecular mechanisms of nitrate accumulation with the goal of molecular breeding.【Method】Nitrate content of non-heading Chinese cabbage leaves and petioles were measured under treatments of 4 forms of sulphur with 3 concentrations. The expression of 30 nitrogen and sulphur assimilation related genes were analyzed by semi-quantitative RT-PCR. 【Result】Sulphur treatments were significantly increased the aboveground biomass of non-heading Chinese cabbage, and the treatments of Na₂SO₄ had the best effect, and 30 mg·kg^-1Na₂SO₄ was the most effective treatment as the aboveground biomass increased by 49.76% compared with the control. Na₂SO₄and Na₂S₂O₃ reduced nitrate content of non-heading Chinese cabbage more significantly than others. Na₂SO₄decreased nitrate content by 12.23%-23.55% in leaves and by 33.08%-41.98% in petioles compared with the control, and it was also found a positive correlation between the reduction and concentration of Na₂SO₄, and 30 mg·kg^-1Na₂SO₄ also had the best effect. Na₂S₂O₃ decreased nitrate content by 15.34%-33.08% compared with the control in leaves and by 11.95%-19.68% in petioles. Sulphur promoted nitrogen assimilation, and the expression of NR-1, NADH-GOGAT-1, NADH-GOGAT-2, Cytoplasm-GS-4, Cytoplasm-GS-5, and GDH-3 were higher than the control in leaves, and the expression of NR-1, NADH-GOGAT-2, Cytoplasm-GS-1, and GDH-2 were higher than the control in petioles. The expression of NADH-GOGAT-2 was correlated with nitrate levels. Sulphur treatments also had an effect on sulphur assimilation genes, and the expressions of ATPS-2, ATPS-3, ATPS-4, APSR-3, SIR, SAT1.1, and SAT2.1 were higher than the control in leaves, and only the expressions of SIR and OASTL-A were higher than the control in petioles. 【Conclusion】 Na₂SO₄ had the best effect in reducing nitrate content and increasing yield of non-heading Chinese cabbage, and 30 mg·kg^-1 Na₂SO₄was the most effective treatment. The expression of NADH-GOGAT-2 was correlated with nitrate levels. The data suggested that NADH-GOGAT-2 may be the key gene in nitrogen assimilation.

Key words: non-heading Chinese cabbage, sulphur, nitrate, nitrogen and sulphur assimilation, gene expression

徐瑶，牟建梅，张国芹，马佳佳，徐君，李军，刘凤军，佘旭东. 施硫对不结球白菜硝酸盐累积及氮硫同化关键基因表达的影响[J]. 中国农业科学, 2016, 49(11): 2222-2233.

XU Yao, MU Jian-mei, ZHANG Guo-qin, MA Jia-jia, XU Jun, LI Jun, LIU Feng-jun, SHE Xu-dong. Effect of Sulphur Availability on Nitrate Accumulation and Expression of Nitrogen and Sulphur Assimilation Related Genes in Non-Heading Chinese Cabbage[J]. Scientia Agricultura Sinica, 2016, 49(11): 2222-2233.

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