Photosystems and antioxidative system of rye, wheat and triticale under Pb stress.

Lead (Pb2+) pollution in the soil sub-ecosystem has been a continuously growing problem due to economic development and ever-increasing anthropogenic activities across the world. In this study, the photosynthetic performance and antioxidant capacity of Triticeae cereals (rye, wheat and triticale) were compared to assess the activities of antioxidants, the degree of oxidative damage, photochemical efficiency and the levels of photosynthetic proteins under Pb stress (0.5 mM, 1 mM and 2 mM Pb (NO3)2). Compared with triticale, Pb treatments imposed severe oxidative damage in rye and wheat. In addition, the highest activity of major antioxidant enzymes (SOD, POD, CAT, and GPX) was also found to be elevated. Triticale accumulated the highest Pb contents in roots. The concentration of mineral ions (Mg, Ca, and K) was also high in its leaves, compared with rye and wheat. Consistently, triticale showed higher photosynthetic activity under Pb stress. Immunoblotting of proteins revealed that rye and wheat have significantly lower levels of D1 (photosystem II subunit A, PsbA) and D2 (photosystem II subunit D, PsbD) proteins, while no obvious decrease was noticed in triticale. The amount of light-harvesting complex II b6 (Lhcb6; CP24) and light-harvesting complex II b5 (Lhcb5; CP26) was significantly increased in rye and wheat. However, the increase in PsbS (photosystem II subunit S) protein only occurred in wheat and triticale exposed to Pb treatment. Taken together, these findings demonstrate that triticale shows higher antioxidant capacity and photosynthetic efficiency than wheat and rye under Pb stress, suggesting that triticale has high tolerance to Pb and could be used as a heavy metal-tolerant plant.

Different Photosynthetic Response to High Light in Four Triticeae Crops.

Photosynthetic capacity is usually affected by light intensity in the field. In this study, photosynthetic characteristics of four different Triticeae crops (wheat, triticale, barley, and highland barley) were investigated based on chlorophyll fluorescence and the level of photosynthetic proteins under high light. Compared with wheat, three cereals (triticale, barley, and highland barley) presented higher photochemical efficiency and heat dissipation under normal light and high light for 3 h, especially highland barley. In contrast, lower photoinhibition was observed in barley and highland barley relative to wheat and triticale. In addition, barley and highland barley showed a lower decline in D1 and higher increase in Lhcb6 than wheat and triticale under high light. Furthermore, compared with the control, the results obtained from PSII protein phosphorylation showed that the phosphorylation level of PSII reaction center proteins (D1 and D2) was higher in barley and highland barley than that of wheat and triticale. Therefore, we speculated that highland barley can effectively alleviate photodamages to photosynthetic apparatus by high photoprotective dissipation, strong phosphorylation of PSII reaction center proteins, and rapid PSII repair cycle under high light.

Salicylic Acid Protects Photosystem II by Alleviating Photoinhibition in Arabidopsis thaliana under High Light.

Salicylic acid (SA) is considered to play an important role in plant responses to environmental stresses. However, the detailed protective mechanisms in photosynthesis are still unclear. We therefore explored the protective roles of SA in photosystem II (PSII) in Arabidopsis thaliana under high light. The results demonstrated that 3 h of high light exposure resulted in a decline in photochemical efficiency and the dissipation of excess excitation energy. However, SA application significantly improved the photosynthetic capacity and the dissipation of excitation energy under high light. Western blot analysis revealed that SA application alleviated the decrease in the levels of D1 and D2 protein and increased the amount of Lhcb5 and PsbS protein under high light. Results from photoinhibition highlighted that SA application could accelerate the repair of D1 protein. Furthermore, the phosphorylated levels of D1 and D2 proteins were significantly increased under high light in the presence of SA. In addition, we found that SA application significantly alleviated the disassembly of PSII-LHCII super complexes and LHCII under high light for 3 h. Overall, our findings demonstrated that SA may efficiently alleviate photoinhibition and improve photoprotection by dissipating excess excitation energy, enhancing the phosphorylation of PSII reaction center proteins, and preventing the disassembly of PSII super complexes.

Different response of photosystem II to short and long-term drought stress in Arabidopsis thaliana.

Short- and long-term drought stress on photosystem II (PSII) and oxidative stress were studied in Arabidopsis thaliana. Under drought stress, chlorophyll (Chl) content, Chl fluorescence, relative water content and oxygen evolution capacity gradually decreased, and the thylakoid structure was gradually damaged. Short-term drought stress caused a rapid disassembly of the light-harvesting complex II (LHCII). However, PSII dimers kept stable under the short-term drought stress and significantly decreased only after 15 days of drought stress. Immunoblotting analysis of the thylakoid membrane proteins showed that most of the photosystem proteins decreased after the stress, especially for Lhcb5, Lhcb6 and PsbQ proteins. However, surprisingly, PsbS significantly increased after the long-term drought stress, which is consistent with the substantially increased non-photochemical quenching (NPQ) after the stress. Our results suggest that the PSII-LHCII supercomplexes and LHCII assemblies play an important role in preventing photo-damages to PSII under drought stress.

The significance of CP29 reversible phosphorylation in thylakoids of higher plants under environmental stresses.

Reversible phosphorylation of proteins is a key event in many fundamental cellular processes. Under stressful conditions, many thylakoid membrane proteins in photosynthetic apparatus of higher plants undergo rapid phosphorylation and dephosphorylation in response to environmental changes. CP29 is the most frequently phosphorylated protein among three minor antennae complexes in higher plants. CP29 phosphorylation in dicotyledons has been known for several decades and is well characterized. However, CP29 phosphorylation in monocotyledons is less studied and appears to have a different phosphorylation pattern. In this review, we discuss recent advancements in CP29 phosphorylation and dephosphorylation studies and its physiological significance under environmental stresses in higher plants, especially in the monocotyledonous crops. Physiologically, the phosphorylation of CP29 is likely to be a prerequisite for state transitions and the disassembly of photosystem II supercomplexes, but not involved in non-photochemical quenching (NPQ). CP29 is phosphorylated in monocots exposed to environmental cues, with its subsequent lateral migration from grana stacks to stroma lamellae. However, neither CP29 phosphorylation nor its lateral migration occurs in dicotyledonous plants after drought, cold, or salt stress. Since the molecular mechanisms of differential CP29 phosphorylation under stresses are not fully understood, this review provides insights for future studies regarding the physiological function of CP29 reversible phosphorylation.

Melatonin improves the removal and the reduction of Cr(VI) and alleviates the chromium toxicity by antioxidative machinery in Rhodobacter sphaeroides.

Bioremediation with photosynthetic bacteria (PSB) is thought to be a promising removal method for hexavalent chromium [Cr(VI)]-containing wastewater. In the present study, Rhodobacter sphaeroides (R. sphaeroides) SC01 was used for the investigation of Cr(VI) removal in Cr(VI)-contaminated solution in the presence of melatonin. It was found that exogenous melatonin alleviated oxidative damage to R. sphaeroides SC01, increased Cr (VI) absorption capacity of cell membrane, and improved the reduction efficiency of Cr(VI) via the activation of chromate reductants. The results showed that melatonin could further promote the increase in Cr(VI) removal efficiency, reaching up to 97.8%. Furthermore, melatonin application resulted in 296.9%, 44.4%, and 69.7% upregulation of ascorbic acid (AsA), glutathione (GSH), and cysteine (Cys) relative to non-melatioin treated R. sphaeroides SC01 at 48 h. In addition, the resting cells, cell-free supernatants (CFS), and cell-free extracts (CFE) with melatonin had a higher Cr(VI) removal rate of 18.6%, 82.0%, and 15.2% compared with non-melatonin treated R. sphaeroides SC01. Fourier transform infrared spectroscopy (FTIR) revealed that melatonin increased the binding of Cr(III) with PO43- and CO groups on cell membrane of R. sphaeroides SC01. X-ray diffractometer (XRD) analysis demonstrated that melatonin remarkably bioprecipitated the production of CrPO4·6H2O in R. sphaeroides SC01. Hence, these results indicated that melatonin plays the important role in the reduction and uptake of Cr(VI), demonstrating it is a great promising strategy for the management of Cr(VI) contaminated wastewater in photosynthetic bacteria.

Exogenous melatonin improved photosynthetic efficiency of photosystem II by reversible phosphorylation of thylakoid proteins in wheat under osmotic stress.

It has been well demonstrated that melatonin plays an important protective role in photosynthesis of plants under various environmental stresses, while the detailed mechanisms by which melatonin protects photosystem II (PSII) under environmental stress are still unclear. In the study, the effects of melatonin on photosynthetic efficiency, energy dissipation, PSII protein composition, and reversible phosphorylation of thylakoid proteins were investigated in wheat plants under osmotic stress. The results showed that osmotic stress significantly reduced pigment content, photochemical efficiency of PSII, oxygen-evolving activity, and dissipation of excess excitation energy, while 25 µM melatonin applications greatly alleviated their decline under osmotic stress. Western blot data of PSII proteins revealed that melatonin upregulated the levels of D1, Lhcb5, Lhcb6, PsbQ, and PsbS proteins in wheat exposed to osmotic stress. In addition, thylakoid membrane proteins were strongly phosphorylated in wheat under osmotic stress with or without melatonin. Furthermore, the results from PSII protein dephosphorylation showed that exogenous melatonin promoted the dephosphorylation of LCHII, CP43, and D1 under osmotic stress. Therefore, our findings suggest that melatonin can provide an effective protection for the photosynthetic apparatus by the regulation of PSII proteins and the reversible phosphorylation of thylakoid proteins under drought stress.

Diversity and evolution of four dispersed repetitive DNA sequences in the genus Secale.

We present the first characterization of 360 sequences in six species of the genus Secale of both cultivated and wild accessions. These include four distinct kinds of dispersed repetitive DNA sequences named pSc20H, pSc119.1, pSaO5(411), and pSaD15(940) belonging to the Revolver family. During the evolution of the genus Secale from wild to cultivated accessions, the pSaO5(411)-like sequences became shorter mainly because of the deletion of a trinucleotide tandem repeating unit, the pSc20H-like sequences displayed apparent homogenization in cultivated rye, and the second intron of Revolver became longer. In addition, the pSc20H-, pSc119.1-, and pSaO5(411)-like sequences cloned from wild rye and cultivated rye could be divided into two large clades. No single case of the four kinds of repetitive elements has been inherited by each Secale accession from a lone ancestor. It is reasonable to consider the vertical transmission of the four repetitive elements during the evolution of the genus Secale. The pSc20H- and pSaO5(411)-like sequences showed evolutionary elimination at specific chromosomal locations from wild species to cultivated species. These cases imply that different repetitive DNA sequences have played different roles in the chromosome development and genomic evolution of rye. The present study adds important information to the investigations dealing with characterization of dispersed repetitive elements in wild and cultivated rye.

The roles of Arabidopsis proteins of Lhcb4, Lhcb5 and Lhcb6 in oxidative stress under natural light conditions.

Under light conditions, highly reactive oxygen species (ROS) can be generated in the antenna systems and the reaction center of photosystems (PS). The protective roles of Lhcb4 (CP29), Lhcb5 (CP26) and Lhcb6 (CP24), three minor chlorophyll binding antenna proteins during photoinhibition have been well studied. However, their regulatory mechanisms against oxidative damages under natural light conditions remain unknown. Here we investigated their specific roles in oxidative stress responses and photosynthetic adaptation by using the Arabidopsis thaliana knockout lines grown in the field condition. All three mutant lines exhibited decreased energy-transfer efficiency from the LHCII (light-harvesting complex II) to the PSII reaction center. Oxygen evolution capacity decreased slightly in the plants lacking Lhcb4 (koLHCB4) and Lhcb6 (koLHCB6). Photosynthetic rates and fitness for the plants lacking Lhcb5 (koLHCB5) or koLHCB6 grown in the field were affected, but not in the plants lacking Lhcb4. Antioxidant analysis indicated the lowest antioxidant enzyme activities and the lowest levels of non-enzymatic antioxidants in koLHCB6 plants. In addition, koLHCB6 plants accumulated much higher levels of superoxide and hydrogen, and suffered more severe oxidative-damages in the field. Our results clearly demonstrate that Lhcb6 may be involved in alleviating oxidative stress and photoprotection under natural conditions.

Biomonitoring chromium III or VI soluble pollution by moss chlorophyll fluorescence.

We systematically compared the impacts of four Cr salts (chromic chloride, chromic nitrate, potassium chromate and potassium bichromate) on physiological parameters and chlorophyll fluorescence in indigenous moss Taxiphyllum taxirameum. Among the four Cr salts, K2Cr2O7 treatment resulted in the most significant decrease in photosynthetic efficiency and antioxidant enzymes, increase in reactive oxygen species (ROS), and obvious cell death. Different form the higher plants, although hexavalent Cr(VI) salt treatments resulted in higher accumulation levels of Cr and were more toxic than Cr(III) salts, Cr(III) also induced significant changes in moss physiological parameters and chlorophyll fluorescence. Our results showed that Cr(III) and Cr(VI) could be monitored distinguishably according to the non-photochemical quenching (NPQ) fluorescence of sporadic purple and sporadic lavender images respectively. Then, the valence states and concentrations of Cr contaminations could be evaluated according to the image of maximum efficiency of PSII photochemistry (Fv/Fm) and the quantum yield of PSII electron transport (ΦPSII). Therefore, this study provides new ideas of moss's sensibility to Cr(III) and a new method to monitor Chromium contaminations rapidly and non-invasively in water.

Comparison of phosphorylation and assembly of photosystem complexes and redox homeostasis in two wheat cultivars with different drought resistance.

Reversible phosphorylation of proteins and the assembly of thylakoid complexes are the important protective mechanism against environmental stresses in plants. This research was aimed to investigate the different responses of the antioxidant defense system and photosystem II (PSII) to osmotic stress between drought-resistant and drought-susceptible wheat cultivars. Results showed that the decrease in PSII photochemistry and six enzyme activities was observed in drought-susceptible wheat compared with drought-resistant wheat under osmotic stress. In addition, a lower accumulation of reactive oxygen species (ROS) and cell death were found in the resistant wheat compared with the susceptible wheat under osmotic stress. Western blot analysis revealed that osmotic stress led to a remarkable decline in the steady state level of D1 protein in drought-susceptible wheat. However, the CP29 protein was strongly phosphorylated in drought-resistant wheat compared with the susceptible wheat under osmotic stress. Our results also showed that drought-resistant wheat presented higher phosphorylated levels of the light-harvesting complex II (LHCII), D1, and D2 proteins and a more rapid dephosphorylated rate than drought-susceptible wheat under osmotic stress. Furthermore, the PSII-LHCII supercomplexes and LHCII trimers were more rapidly disassembled in drought-susceptible wheat than the drought-resistant wheat under osmotic stress. These findings provide that reversible phosphorylation of thylakoid membrane proteins and assembly of thylakoid membrane complexes play important roles in plant adaptation to environmental stresses.

Influence of stripe rust infection on the photosynthetic characteristics and antioxidant system of susceptible and resistant wheat cultivars at the adult plant stage.

Wheat stripe rust (Puccinia striiformis f. sp. tritici, Pst), is one of the most serious diseases of wheat (Triticum aestivum L.) worldwide. To gain a better understanding of the protective mechanism against stripe rust at the adult plant stage, the differences in photosystem II and antioxidant enzymatic systems between susceptible and resistant wheat in response to stripe rust disease (P. striiformis) were investigated. We found that chlorophyll fluorescence and the activities of the antioxidant enzymes were higher in resistant wheat than in susceptible wheat after stripe rust infection. Compared with the susceptible wheat, the resistant wheat accumulated a higher level of D1 protein and a lower level of reactive oxygen species after infection. Furthermore, our results demonstrate that D1 and light-harvesting complex II (LHCII) phosphorylation are involved in the resistance to stripe rust in wheat. The CP29 protein was phosphorylated under stripe rust infection, like its phosphorylation in other monocots under environmental stresses. More extensive damages occur on the thylakoid membranes in the susceptible wheat compared with the resistant wheat. The findings provide evidence that thylakoid protein phosphorylation and antioxidant enzyme systems play important roles in plant responses and defense to biotic stress.

Biomonitoring heavy metal contaminations by moss visible parameters.

Traditional sampling for heavy metal monitoring is a time-consuming and inconvenient method, which also does not indicate contaminants non-invasively and instantaneously. Moss is sensitive to heavy metals and is therefore considered a pollution indicator. However, it is unknown what kind physiological parameters can indicate metal contaminations quickly and non-invasively. Here, we systematically examined the effects of six heavy metals on physiological parameters and photosynthetic activities of two moss species grown in aquatic media or moist soil surface. We suggest that a phenotype with anthocyanin accumulation pattern and chlorosis pattern and two chlorophyll fluorescence parameters with their images can roughly reflect metal species groups, concentrations and differences between the two moss species. In other words, metal contaminations could be roughly estimated visually using the naked eye. Enzymatic and non-enzymatic anti-oxidative abilities and photosynthetic protein contents of Eurhynchium eustegium were higher than those of Taxiphyllum taxirameum, indicating their differential metal tolerance. Neither anti-oxidative abilities nor photosynthetic proteins were found to be ideal indicators. This study provides new ideas to monitor heavy metals rapidly and non-invasively in water or on wetland and moist soil surface.

[Analyses on antigen epitopes and drug resistance mutations of HIV-1 gag and pol genes].

To study the CTL antigen epitopes and drug resistance mutations of HIV-1 gag and pol genes through analyzing gag and pol gene sequences. The HIV-1 gag and pol gene fragments were amplified using nested polymerase chain reaction. A total of 23 PCR sequences, 449 cloned gag sequences and 402 cloned pol sequences were obtained. Sequence analyses showed the 23 samples were subtype B or B'. A total of 4 in 8 CTL antigen epitopes appeared 8 mutations in consensus sequence of subtype B and B'. There were no mutations found in the PCR sequences, whereas a few mutations were found in clone sequences (9.80%) in 5 antigen epitopes in p24 region. Eighteen PIs-related mutations and 24 RTIs-related mutations were found in PCR sequences and clone sequences in pol gene region, in which 17 (94.44%) PIs-related mutations and 15 (62.50%) RTIs-related mutations were found only in the clone sequences, respectively. The results showed that the prevalence of HIV-1 drug resistance strains in this study was at a higher level (17.39%), suggesting that some samples were resistant.to existing antiviral drugs.

Study on powdery mildew resistance transfer from S. cereale L.cv. Weiling rye into wheat.

Weiling rye (S. cereale L.cv.), a Chinese dwarf rye, confers high powdery mildew (Erysiphe gramininis f.sp.tritici) in China. My8443, a wheat cultivars infecting seriously powdery mildew disease, was used as the female parent and Weiling rye was used as the donor of powdery mildew resistance in the study. A new wheat-rye translocation line,named No.147,was developed from BC2F6 progenies of wheat cultivars My8443 and Weiling rye to transfer the resistance from Weiling rye to common wheat. The powdery mildew resistance of No.147 and its parents were investigated in seedling and adult stages by artificially inoculating the mixture of advanced pathogenic races in room and field and the single pathogenic race in room. Improved Giemsa C-banding technique and genomic in situ hybridization (GISH,Genomic in situ hybridization) were used to identify wheat and rye chromosomes. Acid polyacylamide gel electrophoresis(APAGE) separation of endosperm gliadin and simple sequence repeat(SSR) PCR amplification of 11 SCM-Secale cereale markers also were employed for 1RS confirmation in the study. The results showed that No.147 was a new 1BL/1RS wheat-rye chromosome translocation with high powdery mildew resistance derived from Weiling rye. The reason on the formation of the new wheat-rye chromosome translocation was analyzed. The utilizations of resistance gene resource derived from Chinese Weiling rye and the new 1BL/1RS translocation line in wheat genetics and breeding improvement were discussed in the paper.

Diagnostic detection and genetic analysis of wheat stripe rust resistant gene YrCN19.

DNA samples of 19 wheat cultivars or lines were screened with the primer pair Xgwm410, a diagnostic marker of wheat stripe rust resistant gene YrCN19. The results showed that the wheat cultivars or lines CN19, XK5, AIM5 and AIM6 produced the amplicon Xgwm410/391, which co-segregated with the resistance of YrCN19, but other cultivars or lines did not produced it. Pedigree analysis and resistance test showed that CN19, XK5, AIM5 and AIM6 carried wheat stripe rust gene YrCN19. Genetic analysis proved that the resistance to wheat stripe rust of CN19, XK5, and AIM5 inherited in single dominant gene genetic law (3 resistant: 1 susceptible). The results also proved that the resistance descent in the cross combination Yanfu188/AIM6 complied with the single dominant gene genetic law (3 resistant: 1 susceptible), while the resistance inheritance in other cross combinations (CN18/AIM6, Lu955159/AIM6 and Shu3110/ AIM6) complied with two gene complementary genetic law (9 resistant: 7 susceptible). The results suggested that the resistance expression and separation of YrCN19 was distinct in the various genetic backgrounds or cross combinations. This study would expedite the exploitation and utilization of YrCN19 in wheat resistant breeding.

A complete mitochondrial genome sequence of Asian black bear Sichuan subspecies (Ursus thibetanus mupinensis).

We obtained the complete mitochondrial genome of U.thibetanus mupinensis by DNA sequencing based on the PCR fragments of 18 primers we designed. The results indicate that the mtDNA is 16,868 bp in size, encodes 13 protein genes, 22 tRNA genes, and 2 rRNA genes, with an overall H-strand base composition of 31.2% A, 25.4% C, 15.5% G and 27.9% T. The sequence of the control region (CR) located between tRNA-Pro and tRNA-Phe is 1422 bp in size, consists of 8.43% of the whole genome, GC content is 51.9% and has a 6bp tandem repeat and two 10bp tandem repeats identified by using the Tandem Repeats Finder. U. thibetanus mupinensis mitochondrial genome shares high similarity with those of three other Ursidae: U. americanus (91.46%), U. arctos (89.25%) and U. maritimus (87.66%).

Regulation of a gamma-aminobutyric acid transporter by reciprocal tyrosine and serine phosphorylation.

A feature of the rat brain gamma-aminobutyric acid transporter GAT1, and other members of the neurotransmitter transporter family, is its regulated redistribution between intracellular locations and the plasma membrane. Recent studies have focused upon defining the signaling molecules that facilitate this redistribution. Agents that promote direct tyrosine phosphorylation of GAT1 promote a relative increase in surface GAT1 levels, and this results from a slowing of the transporter internalization rate. Agents that act to increase protein kinase C (PKC) activity promote a relative decrease in surface GAT1 levels; whether this effect is caused by direct transporter phosphorylation is unknown. The opposing actions of tyrosine kinase activity and PKC activity raise the possibility that the subcellular distribution of GAT1 is associated with mutually exclusive transporter phosphorylation events. The present experiments show that GAT1 is phosphorylated on serine residues in a PKC-dependent manner, but this state is only revealed when GAT1 tyrosine phosphorylation is eliminated or greatly reduced. The relative levels of serine phosphorylation and tyrosine phosphorylation are negatively correlated. The amount of serine phosphorylation is regulated by agents that affect tyrosine phosphorylation, and vice versa. In addition, the ability of agents that affect tyrosine kinase activity to regulate GAT1 serine phosphorylation requires a change in its tyrosine phosphorylation state. These data support the ideas that GAT1 can exist in either of two mutually exclusive phosphorylation states and that the relative abundance of these states determines in part the relative subcellular distribution of the transporter.

Increased p75(NTR) expression in hippocampal neurons containing hyperphosphorylated tau in Alzheimer patients.

p75(NTR), a low-affinity neurotrophin receptor, may be involved in the pathogenesis of Alzheimer's disease (AD). The aim of present study was to detect the relationship between p75(NTR)-containing neurons and the pretangles consisting of hyperphosphorylated tau stained by Alz-50 in the hippocampus of AD patients. Samples of hippocampus of 10 female AD patients and 10 nondemented female controls matched for age and postmortem delay were investigated immunocytochemically, and the stainings were quantified using an image analysis system. The results showed that: (i) p75(NTR) was present in about half of the pyramidal neurons of the CA1-CA4 subfields of hippocampus. No difference was observed in the number of p75(NTR) immunoreactive neurons in the CA1-CA4 subfields between the two groups. Interestingly, the ratio of p75(NTR) expressing neurons to the total number of neurons as staining with thionin was significantly higher in the CA1 and CA2 subfields of AD hippocampus than in controls. (ii) There were a large number of Alz-50 neurons and double-labeled neurons containing p75(NTR) and Alz-50 in the CA1 and CA2 subfields of AD patients. These results suggest that p75(NTR) may be involved in the formation of tangles in the Alzheimer process.