Microsatellite analysis of genetic structure in the mangrove species Avicennia marina (Forsk.) Vierh. (Avicenniaceae)

The level of genetic variation throughout the entire worldwide range of the mangrove species Avicennia marina (Forsk.) Vierh. was examined using microsatellite markers. Three microsatellite loci detected high levels of allelic diversity (70 alleles in total), essential for an accurate estimation of population genetic parameters. The informativeness of the microsatellite loci tended to increase with increasing average number of repeats. The levels of heterozygosity detected for each population, over all loci, ranged from 0.0 to 0.8, with an average of 0.407, indicating that some populations had little or no genetic variation, whereas others had a large amount. Populations at the extremes of the distribution range showed reduced levels of heterozygosity, and significant levels of inbreeding. This is not unexpected as these populations may be subject to founder effects and environmental constraints. The presence of genetic structure was tested in A. marina populations using three models: (i) a single panmictic model; (ii) the discrete subpopulation model; and (iii) the isolation by distance model. The discrete subpopulations model was supported by the overall measures of population differentiation based on the infinite alleles model (F-statistics), and the stepwise mutation model (R statistics). In addition, an analysis of molecular variance (AMOVA), using both theoretical models, found that most of the variation was between populations (41-71%), and within individuals in the total population (31-49%). There was little variation among individuals within populations (0-10%). There was no significant isolation by distance. The high levels of genetic differentiation observed among populations of A. marina may be due to environmental and ecological factors, particularly past sea level and climatic changes.     

Characterisation and analysis of microsatellite loci in a mangrove species, Avicennia marina (Forsk.) Vierh. (Avicenniaceae)

An enriched microsatellite library of the mangrove species Avicennia marina was constructed, in which 85.8% of the clones contained microsatellite sequences. Of the microsatellite repeat sequences isolated, 55.0% were di-nucleotides, 34.2% were tri-nucleotides, 50.0% were perfect, 24.2% were imperfect, and 15.0% were compound. Four different di-nucleotide repeats were isolated with repeat lengths ranging from 5 to 33; ten different tri-nucleotide repeats were isolated with repeat lengths ranging from 3 to 25. The most common di-nucleotide was the AC/TG repeat; the most common tri-nucleotide was the CCG/GGC repeat. Sixteen microsatellite sequences were selected for primer design, and 6 primers were selected to investigate the polymorphism detected among 15 individuals of A. marina from three natural populations in Australia. A total of 40 alleles were detected at 6 microsatellite loci. The number of alleles per microsatellite locus ranged from 5 to 13. On average, 7 alleles were detected per locus. All microsatellite loci showed high levels of gene diversity (heterozygosity), with values ranging from 0.53 to 0.88; the mean value of gene diversity was 0.70. Microsatellite loci were also tested for conservation across Avicennia species. There was a decline in amplification success with increasing divergence between Avicennia species. The results indicate that microsatellites are abundant in the Avicennia genome and can be valuable genetic markers for assessing the effects of deforestation and forest fragmentation in mangrove communities, which is an important issue for mangrove conservation and afforestation schemes. Received: 8 June 1999 / Accepted: 21 September 1999     

Comparative analysis of genetic diversity in the mangrove species Avicennia marina (Forsk.) Vierh. (Avicenniaceae) detected by AFLPs and SSRs

Avicennia marina is an important mangrove species with a wide geographical and climatic distribution which suggests that large amounts of genetic diversity are available for conservation and breeding programs. In this study we compare the informativeness of AFLPs and SSRs for assessing genetic diversity within and among individuals, populations and subspecies of A. marina in Australia. Our comparison utilized three SSR loci and three AFLP primer sets that were known to be polymorphic, and could be run in a single analysis on a capillary electrophoresis system, using different- colored fluorescent dyes. A total of 120 individuals representing six populations and three subspecies were sampled. At the locus level, SSRs were considerably more variable than AFLPs, with a total of 52 alleles and an average heterozygosity of 0.78. Average heterozygosity for AFLPs was 0.193, but all of the 918 bands scored were polymorphic. Thus, AFLPs were considerably more efficient at revealing polymorphic loci than SSRs despite lower average heterozygosities. SSRs detected more genetic differentiation between populations (19 vs 9%) and subspecies (35 vs 11%) than AFLPs. Principal co-ordinate analysis revealed congruent patterns of genetic relationships at the individual, population and subspecific levels for both data sets. Mantel testing confirmed congruence between AFLP and SSR genetic distances among, but not within, population comparisons, indicating that the markers were segregating independently but that evolutionary groups (populations and subspecies) were similar. Three genetic criteria of importance for defining priorities for ex situ collections or in situ conservation programs (number of alleles, number of locally common alleles and number of private alleles) were correlated between the AFLP and SSR data sets. The congruence between AFLP and SSR data sets suggest that either method, or a combination, is applicable to expanded genetic studies of mangroves. The codominant nature of SSRs makes them ideal for further population-based investigations, such as mating-system analyses, for which the dominant AFLP markers are less well suited. AFLPs may be particularly useful for monitoring propagation programs and identifying duplicates within collections, since a single PCR assay can reveal many loci at once. Received: 3 October 2000 / Accepted: 19 February 2001     

Characterization of microsatellite markers for the mangrove tree Avicennia germinans L. (Avicenniaceae)

We describe 10 primers for amplification of microsatellite loci for the mangrove, Avicennia germinans. Eight loci were isolated from a DNA sample from the Pacific coast of Baja California, Mexico and two loci were isolated from a DNA sample from the Atlantic coast of Bermuda. Polymorphism was investigated in a population from the Mexican Pacific coast (n = 15) and in four samples scattered throughout the range of the species. Total number of alleles for the species ranged from two to 10. Observed heterozygosity in the Mexican Pacific coast population ranged from 0.27 to 0.60, with two loci having fixed alleles.     

Development of microsatellite markers in a mangrove tree species Aegiceras corniculatum (Myrsinaceae)

Aegiceras corniculatum is an ecologically important mangrove tree species. We isolated 18 polymorphic microsatellite loci from this species. These loci provided microsatellite markers with polymorphism of two to eight alleles per locus. The observed and expected heterozygosities ranged from 0.050 to 0.550, and from 0.097 to 0.736, respectively. These markers will contribute to research on the conservation, genetic diversity and mating patterns of A. corniculatum.     

Ventilation and respiration in roots of one-year-old seedlings of grey mangrove Avicennia marina (Forsk.) Vierh.

Avicennia marina (Forsk.) Vierh. was grown from seed for 12 months in artificially tidal tanks providing a range of duration and depth of inundation. Plant growth characteristics were measured at harvest. Root aerenchyma development was estimated by pycnometry, root respiration rates by manometry, and the oxygen supply capacity of the above-ground portions of the plant was determined using oxygen electrode chambers. The mass per plant at harvest was influenced by the extent of inundation during growth with maximal growth at intermediate-length (1.5 to 6.5 h per tide) inundation periods. Those plants that had been submerged the longest (8.5 h per tide) had the least root tissue. The oxygen conductance of the stem base plus any pneumatophores showed a maximum in plants grown under intermediate inundation. Oxygen demand and internal gas space per unit dry weight of root were independent of extent of inundation. During high tide the plants grown at inundation periods of more than about 3–5 hours per tide were likely to become anaerobic. This may constitute a physiological limit for this species at the bottom of the tidal range.     

Development and characterization of ten new microsatellite markers in a mangrove tree species Bruguiera gymnorrhiza (L.)

Bruguiera gymnorrhiza is an ecologically and somewhat economically important mangrove tree species. We isolated 10 polymorphic microsatellite loci from B. gymnorrhiza using a dual‐suppression polymerase chain reaction (PCR) technique. These loci provided microsatellite markers with polymorphism of two to five alleles per locus within 216 individuals from nine natural populations of B. gymnorrhiza on Iriomote Island, the Sakishima Islands, Japan. The expected and observed heterozygosities ranged from 0.220 to 0.720 and from 0.104 to 0.447, respectively.     

Genetic variation of Avicennia marina (Forsk.) Vierh. (Avicenniaceae) in Vietnam revealed by microsatellite and AFLP markers

Genetic variation of Avicennia marina in the costal area of Vietnam was examined using microsatellite and AFLP markers. By using five microsatellite loci a total of 21 alleles were detected. The average number of alleles per locus per population ranged from 1.667 to 3.000. The observed heterozygosity varied from 0.180 to 0.263, with an average of 0.210 indicating relatively low level of genetic variation comparing to the previous studies on A. marina in the worldwide range. The expected heterozygosity was larger than the observed heterozygosity leading to positive inbreeding coefficients in all the six populations. Highly significant departures from Hardy-Weinberg Equilibrium were detected in four populations. AFLP analysis revealed a total of 386 loci, of which 232 (60.1%) were polymorphic. In congruent with microsatellite markers relatively low levels of genetic variation were detected at both gene and nucleotide levels (H = 0.086; pi = 0.0054). Reduced level of genetic variation was found in the central population, and in the southern populations. Both microsatellite and AFLP markers revealed large genetic differentiation (F(ST) = 0.262 and 0.338, respectively) indicating strong genetic structure among regional populations. Pairwise genetic distance by AFLP showed two populations in the north and the other two in the south are closely related each other.     

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.     

Water relations determine short time leaf growth patterns in the mangrove Avicennia marina (Forssk.) Vierh.

High‐resolution leaf growth is rarely studied despite its importance as a metric for plant performance and resource use efficiency. This is in part due to methodological challenges. Here, we present a method for in situ leaf growth measurements in a natural environment. We measured instantaneous leaf growth on a mature Avicennia marina subsp. australasica tree over several weeks. We measured leaf expansion by taking time‐lapse images and analysing them using marker tracking software. A custom‐made instrument was designed to enable long‐term field studies. We detected a distinct diel growth pattern with leaf area shrinkage in the morning and leaf expansion in the afternoon and at night. On average, the observed daily shrinkage was 37% of the net growth. Most of the net growth occurred at night. Diel leaf area shrinkage and recovery continued after growth cessation. The amount of daily growth was negatively correlated with shrinkage, and instantaneous leaf growth and shrinkage were correlated with changes in leaf turgor. We conclude that, at least in this tree species, instantaneous leaf growth patterns are very strongly linked to, and most likely driven by, leaf water relations, suggesting decoupling of short‐term growth patterns from carbon assimilation.     

PERMANENT GENETIC RESOURCES: Development of microsatellite markers for two nonviviparous mangrove species, Acanthus ilicifolius and Lumnitzera racemosa

Eight and nine of microsatellite loci were isolated from two nonviviparous mangrove species, Acanthus ilicifolius and Lumnitzera racemosa, respectively. The number of alleles per locus ranged from two to eight in A. ilicifolius and two to nine in L. racemosa. The observed heterozygosities ranged from 0.200 to 0.875 in A. ilicifolius and from 0.025 to 0.350 in L. racemosa. These loci would be effective for analysing genetic diversity and population genetic structure of these two mangrove species.     

Trace metals complexation behavior with root exudates induced by salinity from a mangrove plant Avicennia marina (Forsk.) Vierh

This study investigated the characteristics of exudates from mangrove plant Avicennia marina seedling roots under 0, 200 and 600?mM NaCl treatments and their complexation behavior with trace metals using excitation emission matrix (EEM) fluorescence spectrometry. Two fulvic-like fluorescence peaks, namely peak A (Em = 440?nm, Ex = 250?nm, UV fulvic-like compounds) and peak B (Em = 440?nm, Ex = 340?nm, visible fulvic-like compounds) were identified. The fluorescence intensities of peak A and peak B were enhanced by increasing salinity. Furthermore, the fluorescence of both peaks could be quenched by the ions of copper (Cu²⁺), manganese (Mn²⁺) and cadmium (Cd²⁺). Conditional stability constant (logK_a) exhibited that binding capacity of both peak A and peak B with trace metals are Cu²⁺?>?Mn²⁺?>?Cd²⁺ in the range from 2.21 to 4.01. Besides, Hill coefficient (n) >1 for Cu²⁺ but n?<?1 for Mn²⁺ and Cd²⁺. The results of high n and high logK_a for Cu²⁺ rather than Mn²⁺ and Cd²⁺ indicate that the fulvic-like compounds in root exudates of A. marina have maximum potential for Cu²⁺ complexation compared to Mn²⁺ and Cd²⁺, suggesting the fulvic acids in root exudates of A. marina have strong complexation with Cu²⁺ rather than Mn²⁺ and Cd²⁺.     

Aerenchyma tissue development and gas-pathway structure in root of Avicennia marina (Forsk.) Vierh.

The aerenchyma differentiation in cable roots, pneumatophores, anchor roots, and feeding roots of the mangrove plant, Avicennia marina (Verbenaceae) was analyzed using a light microscope and scanning electron microscope. In all types, cortex cells were arranged in longitudinal columns extending from the endodermis to the epidermis. No cells in the cortex had intercellular spaces at the root tip (0–150 m), and aerenchyma started developing at 200 m from the root apex. The aerenchyma formation was due to cell separation (schizogeny) rather than cell lysis. The cell separation occurred between the longitudinal cell columns, forming long intercellular spaces along the root axis. During aerenchyma formation, the cortex cells enlarged longitudinally by 1.8–3.9 times and widened horizontally by 2.2–2.9 times. As a result, the aerenchyma had a pronounced tubular structure that was radially long, elliptical or oval in cross section and that ran parallel to the root axis. The tube had tapering ends, as did vessel elements, although there were no perforated plates. The interconnection between neighboring tubes was made by abundant small pores or canals that were schizogenous intercellular spaces between the wall cells. All aerenchyma tubes in the root were interconnected by these small pores serving as a gas pathway.     

Gas exchange responses of a mangrove species, Avicennia marina, to waterlogged and drained conditions

This study was undertaken in summer on fully expanded leaves of Avicennia marina trees in the Beachwood Mangroves Nature Reserve, Durban,South Africa. Data sets were obtained over 5–7 days of relatively dry conditions and over two periods of 5 days during which the swamp was continuously inundated with dilute seawater (< 150 mol m⁻³NaCl). Gas exchange responses were strongly influenced by photosynthetic photon flux density (PPFD), leaf temperature and leaf to air vapour pressure deficit (Δw). Carbon dioxide exchange was saturated at a PPFD of about 800 μmol m⁻² s⁻¹. Maximal CO₂ exchange rates ranged from 8.5 to 9.9 μmol m⁻² s⁻¹ with no differences between drained and waterlogged conditions. Under drained conditions, leaf conductance,transpiration and internal CO₂ concentrations were generally lower, and water use efficiencies higher, than during waterlogging. Continuous waterlogging for 5 days had no adverse effect on CO₂ exchange. Xylem water potentials ranged from −1.32to −3.53 MPa during drained and from −1.02 to −2.65 Mpa during waterlogged conditions. These results are discussed in relation to anatomical and metabolic adaptations of A. marina to waterlogging stress. This revised version was published online in August 2006 with corrections to the Cover Date.     

Development of six microsatellite loci for black mangrove (Avicennia germinans)

We developed six new microsatellite loci for the black mangrove (Avicennia germinans), an important member of wetland communities worldwide. Loci showed moderate to high polymorphism and a survey of four locations [Puerto Rico (Jobos Bay and Luquillo), Mexico, French Guyana] revealed clear regional (and local) population structure. All populations were genetically distinct and the two continental populations showed much higher diversity than the two insular Puerto Rican locations. These loci complement those recently published by Nettel et al. (2005 ) and promise to be valuable for characterizing local and regional population dynamics in the black mangrove.     

Genomic variation patterns of subspecies defined by phenotypic criteria: Analyses of the mangrove species complex,Avicennia marina

Subspecies is used to designate taxa below species but above geographical populations. What patterns of genomic variation are expected if taxa are designated as subspecies? In this study, we carry out such a survey on the mangrove tree Avicennia marina (Forssk.) Vierh. of the Indo-West Pacific coasts. This species has three subspecies, distinguished by morphological traits and geographical distribution. We collected samples from 16 populations (577 individuals) covering all three subspecies and sequenced 94 nuclear genes. We reveal comprehensive genetic divergence among subspecies, generally higher than among geographical populations within subspecies. The level of genetic diversity differs among the three subspecies, possibly hinting at a degree of separation among their gene pools. We observed that divergence varies from locus to locus across the genome. A small portion of the genome is most informative about subspecies delineation, whereas the rest is undifferentiated or slightly differentiated, hinting at uneven gene flow and incomplete isolation. The three subspecies likely split simultaneously with gene flow among lineages. This reticulate evolution results in some discordance between morphology and genetics in areas of population contact. In short, A. marina subspecies show species-like patterns in some respects and population-like patterns in others. We propose that the subspecies designated in A. marina are informative in predicting genetic divergences and useful in making conservation decisions.     

挖捕泥丁对红树植物白骨壤幼苗生长影响的模拟

沿海群众为了增加收入挖掘林下滩涂海洋经济动物可口革囊星虫,这是导致中国红树林退化的一个重要原因.本文研究了挖掘深度、弧度和频率3个因素对1年生白骨壤幼苗的苗高、基径、单叶面积、比叶面积、生物量和死根干质量等生长指标的影响,了解挖掘活动对红树林生长的影响机理,并筛选出红树林健康状况的评估指标.结果表明:挖掘活动使白骨壤幼苗的苗高和基径增长量、单叶面积、比叶面积和生物量显著减少,而死根干质量显著增加;挖掘深度和弧度对幼苗生长具有明显影响,但频率的影响不显著;当挖掘深度＜5 cm、弧度＜240°、每月2次以下时,对幼苗的伤害较轻,而挖掘深度＞5 cm则会对幼苗造成严重影响.     

白骨壤模拟湿地系统中Pb的分布、迁移及其净化效应

在温室中建立红树林植物白骨壤模拟湿地系统,分别用正常(NW)、5倍(FW)和10倍(TW)浓度人工配置的城市污水每周定时定量对模拟系统污灌2次,用海水作对照(CL),持续1年。研究重金属Pb在湿地系统中的分布、迁移及其被净化的效果。结果表明：加入系统中的Pb主要存留于土壤子系统中,很少迁移到植物体和凋落物中：植物各器官中的Pb含量在根中最高;模拟系统对重金属。Pb的净化效果显著,各污水处理组分别为：NW组97．97％、FW组97．86％、TW组98．06％。根据物质平衡模型计算得土壤子系统。Pb的环境容量较大,因此整个模拟系统对Pb污染的承受力较大。     

白骨壤模拟湿地系统中磷的分配循环及其净化效应

在温室中建立红树林植物白骨壤模拟湿地系统 ,分别用正常、5倍和 10倍 3种不同浓度的人工配置的污水每周定时定量对模拟系统进行污灌 1a。研究营养污染物 P在系统中的分配、循环及其被净化的效果。结果表明 :随污水处理浓度的升高 ,被更换的潮汐水中的含 P量和土壤含 P量都有明显提高 ;植物体各部分含 P量的高低顺序排列为 :叶 >茎 >根 ;在植物 -土壤系统中4个组分 CA0 、CA1 、CA5和 CA1 0 植物中 P元素的周转期依次为 4 .70 a、4 .6 1a、3.80 a和 5 .4 4 a;模拟植物 -土壤系统对污水中 P的净化效果显著 ,CA1 、CA5和 CA1 0 组的净化率分别为 81.2 3%、94 .4 0 %和 95 .6 3%。