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.     

Flower morphometry of Rhizophora mangle (Rhizophoraceae): geographical variation in Mexican populations

We explored the patterns of intra- and interpopulation variation in flower morphology of the red mangrove, Rhizophora mangle. Twelve populations in Mexico were studied: five from the Gulf of Mexico and the Caribbean Sea, and seven from the Pacific Coast. Six metric floral attributes were measured from a sample of 1370 flowers. Significant differences among populations were found for all six attributes. Because floral attributes were all correlated, scores derived from principal factor analysis were used to describe the variation in flower morphology. Two factors explained essentially all of the variance in flower morphology. Corolla and calyx size had a strong effect on factor 1, while gynoecium size had the higher effect on factor 2. Nested analyses of variance on the scores from both factors revealed significant differences among coasts, among populations within coasts, and among plants within populations. Nonetheless, this variation cannot be explained as a result of clinal variation, as indicated by a series of regression analyses. Cluster analysis (UPGMA) showed that a population from the Pacific coast was clustered together with those of the Atlantic, and the arrangement of populations within each coast showed no evident geographical pattern. We propose that frequent events of extinction and recolonization by a few individuals, followed by selfing, may produce differentiation among populations of red mangrove.     

国产红树林植物的染色体计数

本文对12种国产红树林植物和红树科、海桑科中2种非红树林植物进行了染色体计数,其中7种为染色体新计数,补充了中国红树林植物染色体资料的缺乏。通过调查,确认我国红树林植物有44种,占世界种类总数的44．9％。     

Molecular cloning and functional expression of a multifunctional triterpene synthase cDNA from a mangrove species Kandelia candel (L.) Druce

Homology based PCRs with degenerate primers designed from the conserved sequences among the known oxidosqualene cylases (OSCs) have resulted in cloning of a triterpene synthase (KcMS) from the young roots of Kandelia candel (L.) Druce (Rhizophoraceae). KcMS consists of a 2286 bp open reading frame, which codes for 761 amino acids. The deduced amino acid sequence showed 79% homology to a lupeol synthase from Ricinus communis suggesting it to be a lupeol synthase of K. candel. KcMS was expressed in a lanosterol synthase deficient yeast with the expression vector pYES2 under the control of GAL1 promoter. GC-MS analysis showed that the transformant accumulated a mixture of lupeol, beta-amyrin and alpha-amyrin in a 2:1:1 ratio, indicating that KcMS encodes a multifunctional triterpene synthase, although it showed high sequence homology to a R. communis lupeol synthase. This is the first OSC cloning from mangrove tree species.     

Photosynthesis and photoprotection in mangroves under field conditions

Net CO₂ exchange and in vivo chlorophyll fluorescence were studied in mangrove (Rhizophora stylosa) leaves at a field site in Western Australia, and leaf samples were collected for the analysis of enzymes and substrates potentially involved in anti-oxidant photoprotection. Photosynthesis saturated at 900 μmol quanta m^?2 s^?1 and at no more than 7.5 μmol CO₂ m^?2 s^?1. However, fluorescence analysis indicated no chronic photoinhibition: F_v:F_m was 0.8 shortly after sunset, and quantum efficiencies of PSII were high up to 500 μmol quanta m^?2 s^?1. Electron flow through PSII was more than 3 times higher than electron consumption through Calvin cycle activity, however, even with photorespiration and temperature-dependent Rubisco specificities taken into account. Acknowledging the growing body of literature attributing a role to antioxidant systems in photoprotection, we also assayed the activities of superoxide dismutase (SOD) and several enzymes potentially involved in H₂O₂ metabolism. Their levels of maximal potential activity were compared with those in greenhouse-grown mangroves (R. mangle), and growth chamber-grown peas. Monodehydroascorbate reductase activities were similar in all species, and glutathione reductase was lower, and ascorbate peroxidase ～40% higher, in the mangroves. SOD activities in field-grown mangroves were more than 40 times those in peas. Our results support the hypothesis that O₂ may be a significant sink for photochemically derived electrons under field conditions, and suggest an important role for O₂^? scavenging in photoprotection. However, when relative patterns are compared between species, imbalances between SOD and the other enzymes in the mangroves suggest that more components of the system (e.g. phenolics or peroxidases) are yet to be identified.     

Pentacyclic triterpenoids and sterols from seven species of mangrove

The isolation of pentacyclic triterpenoids from seven species of fresh mangrove leaves using a simple and rapid method is described. The leaves were homogenized using chloroform—methanol and the extract was diluted with water to precipitate out triterpenoids which were separated into neutral and acidic fractions. These were analysed by gas-liquid chromatography as acetyl and trimethylsilyl ether derivatives on a 3% OV-17 column. Sterols were isolated from the chloroform layer by preparative thin layer chromatography and were analysed by gas-liquid chromatography as their trimethylsilyl ether derivatives on a 3% OV-17 column. The triterpenoids found were α-amyrin, β-amyrin, lupeol, oleanolic acid and ursolic acid in most of the samples. Sterols found in all the samples were cholesterol, campesterol, stigmasterol, sitosterol and stigmast-7-en-3β-ol. Retention indices of the triterpenoids and sterols have been determined.     

Development and polymorphism of simple sequence repeat DNA markers for Kandelia candel (L.) Druce

Microsatellite markers of Kandelia candel were developed. Forty‐nine clones yielded strong positive signals among 331 clones hybridized to repetitive sequence probes. Primer pairs were designed for 19 of these positive clones according to their sequence data. Five of the primer pairs showed polymorphism for 16 individuals from Amami‐O‐Shima Island, Japan. Three to nine alleles per locus were detected, and the observed heterozygosities ranged from 0.250 to 0.938. Because our previous study reported very low level of genetic diversity for allozymes in the same Amami population, these microsatellite markers should be powerful tools for the analysis of genetic structure.     

Seed coat anatomy ofCrossostylis (Rhizophoraceae): its evolutionary and systematic implications

The seed coat structure of all 13 species ofCrossostylis was studied to contribute to an understanding of species delimitation and relationships within the genus. The mature seed coat is relatively uniform and consistently constructed mainly by a well-developed exotesta and a well-developed fibrous exotegmen. The species differ in the thickness of the exotesta and exotegmen, the anatomy of exotestal cells, the presence and absence of persistent mesotesta, and so forth. On the basis of comparisons of these characters, close relationships are suggested in the species groups such as:Crossostylis banksiana andC. cominsii; C. biflora, C. raiateensis andC. multiflora; C. gandiflora, C. sebertii andC. imera; and five species in the Fiji Islands. These relationships except for those of Fijian five species are also supported by cladistics as their common characters are evaluated as synapomorphy. Species-level separation ofC. banksiana, C. pedunculata andC. raiateensis each from the closest species is doubted based on the results of seed coat structure.     

Upstream Petroleum Degradation of Mangroves and Intertidal Shores: The Niger Delta Experience

This article was inspired by a field reconnaissance survey of outcrops along the Nembe‐Brass axis of the petroliferous Niger Delta. It reviews various tradeoffs of the impact of upstream petroleum (seismic and production) operations on the mangrove ecosystems in that region, the largest in Sub‐Saharan Africa. Mangroves and intertidal shores are considered critical to the economic well‐being of this region owing to the people's dual occupation in fishing and farming. The mangrove ecosystem provides a nutrient medium, which serves as a nursery and spawning ground for many fish species and other biota. Oil and gas activities might destroy these spawn areas, causing reduction in resource output and community pressure. Devegetation of the mangrove forest as a result of seismic delineation leaves the fragile soil exposed, unprotected, and susceptible to erosion. Again, loss of vegetation might discourage the natural role of plants in air purification (CO₂ utilization and O₂ production). The release of nutrients (organic N₂, NH₃, and NO$\rm{{_{3}^{-}}}$ ) and polyaromatic hydrocarbons (PAHs) to the environment, with the attendant increase in microbial load, increases biochemical O₂ demand (BOD) and depletes dissolved O₂ (DO) in H₂O to a level that is beyond the tolerance limit of organisms. This anoxic situation leads to asphyxiation and subsequent fish kill in affected areas. In order of increasing vulnerability, the mangroves and intertidal shores of the Niger Delta fall under categories 8 to 10 on the environmental sensitivity index (ESI) scale, which predisposes the areas to serious long‐term effects and clean‐up complexity. Thus, there is need to monitor mangrove systems and shoreline changes in the areas of considerable seismic and production activities, especially in the coastal Niger Delta, where pipeline corrosion due to salt intrusion into the swampy environment and ‘unsighted fingers’ of sabotage have increased the prevalence of oil spills.     

Reproductive biology and population genetic structure of Kandelia candel (Rhizophoraceae), a viviparous mangrove species

The pollination biology, mating system, and population genetic structure of Kandelia candel were investigated. Field observations on its pollination and reproductive biology suggested that this species is pollinator dependent for fruit set, and bee activities can lead to substantial geitonogamous selfing. Quantitative analysis of the mating system parameters was performed using progeny arrays assayed for allozyme markers. Multilocus outcrossing rates (t(m)) were estimated to be 0.697 ± 0.091 and 0.797 ± 0.062 in two populations. In comparison to other plant species with mixed-mating system, the level of allozyme variation was very low in the 13 populations sampled along the coastlines of Hong Kong. At the species level, the proportion of polymorphic loci was 20%, number of alleles per locus was 1.2, and heterozygosity was 0.0362. The total gene diversity was primarily distributed within populations (H(S )= 0.0339), and the coefficient of genetic differentiation among populations was low (G(ST )= 0.064). This pattern of population genetic structure suggests that gene flow, primarily in the form of water-dispersed seedlings in viviparous mangrove species, is not as limited as previously thought. However, microgeographic pattern in allele frequency at the marker loci could still be detected between the western and eastern coastal populations.