强潮区较高纬度移植红树植物秋茄的生理生态特性

以中国红树林引种最北界的乐清湾西门岛海域为研究地点,选择黄零为1.96、1.66和1.35 m 3个高程断面(每个潮水周期平均淹水时间分别为3.18、3.65、4.07 h),研究了1年生和4年生秋茄的生长、叶片的光合色素和保护酶活性、植株可溶性糖含量、游离氨基酸含量、离子含量等指标的变化。结果表明,当滩涂高程为黄零1.66 m以上时,每个潮水周期平均淹水时间低于3.65 h,1年生和4年生秋茄幼苗均可生长;当滩涂高程为低于黄零1.35 m时,每个潮水周期平均淹水时间高于4.07 h,1年生秋茄幼苗仍能正常生长,而4年生秋茄叶片叶绿素、类胡萝卜素总量、茎叶中的可溶性总糖和游离氨基酸含量、过氧化物酶(POD)活性以及K+/Na+比均显著降低,同时,叶片中的丙二醛(MDA)和茎、叶中Na+含量均快速上升,导致了生长缓慢。总体上说,较大树龄秋茄可生长在高程为黄零1.66 m以上,其原因是植株在每个潮水周期平均淹水时间少于3.65 h时仍能够维持正常的碳氮代谢。     

Accumulation,distribution and toxicological effects induced by cadmium on the development of mangrove plant <Emphasis Type="Italic">Kandelia candel</Emphasis> (L.)

Accumulation and distribution pattern of cadmium (Cd) and its toxic effect on growth of the mangrove plant of young Kandelia candel seedlings have been examined. This study demonstrated that under high concentration of Cd stress, the total biomass of K. candel decreased 41.57% compare to control (CK). At the end of 90 days exposure to 25 mg/l Cd, the average seedlings stem height and leaf number of the K. candel decreased by 30.54 and 42.68%, respectively. The results showed that K. candel seedlings, under the experimental condition, accumulated higher concentration of Cd in their roots (411.29 ± 3.60 mg/kg) when compared to hypocotyls, stems and leaves. More than 95% of Cd was accumulated mainly in roots. The distribution pattern of Cd concentration in K. candel seedlings was found in the following order: roots > hypocotyls > stems > leaves. Based on the leaf symptoms and morphological change of K. candel seedlings under heavy metal stress, this study showed that Cd is phytotoxic to K. candel.     

Proteomic Analysis of Salt-Responsive Proteins in the Leaves of Mangrove Kandelia candel during Short-Term Stress

Salt stress is a major abiotic stress that limits crop productivity in many regions of the world. A comparative proteomic approach to identify salt stress-responsive proteins and to understand the molecular mechanisms was carried out in the woody halophyte Kandelia candel. Four-leaf-old K. candel seedlings were exposed to 150 (control), 300, 450, and 600 mM NaCl for 3 days. Proteins extracted from the leaves of K. candel seedlings were separated by two-dimensional gel electrophoresis (2-DE). More than 900 protein spots were detected on each gel, and 53 differentially expressed protein spots were located with at least two-fold differences in abundance on 2-DE maps, of which 48 were identified by matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF-TOF/MS). The results showed that K. candel could withstand up to 450 mM NaCl stress by up-regulating proteins that are mainly involved in photosynthesis, respiration and energy metabolism, Na⁺ compartmentalization, protein folding and assembly, and signal transduction. Physiological data, including superoxide dismutase (SOD) and dehydroascorbate reductase (DHAR) activities, hydrogen peroxide (H₂O₂) and superoxide anion radicals (O₂ ⁻) contents, as well as Na⁺ content and K⁺/Na⁺ ratios all correlated well with our proteomic results. This study provides new global insights into woody halophyte salt stress responses. Identification of differentially expressed proteins promotes better understanding of the molecular basis for salt stress reduction in K. candel.     

Growth and physiological responses of Kandelia candel and Bruguiera gymnorrhiza to livestock wastewater

Growth and physiological responses of two mangrove species (Kandelia candel and Bruguiera gymnorrhiza) to livestock wastewater under two salinity conditions (seawater with salinity of 30þ and freshwater) were examined in greenhouse pot-cultivation systems for 144 days. Wastewater treatment significantly enhanced growth of Kandelia candel and Bruguiera gymnorrhiza in terms of stem height, stem basal diameter, leaf production, maximum unit leaf area and relative growth rate. Wastewater discharges and salinity levels did not significantly change biomass partitioning of Kandelia candel, however, more biomass of Bruguiera gymnorrhiza was allocated to leaf due to wastewater discharges. In Bruguiera gymnorrhiza, contents of chlorophyll a and chlorophyll b increased with wastewater discharges but such increase was not observed in Kandelia candel. On the other hand, livestock wastewater increased leaf electric conductance in Kandelia candel but not in Bruguiera gymnorrhiza. The peroxidase activity in stem and root of Kandelia candel under both salinity conditions increased due to wastewater discharges, while the activity in root of the treated Bruguiera gymnorrhiza seedlings decreased under freshwater condition but increased at seawater salinity. The superoxide dismutase activity in treated Bruguiera gymnorrhiza decreased but did not show any significant change in Kandelia candel receiving livestock wastewater.     

Growth and physiological responses of two mangrove species (Bruguiera gymnorrhiza and Kandelia candel) to waterlogging

Effects of duration of waterlogging on growth and physiological responses of two mangrove species, Bruguiera gymnorrhiza and Kandelia candel, were investigated. The relative growth rate of B. gymnorrhiza decreased significantly with waterlogged time, with the highest value found for drained plants and the lowest in plants under 12 weeks waterlogging. On the contrary, no significant difference was found between waterlogged and drained K. candel plants. The shoot to root biomass ratio of K. candel increased when subjected to 8 or 12 weeks waterlogging but little change was recorded in B. gymnorrhiza, indicating a shift in biomass allocation from roots to shoots in K. candel under prolonged waterlogging but not in B. gymnorrhiza. These different growth responses between the two mangrove species supported the hypothesis that K. candel is more tolerant to waterlogging than B. gymnorrhiza. Under 12 weeks waterlogged treatment, root oxidase activity significantly decreased in B. gymnorrhiza but increased in K. candel. Chlorophyll contents of K. candel increased more rapidly in response to waterlogging than B. gymnorrhiza. Activities of both peroxidase and superoxide dismutase increased significantly in leaves of K. candel when the waterlogging period was longer than 8 weeks, while only the peroxidase activity of B. gymnorrhiza showed a significant increase, indicating that K. candel had stronger resistance to the oxidant damage resulting from waterlogging. These physiological indicators further supported the hypothesis that K. candel is more tolerant to waterlogging than B. gymnorrhiza.     

The growth of Kandelia candel seedlings in mangrove habitats of the Zhangjiang estuary in Fujian, China

Propagules of Kandelia candel collected from the Zhangjiang estuary were planted in mangrove habitats along the intertidal gradient. The rooting rates of K. candel propagules varied spatially. The lowest rate occurred in Avicennia marina forest (69.7%). The rates were higher in K. candel forest (90.0%), at the fringe of the mangrove forest (89.3%) and on the bare tidal flat outside the mangrove forest (82.7%). After one year, the survival rates of seedlings planted under A. marina forest, K. candel forest, at the fringe of the mangrove forest, and on the bare tidal flat were 13.7%, 54.7%, 76.0%, and 34.7%, respectively. Among the surviving K. candel seedlings, those at the fringe of the mangrove forest and on the bare tidal flat had greater height, stem diameter, leaf number, leaf area, and biomass than those under A. marina and K. candel forests. These results demonstrated that establishment and growth of K. candel seedlings occurred successfully at the fringe of the mangrove forest, but were the worst under A. marina forest. The performance of K. candel seedlings was independent of physico-chemical characters of sediment. However, interspecies competition, propagule predation by insects and crabs, and the incident light had significant effects on seedling survival and growth.     

红树植物功能性状对盐和铜胁迫的响应

目前,室内通过人工控制方式开展红树林对环境胁迫的响应研究,忽视了不同红树植物物种的生长习性,且以单一胁迫为主。通过野外生长与人工控制相结合方式,研究5个时期(2018.6-2020.8)盐和铜胁迫对3种红树植物(秋茄、拉关木和木榄)11种功能性状的影响,使用单因素和双因素方差分析研究不同时期不同树种下盐和铜胁迫对叶片功能性状的影响,回答红树植物功能性状对盐和铜单一及复合胁迫响应的科学问题。结果表明,不同时期下,三种红树植物功能性状具有明显差异。随着时间增长,三种植物的碳含量和冠层高度均呈升高趋势,其他功能性状因物种不同而表现出不同趋势;与乡土红树植物(秋茄和木榄)相比,外来入侵物种(拉关木)具有更快的生长速度。不同时期下,拉关木比秋茄和木榄具有更好耐盐性,三个物种的植物生长发育对铜胁迫的响应均不敏感。大部分时期下,单一盐胁迫对拉关木的植物功能性状无显著影响(P < 0.05),对秋茄和木榄的碳含量、冠层高度和比叶面积有显著影响(P < 0.05);单一铜胁迫对三个物种的绝大多数功能性状无显著影响。另外,盐-铜复合胁迫对秋茄叶片碳、磷、氮磷比及冠层高度有显著影响(P < 0.05),对拉关木叶片碳、铜含量、冠层高度、比叶面积有显著影响(P < 0.05),仅对木榄SPAD值有显著影响(P < 0.05)。在红树林保护修复工作中,研究成果可为红树林退化原因和机理阐释、红树林生长监测评估、生境治理以及树种选择提供一定的理论依据。     

Does sea level rise influence propagule establishment, early growth and physiology of Kandelia candel and Bruguiera gymnorrhiza?

Responses of Kandelia candel and Bruguiera gymnorrhiza in loamy-sandy (coarse) and silty (fine) soils to water level rise and prolonged inundation were investigated under mesocosm and field conditions. Acidity due to simulated sea level rise of 30 cm was more serious in fine soil than that in coarse soil. Propagules of K. candel had more rapid establishments under high water level and prolonged inundation, but establishments of B. gymnorrhiza were not significantly influenced by tide condition. Water level rise of 30 cm and prolonged inundation stimulated early growth especially in stem height increments during the first 2 months for K. candel and B. gymnorrhiza. However, relative growth rates (RGRs) during the second 2 months did not significantly change for K. candel and even decreased for B. gymnorrhiza with water level rise. The root/shoot biomass ratio in both K. candel and B. gymnorrhiza treated with high water level was significantly lower than that treated with low water level, especially in coarse soil. Biomass percentages of coarse roots of K. candel and B. gymnorrhiza seedlings significantly increased with water level rise. Water level rise led to decreases in Chl. a/b ratios in leaves of K. candel seedlings, but the ratios did not significantly change with water level for B. gymnorrhiza. Decreases in RGRs of B. gymnorrhiza were due to decreases in total leaf area resulting from increases in stomatal density. Contents of N and P in roots tended to increase with water level rise and prolonged inundation. Under both high and low water levels, all propagules of K. candel and B. gymnorrhiza successfully established and the seedlings survived throughout the mesocosm experiment. For the field trial, K. candel seedlings had similar high survival rates of over 90%, while B. gymnorrhiza seedlings had much less survival in lower intertidal zones than in upper intertidal zones. These differences were ascribed to stronger tidal action and more sedimentation in the field than under mesocosm conditions. Survived seedlings of K. candel and B. gymnorrhiza from both propagule plantation and seedling transplantation in field trial also showed higher stem increments during the first 4 months in lower intertidal zones.     

Isolation and characterization of eight compound microsatellite markers in a mangrove tree Kandelia candel (L.) Druce

Kandelia candel is an important mangrove tree species of family Rhizophoraceae. Here we isolated eight codominant compound microsatellite simple sequence repeat (SSR) loci from K. candel. Our isolated loci provided compound SSR markers with polymorphism of three to 11 alleles per locus. The expected and observed heterozygosities ranged from 0.230 to 0.887 and from 0.083 to 1.00, respectively. These markers would be the useful tools for analysing questions concerning population genetic structure and mating system of K. candel.     

Over-expression of tobacco <Emphasis Type="Italic">UBC1</Emphasis> encoding a ubiquitin-conjugating enzyme increases cadmium tolerance by activating the 20S/26S proteasome and by decreasing Cd accumulation and oxidative stress in tobacco (<Emphasis Type="Italic">Nicotiana tabacum</Emphasis>)

Ubiquitin (Ub)-conjugating enzyme (UBC, E2) receives Ub from Ub-activating enzyme (E1) and transfers it to target proteins, thereby playing a key role in Ub/26S proteasome-dependent proteolysis. UBC has been reported to be involved in tolerating abiotic stress in plants, including drought, salt, osmotic and water stresses. To isolate the genes involved in Cd tolerance, we transformed WT (wild-type) yeast Y800 with a tobacco cDNA expression library and isolated a tobacco cDNA, NtUBC1 (Ub-conjugating enzyme), that enhances cadmium tolerance. When NtUBC1 was over-expressed in tobacco, cadmium tolerance was enhanced, but the Cd level was decreased. Interestingly, 20S proteasome activity was increased and ubiquitinated protein levels were diminished in response to cadmium in NtUBC1 tobacco. By contrast, proteasome activity was decreased and ubiquitinated protein levels were slightly enhanced by Cd treatment in control tobacco, which is sensitive to Cd. Moreover, the oxidative stress level was induced to a lesser extent by Cd in NtUBC1 tobacco compared with control plants, which is ascribed to the higher activity of antioxidant enzymes in NtUBC1 tobacco. In addition, NtUBC1 tobacco displayed a reduced accumulation of Cd compared with the control, likely due to the higher expression of CAX3 (Ca²⁺/H⁺ exchanger) and the lower expression of IRT1 (iron-responsive transporter 1) and HMA-A and -B (heavy metal ATPase). In contrast, atubc1 and atubc1atubc2 Arabidopsis exhibited lower Cd tolerance and proteasome activity than WT. In conclusion, NtUBC1 expression promotes cadmium tolerance likely by removing cadmium-damaged proteins via Ub/26S proteasome-dependent proteolysis or the Ub-independent 20S proteasome and by diminishing oxidative stress through the activation of antioxidant enzymes and decreasing Cd accumulation due to higher CAX3 and lower IRT1 and HMA-A/B expression in response to 50 µM Cd challenge for 3 weeks.     

Macrophage Interaction with Paracoccidioides brasiliensis Yeast Cells Modulates Fungal Metabolism and Generates a Response to Oxidative Stress

Macrophages are key players during Paracoccidioides brasiliensis infection. However, the relative contribution of the fungal response to counteracting macrophage activity remains poorly understood. In this work, we evaluated the P. brasiliensis proteomic response to macrophage internalization. A total of 308 differentially expressed proteins were detected in P. brasiliensis during infection. The positively regulated proteins included those involved in alternative carbon metabolism, such as enzymes involved in gluconeogenesis, beta-oxidation of fatty acids and amino acids catabolism. The down-regulated proteins during P. brasiliensis internalization in macrophages included those related to glycolysis and protein synthesis. Proteins involved in the oxidative stress response in P. brasiliensis yeast cells were also up-regulated during macrophage infection, including superoxide dismutases (SOD), thioredoxins (THX) and cytochrome c peroxidase (CCP). Antisense knockdown mutants evaluated the importance of CCP during macrophage infection. The results suggested that CCP is involved in a complex system of protection against oxidative stress and that gene silencing of this component of the antioxidant system diminished the survival of P. brasiliensis in macrophages and in a murine model of infection.     

Effect of NaCl on photosynthesis,salt accumulation and ion compartmentation in two mangrove species, Kandelia candel and Bruguiera gymnorhiza

In a 4-week study, we investigated the effects of increasing soil NaCl (100–400 mM) on photosynthesis, salt uptake and transport, and intracellular compartmentation of Na⁺ and Cl⁻ in 1-year-old seedlings of Kandelia candel (L.) Druce and Bruguiera gymnorhiza (L.) Savigny. Increasing NaCl stress significantly elevated Na⁺ and Cl⁻ in root and shoot tissues (stem + leaf) of both species, but B. gymnorhiza showed a rapid Na⁺ accumulation upon the initiation of salt stress and leaves contained 90% more Na⁺ and 40% more Cl⁻ than K. candel at the end of experiment. Net photosynthetic rate (Pn) declined with increasing salinity, and the most marked reduction occurred after exposure of mangrove seedlings to a severe salinity, 400 mM NaCl. However, the inhibitory effects of severe stress varied with species: Pn decreased by 80% in K. candel whereas in B. gymnorhiza the decline was 60%. The quantum yield (AQY) and carboxylation efficiency (CE) response to severe salinity showed a trend similar to Pn, in which a lesser reduction of AQY and CE was observed in B. gymnorhiza (33–35%), as compared to K. candel (43–52%). X-ray microanalysis of leaf mesophyll cells showed evidence of distinct vacuolar compartmentation of Na⁺ in K. candel but Cl⁻ in B. gymnorhiza after seedlings were subjected to 100 mM NaCl for 7 d. Moreover, Na⁺ within cell wall, cytoplasm, vacuole and chloroplast remained 23–72% lower in stressed B. gymnorhiza as compared to K. candel. In conclusion, B. gymnorhiza exhibited effective salt exclusion from chloroplasts although increasing salt stress caused a rapid and higher build up of Na⁺ and Cl⁻ in the leaves. We suggest that the salt-induced Pn reduction in the two mangrove species is correlated with the ability to exclude Na⁺ and Cl⁻ from the chloroplast, rather than with the bulk leaf salt concentration.     

Short- and long-term effects of NaCl on physiological and biochemical characteristics in leaves of a true mangrove, <Emphasis Type="Italic">Kandelia candel</Emphasis>

Changes in the physiological and biochemical characteristics of the leaves of Kandelia candel (L.) Druce seedlings in response to short-term (7 days) and long-term (60 days) treatments with two NaCl concentrations (250 and 500 mM) were studied. The growth rates were measured in terms of plant height, leaf area, and dry weight and were greater in the culture treated with 250 mM NaCl. Photosynthetic pigments also showed a preference for salinity growth conditions. The content of soluble sugars increased under any salinity during continuous treatments, whereas the proline level increased by the end of long-term culture. Further, during the treatment with 500 mM NaCl, the contents of hydrogen peroxide increased dramatically, whereas the levels of MDA, a measure of lipid peroxidation, decreased. The intactness of membrane integrity under this salinity condition may be explained by the activities of superoxide dismutase (SOD) and peroxidase (POD) which increased during the long-term experiment. It is concluded that the ability of K. candel to tolerate salt may occur mainly by inducing biosynthesis of soluble sugars and proline and increasing the activities of SOD and POD. The results imply that K. candel can survive well at 250 mM NaCl conditions and become acclimated to seawater salinity (∼500 mM) for 60 days of exposure. Published in Russian in Fiziologiya Rastenii, 2009, vol. 56, No. 3, pp. 403–409. This text was submitted by the authors in English.     

Geographical and seasonal patterns of epiphytic diatoms on a subtropical mangrove (Kandelia candel) in southern China

Geographical and seasonal patterns of epiphytic diatoms on a subtropical mangrove (Kandelia candel (L.) Druce) were examined at four sites in southern China. A total of one hundred and three epiphytic diatom species and varieties belonging to 40 genera were identified. Dominant species included Nitzschia fasciculata, Luticola mutica and Achnanthes javanica var. subcontricta. Further analysis using canonical correspondence analysis revealed three diatom associations which could be closely related to environmental variables such as salinity and nutrients resources of nitrogen and phosphate. Epiphytic diatom assemblages on roots would be a good indicator for the evaluation of water quality in mangroves.     

广西珍珠湾三种红树林林分土壤碳氮储量的研究

为了探讨不同红树林林分土壤有机碳(soil organic carbon,SOC)和全氮(total nitrogen,TN)储量空间的分布特征以及与C/N的相关性,该研究以广西防城珍珠湾红树林湿地为对象,通过样地调查取样和实验室分析,测定了SOC和TN的含量以及土壤碳储量的计量,揭示了广西北仑河珍珠湾秋茄、木榄和混交林三种红树林林分SOC和TN储量空间的分布特征以及C/N与SOC和TN的相关性。结果表明:(1)秋茄、木榄和混交林的SOC储量分别为140.73、124.94、144.71 t·hm^-2,三者无显著性差异(P>0.05);木榄和混交林垂直分布特征表现为20～40 cm>0～20 cm>40～60 cm,秋茄表现为随着土层深度的增加而递减。(2)秋茄、木榄和混交林的TN储量分别为6.49、5.01、5.87 t·hm^-2,表现为随着土层深度的增加而减少的趋势。(3)秋茄、木榄和混交林的SOC与TN储量之间的相关性极显著(P<0.01),相关系数分别为0.924、0.971和0.844,说明SOC与TN之间存...     

红树植物秋茄类黄酮代谢及其抗氧化活性对高盐胁迫的响应

以红树植物秋茄为试验材料,设置不同浓度NaCl(0、200和500mmol·L-1)处理的砂培实验,应用qRTPCR分析秋茄叶片中类黄酮物质合成上游的4个关键酶基因——苯丙氨酸解氨酶基因(PAL)、肉桂酸羟化酶基因(C4 H)、4-香豆酰辅酶A连接酶基因(4CL)和查尔酮合成酶基因(CHS)的转录水平,并对关键酶活性进行了分析,同时测定了幼苗生物量、钾钠离子含量、类黄酮含量及其抗氧化活性,以探讨秋茄耐盐性与类黄酮物质的关系,为揭示木本植物耐盐机制奠定基础。结果显示:(1)在盐处理条件下,秋茄叶片中PAL、4CL、C4 H和CHS4个关键酶基因的转录水平显著上调,PAL、4CL、C4H酶活性和CHS含量随着盐浓度的增加而明显上升。(2)与对照相比,秋茄根、茎、叶干重在盐处理3d和15d后均无显著变化,而秋茄株高仅在200mmol·L-1盐处理15d后显著增加,其余浓度和时间均未发现有显著性变化。(3)随着盐浓度的升高,秋茄叶片类黄酮含量显著增加,K+/Na+明显下降,丙二醛含量显著降低,活性氧自由基清除率显著增加。研究表明,盐处理加强了秋茄叶片中类黄酮代谢过程中相关酶基因的表达,类黄酮物质的累积有助于其抗氧化能力的提高,进而提高秋茄的抗盐性,维持盐胁迫下秋茄的正常生长。     

Use of 2-D electrophoresis and ESI mass spectrometry techniques to characterize Fusobacterium nucleatum proteins up-regulated after oxidative stress

The genus Fusobacterium belongs to the Fusobacteriaceae family and is a Gram-negative obligate anaerobic bacterium found in the human oral microbiota. Even that Fusobacterium nucleatum cannot grow under aerobic conditions, they may exhibit aerotolerance as an adaptive response which could figure as an important virulence factor, during the stages of infection, when these anaerobes are shifted to aerobic conditions. In this regard, little is known about bacterial oxidative stress adaptive response and the influence of this adaptation on the host-bacteria relationship. We aimed to use both techniques 2-DE and Electrospray Ionization Mass Spectrometry (ESI-MS) to characterize proteins in F. nucleatum, after oxidative stress. We related three different proteins which were up-regulated by oxidative stress. As its genome is already sequenced, these proteins were found in data base search, by homology. Thus, by using techniques as ESI-Q/TOF-MS, in addition to 2-DE, the opportunity exists to gain a more holistic view of the bacterial proteome of human pathogens, to achieve a better understanding of species diversity and to elucidate the role of specific proteins in disease. This work represents one of the first studies using genetic and physiological approaches to understand the phenomenon of oxidative stress in F. nucleatum.     

Enzymatic response to cadmium by Impatiens glandulifera: A preliminary investigation

This paper aims to develop our understanding of the effect of cadmium (Cd) on Impatiens glandulifera, a recently identified potential Cd hyperaccumulator. Impatiens glandulifera plants were exposed to three concentrations of Cd (20, 60 and 90 mg/kg) and were sampled at two timepoints (one and seven days) to investigate the stress response of I. glandulifera to Cd. Cd can induce oxidative stress in plants, triggering overproduction of reactive oxygen species (ROS). The level of activity of catalase (CAT) and ascorbate peroxidase (APX), two crucial antioxidant enzymes responsible for detoxifying ROS, were found to increase in a concentration dependent manner. Though there was no change observed in the level of superoxide dismutase (SOD) activity, the activity of glutathione S-transferase (GST), involved in detoxifying and sequestering Cd, increased after exposure to Cd. Cd did not appear to impact the levels of proline and photosynthetic pigments, indicating the plants weren't stressed by the presence of Cd. These results suggest that the rapid response observed in enzyme activity aid the efficacious mitigation of the toxic effects of Cd, preventing significant physiological stress in I. glandulifera.