不同氮素来源对烤烟上部叶碳氮代谢及其基因表达的影响
Influence of different nitrogen sources on carbon and nitrogen metabolism and gene expression in tobacco upper leaves
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摘要: 目的研究不同氮素来源(有机氮肥和无机氮肥)对烤烟上部烟叶碳氮代谢及关键基因表达的影响,为进一步调控肥料施用、提高烟叶品质提供理论依据.方法在保证用氮量一致的基础上,设无机氮肥和有机氮肥两个处理,烟苗移栽后60、70、80和90 d各采样1次,观察不同处理的细胞超微结构,测定烟叶的总糖、总氮和烟碱含量,并对比分析影响碳氮代谢的关键基因表达.结果随着移栽时间的延长,有机氮肥处理上部烟叶细胞中的淀粉颗粒增多,嗜锇颗粒含量增加且体积增大.两种氮肥处理成熟期上部烟叶总糖含量均呈先降低再升高的变化趋势,其中有机氮肥处理上部烟叶的总糖含量相对较高;两处理上部烟叶总氮含量随着生育期的延长呈下降趋势,有机氮肥处理在成熟前期显著低于无机氮肥处理(P<0.05,下同),但移栽后90 d二者趋于一致;两处理上部烟叶烟碱含量在成熟期呈上升趋势,有机氮肥处理的烟碱含量除移栽后80 d显著较低外,其他取样时期均高于无机氮肥处理.碳代谢途径关键基因表达分析结果显示,蔗糖合成酶基因(SuS1)、淀粉合成酶基因(SS1)、颗粒结合型淀粉合成酶基因(GBSSI)、支链淀粉合成酶基因(SBE1)和α-淀粉酶基因(AMML)在成熟后期(移栽后90 d)高表达是导致上部烟叶总糖积累较多的主要原因.氮代谢途径关键基因表达分析结果显示,硝酸盐转运蛋白基因(NRT1)和鸟氨酸脱羧酶基因(ODC)在有机氮源条件下高表达是导致上部烟叶烟碱含量显著升高的主要原因.结论施用有机氮肥有利于成熟期上部烟叶碳氮代谢途径关键基因的表达,从而促进上部烟叶总糖和烟碱含量增加,同时降低总氮含量.Abstract: ObjectiveEffects of different nitrogen sources(organic nitrogen fertilizer and inorganic nitrogen fertili-zer)on carbon and nitrogen metabolism and key gene expression in the upper leaves of flue-cured tobacco were investiga-ted to provide theoretical basis for further regulating the application of fertilizer and improving the quality of tobacco leaves.MethodOrganic nitrogen fertilization and inorganic nitrogen fertilization were set as two treatments on the basis of the same nitrogen amount.The tobacco seedlings were sampled 60,70,80 and 90 d after transplanting.The ultrastruc-ture of the cells in different treatments were observed,and contents of total sugar,total nitrogen and nicotine in tobacco leaves were determined,then expression of key genes affecting carbon and nitrogen metabolism were analyzed.ResultThe starch granules treated by organic nitrogen fertilizer increased in the tobaccoupper leaf cells,the content and volume of osmiophilic granules also increased with the prolongation of transplanting time.Total sugar content in the two nitrogen fertilizer treatments decreased first and then increased in the upper leaves at mature period,and the one treated with or-ganic nitrogen fertilizer was higher than the other. The total nitrogen content in the two treatments decreased with the prolongation of growth period. Compared with the inorganic nitrogen fertilizer treatment,total nitrogen content in the treatment of organic nitrogen fertilizer was significantly lower(P<0.05,the same below)at the mature period.However, the total nitrogen contents in the two treatments tended to be consistent 90 d after transplanting.The nicotine content in the two nitrogen treatments showed an upward trend at mature period.Compared with the inorganic nitrogen fertilizer treat-ment,the nicotine content treated by organic nitrogen fertilizer was significantly lower 80 d after transplanting,but the nicotine content at other sampling periods were all higher than the other treatment.The analysis for key gene expression in carbon metabolism pathway showed that sucrose synthase gene(SuS1),starch synthase gene(SS1),granule bound starch synthase gene(GBSSI),amylopectin synthase gene(SBE1)and alpha amylase gene(AMML)were highly expressed,and this was the main reason for the large amount of accumulation of total sugar in tobacco upper leaves.The analysis for key gene expression in nitrogen metabolism pathway showed that the high expression level of nitrate transporter gene(NRT1) and ornithine decarboxylase gene(ODC)under organic nitrogen source was the main reason for the increase of nicotine content in upper leaves.ConclusionThe application of organic nitrogen fertilizer is beneficial for the expression of key genes in carbon and nitrogen metabolism pathway of tobacco upper leaf at mature stage,which may increasethe content of total sugar and nicotine in tobacco upper leaf and decrease the total nitrogen content.