Physiology and Expression of PIP of Hippophae rhamnoides under Salt Stress
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摘要:
目的 探讨水通道蛋白（Plasma membrane intrinsic proteins，PIPs）在中国沙棘盐胁迫响应中的作用，为中国沙棘应对盐胁迫机理研究提供参考。 方法 以中国沙棘（Hippophae rhamnoides subsp. sinensis）为试验材料，对中国沙棘质膜内在蛋白基因（HrPIP）序列进行生物信息学分析，并研究不同程度盐胁迫（200、400、600、800、1000 mmol·L−1）对中国沙棘生理指标及HrPIP基因表达模式的影响。 结果 中国沙棘HrPIP基因编码110个氨基酸，其编码蛋白质位于细胞膜上，无信号肽，无跨膜螺旋区，为疏水性蛋白。中国沙棘相对含水量随着盐胁迫的加深逐渐下降；叶绿素含量总体呈先下降后升高的趋势；质膜透性和丙二醛含量在低浓度盐胁迫下变化不大，在高盐浓度下急剧上升，在1000 mmol·L−1时丙二醛含量有所降低而膜透性继续升高。HrPIP基因的表达随着盐胁迫程度的加剧而变化，其在根中的表达量呈先上升后下降再上升的趋势，在叶中呈先下调后急剧上升而后又下降的趋势，在茎中则呈“M”型表达模式。 结论 中国沙棘在盐胁迫下具有一定的抗盐性，胁迫下HrPIP基因的表达量变化促进根的吸水、茎的运输、叶的保水等，从而提高水分吸收，调节自身的耐盐性，说明该基因在中国沙棘应对盐胁迫的过程中起着重要作用。 Abstract:Objective Mechanism of aquaporins relating to the plasma membrane intrinsic proteins (PIPs) of Hippophae rhamnoides subsp. sinensis in response to salt stress was studied. Methods Bioinformatics of HrPIP was analyzed. Physiological indexes and expressions of the gene in tissues of H. rhamnoides under normal and imposed salt stresses of 200, 400, 600, 800, or 1,000 mmol ·L−1 NaCl were determined. Results Located in the cell membrane, HrPIP encoded 110 amino acids. The hydrophobic protein had no signal peptide or transmembrane helix region. As salt stress intensified, the RWC decreased gradually, and the chlorophyll content declined initially followed by an incline. The permeability of the plasma membrane and the content of MDA changed slightly when the salt concentration was low but increased significantly when the concentration was high. The 1000 mmol·L−1 salt stress induced declined MDA but continuously rising membrane permeability. The expressions of HrPIP in different organs varied by the increasing salt stress. In roots, it rose at lower salt concentrations then declined but rose again at high salt levels. In the leaves, the opposite trend was observed, whereas it was in an “M” pattern in the stems. Conclusion H.rhamnoides subsp. sinensis was salt resistant to a certain degree. Under the stress, by altering the HrPIP expression to increase water absorption in roots, transport in stems, and retention in leaves the plant manipulated the cellular salt concentration to achieve an improved stress tolerance. -
Key words:
- Hippophae rhamnoides subsp. sinensis /
- HrPIP /
- salt stress /
- expression pattern
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图 3 HrPIP蛋白二级结构预测
蓝色区域：α-螺旋；红色区域：延伸链；紫色区域：无规则卷曲。
Figure 3. Predicted secondary structure of HrPIP protein
Blue regions: α-helix; red regions: extended strand; purple regions: random coil.
图 4 HrPIP蛋白磷酸化位点预测
红色为丝氨酸磷酸化位点；绿色为苏氨酸磷酸化位点；紫色为酪氨酸磷酸化位点；粉色线为阈值。
Figure 4. Predicted phosphorylation site in HrPIP protein
Red indicates serine phosphorylation site; green, tyrosine phosphorylation site; purple, threonine phosphorylation site; pink line, threshold value.
图 5 盐胁迫下中国沙棘生理指标的变化
不同大写字母表示不同处理在0.01水平差异极显著。
Figure 5. Changes on physiological indexes of H. rhamnoides under salt stress
Different uppercase letters indicate significant difference at 0.01 level.
图 6 HrPIP基因在中国沙棘根茎叶中的表达模式
不同大写字母表示不同胁迫处理下差异极显著（P＜0.01），不同小写字母表示不同组织之间差异显著（P＜0.05）。
Figure 6. Expressions of HrPIP in roots and leaves of H. rhamnoides
Data with different capital letters indicate extremely significant difference under treatments (P＜0.01); those with different lowercase letters, significant difference between tissues (P＜0.05).
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