Aluminum tolerance associated with enhancement of plasma membrane H+-ATPase in the root apex of soybean

Abstract

Seventeen soybean cultivars were screened to discern differences in aluminum (Al) sensitivity. The Sowon (Al-tolerant) and Poongsan (Al-sensitive) cultivars were selected for further study by simple growth measurement. Aluminum-induced root growth inhibition was significantly higher in the Poongsan cultivar than in the Sowon cultivar, although the differences depended on the Al concentration (0, 25, 50, 75 or 100?μmol?L^–1) and the amount of exposure (0, 3, 6, 12 or 24?h). Damage occurred preferentially in the root apex. High-sensitivity growth measurements using India ink implicated the central elongation zone located 2–3?mm from the root apex. The Al content was lower 0–5?mm from the root apices in the Sowon cultivar than in the apices of the Poongsan cultivar when exposed to 50?μmol?L^–1 Al for 12?h. Furthermore, the citric acid exudation rate was more than twofold higher in the Sowon cultivar. Protein production of plasma membrane (PM) H⁺-ATPase from the root apices (0–5?mm) was upregulated in the presence of Al for 24?h in both cultivars. This activity, however, decreased in both cultivars treated with Al and the Poongsan cultivar was more severely affected. We propose that Al-induced growth inhibition is correlated with changes in PM H⁺-ATPase activity, which is linked to the exudation of citric acid in the root apex.     

水稻根系细胞膜H+-ATPase对缺磷的反应

用两相法分离了供磷（+P）和缺磷（-P）营养下水稻苗期根系的细胞膜,并测定了细胞膜上H+-ATPase的水解活性,以期阐明水稻根系细胞质膜上H+-ATPase对不同缺磷的反应机制。结果表明,缺磷的水稻根系细胞膜H+-ATPase的水解活性和H+-ATPase的Vmax, Km均低于正常供磷的植物;缺磷的水稻根系细胞膜H+-ATPase最佳pH值为 6.0,而正常供磷植物的为pH 6.4左右;Western Blot结果说明,缺磷水稻根系细胞膜H+-ATPase酶浓度与正常供磷植物相似。本试验结果还说明,缺磷水稻根系细胞膜H+-ATPase活性低的原因并不是因为其单位细胞膜上的H+-ATPase酶分子数量小于正常供磷的植物,而是缺磷水稻根系细胞膜上H+-ATPase的同工酶的组成供磷植物相比发生了变化。这很可能是缺磷胁迫下水稻根系细胞膜H+-ATPase的一种适应机制。     

铵态氮营养下水稻根系分泌氢离子与细胞膜电位及质子泵的关系

为了研究水稻在铵态氮营养下分泌氢离子的机理,采用不同浓度的铵态氮（0.1～1.0 mol/L）处理水稻幼苗根系,4h后用1 mol/L NaOH滴定培养液,计算氢离子的分泌量;同时,将水稻根系用多聚糖PEG-DEXTRAN两相系统分离出细胞膜囊体,并测定细胞膜H+-ATPase的水解活性和质子泵活性。另外,利用毛细管微电极测定水稻根细胞在上述不同铵浓度下膜电位的变化,以阐明水稻根系吸收铵态氮后分泌氢离子与细胞膜电位及细胞膜质子泵之间的关系。结果表明,随着培养液中铵离子浓度的升高,根系分泌氢离子的量随之增加;分离细胞膜后,离体细胞膜囊体H+-ATPase的水解活性和质子泵活性也相应增强。原位测定细胞膜电位时,膜电位去极化程度亦随NH4+浓度的升高而升高;氢离子分泌量与细胞膜电位、细胞膜H+-ATPase水解活性及质子泵活性之间的均具有一定的相关性。说明根系在NH4+-N营养下分泌氢离子是由于细胞膜上H+-ATPase主动泵出氢离子造成的,这与根系吸收NH4+后引起细胞膜去极化,需要通过提高质子泵活性来维持膜电位有关。     

小麦根尖细胞壁对铝的吸附/解吸特性及其与耐铝性的关系

研究了耐铝性明显差异的2个小麦基因型西矮麦1号(耐性)和辐84系(敏感)根系对铝毒胁迫的反应与根尖细胞壁组分以及细胞壁对铝的吸附和解吸的关系。结果表明,30mol/L.AlCl3可迅速抑制小麦根系伸长,但对辐84系根系伸长的抑制更为明显,且小麦根系相对伸长率随着铝浓度的提高而急剧降低。在30mol/L.AlCl3处理24h后,西矮麦1号根系伸长的抑制率为33.3%,而辐84系根系伸长的抑制率高达70.9%。小麦距根尖0～10.mm根段的铝含量和细胞壁中果胶糖醛酸含量显著高于10～20.mm根段,且前者对铝的累积吸附量明显大于后者;在0～10.mm根段,敏感基因型果胶含量高于耐性基因型,其根尖含铝量及根尖细胞壁对铝的吸附量都要大于后者。采用1.0.mol/L.NH3.H2O对细胞壁预处理2.h降低果胶甲基酯化程度后,耐性和敏感基因型根尖细胞壁对铝的累积吸附量分别降低了17.1%和20.9%,但对铝的累积解吸率没有影响。由此可见,小麦根尖是铝毒的主要位点,细胞壁果胶含量和果胶甲基酯化程度可能是导致不同小麦基因型根尖细胞壁对铝吸附量、铝积累量的差异及其对铝毒胁迫反应的差异的重要原因。     

Higher ROS scavenging ability and plasma membrane H+‐ATPase activity are associated with potassium retention in drought tolerant tea plants

Drought is one of the main limiting factors resulting in the reduction of yield and quality of tea production. Our previous studies indicated that K⁺ retention was a key factor associated with alleviating drought‐induced damage in tea plants. However, the underlying mechanisms are still unclear. In the present study, drought tolerant tea plant (variety Zhongcha108) exhibited higher plasma membrane (PM) H⁺‐ATPase activity and lower negative membrane potential compared with the drought sensitive tea plant (variety Ruanzhiwulong). In addition to superior mesophyll K⁺ retention ability and higher PM H⁺‐ATPase activity, the drought tolerant tea plant exhibited lower accumulation of reactive oxygen species (ROS) than the sensitive one. Moreover, exogenous supply of Na₃VO₄ (plasma membrane H⁺‐ATPase inhibitor) and H₂O₂ (mimicking apoplastic ROS) resulted in a significantly higher K⁺ efflux in the mesophyll of the drought sensitive tea plant than the non‐treated one. Taken together, the higher ROS scavenging ability and plasma membrane H⁺‐ATPase activity were major factors contributing to the higher K⁺ retention in drought tolerant tea plants.     

Comparative toxicity of Al3+, Yb3+, and La3+ to root-tip cells differing in tolerance to high Al3+ in terms of ionic potentials of dehydrated trivalent cations

Abstract

Green manure legumes are often used to compare biomass production as well as nitrogen-fixing capacity. Mineral deficiency often limits the symbiotic nitrogen fixation of many legumes, thus limiting their productivity despite their high yielding potential (O’Hara et al. 1988; Flis et al. 1993). Leguminous species require large amounts of P for growth, nodulation, and nitrogen fixation. Consequently, they are often unable to grow in acid soils with low available P. The low P availability in tropical acid soils often arises from fixation of P by Al and Fe in soil. Generally, Al and Fe-phosphates are relatively unavailable to plants (McLachlan 1976; Ae et al. 1990).     

铵、硝营养对水稻叶细胞膜H+-ATPase和质子泵活性的影响

用两相法分离铵态氮(NH4+-N)和硝态氮(NO3--N)培养的水稻苗期叶细胞膜,并测定了细胞膜H+-ATPase水解活性和质子泵活性,以期阐明铵、硝营养对水稻叶细胞膜H+-ATPase的影响。结果表明,叶细胞膜H+-ATPase活性最佳pH值均为6.2。 NO3--N培养的水稻叶细胞膜H+-ATPase的水解活性、Vmax和Km均显著高于NH4+-N培养的水稻叶;Western Blot分析结果看出,NO3--N培养的水稻叶细胞膜H+-ATPase酶浓度也高于NH4+-N培养的水稻叶,说明NO3--N培养的水稻叶中单位细胞膜上的H+-ATPase酶分子数量大于NH4+- N培养的水稻叶,这与细胞膜上H+-ATPase蛋白的表达量升高有关。此外,NO3--N培养的水稻叶质子泵初速度和膜囊体内外H+浓度梯度均高于NH4+- N培养。由于NO3-的跨膜运输是与细胞膜上H+-ATPase紧密联系的主动运输过程,NO3--N培养的水稻叶片细胞膜H+-ATPase活性和质子泵活性高可能与水稻叶细胞吸收大量NO3-有关。     

Real-time imaging of nitrogen fixation in an intact soybean plant with nodules using 13N-labeled nitrogen gas

Real-time images of nitrogen fixation in an intact nodule of hydroponically cultured soybean ( Glycine max [L] Merr.) were obtained. In the present study, we developed a rapid method to produce and purify ¹³N-labeled radioactive nitrogen gas (half life: 9.97 min). ¹³N was produced from a ¹⁶O (p, α) ¹³N nuclear reaction. The target chamber was filled with CO₂ and irradiated for 10 min with protons at an energy of 18.3 MeV and an electric current of 5 μA, which was delivered from a cyclotron. All CO₂ in the collected gas was absorbed and removed with powdered soda-lime in a syringe and replaced with helium gas. The resulting gas was injected into gas chromatography and separated and a 35 mL fraction, including the peak of [¹³N]-nitrogen gas, was collected by monitoring the chromatogram. The obtained gas was mixed with 10 mL of O₂ and 5 mL of N₂ and used in the tracer experiment. The tracer gas was fed into the underground part of intact nodulated soybean plants and serial images of the distribution of ¹³N were obtained non-invasively using a positron-emitting tracer imaging system (PETIS). The rates of nitrogen fixation of the six test plants were estimated to be 0.17 ± 0.10 μmol N₂ h⁻¹ from the PETIS image data. The decreasing rates of assimilated nitrogen were also estimated to be 0.012 ± 0.011 μmol N₂ h⁻¹. In conclusion, we successfully observed nitrogen fixation in soybean plants with nodules non-invasively and quantitatively using [¹³N]N₂ and PETIS.     

水稻根尖边缘细胞对铁毒的形态生理响应

以水稻(Oryza sativa L.)品种Azucena（铁耐性）和IR64（铁敏感）为材料,研究了Fe2+毒胁迫下附着于根尖边缘细胞(即原位边缘细胞)的数目、存活率,根尖细胞形态结构、根尖保护酶活性的变化。结果显示,Fe2+ 毒对根边缘细胞的产生有抑制作用。相对于敏感性品种而言,一定浓度Fe2+(100~200 μmol/L)有利于耐性品种边缘细胞的产生;Fe2+ 毒对边缘细胞有致死效应,随Fe2+浓度的提升,边缘细胞的存活率呈下降趋势,根尖外围细胞壁增厚,并出现细胞程序性死亡特征(敏感性品种)。同时,Fe2+ 毒对根尖保护酶活性有一定的影响,200~400 μmol/L Fe2+处理下,耐性品种POD、CAT、SOD活性都超过对照;敏感品种只有SOD活性超过对照。说明Fe2+毒胁迫下,水稻根尖通过增加边缘细胞数目、提高细胞拒铁作用,维持较高水平的POD、CAT和SOD活性来对抗Fe2+毒,缓解铁毒害。     

Positive correlation between the number of root nodule primordia and seed sugar secretion in soybean (Glycine max L.) seedlings inoculated with a low density of Bradyrhizobium japonicum

Sufficient rhizobium population in the rhizosphere of legume seedlings is required for early and enough setting of root nodules. Potential of seed and seedling root exudates for proliferation of Bradyrhizobium japonicum PNT119 was evaluated in the soybean cvs. Enrei and Tachinagaha. In both cultivars, seed exudates showed a higher potential than root exudates. In the seed exudates, a low-molecular high-polarity fraction including sugars and amino acids showed a higher potential than both high-molecular fraction and low-molecular low-polarity fraction. The correlations between the sugar or amino acid contents of seed exudates and the number of root nodule primordia at 7 d after sowing were investigated among 12 soybean cultivars. When seeds were inoculated with a low rhizobium density, a high correlation coefficient was detected between them. However, there was no positive correlation in the plants inoculated with a high rhizobium density. These results suggest that the amount of low molecular substances in the seed exudates determines the number of root nodule primordia through rhizobium proliferation around seed and young roots when the rhizobium density is the limiting factor. The genetic trait relating to seed coat secretion should be considered as a possible key factor contributing to adequate root nodulation in soybean seedlings cultivated in fields with a low rhizobium density.     

NO-3胁迫下K+、Ca2+对黄瓜幼苗膜质过氧化及活性氧清除酶系统的影响

通过水培试验探讨了NO^-₃胁迫下K⁺、Ca²⁺对黄瓜幼苗膜质过氧化及活性氧清除酶系统的影响。结果表明，在相同NO^-₃浓度胁迫7d后， Ca²⁺浓度越大，膜脂过氧化产物丙二醛(MDA)含量越高，而K⁺浓度越大，电解质相对渗透率越高，由此说明K⁺、Ca²⁺对细胞膜造成伤害的机理不同。黄瓜幼苗活性氧清除酶系统对K⁺、Ca²⁺的响应亦不同，在一定程度上，K⁺和Ca²⁺ 可提高SOD、POD和CAT活性，保护植物免受自由基伤害，继而可增强植物对逆境的适应能力。