• ISSN 1008-505X
  • CN 11-3996/S

盐及盐碱胁迫对葡萄砧木幼苗氮素代谢关键酶活性及相关基因表达的差异影响

Disparate impact of salt stress and saline-alkali stress on key nitrogen metabolism enzyme activities and related gene expression in grape rootstock seedlings

  • 摘要:
    目的 探究盐胁迫与盐碱复合胁迫对葡萄砧木幼苗氮代谢关键酶活性以及相关基因表达的差异影响。
    方法 沙培试验以‘5BB’葡萄砧木幼苗为材料,设置3个营养液处理,分别为对照(不添加盐、碱),盐胁迫(0.20 mol/L NaCl溶液)和盐碱复合胁迫(0.10 mol/L NaCl溶液+0.10 mol/L NaHCO3溶液)。幼苗处理14天时,取样测定干鲜重、根系形态、SPAD值、钠钾离子含量、氮代谢关键酶活性、硝态氮、铵态氮和总氮含量,分析15N利用率和氮吸收同化相关基因的表达量。
    结果 与对照相比,盐碱复合胁迫显著降低了葡萄砧木幼苗总鲜重和SPAD值。盐胁迫与盐碱复合胁迫叶片中Na+含量分别是对照的1.50、2.50倍,根系中分别是对照的1.68、1.79倍,且根中Na+含量高于叶片中Na+含量。盐碱复合胁迫处理葡萄叶片的K+含量显著高于对照,又显著高于盐胁迫处理,对照根中K+含量显著高于盐胁迫和盐碱复合胁迫。盐胁迫和盐碱复合胁迫处理降低了叶片和根部氮代谢关键酶硝酸还原酶、亚硝酸还原酶、谷氨酸合成酶、谷氨酰胺合成酶活性,盐胁迫和盐碱复合胁迫下叶片和根部NO3含量、根部NH4+含量、叶片全氮含量及盐碱复合胁迫叶片NH4+含量、根系全氮含量均显著降低。盐胁迫和盐碱复合胁迫下调了叶片和根系中9个氮代谢相关基因的表达量,其中,盐胁迫和盐碱复合胁迫处理叶片VvNIA 基因的表达水平分别较对照降低了68.61%、92.55%,VvNIR基因的表达水平分别降低了66.26%、81.61%,盐碱复合胁迫处理的降幅大于盐胁迫;盐胁迫和盐碱复合胁迫下葡萄叶片的VvNRT1.1VvNRT1.2VvNRT2.1VvNRT2.2 表达量显著下降,且盐胁迫和盐碱胁迫间4个基因的表达量差异显著;盐碱复合胁迫下的葡萄根系VvNRT1.1VvNRT1.2表达水平以及盐胁迫与盐碱复合胁迫下根系VvNRT2.1VvNRT2.2表达水平均显著下降。 K15NO3同位素标记试验结果表明,盐胁迫和盐碱复合胁迫条件下葡萄砧木幼苗15N利用率分别较CK显著降低了63.83%、75.95%。
    结论 盐胁迫和盐碱复合胁迫降低叶片和根系中氮代谢关键酶活性,下调氮吸收同化相关基因的表达水平,减少叶片和根系中NO3、NH4+及总氮的含量,影响葡萄砧木幼苗的15N利用率和葡萄的生长发育。盐碱复合胁迫对葡萄幼苗氮素代谢的不利影响,以及对葡萄幼苗生长的抑制作用甚于盐胁迫。

     

    Abstract:
    Objectives We investigated the differences in the effects of salt stress and saline-alkali stress on the activities of nitrogen metabolism enzymes, and the expression of genes related to nitrogen absorption and assimilation in grape root stock seedlings.
    Methods Sand culture experiment was conducted using ‘5BB’ rootstock seedlings as materials, which were under three treatments for 14 days, CK (normal nutrient solution), salt stress (nutrient solution containing 0.20 mol/L NaCl), and saline-alkali stress (nutrient solution containing 0.10 mol/L NaCl & 0.10 mol/L NaHCO3). Plants were sampled for the measurement of dry and fresh weight, root morphology, SPAD value, Na+ and K+ content, activities of five nitrogen metabolism enzymes, 15N content, and the expression levels of nine genes.
    Results Compared with CK, saline-alkali stress significantly reduced the fresh weight of whole plants, and markedly decreased the SPAD values. Salt and saline-alkali stress increased Na+ content to 1.50 and 2.50 fold in leaves, and 1.68 and 1.79 fold in roots, compared to CK, and were recorded higher Na+ content in roots than in leaves. Saline-alkali stress was tested the highest leaf K+ content, while CK was tested the highest root K+ content. Compare with CK, salt and saline-alkali stress decreased the activities of enzymes in leaves and roots, and decreased NO3, NH4+ and total nitrogen content in different extent. Compare with CK, salt and saline-alkali stress decreased the NO3 and total N contents in leaves and roots, saline-alkali decreased the NH4+ contents in leaves, salt and saline-alkali stress decreased the NH4+ contents in roots. Salt and saline-alkali stress down-regulated the expression levels of nine nitrogen metabolism-related genes in leaves and roots. Compared with CK, the expression levels of leaf VvNIA decreased by 68.61%, 92.55%, and VvNIR decreased by 66.26%, 81.61%, respectively; both salt and saline-alkali stress decreased the expression levels of VvNRT1.1, VvNRT1.2, VvNRT2.1 and VvNRT2.2 in leaves, and the difference in the expression levels of the four genes was significant between salt and saline-alkali stress. Saline-alkali stress down-regulated the expression levels of VvNRT1.1 and VvNRT1.2 in roots, and salt and saline-alkali stress down-regulated the expression levels of VvNRT2.1 and VvNRT2.2 in roots. The result of K15NO3 isotope labeling showed that 15N use efficiency of grape rootstock seedlings under salt and saline-alkali stress were decreased by 63.83% and 75.95%, respectively.
    Conclusions Under salt and saline-alkali stress, grape seedlings would reduce the activities of key nitrogen metabolism enzymes in leaves and roots, down-regulate the expression levels of genes related to nitrogen absorption and assimilation to some extent, thus reduce the NO3, NH4+ and total N content in leaves and roots, and result in low 15N use efficiency and growth of grape root stock seedlings. The adverse effect of saline-alkali stress is worse than salt stress on grape root stock seedlings.

     

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