文献类型：期刊文献

中文题名：菠萝硝态氮的吸收积累及其转运蛋白基因表达模式研究

英文题名：Study on the absorption and accumulation of nitrate and the expression of nitrate transporter genes in pineapple

作者：杨晓雪[1];刘胜辉[2];魏茜雅[1];林欣琪[1];叶春海[1];张秀梅[2];李映志[1]

机构：[1]广东海洋大学滨海农业学院园艺系,广东湛江524088;[2]中国热带农业科学院南亚热带作物研究所,广东湛江524091

年份：2022

卷号：39

期号：10

起止页码：1774

中文期刊名：果树学报

外文期刊名：Journal of Fruit Science

收录：CSTPCD、、CSCD2021_2022、Scopus、北大核心、CSCD、北大核心2020

基金：广东海洋大学创新强校工程项目(GDOU2016050256)。

语种：中文

中文关键词：菠萝;硝态氮转运蛋白;基因表达;氮营养

外文关键词：Pineapple;Nitrate transporter;Gene expression;Nitrogen

中文摘要：【目的】了解菠萝硝态氮转运蛋白基因在菠萝硝态氮吸收与同化中的作用。【方法】以金菠萝为材料,通过2个试验,分别研究了施氮对菠萝各组织硝态氮含量和40个硝态氮转运蛋白基因表达日变化的影响以及不同施氮处理对菠萝植株生长、叶片硝态氮含量和其转运蛋白基因表达的影响。【结果】菠萝根、茎、叶硝态氮含量均在10:00和16:00具有最高值;施氮后2 h,根系硝态氮含量显著增加;施氮增加了叶中18:00的硝态氮含量,降低了茎中14:00和18:00时及根中16:00的硝态氮含量。根中高表达的硝态氮转运蛋白基因最多,峰值多在12:00和14:00;叶中高表达基因的峰值多在18:00;茎中高表达基因的峰值多在12:00;施氮后,多数基因的表达峰值被延后,有些基因的表达峰值升高、降低或提前。不同施氮处理对菠萝植株的生长效应不同,MS营养液处理的植株质量增加幅度最大,其次为纯氮处理,无氮MS营养液和清水处理的植株质量增加幅度最小,但前者促进了根系生长;处理后3 d,叶片硝态氮含量增加;处理后6 d,叶片硝态氮同化加强;处理后26 d,叶片进入缺氮状态;绝大多数基因在清水和无氮MS营养液处理后的6 d达到表达峰值,但两者仅共有3个基因;处理后26 d,高表达基因主要分布在30 mmol·L^(-1)氮、无氮MS和清水处理中,三者无共有高表达基因。【结论】硝态氮转运蛋白基因表达受植株氮营养状态调控,AcNRT 1.13、AcNRT 2.1和AcNRT 1.12基因可能与叶片氮吸收有关,AcNRT 1.14、AcNRT 1.21和AcNRT 1.22基因可能与叶片氮再分配有关。

外文摘要：【Objective】Reducing nitrogen use and improving nitrogen use efficiency are two of the central concerns in pineapple cultivation.Nitrate transporters(NRT)play important roles in the absorption,transportation,redistribution and signaling of nitrate and other nutrients,but their roles in nitrate absorption and redistribution in pineapple remain unclear.【Methods】Two experiments were carried out.The first was application of 30 mmol·L^(-1) NH_(4)NO_(3) to pineapple plantlet,and changes of nitrate contents and expressions of 40 nitrate nitrogen transporter genes in pineapple roots,stems and leaves were analyzed every 2 h from 10:00 to 18:00;The second was application of six treatments(30 mmol·L^(-1) NH_(4)NO_(3) ,60 mmol·L^(-1) NH_(4)NO_(3) ,MS nutrient solution containing 60 mmol·L^(-1) NH_(4)NO_(3) ,nitrogen-deficient MS nutrient solution,MS nutrient solution and water)to pineapple plantlet,and changes of nitrate contents and expressions of 40 nitrogen transporter genes in leaves were analyzed in 3rd d,6th d,9th d and 26th d after treatment,and root,stem and leaf weights were analyzed 60th d after treatment.【Results】The nitrate contents of pineapple roots,stems and leaves all showed higher values at both 10:00 and 16:00;2 h after nitrate application(30 mmol·L^(-1) NH_(4)NO_(3) ),the nitrate content in roots increased significantly;nitrate application also increased the nitrate content in leaves at 18:00,while it decreased the nitrate content in stems at 14:00 and 18:00 as well as that in roots at 16:00.Twenty-five NRT genes were highly expressed in roots.Among them,the highest expression levels across all sampling time points appeared at 12:00 for 2 genes that were postponed(AcNRT1.14)to 14:00 or weakened(AcNRT1.12)after nitrate application;expression levels of 13 genes peaked at 14:00 that were delayed to 14:00(AcNRT1.25,AcNRT1.17,AcNRT1.20,AcNRT1.8,AcNRT1.19,AcNRT1.21,AcNRT1.32,AcNRT1.24,AcNRT1.23,AcNRT1.35,AcNRT1.22,AcNRT3.1)or to 16:00(AcNRT1.42)after nitrate application;expression levels of AcNRT1.31 peaked at 18:00,which were advanced to 14:00 after nitrate application;for other genes,the times when the expression peaked were not altered but the expression level was enhanced(AcNRT1.9,AcNRT1.17,AcNRT1.18,AcNRT1.11,AcNRT1.33)or weakened(AcNRT2.3,AcNRT1.35,AcNRT1.38,AcNRT1.12)by nitrate application.Four NRT genes were highly expressed in stems.Among them,expression levels of AcNRT1.7 peaked at 14:00 and were enhanced till 18:00 by nitrate application;expression levels of AcNRT1.17 peaked at 12:00 and 18:00,and both peaks were enhanced by nitrate application;expression levels of AcNRT1.5 and AcNRT1.37 peaked at 18:00 and were weakened by nitrate application.Twelve NRT genes were highly expressed in leaves.After nitrate application,AcNRT1.31,AcNRT1.41,AcNRT1.2,AcNRT2.1 and AcNRT1.36 were up-regulated at 12:00;AcNRT1.34 was up-regulated at 14:00;AcNRT1.27 and AcNRT1.10 were up-regulated at 18:00;AcNRT1.44 were up-regulated at both 12:00 and 18:00;AcNRT1.38 were up-regulated at both 12:00 and 14:00;AcNRT1.26 were up-regulated at 12:00,16:00 and 18:00;AcNRT1.4 was down-regulated at 18:00.The six different nitrogen fertilization treatments had different effects on the growth of pineapple plantlet.The MS nutrient solution treatment had the largest weight gain,followed by the pure nitrogen treatment(30 and 60 mmol·L^(-1) NH_(4)NO_(3) ).The plant weight gain of treatment with nitrogen-deficient MS nutrient solution and water was the least,but the former promoted root growth.At 3rd d after treatments,except for the treatment of water(control),the nitrate content in leaves of pineapple plantlet with all other treatments increased.At 6th d after treatments,except for the treatment of MS nutrient solution containing 60 mmol·L^(-1) NH_(4)NO_(3) ,the nitrate content in leaves of plants with all other treatments decreased,indicating that the assimilation of nitrate was enhanced.At 9th d after treatment,except for the treatments of pure nitrogen(30 and 60 mmol·L^(-1) NH_(4)NO_(3)),the nitrate content in leaves of plants with all other treatments decreased,indicating that plants treated with pure nitrogen-initiated accumulation of nitrate.At 26th d after the treatment,the nitrate content of leaves with all treatments decreased significantly,indicating that the leaves were undergoing a nitrogen-deficient status.Three days after treatment,only a few genes were up-regulated by the treatments(e.g.AcNRT 1.2,AcNRT 1.43 and AcNRT 1.3 by treatment of nitrogen-deficient MS nutrient solution;AcNRT 1.18,AcNRT1.5 and AcNRT 1.37 by water control),and there was no common up-regulated gene among all nitrogen-containing treatments;most genes reached their peak expression at 6th day in leaves with water control(AcNRT 1.17,AcNRT 1.42,AcNRT 1.20,AcNRT 1.34,AcNRT 1.41,AcNRT 1.44,AcNRT 1.13,AcNRT 1.30,AcNRT 1.38,AcNRT 2.1,AcNRT1.43,AcNRT 3.1,AcNRT 1.24,AcNRT 2.3,AcNRT1.25,AcNRT 1.23,AcNRT 1.31,AcNRT 1.37,AcNRT 1.8,AcNRT 1.9 and AcNRT 1.12)and nitrogen-deficient MS nutrient solution treatment(AcNRT 1.7,AcNRT 2.1,AcNRT 1.33,AcNRT1.36,AcNRT 1.12,AcNRT 1.2),but only 3 genes were shared by both;6 days after treatment,8 genes(AcNRT 1.11,AcNRT 1.19,AcNRT 1.24,AcNRT 2.3,AcNRT 1.25,AcNRT 1.37,AcNRT 1.8 and AcNRT 1.32)were highly expressed in leaves of plants treated with 30 mmol·L^(-1) NH_(4)NO_(3) but not in those treated with 60 mmol·L^(-1) NH_(4)NO_(3);Except for AcNRT 1.11,AcNRT 1.19 and AcNRT 1.32,these genes were also highly expressed in those with water control.26th day after treatment,the highly expressed genes were mainly found with 30 mmol·L^(-1) NH_(4)NO_(3) treatment(AcNRT 1.5,AcNRT 1.11,AcNRT 1.19 and AcNRT 1.35),nitrogen-deficient MS treatment(AcNRT 1.14,AcNRT 1.21 and AcNRT 1.22)and water control(AcNRT 1.6 and AcNRT 1.10),but no genes were shared by these three treatments.【Conclusion】The contents of nitrate and the expressions of nitrate transporter genes in various tissues of pineapple plantlet showed diurnal variation patterns,and nitrate application could change the time when the peaks of these two events occurred;the expression of nitrate transporter genes was affected by the nitrogen status of the plant.AcNRT 1.13,AcNRT 2.1 and AcNRT 1.12 genes may be related to leaf nitrogen uptake,and AcNRT 1.14,AcNRT 1.21 and AcNRT 1.22 genes may be related to leaf nitrogen redistribution.