Microbial oil production from sugarcane bagasse hydrolysates by oleaginous yeast and filamentous fungi

This study investigated the potential use of sugarcane bagasse as a feedstock for oil production through microbial cultivation. Bagasse was subjected to dilute acid pretreatment with 0.4 wt% H2SO4 (in liquid) at a solid/liquid ratio of 1:6 (wt/wt) at 170 °C for 15 min, followed by enzymatic hydrolysis of solid residue. The liquid fractions of the pretreatment process and the enzymatic hydrolysis process were detoxified and used as liquid hydrolysate (SCBLH) and enzymatic hydrolysate (SCBEH) for the microbial oil production by oleaginous yeast (Rhodotorula mucilaginosa) and filamentous fungi (Aspergillus oryzae and Mucor plumbeus). The results showed that all strains were able to grow and produce oil from bagasse hydrolysates. The highest oil concentrations produced from bagasse hydrolysates were by M. plumbeus at 1.59 g/L (SCBLH) and 4.74 g/L (SCBEH). The microbial oils obtained have similar fatty acid compositions to vegetable oils, indicating that the oil can be used for the production of second generation biodiesel. On the basis of oil yields obtained by M. plumbeus, from 10 million t (wet weight) of bagasse generated annually from sugar mills in Australia, it is estimated that the total biodiesel that could be produced would be equivalent to about 9% of Queensland’s diesel consumption.

On the ecology of cold-water phytoplankton in the Baltic Sea

Increased anthropogenic loading of nitrogen (N) and phosphorus (P) has led to an eutrophication problem in the Baltic Sea, and the spring bloom is a key component in the biological uptake of increased nutrient concentrations. The spring bloom in the Baltic Sea is dominated by both diatoms and dinoflagellates. However, the sedimentation of these groups is different: diatoms tend to sink to the sea floor at the end of the bloom, while dinoflagellates to a large degree are been remineralized in the euphotic zone. Understanding phytoplankton competition and species specific ecological strategies is thus of importance for assessing indirect effects of phytoplankton community composition on eutrophication problems. The main objective of this thesis was to describe some basic physiological and ecological characteristics of the main cold-water diatoms and dinoflagellates in the Baltic Sea. This was achieved by specific studies of: (1) seasonal vertical positioning, (2) dinoflagellate life cycle, (3) mixotrophy, (4) primary production, respiration and growth and (5) diatom silicate uptake, using cultures of common cold-water diatoms: Chaetoceros wighamii, C. gracilis, Pauliella taeniata, Thalassiosira baltica, T. levanderi, Melosira arctica, Diatoma tenuis, Nitzschia frigida, and dinoflagellates: Peridiniella catenata, Woloszynskia halophila and Scrippsiella hangoei. The diatoms had higher primary production capacity and lower respiration rate compared with the dinoflagellates. This difference was reflected in the maximum growth rate, which for the examined diatoms range from 0.6 to 1.2 divisions d-1, compared with 0.2 to 0.3 divisions d-1 for the dinoflagellates. Among diatoms there were species specific differences in light utilization and uptake of silicate, and C. wighamii had the highest carbon assimilation capacity and maximum silicate uptake. The physiological properties of diatoms and dinoflagellates were used in a model of the onset of the spring bloom: for the diatoms the model could predict the initiation of the spring bloom; S. hangoei, on the other hand, could not compete successfully and did not obtain positive growth in the model. The other dinoflagellates did not have higher growth rates or carbon assimilation rates and would thus probably not perform better than S. hangoei in the model. The dinoflagellates do, however, have competitive advantages that were not included in the model: motility and mixotrophy. Previous investigations has revealed that the chain-forming P. catenata performs diurnal vertical migration (DVM), and the results presented here suggest that active positioning in the water column, in addition to DVM, is a key element in this species' life strategy. There was indication of mixotrophy in S. hangoei, as it produced and excreted the enzyme leucine aminopeptidase (LAP). Moreover, there was indirect evidence that W. halophila obtains carbon from other sources than photosynthesis when comparing increase in cell numbers with in situ carbon assimilation rates. The results indicate that mixotrophy is a part of the strategy of vernal dinoflagellates in the Baltic Sea. There were also indications that the seeding of the spring bloom is very important for the dinoflagellates to succeed. In mesocosm experiments dinoflagellates could not compete with diatoms when their initial numbers were low. In conclusion, this thesis has provided new information about the basic physiological and ecological properties of the main cold-water phytoplankton in the Baltic Sea. The main phytoplankton groups, diatoms and dinoflagellates, have different physiological properties, which clearly separate their life strategies. The information presented here could serve as further steps towards better prognostic models of the effects of eutrophication in the Baltic Sea.

Herbal medicines used in the treatment of diabetes mellitus in Arunachal Himalaya, northeast, India

Ethnopharmacological relevance: Medicinal plants have played an important role in treating and preventing a variety of diseases throughout the world. Khampti tribal people living in the far-flung Lohit district of the Eastern Arunachal Himalaya, India still depend on medicinal plants and most of them have a general knowledge of medicinal plants which are used for treating a variety of ailments. This survey was undertaken in Lohit district in order to inventory the medicinal plants used in folk medicine to treat diabetes mellitus. Materials and methods: Field investigations were conducted in seventeen remote villages of Lohit district starting from April 2002 to May 2004 through interviews among 251 key informants who were selected randomly during our household survey. To elucidate community domains and determine differences in indigenous traditional knowledge of medicinal plants with anti-diabetic efficacy, we repeated our field survey starting from April 2008 to May 2010 with one hundred traditional healers locally called as ``Chau ya'' in Khampti of Lohit district. ``Chau ya'' traditional healers who know and use medicinal plants for treating diabetes mellitus were interviewed using a semi-structured questionnaire. Results: This study reports an ethnobotanical survey of medicinal plants in Lohit district of Arunachal Pradesh reputed for the treatment of diabetes mellitus. Forty-six plant species were identified in the study area to treat diabetes mellitus by the Khamptis ``Chau ya'' traditional healers. Comparative published literature survey analysis of this study with other ethnobotanical surveys of plants used traditionally in treating diabetes mellitus suggests that eleven plant species make claims of new reports on antidiabetic efficacy. These plant species are Begonia roxburghii, Calamus tenuis, Callicarpa arborea, Cuscuta reflexa, Dillenia indica, Diplazium esculentum, Lectuca gracilis, Millingtonia hortensis, Oxalis griffithii, Saccharum spontaneum, and Solanum viarum. Some of the plants reported in this study have an antidiabetic effect on rodent models but none have sufficient clinical evidence of effectiveness. Conclusions: The wide variety of medicinal plants that are used to treat diabetes mellitus in this area supports the importance of plants in the primary healthcare system of the rural people of Lohit district of Arunachal Pradesh. The finding of new plant uses in the current study reveals the importance of the documentation of such ethnobotanical knowledge. (C) 2012 Elsevier Ireland Ltd. All rights reserved.

Ecological condition of coastal ocean waters along the U.S. Western Continental Shelf: 2003

Executive Summary: The western National Coastal Assessment (NCA-West) program of EPA, in conjunction with the NOAA National Ocean Service (NOS), conducted an assessment of the status of ecological condition of soft sediment habitats and overlying waters along the western U.S. continental shelf, between the target depths of 30 and 120 m, during June 2003. NCA-West and NOAA/NOS partnered with the West Coast states (Washington (WA), Oregon (OR), and California (CA)), and the Southern California Coastal Water Research Project (SCCWRP) Bight ’03 program to conduct the survey. A total of 257 stations were sampled from Cape Flattery, WA to the Mexican border using standard methods and indicators applied in previous coastal NCA projects. A key study feature was the incorporation of a stratified-random sampling design with stations stratified by state and National Marine Sanctuary (NMS) status. Each of the three states was represented by at least 50 random stations. There also were a total of 84 random stations located within NOAA’s five NMSs along the West Coast including the Olympic Coast NMS (OCNMS), Cordell Bank NMS (CBNMS), Gulf of Farallones NMS (GFNMS), Monterey Bay NMS (MBNMS), and Channel Islands NMS (CINMS). Collection of flatfish via hook-and-line for fish-tissue contaminant analysis was successful at 50 EMAP/NCA-West stations. Through a collaboration developed with the FRAM Division of the Northwest Fisheries Science Center, fish from an additional 63 stations in the same region and depth range were also analyzed for fish-tissue contaminants. Bottom depth throughout the region ranged from 28 m to 125 m for most stations. Two slightly deeper stations from the Southern California Bight (SCB) (131, 134 m) were included in the data set. About 44% of the survey area had sediments composed of sands (< 20% silt-clay), about 47% was composed of intermediate muddy sands (20-80% silt-clay), and about 9% was composed of muds (> 80% silt-clay). The majority of the survey area (97%) had relatively low percent total organic carbon (TOC) levels of < 2%, while a small portion (< 1%) had high TOC levels (> 5%), in a range potentially harmful to benthic fauna. Salinity of surface waters for 92% of the survey area were > 31 psu, with most stations < 31 psu associated with the Columbia River plume. Bottom salinities ranged only between 31.6 and 34.4 psu. There was virtually no difference in mean bottom salinities among states or between NMS and non-NMS stations. Temperatures of surface water (range 8.5 -19.9 °C) and bottom water (range 5.8 -14.7 °C) averaged several degrees higher in CA in comparison to WA and OR. The Δσt index of watercolumn stratification indicated that about 31% of the survey area had strong vertical stratification of the water column. The index was greatest for waters off WA and lowest for CA waters. Only about 2.6 % of the survey area had surface dissolved oxygen (DO) concentrations ≤ 4.8 mg/L, and there were no values below the lower threshold (2.3 mg/L) considered harmful to the survival and growth of marine animals. Surface DO concentrations were higher in WA and OR waters than in CA, and higher in the OC NMS than in the CA sanctuaries. An estimated 94.3% of the area had bottom-water DO concentrations ≤ 4.8 mg/L and 6.6% had concentrations ≤ 2.3 mg/L. The high prevalence of DO from 2.3 to 4.8 mg/L (85% of survey area) is believed to be associated with the upwelling of naturally low DO water across the West Coast shelf. Mean TSS and transmissivity in surface waters (excluding OR due to sample problems) were slightly higher and lower, respectively, for stations in WA than for those in CA. There was little difference in mean TSS or transmissivity between NMS and non-NMS locations. Mean transmissivity in bottom waters, though higher in comparison to surface waters, showed little difference among geographic regions or between NMS and non-NMS locations. Concentrations of nitrate + nitrite, ammonium, total dissolved inorganic nitrogen (DIN) and orthophosphate (P) in surface waters tended to be highest in CA compared to WA and OR, and higher in the CA NMS stations compared to CA non-sanctuary stations. Measurements of silicate in surface waters were limited to WA and CA (exclusive of the SCB) and showed that concentrations were similar between the two states and approximately twice as high in CA sanctuaries compared to OCNMS or nonsanctuary locations in either state. The elevated nutrient concentrations observed at CA NMS stations are consistent with the presence of strong upwelling at these sites at the time of sampling. Approximately 93% of the area had DIN/P values ≤ 16, indicative of nitrogen limitation. Mean DIN/P ratios were similar among the three states, although the mean for the OCNMS was less than half that of the CA sanctuaries or nonsanctuary locations. Concentrations of chlorophyll a in surface waters ranged from 0 to 28 μg L-1, with 50% of the area having values < 3.9 μg L-1 and 10% having values > 14.5 μg L-1. The mean concentration of chlorophyll a for CA was less than half that of WA and OR locations, and concentrations were lowest in non-sanctuary sites in CA and highest at the OCNMS. Shelf sediments throughout the survey area were relatively uncontaminated with the exception of a group of stations within the SCB. Overall, about 99% of the total survey area was rated in good condition (<5 chemicals measured above corresponding effect range low (ERL) concentrations). Only the pesticides 4,4′-DDE and total DDT exceeded corresponding effect range-median (ERM) values, all at stations in CA near Los Angeles. Ten other contaminants including seven metals (As, Cd, Cr, Cu, Hg, Ag, Zn), 2-methylnaphthalene, low molecular weight PAHs, and total PCBs exceeded corresponding ERLs. The most prevalent in terms of area were chromium (31%), arsenic (8%), 2-methylnaphthalene (6%), cadmium (5%), and mercury (4%). The chromium contamination may be related to natural background sources common to the region. The 2-methylnaphthalene exceedances were conspicuously grouped around the CINMS. The mercury exceedances were all at non-sanctuary sites in CA, particularly in the Los Angeles area. Concentrations of cadmium in fish tissues exceeded the lower end of EPA’s non-cancer, human-health-risk range at nine of 50 EMAP/NCA-West and nine of 60 FRAM groundfish-survey stations, including a total of seven NMS stations in CA and two in the OCNMS. The human-health guidelines for all other contaminants were only exceeded for total PCBs at one station located in WA near the mouth of the Columbia River. Benthic species richness was relatively high in these offshore assemblages, ranging from 19 to 190 taxa per 0.1-m2 grab and averaging 79 taxa/grab. The high species richness was reflected over large areas of the shelf and was nearly three times greater than levels observed in estuarine samples along the West Coast (e.g NCA-West estuarine mean of 26 taxa/grab). Mean species richness was highest off CA (94 taxa/grab) and lower in OR and WA (55 and 56 taxa/grab, respectively). Mean species richness was very similar between sanctuary vs. non-sanctuary stations for both the CA and OR/WA regions. Mean diversity index H′ was highest in CA (5.36) and lowest in WA (4.27). There were no major differences in mean H′ between sanctuary vs. nonsanctuary stations for both the CA and OR/WA regions. A total of 1,482 taxa (1,108 to species) and 99,135 individuals were identified region-wide. Polychaetes, crustaceans and molluscs were the dominant taxa, both by percent abundance (59%, 17%, 12% respectively) and percent species (44%, 25%, 17%, respectively). There were no major differences in the percent composition of benthic communities among states or between NMSs and corresponding non-sanctuary sites. Densities averaged 3,788 m-2, about 30% of the average density for West Coast estuaries. Mean density of benthic fauna in the present offshore survey, averaged by state, was highest in CA (4,351 m-2) and lowest in OR (2,310 m-2). Mean densities were slightly higher at NMS stations vs. non-sanctuary stations for both the CA and OR/WA regions. The 10 most abundant taxa were the polychaetes Mediomastus spp., Magelona longicornis, Spiophanes berkeleyorum, Spiophanes bombyx, Spiophanes duplex, and Prionospio jubata; the bivalve Axinopsida serricata, the ophiuroid Amphiodia urtica, the decapod Pinnixa occidentalis, and the ostracod Euphilomedes carcharodonta. Mediomastus spp. and A. serricata were the two most abundant taxa overall. Although many of these taxa have broad geographic distributions throughout the region, the same species were not ranked among the 10 most abundant taxa consistently across states. The closest similarities among states were between OR and WA. At least half of the 10 most abundant taxa in NMSs were also dominant in corresponding nonsanctuary waters. Many of the abundant benthic species have wide latitudinal distributions along the West Coast shelf, with some species ranging from southern CA into the Gulf of Alaska or even the Aleutians. Of the 39 taxa on the list of 50 most abundant taxa that could be identified to species level, 85% have been reported at least once from estuaries of CA, OR, or WA exclusive of Puget Sound. Such broad latitudinal and estuarine distributions are suggestive of wide habitat tolerances. Thirteen (1.2%) of the 1,108 identified species are nonindigenous, with another 121 species classified as cryptogenic (of uncertain origin), and 208 species unclassified with respect to potential invasiveness. Despite uncertainties of classification, the number and densities of nonindigenous species appear to be much lower on the shelf than in the estuarine ecosystems of the Pacific Coast. Spionid polychaetes and the ampharetid polychaete Anobothrus gracilis were a major component of the nonindigenous species collected on the shelf. NOAA’s five NMSs along the West Coast of the U.S. appeared to be in good ecological condition, based on the measured indicators, with no evidence of major anthropogenic impacts or unusual environmental qualities compared to nearby nonsanctuary waters. Benthic communities in sanctuaries resembled those in corresponding non-sanctuary waters, with similarly high levels of species richness and diversity and low incidence of nonindigenous species. Most oceanographic features were also similar between sanctuary and non-sanctuary locations. Exceptions (e.g., higher concentrations of some nutrients in sanctuaries along the CA coast) appeared to be attributable to natural upwelling events in the area at the time of sampling. In addition, sediments within the sanctuaries were relatively uncontaminated, with none of the samples having any measured chemical in excess of ERM values. The ERL value for chromium was exceeded in sediments at the OCNMS, but at a much lower percentage of stations (four of 30) compared to WA and OR non-sanctuary areas (31 of 70 stations). ERL values were exceeded for arsenic, cadmium, chromium, 2- methylnaphthalene, low molecular weight PAHs, total DDT, and 4,4′-DDE at multiple sites within the CINMS. However, cases where total DDT, 4,4′-DDE, and chromium exceeded the ERL values were notably less prevalent at CINMS than in non-sanctuary waters of CA. In contrast, 2-methylnaphthalene above the ERL was much more prevalent in sediments at the CINMS compared to non-sanctuary waters off the coast of CA. While there are natural background sources of PAHs from oil seeps throughout the SCB, this does not explain the higher incidence of 2-methylnaphthalene contamination around CINMS. Two stations in CINMS also had levels of TOC (> 5%) potentially harmful to benthic fauna, though none of these sites exhibited symptoms of impaired benthic condition. This study showed no major evidence of extensive biological impacts linked to measured stressors. There were only two stations, both in CA, where low numbers of benthic species, diversity, or total faunal abundance co-occurred with high sediment contamination or low DO in bottom water. Such general lack of concordance suggests that these offshore waters are currently in good condition, with the lower-end values of the various biological attributes representing parts of a normal reference range controlled by natural factors. Results of multiple linear regression, performed using full model procedures to test for effects of combined abiotic environmental factors, suggested that latitude and depth had significant influences on benthic variables regionwide. Latitude had a significant inverse influence on all three of the above benthic variables, i.e. with values increasing as latitude decreased (p< 0.01), while depth had a significant direct influence on diversity (p < 0.001) and inverse effect on density (p <0.01). None of these variables varied significantly in relation to sediment % fines (at p< 0.1), although in general there was a tendency for muddier sediments (higher % fines) to have lower species richness and diversity and higher densities than coarser sediments. Alternatively, it is possible that for some of these sites the lower values of benthic variables reflect symptoms of disturbance induced by other unmeasured stressors. The indicators in this study included measures of stressors (e.g., chemical contaminants, eutrophication) that are often associated with adverse biological impacts in shallower estuarine and inland ecosystems. However, there may be other sources of humaninduced stress in these offshore systems (e.g., bottom trawling) that pose greater risks to ambient living resources and which have not been captured. Future monitoring efforts in these offshore areas should include indicators of such alternative sources of disturbance. (137pp.) (PDF contains 167 pages)

A comparison of seafloor habitats and associated benthic fauna in areas open and closed to bottom trawling along the central California Continental Shelf

Executive Summary: A number of studies have shown that mobile, bottom-contact fishing gear (such as otter trawls) can alter seafloor habitats and associated biota. Considerably less is known about the recovery of these resources following such disturbances, though this information is critical for successful management. In part, this paucity of information can be attributed to the lack of access to adequate control sites – areas of the seafloor that are closed to fishing activity. Recent closures along the coast of central California provide an excellent opportunity to track the recovery of historically trawled areas and to compare recovery rates to adjacent areas that continue to be trawled. In June 2006 we initiated a multi-year study of the recovery of seafloor microhabitats and associated benthic fauna inside and outside two new Essential Fish Habitat (EFH) closures within the Cordell Bank and Gulf of the Farallones National Marine Sanctuaries. Study sites inside the EFH closure at Cordell Bank were located in historically active areas of fishing effort, which had not been trawled since 2003. Sites outside the EFH closure in the Gulf of Farallones were located in an area that continues to be actively trawled. All sites were located in unconsolidated sands at equivalent water depths. Video and still photographic data collected via a remotely operated vehicle (ROV) were used to quantify the abundance, richness, and diversity of microhabitats and epifaunal macro-invertebrates at recovering and actively trawled sites, while bottom grabs and conductivity/temperature/depth (CTD) casts were used to quantify infaunal diversity and to characterize local environmental conditions. Analysis of still photos found differences in common seafloor microhabitats between the recovering and actively trawled areas, while analysis of videographic data indicated that biogenic mound and biogenic depression microhabitats were significantly less abundant at trawled sites. Each of these features provides structure with which demersal fishes, across a wide range of size classes, have been observed to associate. Epifaunal macro-invertebrates were sparsely distributed and occurred in low numbers in both treatments. However, their total abundance was significantly different between treatments, which was attributable to lower densities at trawled sites. In addition, the dominant taxa were different between the two sites. Patchily-distributed buried brittle stars dominated the recovering site, and sea whips (Halipteris cf. willemoesi) were most numerous at the trawled site though they occurred in only five of ten transects. Numerical classification (cluster analysis) of the infaunal samples also revealed a clear difference between benthic assemblages in the recovering vs. trawled areas due to differences in the relative abundances of component species. There were no major differences in infaunal species richness, H′ diversity, or J′ evenness between recovering vs. trawled site groups. However, total infaunal abundance showed a significant difference attributable to much lower densities at trawled sites. This pattern was driven largely by the small oweniid polychaete Myriochele gracilis, which was the most abundant species in the overall study region though significantly less abundant at trawled sites. Other taxa that were significantly less abundant at trawled sites included the polychaete M. olgae and the polychaete family Terebellidae. In contrast, the thyasirid bivalve Axinopsida serricata and the polychaetes Spiophanes spp. (mostly S. duplex), Prionospio spp., and Scoloplos armiger all had significantly to near significantly higher abundances at trawled sites. As a result of such contrasting species patterns, there also was a significant difference in the overall dominance structure of infaunal assemblages between the two treatments. It is suggested that the observed biological patterns were the result of trawling impacts and varying levels of recovery due to the difference in trawling status between the two areas. The EFH closure was established in June 2006, within a month of when sampling was conducted for the present study, however, the stations within this closure area are at sites that actually have experienced little trawling since 2003, based on National Marine Fishery Service trawl records. Thus, the three-year period would be sufficient time for some post-trawling changes to have occurred. Other results from this study (e.g., similarly moderate numbers of infaunal species in both areas that are lower than values recorded elsewhere in comparable habitats along the California continental shelf) also indicate that recovery within the closure area is not yet complete. Additional sampling is needed to evaluate subsequent recovery trends and persistence of effects. Furthermore, to date, the study has been limited to unconsolidated substrates. Ultimately, the goal of this project is to characterize the recovery trajectories of a wide spectrum of seafloor habitats and communities and to link that recovery to the dynamics of exploited marine fishes. (PDF has 48 pages.)

Bright-field and fluorescence chip-scale microscopy for biological imaging

Optical microscopy is an essential tool in biological science and one of the gold standards for medical examinations. Miniaturization of microscopes can be a crucial stepping stone towards realizing compact, cost-effective and portable platforms for biomedical research and healthcare. This thesis reports on implementations of bright-field and fluorescence chip-scale microscopes for a variety of biological imaging applications. The term “chip-scale microscopy” refers to lensless imaging techniques realized in the form of mass-producible semiconductor devices, which transforms the fundamental design of optical microscopes.

Our strategy for chip-scale microscopy involves utilization of low-cost Complementary metal Oxide Semiconductor (CMOS) image sensors, computational image processing and micro-fabricated structural components. First, the sub-pixel resolving optofluidic microscope (SROFM), will be presented, which combines microfluidics and pixel super-resolution image reconstruction to perform high-throughput imaging of fluidic samples, such as blood cells. We discuss design parameters and construction of the device, as well as the resulting images and the resolution of the device, which was 0.66 µm at the highest acuity. The potential applications of SROFM for clinical diagnosis of malaria in the resource-limited settings is discussed.

Next, the implementations of ePetri, a self-imaging Petri dish platform with microscopy resolution, are presented. Here, we simply place the sample of interest on the surface of the image sensor and capture the direct shadow images under the illumination. By taking advantage of the inherent motion of the microorganisms, we achieve high resolution (~1 µm) imaging and long term culture of motile microorganisms over ultra large field-of-view (5.7 mm × 4.4 mm) in a specialized ePetri platform. We apply the pixel super-resolution reconstruction to a set of low-resolution shadow images of the microorganisms as they move across the sensing area of an image sensor chip and render an improved resolution image. We perform longitudinal study of Euglena gracilis cultured in an ePetri platform and image based analysis on the motion and morphology of the cells. The ePetri device for imaging non-motile cells are also demonstrated, by using the sweeping illumination of a light emitting diode (LED) matrix for pixel super-resolution reconstruction of sub-pixel shifted shadow images. Using this prototype device, we demonstrate the detection of waterborne parasites for the effective diagnosis of enteric parasite infection in resource-limited settings.

Then, we demonstrate the adaptation of a smartphone’s camera to function as a compact lensless microscope, which uses ambient illumination as its light source and does not require the incorporation of a dedicated light source. The method is also based on the image reconstruction with sweeping illumination technique, where the sequence of images are captured while the user is manually tilting the device around any ambient light source, such as the sun or a lamp. Image acquisition and reconstruction is performed on the device using a custom-built android application, constructing a stand-alone imaging device for field applications. We discuss the construction of the device using a commercial smartphone and demonstrate the imaging capabilities of our system.

Finally, we report on the implementation of fluorescence chip-scale microscope, based on a silo-filter structure fabricated on the pixel array of a CMOS image sensor. The extruded pixel design with metal walls between neighboring pixels successfully guides fluorescence emission through the thick absorptive filter to the photodiode layer of a pixel. Our silo-filter CMOS image sensor prototype achieves 13-µm resolution for fluorescence imaging over a wide field-of-view (4.8 mm × 4.4 mm). Here, we demonstrate bright-field and fluorescence longitudinal imaging of living cells in a compact, low-cost configuration.

Methods of feeding of some macrothricids (Cladocera, Macrothricidae). [Translation from: Informatsionnyi Byulleten Biologiya Vnutrennikh Vod 16, 29-34, 1973.]

The feeding methods of macrothricids was investigated in three species — Ophryoxus gracilis G.O. Sars, Ilyocryptus sordidus (Lievin) and Lathonura rectirostris O.F.Muller. The procuring of food in those species follows two methods: filtration of dispersed particles or scraping off and collecting food from the surface of the substrate. When collecting food by scraping off is the sole method of feeding (as in Lathonura rectirostris), movement along the substrate and feeding are combined so intimately that they appear as parts of one and the same mechanism.

辽西中生代银杏目和茨康目植物生殖器官研究

本文研究了采自辽宁西部下白垩统义县组和上三叠统羊草沟组的三种植物的生殖器官，它们分别属于银杏目 (Ginkgoales) 和茨康目 (Czekanowskiales)。辽宁银杏 (G. liaoningensis Liu, Crane, Li et Wang sp. nov.) 和纤细堆囊穗 (Sorosaccus gracilis Harris 1935 emend. Liu, Hueber, Li et Wang) 是银杏目植物的雄球花，通过将这两种雄球花与现存银杏 (Ginkgo biloba L.) 雄球花的对比研究，讨论了自晚三叠世至今银杏目雄性生殖器官可能的演化途径。蟹形薄果穗 (Leptostrobus cancer Harris 1951 emend. Liu, Li et Wang) 是茨康目植物的雌性生殖器官，我们根据辽西标本对该种的特征集要做了修订，并对薄果穗属 (Leptostrobus Heer 1876) 的种进行了讨论，将三个种归并到蟹形薄果穗薄。主要内容包括： 1. 银杏属植物雄球花——辽宁银杏 采自中国辽宁下白垩统义县组的辽宁银杏，为葇荑花序状，主轴基部有鳞片。小孢子叶螺旋排列在主轴上，小孢子叶上着生3—4个 (少数为2个) 卵形到椭圆形的小孢子囊。小孢子囊下垂，纵状开裂。成熟花粉粒单沟，长椭圆形;而未成熟的花粉粒处在四分体阶段。辽宁银杏的特征和现存银杏十分相似。主要差别在于辽宁银杏的小孢子叶着生3—4个 (少数为2个) 小孢子囊，而现存银杏的小孢子叶绝大多数着生2个小孢子囊。辽宁银杏在大小及小孢子囊数目上区别于其它的银杏属雄球花化石种，如：英国约克郡侏罗纪的胡顿银杏 (Ginkgo huttoni (Sternberg) Heer) 雄球花，加拿大阿尔贝塔晚白垩世一个未定名的银杏属雄球花。辽宁银杏 (化石) 和银杏 (现存) 的比较支持了根据银杏属胚珠器官而提出的缩减假说，因为自早白垩世至今，银杏属雄球花小孢子叶着生的小孢子囊的数目经历了从3—4个到2个的减少。 2. 银杏目植物雄球花——纤细堆囊穗 本文研究的纤细堆囊穗采自中国东北上三叠统羊草沟组。这种雄球花小孢子 叶末端的裂片在形态上差异很大，这是在以前对这种植物的研究中没有发现的重要特征。这些保存精美的化石对于纤细堆囊穗特征的阐明和修订以及种的复原很有帮助。将中国的标本与西伯利亚堆囊穗 (Sorosaccus sibiricus Prynada 1962)、小堆囊穗 (S. minor Harris 1935)、乌马尔堆囊穗 (S. umaltensis Krassilov 1972) 和被定为长叶拜拉 (Baiera longifolia (Pom.) Heer 1876) 的雄球花进行了比较，发现它们与纤细堆囊穗在特征上一致，故将它们处理为纤细堆囊穗的异名。同时，也讨论了堆囊穗属可能的演化意义。它可能是银杏属的远祖，经过小孢子囊数目的减少和小孢子叶长度的缩短而演化到现在的银杏，而辽西早白垩世的辽宁银杏可能代表了堆囊穗和现代银杏在演化上的一个中间阶段。 3. 茨康目雌性生殖器官——蟹形薄果穗 本文研究的蟹形薄果穗采自辽西上三叠统羊草沟组和下白垩统义县组。羊草沟组的材料是蟹形薄果穗属最早的纪录，从而该种的分布在时间上有了新的资料：从晚三叠世到早白垩世。据标本穗轴顶端的完整性，我们修订了蟹形薄果穗的特征集要和提供了它的复原图，并将三种薄果穗归并到蟹形薄果穗。我们也回顾了薄果穗属自1876年建立以来100多年的研究历史，并列出了该属所有材料，还讨论了该属各种的主要特征和建种依据。

提高生物拮抗菌对果实采后病害的生防效力研究

摘要 　　"随着人们对身体健康和环境污染的日益重视，化学农药作为控制果实采后病害的主要方法受到了很大限制，科学研究者不得不寻求更加安全有效的防治果实采后病害的新方法。生物防治以其对环境和人类健康不造成危害的优点而逐渐受到人们的青睐。然而，由于生物防治是以活菌为基础，有其局限性和时效性，单独使用拮抗菌很难达到化学药剂完全控制果实采后病害的效果，因此，提高拮抗菌的生防效力成为当今生物防治领域的研究重点。本文主要研究了拮抗菌与不同外源物质配合使用的抑病效果及协同抑病机理;拮抗菌对采前田间和采后贮藏环境条件的适应能力;以及采前应用拮抗菌对果实采后贮藏期间病害的生物防治效力。研究结果表明： 　　1、酵母拮抗菌Cryptococcus laurentii与低浓度化学杀菌剂imazalil（25g/ml）和kresoxim-methyl（50g/ml）配合使用可以显著提高对冬枣果实采后黑霉病（Alternaria alternata）和褐腐病（Monilinia fructicola）的防治效果，杀菌剂并不影响拮抗菌在冬枣果实伤口的生长动态。 　　2、酵母拮抗菌Pichia membranefaciens和C. laurentii 与钼酸铵（NH4-Mo，5 mmol/L）和碳酸氢钠（NaHCO3，2%）配合能够显著提高对甜樱桃果实采后褐腐病（M. fructicola）的抑病能力。通过in vitro和扫描电镜观察结果表明，NH4-Mo和NaHCO3能够显著地抑制病原菌M. fructicola在培养基和果实伤口的生长，具有杀菌作用。 　　3、酵母拮抗菌C. laurentii和Rhodotorula glutinis与硅酸钠（Na2SiO3）配合使用对甜樱桃果实采后青霉病（Penicillium expansum）和褐腐病（M. fructicola）以及对冬枣果实青霉病（P. expansum）和黑霉病（A. alternata）的防治效果更好。经in vitro和扫描电镜观察表明，Na2SiO3对病原菌在培养基和果实伤口的生长有明显的抑制作用。同时，Na2SiO3还能诱导果实苯丙氨酸解氨酶（PAL）、多酚氧化酶（PPO）和过氧化物酶（POD）等抗性相关酶活性的提高。 　　4、酵母拮抗菌R. glutinis与水杨酸（SA，0.5mmol/L）配合可显著提高对甜樱桃果实采后青霉病（P. expansum）和黑霉病（A. alternata）的抑病能力。SA不影响拮抗菌在果实伤口的生长，in vitro实验中低浓度的SA对病原菌孢子萌发和芽管伸长也没有抑制作用。SA可能是通过诱导果实产生抗性来协同提高拮抗菌的抑病效果，而不是直接抑制病原菌生长。 　　5、酵母拮抗菌C. laurentii和R. glutinis在气调（Controlled atmospheres, CA）贮藏条件下对樱桃果实采后青霉病（P. expansum）和黑霉病（A. alternata）的防治效果显著提高。气调贮藏不抑制拮抗菌在甜樱桃果实伤口的生长。 　　6、采前应用酵母拮抗菌C. laurentii 和R. glutinis能够显著抑制甜樱桃果实在采后不同贮藏环境下的发病率。拮抗菌能够在田间果实表面生长并一直保持较高的数量。在试验的三种酵母拮抗菌中，C. laurentii的防病效果最好，该菌不仅能在果实表面迅速生长，也能适应低温和CA贮藏环境。"

酵母拮抗菌的抑病机理及生防效力改良的研究

相对于酵母拮抗菌的使用来说，人们对其作用机理了解得还不是很清楚。而了解拮抗菌的抑菌机理却是增强拮抗菌的生防效果以及进行拮抗菌筛选标准的重要前提。本文主要研究了酵母拮抗菌Pichia membranefaciens、Cryptococcus albidus以及Crytococcus laurentii对水果采后软腐病、褐腐病以及青霉病的防治效果，拮抗菌与病原菌之间的相互作用，并对酵母拮抗菌与外源物质配合使用，以及通过遗传改良途径来提高酵母拮抗菌生防能力等进行了初步研究。实验结果如下： 1、酵母拮抗菌P. membranefaciens、C. albidus以及C. laurentii能在果实伤口大量繁殖。采用扫描电镜技术，观察发现在桃果实伤口处P. membranefaciens能紧密地吸附在软腐病菌Rhizopous stolonfier的菌丝体上;C. laurentii与青霉病菌Penicillium expansum在苹果果实伤口处也存在着直接的拮抗作用;但P. membranefaciens和C. albidus对P. expansum的直接作用不明显。 2、酵母拮抗菌P. membranefaciens能够有效地抑制甜樱桃果实在常温和低温贮藏条件下褐腐病的发生。在常温贮藏条件下，P. membranefaciens和褐腐病菌Monilinia fracticola 处理都能够提高果实β-1,3-葡聚糖酶、POD、以及PAL酶的活性，但在低温贮藏条件下，拮抗菌和病原菌处理对甜樱桃果实β-1,3-葡聚糖酶、POD酶活性的升高有促进作用，对PAL和PPO酶活性的诱导作用不明显。 3、梨果实采后经过水杨酸，CaCl2，UV辐射和草酸等各种激发子处理以后，再接种病原菌Alternaria alternata，可以显著降低梨果实的发病率。其中，水杨酸处理的果实发病率最低。不同的激发子均可以诱导梨果实β-1,3-葡聚糖酶、POD、PAL和PPO酶活性的升高，但对果实乙烯含量的影响不明显。 4、氨基糖甙类抗菌素G418能够抑制P. membranefaciens的生长，其最低抑制浓度为100g ml-1。将G418抗性基因Neor插入到酵母-大肠杆菌穿梭表达载体pFL61中，构建PGK启动子驱动的表达载体pFL61-neo，利用醋酸锂转化法转化P. membranefaciens。酵母转化子在非选择性培养条件下连续生长50代后，仍有67.87%的细胞保留该质粒。这表明穿梭表达载体pFL61-neo能稳定地存在于P. membranefaciens中，并且该酵母细胞能有效地识别PGK启动子和终止子指导Neor的表达。 5、酵母拮抗菌C. laurentii和Rhodotorula glutinis与2%的碳酸氢钠混合使用，对冬枣果实青霉病的防治效果明显比单独使用拮抗菌或化学物质的防病效果好。其中，107CFU ml-1的拮抗菌与238 mmol l-1的碳酸氢钠配合使用可以达到单独使用108CFU ml-1拮抗菌的防病效果。另外，钼酸铵作为一种添加剂也能提高R. glutinis对梨果实青霉病和黑霉病的防治效果，但将钼酸铵与Trichosporon sp.配合使用的防病效果不明显。碳酸氢钠和钼酸铵在果实伤口对酵母拮抗菌的生长都有一定的抑制作用。 6、酵母拮抗菌P. membranefaciens在不同碳源、氮源中生长情况表明：在几种氮源中，大豆蛋白胨、酵母提取物、牛肉浸膏对P. membranefaciens的生长有显著的促进作用，其中，大豆蛋白胨的效果最好。在检测以葡萄糖、果糖和麦芽糖作为碳源的生长实验中，发现这几种碳源都能够被拮抗菌很好的利用，其中葡萄糖的利用率最好。小球藻生长因子（CGF）能够明显地促进了P. membranefaciens的生长。但是，CGF的浓度从0.5%增加到1%并没有促进酵母菌细胞数量的增加。

酵母拮抗菌的抑病效力、规模化培养及相关机理研究

我们实验室从果实表面分离获得的酵母拮抗菌已经证明能有效防治各种果实采后主要病害，为了加快生物拮抗菌的商业化应用，本文在完善拮抗菌抑病机理的基础上，重点研究了拮抗菌规模化培养条件，生物菌剂制品的稳定性，以及拮抗菌对环境胁迫的生理反应。主要研究内容包括：（1）分析酵母菌拮抗菌、病原菌与果实之间的互作效应及其影响因子;（2）筛选酵母拮抗菌规模化培养的最佳营养配方及培养条件;（3）优化酵母拮抗菌干粉与液体剂型的制备方式;（4）研究酵母拮抗菌在不同剂型中生活力下降的可能机理;（5）探讨酵母拮抗菌次生代谢产物的抑菌效果。研究结果如下： 1、单独接种Monilinia fructicola或同时接种M. fructicola和Cryptococcus laurentii均能诱导甜樱桃果实SOD、CAT和POD等抗氧化酶活性升高并加速脂质过氧化，同时伴有PPO同工酶新酶带出现。病原菌M. fructicola和Penicillum expansum在接种初期均显著促进拮抗菌C. laurentii在桃果实伤口处的生长。C. laurentii在接种24h内显著抑制桃果实LOX活性、O2•-产生与H2O2积累。单独接种病原菌能显著诱导桃果实LOX活性升高，促进O2•-产生，但抑制H2O2积累。病菌侵染后果实中 O2•-增加，以及H2O2的降低可能是桃果实对病原菌侵染的一种生理应答方式。 2、抗坏血酸钠能显著提高C. laurentii对甜樱桃果实褐腐病的防治效果，较低浓度的拮抗菌（ 1×107 cells mL-1）与200mM抗坏血酸配合使用可以达到较高浓度拮抗菌（1×108 cells mL-1）单独使用对M. fructicola的防治效果。抗坏血酸钠的协同抑病机理可能是在抑制病原菌生长的同时，也抑制了果实的抗氧化酶活性，从而加速了脂质过氧化过程。 3、酵母菌产业化培养条件的筛选结果表明，不同拮抗菌对培养基中营养物质的需求不一样，培养所需的温度有差异。在120L发酵罐的中试实验表明两种拮抗菌采用筛选出的最佳培养条件均得到浓度大于1× 109 CFU mL-1的菌悬液。 4、保护剂种类是影响Rhodotorula glutinis 和 C. laurentii两种酵母拮抗菌冷冻干燥效果的最主要因素，但保护剂效果的发挥依赖于其浓度与酵母菌生长阶段。无菌水和PBS（100mM, pH5.8）可以作为拮抗菌C. laurentii液体剂型的有效保护剂，而柠檬酸钠（100mM, pH5.8）则诱导拮抗菌C. laurentii的生活力快速丧失。 5、酵母菌拮抗菌冻干制品的研究表明，在胁迫环境下酵母菌生活力快速丧失与大量产生活性氧有关，这暗示活性氧的产生可能是导致酵母菌细胞死亡的主要因素。柠檬酸钠（100mM, pH5.8）对C. laurntii死亡的诱导效应受柠檬酸根浓度和介质酸度的双重影响。活性氧在柠檬酸钠诱导酵母菌生活力快速丧失中大量产生并发挥重要作用。 6、酵母拮抗菌能够产生某些对果实采后病原真菌具有抑制效果的挥发性和不挥发性物质。同一种酵母菌产生的物质对不同病原菌有不同的拮抗效果，而不同酵母菌对同一种病原菌的拮抗效果也不完全相同。但是，不同类型的培养基对拮抗菌产生的抑菌物质有明显的影响。

海藻糖提高酵母拮抗菌生活力和生防效力的作用机制

近年来，利用酵母拮抗菌进行果实采后病害的生物防治已经成为果实采后领域的研究热点。但是，在实际应用中生物拮抗菌制剂的防病效果远不如化学药剂稳定。由于生物拮抗菌是活体，生活力和生防效力易受诸多因素影响。在商品化制剂的剂型加工、销售、以及使用过程中的环境条件往往影响酵母拮抗菌的生活力和抑病能力，成为酵母拮抗菌产业化生产和商品化应用过程中的一个主要障碍。将酵母拮抗菌和其它化学物质配合使用可以提高拮抗菌的生防效力。另外，增加酵母拮抗菌对逆境条件的耐受力也是增加或稳定其防治效果的有效途径。本文研究了海藻糖与酵母拮抗菌生活力和生防效力的关系，通过生理手段提高了酵母拮抗菌内源海藻糖的含量，同时探讨了在多种逆境条件下内源海藻糖含量对酵母拮抗菌生活力和生防效力的影响及其作用机制。主要研究结果如下： 1. 以1 %海藻糖作为碳源培养酵母拮抗菌Cryptococcus laurentii可以提高其内源海藻糖含量。在in vitro试验中，提高内源海藻糖含量可以提高C. laurentii在低温（1 ºC）、气调（1 ºC，5 % O2，5 % CO2）条件下的生活力;在in vivo试验中，提高C. laurentii内源海藻糖含量可以提高其在苹果果实伤口上的种群密度和对苹果青霉病的防治效果。内源海藻糖的积累还能提高C. laurentii冷冻干燥后的生活力，海藻糖对酵母细胞质膜的保护作用可能是一个主要原因。 2. 以1 %海藻糖作为碳源能提高酵母拮抗菌Rhodotorula glutinis的内源海藻糖含量。内源海藻糖含量的增加可以提高C. laurentii和R. glutinis在慢速冷冻处理中的生活力。同时，海藻糖作为外源保护剂可以明显提高两种酵母拮抗菌在冷冻干燥处理后的生活力。在快速冷冻、慢速冷冻和冷冻干燥处理中，提高酵母拮抗菌的内源海藻糖含量并使用海藻糖作为外源保护剂可以获得更高的生活力。同时，这种内、外源保护因子的综合作用也可以提高两种拮抗菌在苹果果实伤口上的种群密度和对苹果青霉病的防治效果。 3. 脱脂牛奶和糖（葡萄糖，半乳糖，蔗糖，海藻糖）作为保护剂可以提高C. laurentii冷冻干燥后在常温（25 ºC）和低温（4 ºC）保存过程中的生活力。脱脂牛奶和糖保护剂的复合使用对C. laurentii的保护效果高于其单独使用的效果。通过对几种常用的碳源进行筛选，发现柠檬酸作为碳源对C. laurentii内源海藻糖的积累有明显的诱导作用。当使用相同的保护剂或保护剂组合时，高内源海藻糖含量的酵母拮抗菌生活力更强。当冷冻干燥前使用半乳糖 + 脱脂牛奶作为保护剂时，高内源海藻糖含量的C. laurentii在4 ºC保存90 天后对苹果青霉病的防治效果和与新鲜培养的酵母拮抗菌相当。 4. 酵母拮抗菌C. laurentii冷冻干燥后在常温(25 ºC)保存期间，细胞生活力下降，细胞膜的完整性降低，胞内活性氧水平增加，同时与抗氧化相关的超氧化物歧化酶（SOD）活性也增加，而过氧化氢酶（CAT）活性则下降。提高内源海藻糖含量和/或使用外源保护剂（5 %脱脂牛奶 + 10 %葡萄糖）可以减缓上述指标下降或上升的速度。内、外源因子的共同作用有利于提高对拮抗菌细胞的保护作用。 5. 利用褐藻酸钠制成的含酵母拮抗菌的胶球在干燥后能有效的保持C. laurentii的生活力。在3种不同粘度的褐藻酸钠中，0.5 % 3500 cp的褐藻酸钠表现出较好的保护效果。内源海藻糖的积累可以提高干胶球中C. laurentii的生活力，作为外源保护剂的4种糖（葡萄糖、半乳糖、蔗糖、海藻糖）中只有海藻糖在低温条件下可以提高C. laurentii的生活力，其它3种糖反而降低了C. laurentii的生活力。

外源物质对酵母拮抗菌生活力的影响

　　近年来，酵母拮抗菌在水果采后病害防治中展示了良好的应用前景。然而，在实际应用中,酵母拮抗菌在逆境条件下会因为发生凋亡或细胞损伤而引起生活力的下降，最终导致拮抗菌抑病能力降低。研究酵母拮抗菌生活力下降的规律，提高酵母拮抗菌的生产效率，减少剂型加工过程中的细胞损伤，增强其对逆境条件的耐受力是增加或稳定生防制剂防治效果的有效途径。本文主要研究酵母拮抗菌正常培养过程中生活力下降的规律，筛选剂型加工过程中对酵母拮抗菌具有保护作用的化学物质，并对酵母拮抗菌的培养条件进行了优化。主要研究结果如下： 　　1. 在正常培养过程中，酵母拮抗菌Rhodotorula glutinis和Cryptococcus laurentii中细胞染色质凝集或细胞膜破损的发生一般在6天以后。外源加入的N-乙酰半胱氨酸及硅酸钠等物质在超过一定浓度时会加速酵母菌的死亡。 　　2. 在不同的液体悬浮制剂中，对R. glutinis而言，使用磷酸缓冲液（PBS）悬浮时保护效果最好；而C. laurentii悬浮在NYDB培养基中或海藻糖、乳糖溶液中时的生活力最高。 　　3. 以10 %葡萄糖 + 5 %脱脂牛奶作保护剂，可以有效地保持酵母拮抗菌C. laurentii冻干制剂的生活力，配合使用的保护效果高于它们单独使用时的保护效果。添加1 mM N-乙酰半胱氨酸能更好地保持拮抗菌制剂在常温保存过程中的生活力，这可能与这种还原性物质缓解了细胞内活性氧的积累有关。 　　4. 不同酵母拮抗菌对不同碳、氮源的利用能力有明显差异。在9种不同的碳源和10种不同的氮源中，Pichia membranefaciens能够最有效利用的碳、氮源是葡萄糖、果糖和多价胨，而Candida guilliermondii的最佳碳源和氮源分别是果糖和肉蛋白胨。

酵母拮抗菌和外源水杨酸对核果类果实抗病相关基因的诱导表达

在果实采后贮藏过程中，病原真菌的侵染会引起果实腐烂，造成巨大的经济损失。利用生物和非生物因子诱导果实抗病性，已经成为采后病害防治领域的一个研究热点。本文主要利用RT-PCR和RACE技术克隆果实抗病相关基因，通过分子杂交和蛋白羰基化免疫检测技术，研究了外源SA和酵母拮抗菌诱导果实抗病性机理，结果表明： 1. 通过优化RNA提取方法，能从含有多糖的冬枣、葡萄、甜樱桃、桃、番茄等果实中提取到质量较好的RNA，用于RT-PCR和Northern杂交。 2. 采用RT-PCR和RACE方法，从甜樱桃果实克隆了两个抗氧化相关基因CAT2（Genbank：EF165590）和GPX（Genbank：EF165591）和两个PR基因GLU-1（Genbank：EF177487）和GLU-3（Genbank：EF177488)。其中CAT2全长cDNA序列为1479 bp，编码492个氨基酸；GPX全长cDNA序列为513 bp，编码170个氨基酸；GLU-1全长cDNA序列为1050 bp，编码349个氨基酸；GLU-3部分cDNA序列为454 bp，编码141个氨基酸。 3. 酵母拮抗菌Pichia membranaefaciens处理不同成熟度的甜樱桃果实，能显著降低果实贮藏期间青霉病（Penicillium expansum）的发生，并且对低成熟度果实的病害防治效果更为明显。酵母拮抗菌的抑病机理与减轻了甜樱桃果实蛋白羰基化程度，诱导了果实抗氧化酶基因（CAT和GPX）和PR基因（GLU-1）的表达和提高了抗氧化酶（CAT和GPX）和β-1,3-葡聚糖酶的活性有关。 4. 四种酵母拮抗菌P. membranaefaciens, Cryptococcus laurentii, Candida guilliermondii和Rhodotorula glutinis处理桃果实，可显著降低贮藏期间的褐腐病（Monilinia fructicola）。这是由于酵母拮抗菌能抑制病原菌侵染造成的氧化胁迫和蛋白羰基化。此外，酵母拮抗菌处理还能显著诱导CAT、POD、几丁质酶、β-1,3-葡聚糖酶活性及相应基因的表达。 5. 水杨酸（SA，2 mM）处理采后不同成熟度的甜樱桃果实，能显著降低青霉病的危害。其抑病机理与SA处理能减轻P. expansum侵染引起的果实蛋白羰基化程度，显著提高CAT、GPX和β-1,3-葡聚糖酶基因的表达和相关的酶的活性有关。而2 mM的SA处理对P. expansum的生长没有直接抑制作用。 6. 水杨酸（SA，2 mM）与P. membranaefaciens（1×108 CFU/ml）配合处理能显著降低低温贮藏期间桃果实的褐腐病，并能提高几丁质酶、β-1,3-葡聚糖酶和POD的活性和相关基因的表达。另外，2 mM的SA对拮抗菌P. membranaefaciens的生长没有影响，但能够抑制病原菌M. fructicola的孢子萌发和菌丝扩展。