Peculiar properties of the PsaF photosystem I protein from the green alga Chlamydomonas reinhardtii: presequence independent import of the PsaF protein into both chloroplasts and mitochondria

It has previously been shown that presequences of nuclear-encoded chloroplast proteins from the green alga Chlamydomonas reinhardtii contain a region that may form an amphiphilic -helix, a structure characteristic of mitochondrial presequences. We have tested two precursors of chloroplast proteins (the PsaF and PsaK photosystem I subunits) from C. reinhardtii for the ability to be imported into spinach leaf mitochondria in vitro. Both precursors bound to spinach mitochondria. The PsaF protein was converted into a protease-protected form with high efficiency in a membrane potential-dependent manner, indicating that the protein had been imported, whereas the PsaK protein was not protease protected. The protease protection of PsaF was not inhibited by a synthetic peptide derived from the presequence of the N. plumbaginifolia mitochondrial F₁ subunit. Furthermore, if the presequence of PsaF was truncated or deleted by in vitro mutagenesis, the protein was still protease-protected with approximately the same efficiency as the full-length precursor. These results indicate that PsaF can be imported by spinach mitochondria in a presequence-independent manner. However, even in the absence of the presequence, this process was membrane potential-dependent. Interestingly, the presequence-truncated PsaF proteins were also protease-protected upon incubation with C. reinhardtii chloroplasts. Our results indicate that the C. reinhardtii chloroplast PsaF protein has peculiar properties and may be imported not only into chloroplasts but also into higher-plant mitochondria. This finding indicates that additional control mechanisms in the cytosol that are independent of the presequence are required to achieve sorting between chloroplasts and mitochondria in vivo.Abbreviations cTP chloroplast transit peptide - mTP mitochondrial targeting peptide - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase - pF₁(1,25) a synthetic peptide derived from the first 25 residues of the Nicotiana plumbaginifolia mitochondrial ATP synthase F₁ subunit - PsaF(2–30) and PsaF(2–61) mutant proteins lacking regions corresponding to residues 2–30 and 2–61 in the PsaF precursor protein, respectively     

A comparison of low temperature growth vs low temperature shifts to induce resistance to photoinhibition in spinach (Spinacia oleracea)

Plants of Spinacia oleracea L. cv. Savoy grown under cold-hardening (5°C) and nonhardening (16°C) conditions were exposed to a photoinhibitory irradiance of 1300 μmol rrr^: m-² S-¹ 5°C for 12 h. Plants grown at 5°C exhibited a greater resistance to photoinhibition at low temperature in comparison to plants grown at 16°C as measured by the photochemical efficiency of photosyslem II. In contrast, tuily expanded leaves of plants grown at 16°C and then shifted to 5°C for 10 days did not exhibit increased resistance to photoinhibition. This was observed irrespective of the phoioperiod experienced during the shift to a lower temperature. Furthermore, spinach grown at 16°C and subsequently exposed to a stepped, daily decrease in temperature from 16 to 1°C over 10 days w ith a concomitant reduction in photoperiod. also did not exhibit any change in susceptibility to photoinhibition. Thus, a decrease in photoperiod accompanied by either an abrupt or stepped low temperature shift cannot induce increased resistance to photoinhibition. This confirms the hypothesis that growth and development at cold-hardening temperature are absolute requirements for the acquisition of resistance to photoinhibition at low temperature.     

Removal of nuclear contaminants and of non-specifically photosystem II-bound copper from photosystem II preparations

In conventional photosystem II preparation high amounts of Cu are found. After fractionation by centrifugation, Cu can be completely removed from photosystem II without affecting either its photosynthetic activity or the composition of its specific proteins. We could demonstrate that the Cu was associated with nuclear contaminants in the starch fraction. Among the contaminants, several histones were identified by specific antisera and by N-terminal sequencing. In order to obtain homogeneous BBY preparations of PSII a procedure is employed that involves a 10000 g centrifugation step and which eliminates non-specifically bound metals, nucleic acids and histones with the starch pellet. The resulting starch-free BBY (BBY_s-), which is free of these nuclear contaminants, is an appropriate preparation for biophysical studies or for those of metal interactions with PSII.     

Primary and secondary induction requirements for flowering of Festuca rubra

Root and shoot temperatures were varied independently to determine the importance of root temperature during cold acclimation. Spinach ( Spinacia oleracea L. cvs Harbin and Bloomsdale) plants were subjected to 20/20°C. 20/5°C, 5/20°C, and 5/5°C (shoot/root) temperature treatments. Leaf freezing tolerance, water potential, stomatal resistance, osmotic potential, and water content were measured at 0.25. 1.25, 3.25, and 7.25 days of treatment. There was no change in freezing tolerance or the water relations of the 20/20°C treated plants during the course of the experiment. Freezing tolerance was increased by the 5°C shoot temperature treatments, but was not enhanced by water stress induced by the low root temperature. Leaf water potential and water content decreased and stomatal resistance increased within 6 h in the 20/5°C plants. By day 3, osmotic potential began decreasing in the 20/5°C plants. Leaf water content, osmotic potential, and water potential decreased more gradually in plants grown with 5°C shoot temperature, irrespective of root temperature. Decreased water content and osmotic potential were not correlated with increased freezing tolerance as reported for other herbaceous crop plants.     

Soil pollution through industrial effluent and its management

During the survey of sewer water/industrial effluent composition, we identified a site at Sonepat that had turned barren due to excessive irrigation with cycle industry effluent. To study the ameliorative effect of farmyard manure, the bulk surface soil sample was brought from the site. Soil was amended with five levels of farmyard manure (0, 0.25, 0.5, 1.0, and 2.0% on a soil weight basis), and carrot, fenugreek, spinach, and wheat crops were grown as test crops in a screen house. The deleterious effect of excessive heavy metals, particularly Ni, on the yield of all the crops was reduced with the application of 2% farmyard manure. The Ni content was highest in carrot, followed by spinach, fenugreek, and wheat. With the application of 2% farmyard manure, Ni content was reduced from 434 to 267 mg/kg in carrot, 167 to 100 mg/kg in fenugreek, 300 to 166 mg/kg in spinach, and 65 to 42 mg/kg in wheat grain.     

Interaction of electron acceptors with thylakoids from halophytic and non-halophytic species

Thylakoid membranes isolated from halophytic species showed differences in their interactions with ionic and lipophilic electron acceptors when compared to thylakoids from non-halophytes. FeCN was considerably less efficient as electron acceptor with halophyte thylakoids, supporting much lower rates of O₂ evolution and having a lower affinity. FeCN accepted electrons at a different, DMMIB insensitive, site with these thylakoids. 1,4-Benzo-quinones with less positive midpoint potentials were less effective in accepting electrons from halophyte thylakoids compared to nonhalophyte thylakoids, also reflected in lower rates of O₂ evolution and lower affinity. Considering the lipolphilic nature and the fact that there was no apparent change in the site donating electrons to the quinones, an alteration in the midpoint potential of this site by about +100mV is postulated for the halophyte thylakoids.Abbreviations AMPD 2-amino-2-methyl-1,3-propanediol - Cyt b₆/f cytochrome b₆/f complex - DBMIB 2,5-dibromo-6-isopropyl-3-methyl-1,4-benzoquinone - DCBQ 2,6-dichloro-1,4-benzoquinone - DCIP 2,6-dichlorophenol-indolphenol - DMBQ 2,5-dimethyl-1,4-benzoquinone - E_m7 midpoint redox potential at pH 7.0, FeCN-K₃Fe(CN)₆ - HNQ 5-hydroxy-1,4-naphthoquinone - MV methylviologen - NQ 1,4-naphthoquinone - PBQ phenyl-1,4-benzoquinone - PC plastocyanin - PQ plastoquinone     

菠菜和菜豆对二氧化氮的反应和抗性

比较了菠菜和菜豆在光下或暗中接触不同浓度NO_2时的反应。菠菜的抗性强于菜豆,是由于它既能忍受NO_2~-的大量积累,又有代谢NO_2~-的较强能力。菠菜在光照下,高浓度NO_2熏气所产生的伤害不是由于NO_2~-的积累,而是与NH_3的大量积累有关。NH_3的积累:一方面由于NiR活性不受熏气影响,另一方面由于GS/GOGAT活性受到阻抑所致。     

不同硝酸盐含量菠菜品种的生理差异分析

采用营养液栽培方式,以4个硝酸盐含量有显著差异的菠菜品种为材料,测定其生长指标（株高、主根长、株幅、柄长、干鲜重和叶面积）、光合色素（叶绿素a、b及类胡萝卜素）含量、光合作用参数(实际光化学量子产率、表观光合电子传递率、光化学猝灭系数、非光化学猝灭系数)、硝酸根吸收速率和硝酸还原酶活性等,研究不同硝酸盐含量基因型品种的生理差异。结果显示：（1）不同基因型菠菜叶片的硝酸盐含量始终存在着巨大差异,且硝酸盐含量与生物量之间没有明显的相关关系,在同样的栽培条件下,生物量大的品种并不代表其硝酸盐含量低。（2）高硝酸盐含量基因型菠菜品种的光合色素含量较低,光合作用效率较差,硝酸吸收速率较高,硝酸还原酶活性较低;而低硝酸盐含量基因型菠菜品种的光合色素含量较高,光合作用较强,硝酸吸收速率较低,硝酸还原酶活性较高。（3）低硝酸盐含量基因型品种SL 06 13的综合评价最好,可以作为选育低硝酸含量菠菜品种的材料。研究表明,菠菜硝酸盐含量的基因型差异产生的原因可能是受硝酸盐吸收和转运的共同影响。     

Effect of some heterocyclic herbicides on CO2 assimilation by spinach chloroplasts

We have studied the inhibitory effect of heterocyclic herbicides simazine, paraquat, pyrazon and amitrole on photosynthetic CO₂ fixation and on the level of intermediates of the CO₂ assimilation cycle in isolated spinach (Spinacia oleracea) chloroplasts, as well as their in vitro activities on ribulose-1,5-bisphosphate carboxylase and fructose-1,6-bisphosphatase. The half inhibitory concentrations (I₅₀) of CO₂ assimilation were about 1 μM for simazine and paraquat, and 10 μM for pyrazon. Amitrole, with an I₅₀ 100 μM, gave only a weak inhibition. In the presence of simazine or pyrazon the triose-phosphates/phosphoglycerate ratio diminished because of a decrease of the triose-phosphates percentage from 47% to 19%, which means an inhibition of the phosphoglycerate reduction step by a low NADPH synthesis. However, there was not a parallel increase of phosphoglycerate, because of collateral pathways leading to phospho-enolpyruvate, amino acids and other non-identified compounds. Paraquat did not give such a decreased ratio, which could be explained as an inhibition of some step of the Calvin cycle later than triose-phosphates by the H₂O₂ formed in a Mehler reaction. Amitrole did not show any effect on the pattern of intermediates. Simazine and pyrazon at 10 μM concentration promote a 20–30% activation of ribulose-1,5-bisphosphate carboxylase activity, whereas paraquat, pyrazon and simazine showed an I₅₀ about 100 μM for the inhibition of the photosynthetic fructose-1,6-bisphosphatase.