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1.
Antibodies against nitrate reductase from Monoraphidium braunii have been used to determine the antigenic relationships of nitrate reductases from different green algae. Nitrate reductases from Chlamydomonas reinhardii, Chlorella fusca, Dunaliella salina, and Scenedesmus obliquus, were inhibited by, and cross-reacted with, antibodies raised against the enzyme from Monoraphidium braunii.
These antibodies were also used to determine, by immunoelectron microscopy, the intracellular location of nitrate reductase in the aforementioned green algae. In all cases, the enzyme was specifically located in the pyrenoid.
相似文献2.
Immunocytochemical Localization of Ribulose-1,5-Bisphosphate Carboxylase in the Pyrenoid and Thylakoid Region of the Chloroplast of Chlamydomonas reinhardtii 总被引:5,自引:4,他引:1
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The distribution of the large and small subunits of ribulose-1,5-bisphosphate carboxylase in the chloroplast of Chlamydomonas reinhardtii was studied by immunoelectron microscopy by labeling Lowicryl-embedded sections with antibody to each subunit followed by protein A-gold. In light-harvested synchronously dividing cells, antibodies to each subunit heavily labeled the pyrenoid, whereas the thylakoid region of the plastid was lightly labeled. By estimating the volume of each chloroplast compartment, it was determined that approximately 40% of the total small subunit in the plastid and 30% of the large subunit are localized in the thylakoid region, presumably in the stroma. In synchronously dividing cells exposed to an extended dark period, the amount of labeling of the pyrenoid region by antibody to the small subunit stayed constant, but the labeling of the thylakoid region decreased. In stationary phase cells, the proportion of the label over the pyrenoid is higher than in synchronously dividing cells suggesting that the pyrenoid may be a storage organelle. 相似文献
3.
When radiolabeled precursors and autoradiography are used to investigate turnover of protein components in photoreceptive cone outer segments (COSs), the labeled components—primarily visual pigment molecules (opsins)—are diffusely distributed along the COS. To further assess this COS labeling pattern, we derive a simplified mass-transfer model for quantifying the contributions of advective and diffusive mechanisms to the distribution of opsins within COSs of the frog retina. Two opsin-containing regions of the COS are evaluated: the core axial array of disks and the plasmalemma. Numerical solutions of the mass-transfer model indicate three distinct stages of system evolution. In the first stage, plasmalemma diffusion is dominant. In the second stage, the plasmalemma density reaches a metastable state and transfer between the plasmalemma and disk region occurs, which is followed by an increase in density that is qualitatively similar for both regions. The final stage consists of both regions slowly evolving to the steady-state solution. Our results indicate that autoradiographic and cognate approaches for tracking labeled opsins in the COS cannot be effective methodologies for assessing new disk formation at the base of the COS.Abbreviations used: A, area (μm2), COS, cone outer segment, D, mass diffusion coefficient (μm2/s), hm, mass transfer coefficient (μm/s), L, cone outer segment length (μm), PDE, partial differential equation, r, radius (μm), t, time (s), T, plasmalemma thickness (μm), u, plasmalemma or disk region (axial) velocity (μm/s), V, volume (μm3), W, plasmalemma width (μm), x, axial direction, v, disk to plasmalemma velocity (μm/s), ρ1, disk label density, ρ2, plasmalemma label density, ϕ, nonvoid fraction 相似文献
4.
Homogeneous nitrate reductase (EC 1.6.6.2) from Monoraphidium braunii was obtained by means of affinity chromatography in blue-Sepharose and gel filtration. After electrophoresis in polyacrylamide, gel slices containing pure nitrate reductase were disrupted and injected into previously unimmunized rabbits. The antiserum produced after several weeks was found to inhibit the different activities of nitrate reductase to a similar degree. Monospecificity of the antiserum was demonstrated by Ouchterlony double diffusion and crossed immunoelectrophoresis. The antibodies were purified by immunoabsorption to Sepharose-bound nitrate reductase.
The intracellular location of nitrate reductase in green algae was examined by applying an immunocytochemical method to M. braunii cells. Ultrathin frozen sections were first treated with immunopurified anti-nitrate reductase monospecific antibodies, followed by incubation with colloidal gold-labeled goat antirabbit immunoglobulin G as a marker. The enzyme was specifically located in the pyrenoid region of the chloroplast.
相似文献5.
Eiko Morita Haruko Kuroiwa Tsuneyoshi Kuroiwa Hisayoshi Nozaki 《Journal of phycology》1997,33(1):68-72
The distribution of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in the chloroplasts of the unicellular green alga Chlamydomonas reinhardtii Dangeard was examined using cryotechnique and conventional fixation for immunogold electron microscopy. Both methods provided essentially identical results, although somewhat higher densities of gold particles indicating Rubisco molecules were recognized in the pyrenoids of cryofixed cells. The gold particles were highly concentrated in the pyrenoid matrix within the chloroplasts. Even when considering the vast difference in volume between the pyrenoid and the rest of the Chloroplast, more than 99% of the total Rubisco labeling in the chloroplast was calculated to be present in the pyrenoid matrix. High localization of Rubisco in the pyrenoid matrix was also recognized regardless of cell age, based on immunofluorescence microscopy of the same en bloc samples. These results are inconsistent with a recent immunocytochemical study employing cryotechnique in which more than 90% of the total Rubisco was recognized in the thylakoid region (thylakoid membranes and stroma) of C. reinhardtii cells. Rubisco highly localized in the pyrenoid matrix may take part in active photosynthetic CO2 fixation and/or the CO2 concentrating mechanism . 相似文献
6.
E. Kessler Michaela Schfer Carola Hümmer Annette Kloboucek V. A. R. Huss 《Plant biology (Stuttgart, Germany)》1997,110(3):244-250
Biochemical and physiological properties of 16 Scenedesmus species representing the three subgenera Scenedesmus, Acutodesmus, and Desmodesmus are not suitable for species differentiation. All Scenedesmus species studied thus far produce secondary carotenoids, e.g. astaxanthin and canthaxanthin, under nitrogen-deficient conditions. In addition, with the exception of one strain, hydrogenase activity under anaerobic conditions is generally present. Sequence analyses of ribosomal 18S RNAs indicate that the subgenus Desmodesmus is phylogenetically well separated from the other subgenera, whereas the separation of Scenedesmus and Acutodesmus appears less convincing and is dismissed in favour of a single subgenus, Scenedesmus. Three taxa formerly assigned to the genus Chlorella are shown to be unicellular species of the genus Scenedesmus. “Chlorella” fusca var. vacuolata and “C.” fusca var. rubescens, which is closely related to S. obliquus, belong to the Scenedesmus/‘Acutodesmus group. “C.” fusca var. fusca is closely related to S. communis and thus belongs to the subgenus Desmodesmus. Inclusion of Kermatia pupukensis into the genus Scenedesmus is also strongly supported by the RNA data which furthermore indicate a relationship with the subgenus Desmodesmus. 相似文献
7.
El-Sayed Salama Hyun-Chul Kim Reda A. I. Abou-Shanab Min-Kyu Ji You-Kwan Oh Seong-Heon Kim Byong-Hun Jeon 《Bioprocess and biosystems engineering》2013,36(6):827-833
Two freshwater microalgae including Chlamydomonas mexicana and Scenedesmus obliquus were grown on Bold Basal Medium (BBM) with different levels of salinity up to 100 mM NaCl. The dry biomass and lipid content of microalgae were improved as the concentration of NaCl increased from 0 to 25 mM. Highest dry weight (0.8 and 0.65 g/L) and lipid content (37 and 34 %) of C. mexicana and S. obliquus, respectively, were obtained in BBM amended with 25 mM NaCl. The fatty acid composition of the investigated species was also improved by the increased NaCl concentration. At 50 mM, NaCl palmitic acid (35 %) and linoleic acid (41 %) were the dominant fatty acids in C. mexicana, while oleic acid (41 %) and α-linolenic acid (20 %) were the major fractions found in S. obliquus. 相似文献
8.
In Situ Association of Calvin Cycle Enzymes,Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Activase,Ferredoxin-NADP+ Reductase,and Nitrite Reductase with Thylakoid and Pyrenoid Membranes of Chlamydomonas reinhardtii Chloroplasts as Revealed by Immunoelectron Microscopy 总被引:2,自引:2,他引:0
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The in situ localization of the chloroplast enzymes ribulose-1,5-bisphosphate carboxylase (Rubisco), Rubisco activase, ribose-5-phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, aldolase, nitrite reductase, ferredoxin-NADP+ reductase, and H+-ATP synthase was studied by immunoelectron microscopy in Chlamydomonas reinhardtii. Immunogold labeling revealed that, despite Rubisco in the pyrenoid matrix, Calvin cycle enzymes, Rubisco activase, nitrite reductase, ferredoxin-NADP+ reductase, and H+-ATP synthase are associated predominantly with chloroplast thylakoid membranes and the inner surface of the pyrenoid membrane. This is in accord with previous enzyme localization studies in higher plants (K.H. Suss, C. Arkona, R. Manteuffel, K. Adler [1993] Proc Natl Acad Sci USA 90: 5514-5518). Pyrenoid tubules do not contain these enzymes. The pyrenoid matrix consists of Rubisco but is devoid of the other photosynthetic enzymes investigated. Evidence for the occurrence of two Rubisco forms differing in their spatial localization has also been obtained: Rubisco form I appears to be membrane associated like other Calvin cycle components, whereas Rubisco form II is confined to the pyrenoid matrix. It is proposed that enzyme form I represents an active Rubisco when assembled into Calvin cycle enzyme complexes, whereas Rubisco form II may be part of a CO2-concentrating mechanism. Pyrenoidal Calvin cycle complexes are thought to be highly active in CO2 fixation and important for the synthesis of starch around the pyrenoid. 相似文献
9.
Manish Shukla Renu Minda Himanshu Singh Srikanth Tirumani Kandala V. R. Chary Basuthkar J. Rao 《PloS one》2012,7(12)
UVI31+ is an evolutionarily conserved BolA family protein. In this study we examine the presence, localization and possible functions of this protein in the context of a unicellular alga, Chlamydomonas reinhardtii. UVI31+ in C. reinhardtii exhibits DNA endonuclease activity and is induced upon UV stress. Further, UVI31+ that normally localizes to the cell wall and pyrenoid regions gets redistributed into punctate foci within the whole chloroplast, away from the pyrenoid, upon UV stress. The observed induction upon UV-stress as well as the endonuclease activity suggests plausible role of this protein in DNA repair. We have also observed that UV31+ is induced in C. reinhardtii grown in dark conditions, whereby the protein localization is enhanced in the pyrenoid. Biomolecular interaction between the purified pyrenoids and UVI31+ studied by NMR demonstrates the involvement of the disordered loop domain of the protein in its interaction. 相似文献
10.
We examined the hypothesis that toxic effects of Microcystis aeruginosa to population dynamics, ephippial production, and resting egg formation of Daphnia were restricted by food quality levels. Four unicellular, toxic M. aeruginosa strains with contrasting concentration of microcystin and two Daphnia species were used in this study. The number of offspring at first reproduction, population densities, and maximum population growth rate of two Daphnia species were lower at the highest toxic M. aeruginosa 7820 treatment compared with other treatments under two concentrations of Scenedesmus obliquus. The maximum number of offspring at first reproduction of two Daphnia species appeared in the higher toxic M. aeruginosa 526 strain at the high concentration of S. obliquus. The effects of M. aeruginosa strains, S. obliquus, and their combination on the number of offspring at first reproduction and the maximum population growth rate of two Daphnia species were significant (P < 0.001). Two Daphnia species could not reproduce in the highest toxic M7820 treatment under two concentrations of S. obliquus. They had lower population size and maximum population growth rate in the higher toxic M526 treatment at the low concentration of S. obliquus, but they were higher at the high concentration of S. obliquus. This result suggests that high S. obliquus concentration could relieve the toxicity of M. aeruginosa to Daphnia, and Daphnia could utilize the lower toxic Microcystis as food. The cumulative ephippia numbers of two Daphnia species were more at the high concentration of S. obliquus than those at the low concentration. The percentage of ephippia containing no resting eggs of two Daphnia species was evidently higher at the low concentration of S. obliquus but was lower at the high concentration of S. obliquus. Our results indicated that the cumulative ephippia numbers of Daphnia were population density dependent at the high-level food, and the productions of ephippia in Daphnia were significantly controlled by microcystin concentration at the low-level food. 相似文献
11.
In order to investigate the possible impacts of increased atmospheric CO2 levels on algal growth and photosynthesis, the influence of CO2 concentration was tested on three planktonic algae (Chlamydomonas reinhardtii, Chlorella pyrenoidosa, and Scenedesmus obliquus). Increased CO2 concentration enhanced significantly the growth rate of all three species. Specific growth rates reached maximal values at 30, 100, and 60 M CO2 in C. reinhardtii, C. pyrenoidosa, and S. obliquus, respectively. Such significant enhancement of growth rate with enriched CO2 was also confirmed at different levels of inorganic N and P, being more profound at limiting levels of N inC. pyrenoidosa and P in S. obliquus. The maximal rates of net photosynthesis, photosynthetic efficiency and light-saturating point increased significantly (p < 0.05) in high-CO2-grown cells. Elevation of the CO2 levels in cultures enhanced the photoinhibition of C. reinhardtii, but reduced that of C. pyrenoidosa and S. obliquus when exposed to high photon flux density. The photoinhibited cells recovered to some extent (from 71% to 99%) when placed under dim light or in darkness, with better recovery in high-CO2-grownC. pyrenoidosa and S. obliquus. Although pH and pCO2 effects cannot be distinguished from this study, it can be concluded that increased CO2 concentrations with decreased pH could affect the growth rate and photosynthetic physiology of C. reinhardtii, C. pyrenoidosa, and S. obliquus. 相似文献
12.
Mature maize leaf tissue (Zea mays L.) was immunolabeled using a pre-embedding protocol with specific antibodies for nitrate reductase and protein A-colloidal gold. Immunogold label was found exclusively in the cytoplasm of mesophyll cells; no reaction was detected in bundle sheath cells. Chloroplasts, which were sliced open during cryosectioning, had no labeling. Thus, it appears nitrate reductase is localized exclusively in the cytoplasm of maize leaf mesophyll cells. No gold labeling was found on tissue sections embedded in L. R. White's or Lowicryl resin, indicating that previous chloroplast localization utilizing these protocols may be artifactual, resulting from shared epitopes or nonspecific antibody binding. 相似文献
13.
Unicellular algae grown under low-CO2 conditions (0.03% CO2) have developed a means of concentrating CO2 at the site of ribulose-1,5-bisphosphate carboxylase/oxygenase. Cells with the CO2-concentrating mechanism (CCM) acquire the ability to accumulate inorganic carbon to a level higher than that obtained by simple diffusion. To identify proteins which are involved in the organization of the CCM, cells of Scenedesumus obliquus and Chlorella vulgaris grown in high CO2 (5% CO2 in air) were transferred to low-CO2 (0.03%) conditions in the presence of 35SO inf4 sup2? and, thereafter, polypeptides labeled with 35S were detected. Under low-CO2 conditions the inducton of 36-, 39-, 94- and 110- to 116kDa polypeptides were particularly observed in S. obliquus and 16-, 19-, 27-, 36-, 38- and 45-kDa polypeptides were induced in C. vulgaris. Western blots with antibodies raised against 37-kDa subunits of the periplasmic carbonic anhydrase (CA) of Chlamydomonas reinhardtii showed immunoreactive bands with the 39-kDa polypeptide in the whole-cell homogenates from S. obliquus and with 36 and 38-kDa polypeptides in both high- and low-CO2grown cells of C. vulgaris. Anti-pea-chloroplast CA antibodies cross-reacted with a single polypeptide of 30 kDa in the whole-cell homogenates but not with thylakoid membranes. The CA activity was associated with soluble and membrane-bound fractions, except thylakoid membranes. 相似文献
14.
Minqi Han Fang Liu Fengli Zhang Zhiyong Li Houwen Lin 《Marine biotechnology (New York, N.Y.)》2012,14(6):701-713
Many biologically active natural products have been isolated from Phakellia fusca, an indigenous sponge in the South China Sea; however, the microbial symbionts of Phakellia fusca remain unknown. The present investigations on sponge microbial community are mainly based on qualitative analysis, while quantitative analysis, e.g., relative abundance, is rarely carried out, and little is known about the roles of microbial symbionts. In this study, the community structure and relative abundance of bacteria, actinobacteria, and archaea associated with Phakellia fusca were revealed by 16S rRNA gene library-based sequencing and quantitative real time PCR (qRT-PCR). The ammonia-oxidizing populations were investigated based on amoA gene and anammox-specific 16S rRNA gene libraries. As a result, it was found that bacterial symbionts of sponge Phakellia fusca consist of Proteobacteria including Gamma-, Alpha-, and Delta-proteobacteria, Cyanobacteria with Gamma-proteobacteria as the predominant components. In particular, the diversity of actinobacterial symbionts in Phakellia fusca is high, which is composed of Corynebacterineae, Acidimicrobidae, Frankineae, Micrococcineae, and Streptosporangineae. All the observed archaea in sponge Phakellia fusca belong to Crenarchaeota, and the detected ammonia-oxidizing populations are ammonia-oxidizing archaea, suggesting the nitrification function of sponge archaeal symbionts. According to qRT-PCR analysis, bacterial symbionts dominated the microbial community, while archaea represented the second predominant symbionts, followed by actinobacteria. The revealed diverse prokaryotic symbionts of Phakellia fusca are valuable for the understanding and in-depth utilization of Phakellia fusca microbial symbionts. This study extends our knowledge of the community, especially the relative abundance of microbial symbionts in sponges. 相似文献
15.
Does microcystin disrupt the induced effect of Daphnia kairomone on colony formation in Scenedesmus?
In natural aquatic system, Scenedesmus and Microcystis species usually coexist. Microcystins are released into water after lysis of Microcystis cells during the collapse of heavy blooms. The released toxins can then come into contact with a wide range of aquatic organisms. In this study, we used filtered Daphnia test water containing kairomone from Daphnia magna to stimulate the inducible colony formation in Scenedesmus obliquus under microcystin-contaminated system, to examine how microcystin affects the induced effect of Daphnia kairomone on colony formation in S. obliquus. The results showed neither microcystin nor Daphnia kairomone affected the growth of S. obliquus. Microcystin neither promoted nor impaired the overall Daphnia-induced colony formation in S. obliquus, except reducing the proportion of eight-celled colonies on day 2, indicating that the effect of microcystin was just short-term and in general did not disrupt grazer-induced colony formation of S. obliquus. 相似文献
16.
Reaction of the affinity-labeling reagent N-bromoacetyl-[14C]phenylalanyl-tRNA with Escherichia coli ribosomes results in covalent labeling of 23 S ribosomal RNA in addition to the previously reported labeling of ribosomal proteins. The reaction with the 23 S RNA is absolutely dependent on the presence of messenger RNA. Covalent attachment of the affinity label to 23 S RNA was demonstrated by its integrity in strongly dissociating solvents, and the conversion of the labeled material to small oligonucleotides by ribonuclease treatment. After digestion of labeled 23 S RNA with T1 ribonuclease, the radioactivity is found mainly in two oligonucleotide fragments. These results support models in which both ribosomal RNA and ribosomal protein contribute to the structure of the region of the ribosome surrounding the peptidyl transferase center. 相似文献
17.
A. G. Markelova M. P. Sinetova E. V. Kupriyanova N. A. Pronina 《Russian Journal of Plant Physiology》2009,56(6):761-768
Localization of lumenal carbonic anhydrase Cah3 in thylakoid membranes of Chlamydomonas reinhardtii was studied using wild-type algae and photosynthetic mutants with different composition of chlorophyll-protein complexes
in the photosystems. In addition, the photosynthetic characteristics of wild-type C. reinhardtii and cia3 mutants lacking the activity of carbonic anhydrase Cah3 were examined. Western blot analysis revealed the lack of cross reaction
with antibodies to Cah3 in the mutant lacking the photosystem II (PSII) reaction center, in contrast to the mutant deficient
in light-harvesting complex of PSII. These data show that the lumenal Cah3 is associated with polypeptides on the donor side
of PSII reaction center. Using immunoelectron microscopy and antibodies to Cah3 from C. reinhardtii, we showed for the first time that the major part of thylakoid Cah3 is localized in the pyrenoid where the bulk of Rubisco
is located. The rate of photosynthetic oxygen evolution and PSII photochemical efficiency were lower in C. reinhardtii cia3 mutant than in the wild type, especially in the cells grown at limiting CO2 concentrations. These observations show that Cah3 takes part in CO2-concentrating mechanism of the chloroplast. The results support our hypothesis [1, 2] that the carboxylation reaction in
microalgae proceeds in the pyrenoid, a specific Rubisco-containing part of the chloroplast, which acquires CO2 from the lumen of intrapyrenoid thylakoids. We discuss significance of the pyrenoid as an autonomous metabolic microcompartment,
in which Cah3 plays a key role in the production and concentration of CO2 for Rubisco. These functions may promote the photosynthetic efficiency owing to the effective CO2 supply for the Calvin cycle. 相似文献
18.
Izabela Krzemińska Barbara Pawlik-Skowrońska Magdalena Trzcińska Jerzy Tys 《Bioprocess and biosystems engineering》2014,37(4):735-741
The effect of different photoperiods: 24 h illumination and a 12:12-h light/dark (12L:12D) cycle on the growth rate and biomass productivity was studied in five algal species: Neochloris conjuncta, Neochloris terrestris, Neochloris texensis, Botryococcus braunii and Scenedesmus obliquus. The green microalgae examined differ in the reproduction mode. Continuous illumination stimulated the growth of B. braunii and S. obliquus more effectively than the growth of the microalgal species from the genus Neochloris. However, under shorter duration of light of the same intensity (12L:12D cycle), the growth of all the three species of Neochloris was stimulated. Under continuous illumination, the specific growth rate in the first phase of B. braunii and S. obliquus cultures was higher than the growth rate of Neochloris, whereas under the 12L:12D cycle, the specific growth rate of all the three Neochloris species was generally higher than that in B. braunii and S. obliquus. As a result, the light regime influenced algal biomass productivity differently. The maximum biomass productivity was obtained in B. braunii and S. obliquus cultures carried out at continuous illumination. All the Neochloris species produced biomass more efficiently at the 12L:12D cycle, which was two–threefold higher than that of B. braunii and S. obliquus. The unicellular species of the green microalgae from the genus Neochloris, examined for the first time in this study, are promising prospective objects for algal biotechnology. 相似文献
19.
To investigate the effects of infochemicals from Daphnia magna reared at different densities on the growth and morphological development of Scenedesmus obliquus, we cultured S. obliquus with filtrates from D. magna with a density of 0, 6, 30, 150, 300, and 600 ind L−1, respectively. Results showed no significant differences in S. obliquus growth rates among the treatments with different Daphnia densities. On days 3 and 5, the proportion of eight-celled colonies in the treatments constituted 30% of the S. obliquus populations. Importantly, the number of cells per particle increased significantly with increasing density of Daphnia, indicating a grazer density-dependent response. Specifically, the density-response relationship followed a rectangular hyperbolic model in the first five days of treatment, and then switched to a linear model, which implies that the relationship between the inducible colony size of S. obliquus and Daphnia density may also be dependent upon incubation time. 相似文献
20.
水芹水浸提液对斜生栅藻的化感效应研究 总被引:3,自引:0,他引:3
宰学明;朱士农;钦佩;袁亚光;吴向华;周文宗 《植物研究》2011,31(6):735-738
采用栅藻在不同浓度的水芹水浸提液中纯培养的方法,研究了水芹水浸提液对斜生栅藻细胞数、叶绿素含量及藻细胞超微结构的影响。结果显示,10 g·L-1水芹水浸提液对斜生栅藻的生长和叶绿素含量的增加具有明显的促进效应;20 g·L-1水芹水浸提液对藻细胞数和叶绿素含量的增加持续到第3 d,3 d后出现显著的抑制效应;高浓度(30~50 g·L-1)水芹水浸提液对藻细胞数和叶绿素含量的明显抑制在第2 d始现,随时间延长而加剧,并具有浓度效应;40 g·L-1水芹水浸提液处理后,斜生栅藻细胞壁断裂甚至消失,细胞中叶绿体片层肿胀甚至解体,核膜破裂,核质外渗。水芹水浸提液对斜生栅藻具有化感效应,低浓度促进生长,高浓度抑制。 相似文献