Spatio-temporal heterogeneity in Arabidopsis thaliana leaves under drought stress

Using chlorophyll (chl) fluorescence imaging, we studied the effect of mild (MiDS), moderate (MoDS) and severe (SDS) drought stress on photosystem II (PSII) photochemistry of 4-week-old Arabidopsis thaliana. Spatio-temporal heterogeneity in all chl fluorescence parameters was maintained throughout water stress. After exposure to drought stress, maximum quantum yield of PSII photochemistry (F(v)/F(m)) and quantum efficiency of PSII photochemistry (Φ(PSΙΙ)) decreased less in the proximal (base) than in the distal (tip) leaf. The chl fluorescence parameter F(v) /F(m) decreased less after MoDS than MiDS. Under MoDS, the antioxidant mechanism of A. thaliana leaves seemed to be sufficient in scavenging reactive oxygen species, as evident by the decreased lipid peroxidation, the more excitation energy dissipated by non-photochemical quenching (NPQ) and decreased excitation pressure (1-q(p)). Arabidopsis leaves appear to function normally under MoDS, but do not seem to have particular metabolic tolerance mechanisms under MiDS and SDS, as revealed by the level of lipid peroxidation and decreased quantum yield for dissipation after down-regulation in PSII (Φ(NPQ)), indicating that energy dissipation by down-regulation did not function and electron transport (ETR) was depressed. The simultaneous increased quantum yield of non-regulated energy dissipation (Φ(NO)) indicated that both the photochemical energy conversion and protective regulatory mechanism were insufficient. The non-uniform photosynthetic pattern under drought stress may reflect different zones of leaf anatomy and mesophyll development. The data demonstrate that the effect of different degrees of drought stress on A. thaliana leaves show spatio-temporal heterogeneity, implying that common single time point or single point leaf analyses are inadequate.     

An insight into functional genomics of transgenic lines of tomato cv Rio Grande harbouring yeast halotolerance genes

Tomato cv Rio Grande plants were transformed with yeast halotolerance genes (HAL I or HAL II) using pPM7HAL I or pJRM16HAL II, with p35GUSINT as control. Transformation efficiency varied in the three constructs, with highest transformation found with p35GUSINT. Final selection of the transgenic plants was made on the basis of PCR. Transgene integration and copy number were assessed by Southern hybridisation. The primary transformants were allowed to self-pollinate and the expected Mendelian ratios were studied in second-generation progeny. Five independent homozygous lines each of HAL I and HAL II, as well as the control, were characterised to study inter-transformant expression variability. The transformants showed considerable variability in expression of the respective genes, as shown by salt tolerance assays, chlorophyll content and peroxidase activity. The transgene expression in transgenic lines was analysed by semi-quantitative RT-PCR. In response to different salt concentrations, transgenic plants over-expressing HAL I and HAL II had significantly (α=0.05) better performance than the control This study presents the comparative responses of the three constructs under the same transformation conditions and suggests possible mechanisms governed by yeast HAL I and HAL II genes, which seem to work in a coordinated manner by relatively decreasing osmotic and oxidative shock at different rates. Our results suggest that the yeast HAL I increases K(+) /Na(+) selectivity and has a more functional role than HAL II in improving salt tolerance of the tomato cv Rio Grande grown in Pakistan.     

强光下高温与干旱胁迫对花生光系统的伤害机制

为探讨高温和干旱胁迫对花生光合系统的不同影响机制,以鲁花14为试材进行高温(42 ℃)强光(1200 μmol · m^-2 · s^-1)(HH)、干旱(PEG6000,30%)强光(1200 μmol · m^-2 · s^-1)(DH)和强光(1200 μmol · m^-2 · s^-1)胁迫(NH)处理,以未处理为对照(CK)的实验。与CK及NH处理相比,HH和DH的最大光化学效率(Fv/Fm)和820 nm光吸收大幅下降,叶绿素荧光动力学曲线上J点相对荧光(Vj)上升,单位面积内吸收的光量子(ABS/CSm)、单位面积内反应中心捕获的光量子(TRo/CSm)和单位面积内有活性的反应中心的数目(RC/CSm)均出现大幅下降,而PSⅡ的关闭程度(1-qP)明显升高,依赖于叶黄素循环的非辐射能量耗散(NPQ)升高,同时超氧化物歧化酶(SOD)活性出现下降,丙二醛(MDA)和膜透性增加,这些结果表明,HH和DH胁迫引起了花生叶片的严重光抑制,但快速叶绿素荧光诱导动力学曲线中均没有出现K点,表明花生叶片光合系统放氧复合体(OEC)对高温和干旱胁迫不敏感,光合系统Ⅱ(PSⅡ)反应中心的受体侧更容易受到高温和干旱的影响,而对花生光系统造成严重破坏的主要原因则是过剩光能的积累,一方面虽然叶黄素循环可以耗散部分能量,但不是全部;另一方面水-水循环受到高温和干旱的影响不能有效起到能量消耗的作用,造成活性氧的大量积累。HH和DH处理对花生光系统造成的伤害相似,但DH处理对花生光系统的伤害程度大一些,强光下,高温和干旱对花生叶片的伤害位点及破坏机制却较为相似。     

夜间低温胁迫对两种生长光强下藤黄幼苗叶片荧光特征和活性氧代谢的影响

对两种不同生长光强下（自然光的8％和50％）西双版纳热带雨林木本植物藤黄（Garcinia han-buryi）幼苗经夜间低温（4℃）处理后荧光特性和活性氧代谢的研究结果表明,低温使藤黄叶片光合机构PSⅡ原初光能转化效率（Fv/Fm）、PSⅡ非环式电子传递的量子效率（ФPSⅡ）、非光化学猝灭系数（NPQ）下降,原初荧光（F0）上升。低温胁迫消除后,生长在50％光强下藤黄叶片的Fv/Fm和F0在3d后仍不能完全恢复,而生长在8％光强下藤黄叶片的Fv/Fm和F0基本恢复,说明低温使生长在8％光强下藤黄的光合机构PSⅡ反应中心受到可逆失活,而生长在50％光强下藤黄的光合机构受到氧化伤害。随着低温胁迫时间的延长,两种生长光强藤黄叶片活性氧保护酶（SOD,CAT,APX）的活性虽升高,但O2^-的生成速率、H2O2和MDA含量积累增加。而在恢复阶段,生长在8％光强比生长在50％光强下藤黄叶片的活性氧含量下降得快,进一步说明生长在高光强的植物比生长在低光强的植物受低温伤害大。     

Competence of the pollen of wild species and the cultivar of tomato to affect fertilization at low temperature

Summary The competitive ability of the pollen of wild tomato species (Solanum pennellii, Lycopersicum hirsutum and L. minutum) and a cultivar of tomato to affect fertilization at low temperature was investigated. Pollen grains of wild species were mixed with those of the cultivar and pollinated onto the stigma of the cultivar. The number of seedlings resembling the wild species, excluding L. minutum, were significantly higher when the plants were maintained at a low temperature (8°–12° C) than when they were maintained at a normal temperature (22°–28° C). This indicates that the pollen of S. pennellii and L. hirsutum wild species can compete better with pollen of the cultivar at a low temperature, while there was no change in the competitive ability of L. minutum pollen under these conditions.     

The lower cell density of leaf parenchyma in the Arabidopsis thaliana mutant lcd1-1 is associated with increased sensitivity to ozone and virulent Pseudomonas syringae

Under optimal growth conditions (120 micro mol photons m-2 sec-1 photosynthetically active radiation (PAR), 16-h photoperiod), the recessive ozone-sensitive Arabidopsis thaliana L. Heynh. mutant lcd1-1 exhibits a pale phenotype compared to the wild type. Confocal and multiphoton microscopy revealed that the paleness of lcd1-1 is because of a lower cell density in the leaf palisade parenchyma, resulting in decreased chlorophyll content. When exposed to ozone, lcd1-1 leaves become paler and contain an increased amount of the lipid peroxidation product malondialdehyde compared to the wild type, suggesting that lcd1-1 suffers from elevated levels of reactive oxygen species (ROS) generated in the apoplast. Infection of leaves with virulent Pseudomonas syringae reveals higher bacterial growth as well as lower pathogenesis-related protein 1 (PR-1) and PR-5 expression in lcd1-1 than in the wild type. When the wild type and lcd1-1 are exposed to short-term high-light stress, leaves do not bleach in lcd1-1 and potential activities of photosystems I (PSI) and II (PSII) decrease to a similar extent in both the genotypes, indicating that the photosynthetic apparatus is not affected by lcd1-1 mutation. The LCD1 gene, found to contain a nonsense mutation in the mutant, has been identified. It is located at the bottom of chromosome 2 of the Arabidopsis genome. However, the function of the protein encoded by LCD1 is not yet known. We hypothesize that LCD1 plays a role in normal leaf development, and that the increased sensitivity to ozone and virulent P. syringae is a secondary effect that presumably results from the lower-cell-density phenotype in lcd1-1.     

低温下的植物光抑制机理

低温是影响许多植物的产量和地理分布的一个主要环境因素。如果低温与光照交互作用,则对植物的伤害会更大。本文论述了低温对植物伤害的原初反应,例如低温对膜的伤害、低温对两个光系统和卡尔文循环中某些酶类的影响,并讨论了植物体内的冷敏感组分以及提高植物防御低温光抑制能力的途径。     

Field assessment of efficacy of nitrogen salts to control the root-knot nematode (Meloidogyne javanica) on tomato

Influence of different nitrogen salts at electrical conductivity levels (EC2, 4 and 8?mmhos/cm) on tomato and root-knot nematode (Meloidogyne javanica) and their interactions was evaluated under field conditions. It was found that both diammonium phosphate ((NH₄)₂HPO₄) and ammonium sulphate ((NH₄)₂SO₄) were more effective than ammonium chloride (NH₄Cl) in causing an obvious suppression of M. javanica infection on tomato through reducing root galling and nematode reproduction and improving tomato growth and yield and their suppressive effect was similar to that of oxamyl or ethoprophos. At higher ECs, the tested nitrogen salts did not greatly affect pH, EC and salinity of rhizospheric soil except NH₄Cl at EC8 that caused higher EC and salinity over the untreated control which makes NH₄Cl less suitable candidate. Therefore, the use of (NH₄)₂HPO₄ and (NH₄)₂SO₄ alone or in combination with other control measures could control M. javanica and improve the growth and yield of tomato under field conditions.     

拟南芥CBF1与植物对低温和干旱的抗性

对冷驯化过程中基因表达差异的认识,使抗冻基因(COR)的克隆及其功能的分析成为研究冷驯化过程的主要目标。在拟南芥和其他抗冻植物中,分离了许多COR基因,这些基因对植物抗冻起着非常重要的作用。在拟南芥COR调控的研究中,发现了CBF转录因子的基因家族,其中CBF1能调控一组COR基因的表达。近年来,在冷敏植物如番茄和玉米中也发现了CBF类似基因,拟南芥CBF1基因在转基因番茄中的过量表达提高了植株的抗寒和抗旱性。这一研究结果展示了拟南芥CBF1类似基因的应用可能为冷敏植物抗寒和抗旱性的品种改良提供一条新的途径。     

Small CAB-like proteins prevent formation of singlet oxygen in the damaged photosystem II complex of the cyanobacterium Synechocystis sp. PCC 6803

The cyanobacterial small CAB-like proteins (SCPs) are single-helix membrane proteins mostly associated with the photosystem II (PSII) complex that accumulate under stress conditions. Their function is still ambiguous although they are assumed to regulate chlorophyll (Chl) biosynthesis and/or to protect PSII against oxidative damage. In this study, the effect of SCPs on the PSII-specific light-induced damage and generation of singlet oxygen ((1)O(2)) was assessed in the strains of the cyanobacterium Synechocystis sp. PCC 6803 lacking PSI (PSI-less strain) or lacking PSI together with all SCPs (PSI-less/scpABCDE(-) strain). The light-induced oxidative modifications of the PSII D1 protein reflected by a mobility shift of the D1 protein and by generation of a D1-cytochrome b-559 adduct were more pronounced in the PSI-less/scpABCDE(-) strain. This increased protein oxidation correlated with a faster formation of (1)O(2) as detected by the green fluorescence of Singlet Oxygen Sensor Green assessed by a laser confocal scanning microscopy and by electron paramagnetic resonance spin-trapping technique using 2, 2, 6, 6-tetramethyl-4-piperidone (TEMPD) as a spin trap. In contrast, the formation of hydroxyl radicals was similar in both strains. Our results show that SCPs prevent (1)O(2) formation during PSII damage, most probably by the binding of free Chl released from the damaged PSII complexes.     

Effect of temperature on ascorbate peroxidase activity and flowering of Arabidopsis thaliana ecotypes under different light conditions

Four Arabidopsis thaliana ecotypes were grown at 14 degrees C and 22 degrees C under two light conditions (300 microE m-2 s-1 or 150 microE m-2 s-1) and the effect of temperature on their growth and flowering time was studied. Flowering occurred within 31 days (experimental period) at 22 degrees C, whereas a decrease in growth temperature resulted in a delay in flowering (63 days) under both light conditions. At 14 degrees C, membrane-bound APX (tAPX) activity decreased and total chlorophyll (Chl) content increased with growth under both light conditions. However, at 22 degrees C, the tAPX activity increased and total Chl content decreased with growth under both light conditions. These results suggest that at 22 degrees C oxidative stress was high under both light conditions and consequently Chl content decreased under stressful conditions or vice versa for all the four A. thaliana ecotypes studied. Under both the temperature and light conditions, soluble APX activity showed an irregular pattern of growth. The increase in tAPX activity, with growth only at 22 degrees C but not at 14 degrees C, suggests increased H2O2 formation in flowering plants at 22 degrees C for all the four A. thaliana ecotypes studied. Before flowering, the tAPX activity showed a significantly negative correlation with flowering time. Higher oxidative stress in the lower-latitude ecotypes might induce earlier flowering than the higher-latitude ecotypes. From these results, we propose a hypothesis that H2O2 is one of the possible factors in flower induction.     

In vivo cytochrome and alternative pathway respiration in leaves of Arabidopsis thaliana plants with altered alternative oxidase under different light conditions

The in vivo activity of the alternative pathway (ν(alt)) has been studied using the oxygen isotope fractionation method in leaves of Arabidopsis thaliana modified for the expression of the AtAOX1a gene by anti-sense (AS-12) or overexpression (XX-2). Under non-stressful conditions, ν(alt) was similar in all plant lines regardless of its different alternative pathway capacities (V(alt)). Total leaf respiration (V(t)) and V(alt) were directly related to growth light conditions while electron partitioning between the cytochrome pathway (CP) and alternative pathway (AP) was unchanged by light levels. Interestingly, the AP functioned at full capacity in anti-sense plants under both growth light conditions. The role of the AP in response to a high light stress induced by short-term high light treatment (HLT) was also studied. In wild type and XX-2, both CP and AP rates increased proportionally after HLT while in AS-12, where the AP was unable to increase its rate, the CP accommodated all the increase in respiration. The results obtained under high light stress suggest that flexibility in the response of the mitochondrial electron transport chain is involved in sustaining photosynthetic rates in response to this stress while the saturated AP in AS-12 plants may contribute to the observed increase in photoinhibition.     

Behavior and biology of the tomato psyllid, Bactericerca cockerelli, in response to the Mi-1.2 gene

The Mi‐1.2 gene, identified from wild varieties of tomato, Solanum peruvianum (Mill) (Solanaceae), has been incorporated into near‐isogenic commercial varieties of tomato and has been shown to confer resistance to three different species of phloem feeders: aphids, whiteflies, and nematodes. The results presented here show that plants bearing Mi‐1.2 were also resistant to the tomato psyllid, Bactericerca [Paratrioza] cockerelli (Sulc) (Homoptera: Psyllidae), a serious pest of tomato, Solanum lycopersicon (Mill), in the western half of North America. In choice studies, tomato psyllids preferred to settle on plants that did not contain the gene [Moneymaker (mi‐1.2)] compared to near‐isogenic plants with the gene [Motelle (Mi‐1.2)]. As a result, total oviposition was higher on the susceptible variety, although no‐choice studies indicated that there were no differences in numbers of eggs laid by individual females on either variety. Survival from egg to adult was higher on plants lacking the gene compared to plants containing the gene. However, there were no differences in total development time of individuals reared from either variety. The results suggest that mechanisms of resistance to the tomato psyllid observed in plants bearing the Mi‐1.2 gene are distinct from the mechanisms of resistance to the three phloem feeders examined in other studies.     

In situ monitoring of chlorophyll fluorescence to assess the synergistic effect of low temperature and high irradiance stresses inSpirulina cultures grown outdoors in photobioreactors

A chlorophyll fluorescence technique was applied to anin situ study on the effects of low temperature and high light stresses onSpirulina cultures grown outdoors in controlled tubular photobioreactors at high (1.1 g L^–1) and low (0.44 g L^–1) biomass concentrations. Diurnal changes in PSII photochemistry (F _v/F _m) after 15 min of darkness, or in the light (dF/F _m), and non-photochemical (qN) quenching were measured using a portable, pulse-amplitude-modulated fluorometer. The depression of theF _v/F _m ratio ofSpirulina cultures grown outdoors at 25°C (i.e. 10°C below optimum for growth) and 0.44 g L^–1, reached 30% at the middle of the day. At the same time of the day thedF/F _m ratio showed a reduction of up to 52%. The depression of bothF _v/F _m anddF/F _m was lower in the cultures grown at 1.1 g L^–1. Photoinhibition reduced the daily productivity of the culture grown at 0.44 g L^–1 and 25°C by 33% with respect to that grown at 35°C. Changes in the growth yields of the cultures grown under different temperatures and growth rates correlate well with analogous changes in photon yield (dF/F _m). Simple measurements of photochemical yield (F _v/F _m) can be used to test the physiological status ofSpirulina cultures. The results indicate that the saturating pulse fluorescence technique, when usedin situ, is a powerful tool for assessment of the photosynthetic characteristics of outdoor cultures ofSpirulina.     

Disruption of the human pathogenic yeast Candida albicans catalase gene decreases survival in mouse-model infection and elevates susceptibility to higher temperature and to detergents

Catalase-deficient strains of the human pathogenic yeast Candida albicans were constructed using the URA-blaster method. The disruptant was viable and grew normally in an ordinary culture condition, but became extremely sensitive to treatment with hydrogen peroxide. No catalase activity was observed in a catalase (CCT)-gene-disrupted strain, 1F5-4-1, suggesting that there were no other catalase or catalase-like enzymes in this yeast. The disruptant was shown to be sensitive to higher temperature and to low concentrations of SDS, NP-40, or Triton X-100. After a wild-type CCT gene was reintroduced into the disruptant, catalase activity was restored and the strain became moderately sensitive to treatment with hydrogen peroxide. However, neither the temperature sensitivity nor the susceptibility to SDS observed in the disruptant was restored in the CCT-reintroduced strain. A model infection experiment using wild-type and dCCT strains showed that the disruptants disappeared more rapidly than the wild-type strain in mouse liver, lung, and spleen. These results suggest that the catalase plays a significant role in survival in the host immune system and thus leads this organism to establish infection in the host.     

钙对花生幼苗生长、活性氧积累和光抑制程度的影响

为探讨钙元素对花生幼苗生长的影响,以花育22为试材,用改良的Hoagland溶液进行培养,培养液钙离子(Ca2+)浓度分别为0、6和12 mmol/L(依次简称为CK、C6和C12),研究了不同Ca2+浓度培养下花生幼苗生长以及根系和叶片活性氧(ROS)的积累情况。结果表明,Ca2+显著提高花生植株的株高和鲜重,并降低根冠比,而且正常培养条件下,Ca2+显著提高根系活力、降低叶片和根系的ROS积累,而且C12幼苗的生理状态要好于C6。花生幼苗功能叶在高温(42℃)强光(1200μmol m-2s-1)胁迫处理下,与CK植株相比,C6和C12叶片的过氧化氢(H2O2)和超氧阴离子(O-2)的积累水平低、其叶片PSⅡ最大光化学效率(Fv/Fm)高、光系统Ⅱ(PSⅡ)的关闭程度低,而且C12幼苗的活性氧积累和光抑制程度都明显低于C6,表明高温强光胁迫下,Ca2+有利于减轻花生幼苗叶片的光抑制和ROS积累。C6和C12叶片的部分ROS清除酶活性以及有关渗透调节物质的含量明显高于CK,丙二醛(MDA)的含量明显低于CK,表明胁迫条件下Ca2+通过提高ROS清除酶活性和渗透调节物质含量降低ROS的积累和危害,保护花生类囊体膜从而保证花生正常生长。