Transgenic overexpression of Leucaena β-carbonic anhydrases in tobacco does not affect carbon assimilation and overall biomass

The role of β-carbonic anhydrases (CAs) in C₃ plant carbon assimilation is not clear. In this study, the primary role of C₃ plant β-CAs in carbon assimilation was investigated for which, a chloroplastic β-CA gene (cacp) and a cytoplasmic β-CA gene (cacyt) from a C₃ tree-legume of tropics, Leucaena leucocephala (leucaena) were overexpressed in Nicotiana tabacum (tobacco). The cacp and cacyt β-CA isoforms from leucaena were overexpressed separately and also together in tobacco resulting in three types of transgenic tobacco plants (i) expressing cacp only (ii) expressing cacyt only and (iii) co-expressing both cacp and cacyt. These transgenic plants exhibited significantly higher activity of β-CAs as compared with wild-type plants. The percent increase in the CA activity of transgenic plants expressing leucaena cacyt or cacp was found to be ～51 and ～55%, respectively. The transgenic tobacco expressing both the leucaena β-CA isoforms exhibited ～63% increase in CA activity as compared with the wild-type. However, despite notable increase in the CA activity of transgenic tobacco plants, no difference was observed in their phenotype, chlorophyll content and the overall dry weight compared with that of wild-type suggesting that C₃ β-CAs are not involved in active accumulation of inorganic carbon.     

Modification of carbonic anhydrase activity by antisense and over-expression constructs in transgenic tobacco

The activity and location of carbonic anhydrase has been modified by transformation of tobacco with antisense and over-expression constructs. Antisense expression resulted in the inhibition of up to 99% of carbonic anhydrase activity but had no significant impact on net CO₂ assimilation. Stomatal conductance and susceptibility to water stress appeared to increase in response to the decline in carbonic anhydrase activity. An over-expression construct designed to increase cytosolic carbonic anhydrase abundance resulted in a significant increase in net activity, a small increase in stomatal conductance but little impact on CO₂ assimilation. Chloroplastic carbonic anhydrase activity was enhanced by the expression of an additional construct which targeted the polypeptide to the organelle. The increase in chloroplastic carbonic anhydrase appeared to be accompanied by a concomitant increase in Rubisco activity.     

Water bird guilds and their feeding connections in the Bodrogzug, Hungary

Thalli of the intertidal Phaeophyte Fucus spiralis L. and the subtidal Chlorophyte Ulva olivascens Dangeard were exposed to artificial UV-A, UV-B and photosynthetically active radiation (PAR) by combination of PAR + UV-A + UV-B (PAB), PAR + UV-A (PA) and PAR (P) treatments. UV-A enhanced photosynthesis and stimulated carbonic anhydrase (CA) and nitrate reductase (NR) in F. spiralis whilst PAR only had an inhibitory effect in this species. U. olivascens suffered chronic photoinhibition in all the treatments as evidenced by reduced maxima photosynthesis (P_max) and photosynthetic efficiency (α). Non stimulatory effect was observed upon CA and NR in this species. Our results showed that artificial UV radiation triggered opposite responses in both species. We suggest that differences shown by both species might be related to their location in the rocky shore and their ability to sense UV. We propose that the ratio UV:PAR acts as an environmental signal involved in the control of photosynthesis as shown by pronounced inhibition in samples exposed to only PAR. We also suggest that UV-regulated photosynthesis would be related to carbon (C) and nitrogen (N) cycles, regulating feedback processes that control C and N assimilation.     

Studies on photoacoustic uptake signals in tobacco leaves under high carbon dioxide levels

Photoacoustic signals were measured in expanded tobacco leaves, exposed to a controlled atmosphere by being only partly enclosed within the photoacoustic cell. It was aimed to corroborate the conjecture of Reising and Schreiber (Photosynthesis Research 42: 65-73, 1994) that under exceptionally high CO2 levels (ca. 1–5%) the photobaric uptake contribution reflects CO2 uptake induced by light dependent stromal alkalinization. This is shown here by: (1) the shallower damping of the uptake signal vs. the modulation frequency, compared to a normal oxygen evolution signal; (2) the partial inhibition of the uptake signal under 5% CO2 by nigericin; (3) the complete absence of uptake signals under 5% CO2 in a carbonic-anhydrase-deficient mutant, which gave rather a normal oxygen evolution signal. The photoacoustic signals from the wild type and the transgenic tobacco in air could not be distinguished, indicating that the CO2 uptake signal is negligible under this condition. Uptake photobaric signals were also measured in modulated far-red light (ca. 715–750 nm), following addition of white background light (in light limiting intensity). In normal tobacco under 5% CO2, the background light induced an uptake transient, lasting about a minute, then declining to a low steady level. Significantly smaller transients were obtained under normal air, and in the carbonic-anhydrase deficient mutant also under 5% CO2. Extrapolation to zero frequency of the signal damping vs. modulation frequency, in both tobacco genotypes, suggests however similar magnitudes of the uptake transients. On the other hand, no proportional steady-state uptake was observed for the last two cases. Presumably, the steady uptake under 5% CO2 in modulated far-red light reflects CO2 solubilization, while it is an open question whether the transient could be partly contributed also by oxygen photoreduction by PS I (Mehler reaction). It is reasoned that, under conditions of low light, the respiratory activity results in accumulation of CO2 in the photoacoustic cell, which is sufficient to induce an uptake phenomenon, giving a more satisfactory interpretation for the so-called 'low light state' [Cananni and Malkin (1984) Biochim Biophys Acta 766: 525–532].     

土壤干旱对水稻叶片光合速率和碳酸酐酶活性的影响

以适应性广的水稻品种“汕优 6 3”为材料 ,具有较高的光合速率但适应性较差的品系“NAU30 3”为对照 ,比较了不同阶段的叶片光合机构对于干旱处理的适应性。结果表明“汕优 6 3”具有较广泛的适应性 ,在逆境条件下仍能维持较高的光合速率 ,这可能和叶片中碳酸酐酶活性对逆境的响应程度高有关。这种对干旱逆境的适应性反应只在叶片光合功能的可逆衰退阶段发生     

干旱对番茄幼苗光合和某些生理指标的影响

干旱缺水已成为植物光合作用和生长发育主要的限制因素,在干旱胁迫下,作物的生长发育受到影响,依据作物的形态变化进行浇灌属于延后性灌溉,未必能完全补偿对作物生长造成的影响。确定灌溉时间点,既确保植物正常生长不受影响,也可以提高水分利用效率,减少水资源浪费,从而达到节水灌溉的目的。该研究以温室土槽栽培番茄幼苗为材料,设定土壤含水量为30.00%(对照)、21.00%、18.00%、15.00%、12.00%、9.00%,研究了干旱胁迫对番茄叶片光合特性、抗氧化酶(超氧化物歧化酶、过氧化物酶、过氧化氢酶)、碳酸酐酶活性变化的影响,并以此表征番茄幼苗需水信息。结果表明:随着干旱胁迫程度的增加,叶片水势逐渐降低。超氧化物歧化酶、过氧化物酶及过氧化氢酶等抗氧化酶在番茄幼苗耐受水分胁迫中起到重要的作用;超氧化物歧化酶、过氧化物酶在干旱胁迫条件下反应更迅速,但过氧化氢酶相对于超氧化物歧化酶、过氧化物酶对干旱胁迫的耐受能力更强;干旱胁迫条件下抗氧化酶活性的转折点在15.00%土壤含水量左右;水分胁迫条件下碳酸酐酶参与了对光合作用的调节,并在15.00%土壤含水量时活性升至最高,使得番茄仍能维持较高的光合速率,以维持正常的生理机能;随着干旱胁迫程度的加剧(12.00%土壤含水量),碳酸酐酶活性与净光合速率都迅速下降。综上分析,当土壤含水量低于15.00%并高于12.00%时,对作物进行灌溉最为合适。抗氧化酶及碳酸酐酶活性可为作物最佳灌溉时间点的预测提供科学依据。     

Inhibition by cupric ions of the hydration of CO2 catalyzed by carbonic anhydrase II

The inhibition by cupric ions of the hydration of CO₂ catalyzed by carbonic anhydrase II is interesting because of the results of Tuet al. obtained at chemical equilibrium, indicating that Cu²⁺ inhibits specifically a proton transfer in the catalytic pathway. We have measured this inhibition at steady state, using stopped-flow methods. The inhibition by Cu²⁺ of the hydration of CO₂ catalyzed by carbonic anhydrase II had aK_I near 1×10^–6 M atpH 7.0 and gave inhibition that is noncompetitive atpH 6.0 and mixed, but close to uncompetitive, atpH 6.8. ThepH dependence of this binding is consistent with a binding site for Cu²⁺ on the enzyme with apK_a near 7. The binding interaction between Cu²⁺ and the fluorescent inhibitor 5-dimethylaminonaphthalene-l-sulfonamide on carbonic anhydrase II was noncompetitive, indicating that the binding site for Cu²⁺ is distinct from the coordination sphere of zinc in which the actual interconversion of CO₂ and HCO₃^– and the binding of sulfonamides takes place.     

贵州玉舍国家森林公园三种造林植物光合生理特征研究

该研究以贵州省玉舍国家森林公园三种造林植物水榆花楸(Sorbus alnifolia)、近轮叶木姜子(Litsea elongata var.subverticillata)、山杨(Populus davidiana)为对象,测定其光合作用日变化和叶绿素荧光、叶片碳酸酐酶活力以及叶绿素含量和稳定碳同位素组成,并测定了植物生长地土壤理化性质,综合分析了三种植物的喀斯特生态适生能力。结果表明:水榆花楸和山杨的净光合速率明显高于近轮叶木姜子,这与其较高的气孔导度和光化学效率有关;而山杨因具有较高碳酸酐酶活力和叶绿素含量,表现出较高的水分利用效率。水榆花楸和山杨的光合生产能力较高,且受光强和温度限制,可在光照较好的地区大范围种植,作为速生植物来加快经济收入并改造当地脆弱生境的优选植物;近轮叶木姜子作为中药型植物,且在低光强下能够保持最大生长能力,可在光照条件不好的地区优先种植,既增加植被覆盖率又增加经济收入。该研究结果为山地森林资源的保护和开发利用提供了科学依据。     

贵州山区3种木本植物无机碳利用特性的比较

以生长在喀斯特高原地区玉舍国家森林公园内的成熟银鹊树(Tapiscia sinensis)、白栎(Quercus fabri)和亮叶桦(Betula luminifera)为实验材料,通过对光合作用、叶绿素含量、叶绿素荧光参数、羧化效率(CE)、呼吸速率(Resp)、碳酸酐酶活性(WA)以及稳定碳同位素组成(δ13 C)等指标的测定,分析3种植物不同的无机碳利用特性,为该区生态环境修复选择合适的建群植物种提供依据。结果显示:(1)银鹊树、白栎和亮叶桦分布都较为广泛,银鹊树生长的最佳土壤pH是4.5~5.5,而白栎更倾向于中性到弱酸性土壤,肥沃的酸性土壤则更有益亮叶桦生长;白栎和亮叶桦都能忍受干旱和贫瘠,但是银鹊树不能忍受干旱和高温。(2)银鹊树叶片的Pn、Tr和Gs显著大于白栎和亮叶桦,亮叶桦和白栎的Pn、Tr和Gs分别是银鹊树的69.5%、48.2%、66.7%和28.6%、21.7%、22.2%;亮叶桦叶绿素含量均为银鹊树和白栎的2倍,但3种植物间的WUE则无显著差异。(3)3个树种叶片净光合速率均随着CO2浓度升高呈持续上升的趋势,但它们之间的CO2补偿点和饱和点明显不同。其中,银鹊树和亮叶桦的CO2补偿点均低于50μmol·mol-1,而白栎的则在250~300μmol·mol-1之间;银鹊树的CO2饱和点在1 200μmol·mol-1左右,亮叶桦则在2 300μmol·mol-1左右,而白栎的CO2饱和点明显高于2 300μmol·mol-1。(4)3个树种的CE、Resp和WA均为银鹊树>亮叶桦>白栎;而δ13 C值则以银鹊树最低,亮叶桦和白栎较高。其中,白栎和亮叶桦的CE、Resp、WA分别为银鹊树的5.1%、25.7%、4.0%和45.3%、54.6%、6.8%,且树种间差异显著;白栎和亮叶桦的δ13 C值显著高于银鹊树。研究表明,银鹊树能够吸收大气中的CO2或者在高活性碳酸酐酶作用下转化利用细胞内的HCO3-,它拥有较高的CO2利用能力及无机碳同化效率,因而能够拥有较高的产能;亮叶桦只能获取大气中的CO2作为无机碳源,但它对CO2的利用能力也较高,其产能仅次于银鹊树;白栎同样只能获取大气中的CO2作为无机碳源,同时它对大气中CO2的捕获、利用能力均低于银鹊树和亮叶桦,因而白栎生长非常缓慢,造成其本身对无机碳的需求也最低,所以其产能最低。