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.     

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.     

Mating system,pollen and propagule dispersal,and spatial genetic structure in a high-density population of the mangrove tree Kandelia candel

Mangrove tree species form ecologically and economically important forests along the tropical and subtropical coastlines of the world. Although low intrapopulation genetic diversity and high interpopulation genetic differentiation have been detected in most mangrove tree species, no direct investigation of pollen and propagule dispersal through paternity and/or parentage analysis and spatial genetic structure within populations has been conducted. We surveyed the mating system, pollen and propagule dispersal, and spatial genetic structure in a natural population of Kandelia candel, one of the typical viviparous mangrove tree species, using nuclear and chloroplast microsatellite markers. High diversity and outcrossing rates were observed. Paternity and parentage analysis and modelling estimations revealed the presence of an extremely short-distance component of pollen and propagule dispersal (pollen: 15.2 ± 14.9 m (SD) by paternity analysis and 34.4 m by modelling; propagule: 9.4 ± 13.8 m (SD) by parentage analysis, and 18.6 m by modelling). Genetic structure was significant at short distances, and a clumped distribution of chloroplast microsatellite genotypes was seen in K. candel adults. We conclude that the K. candel population was initiated by limited propagule founders from outside by long-distance dispersal followed by limited propagule dispersal from the founders, resulting in a half-sib family structure.     

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.     

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.     

Cloning and Functional Expression of Cycloartenol Synthases from Mangrove Species Rhizophora stylosa Griff. and Kandelia candel (L.) Druce

To obtain cDNAs encoding oxidosqualene cyclase (OSC), we cloned two cDNAs, KcCAS and RsCAS, from roots of Kandelia candel (L.) Druce and leaves of Rhizophora stylosa Griff. by homology based PCR method respectively. The deduced amino acid sequences of both OSCs showed 82% homology to cycloartenol synthases from Lotus japonicus (OSC5) and Ricinus cummunis (RcCAS), suggesting that these are cycloartenol synthases of K. candel and R. stylosa. The genes obtained were expressed in a lanosterol synthase deficient Saccharomyces cerevisiae (ERG7) strain, GIL77. GC–MS analysis identified the accumulated reaction product in the yeast transformant to be cycloartenol, indicating that both KcCAS and RsCAS encode cycloartenol synthase.     

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.     

Isolation and characterization of 10 new compound microsatellite markers for a mangrove tree species,Avicennia marina (Forsk.) Vierh. (Avicenniaceae)

Avicennia marina is an ecologically important mangrove tree species. We isolated 10 polymorphic microsatellite loci from this species using an improved technique. Our isolated loci provided compound microsatellite markers with polymorphism of two to six alleles per locus. The observed and expected heterozygosities ranged from 0.025 to 0.625 and from 0.096 to 0.767, respectively. These markers would be the useful tools for researching on the genetic diversity and population genetic structure of A. marina.     

Development and characterization of microsatellite markers for a mangrove tree species Sonneratia caseolaris (L.) Engler (Lythraceae sensu lato)

Sonneratia caseolaris is a typical non-viviparous mangrove species and a key component of mangrove community in the Indo-West Pacific region. Here we isolated nine microsatellite simple sequence repeat (SSR) loci from the genome of S. caseolaris. Our isolated loci provided SSR markers with polymorphism of 2–6 alleles per locus. The expected and observed heterozygosities ranged from 0.242 to 0.745 and from 0.083 to 0.417, respectively. Cross-species amplification in S. alba and S. ovata showed that a subset of these markers holds promise for these congeneric species. These polymorphic SSR markers would be useful tools for population genetics studies on S. caseolaris as well as other congeneric species.     

秋茄(Kandelia candel)幼苗对多氯联苯污染的生理生态响应

通过盆栽实验,研究了4种不同浓度（180、900、1800和2700μg kg^-1）的多氯联苯（PCBs）对红树植物秋茄幼苗的茎高、茎径、生物量、相对生长速率以及叶片的叶绿素含量、水势、丙二醛含量和游离脯氨酸含量等生理生态指标的影响,结果表明：（1）在所设PCBs浓度范围内,PCBs对秋茄幼苗的茎高、茎径、生物量和相对生长速率等生长指标的生长没有产生不利的影响,相反具有促进作用,红树植物秋茄在PCBs污染情况下能旺盛生长;（2）在所设PCBs浓度范围内,秋茄幼苗叶片能保持相对正常的叶绿素水平和相对稳定的叶绿素a／b值,叶绿素a、叶绿素b和叶绿素a＋b的含量虽然有所降低,但均为对照的70％以上;叶绿素a／b值有所升高,但均未超过对照的15％。（3）随着PCBs浓度的升高,秋茄幼苗叶片水势呈上升趋势,而游离脯氨酸含量和膜质过氧化产物MDA含量均有一定的增加,说明PCBs对秋茄幼苗产生了一定的影响。总体来看,秋茄幼苗能在所设浓度的PCBs范围内正常生长,对PCBs有较强的耐受性和适应性,对PCBs污染的沉积物进行修复是可行的。     

Chloroplast microsatellite markers for the mangrove tree species Bruguiera gymnorrhiza, Kandelia candel, and Rhizophora stylosa, and cross-amplification in other mangrove species

Chloroplast microsatellite (cpSSR) markers were developed for three ecologically and economically important tree species in the mangrove family, Rhizophoraceae: Bruguiera gymnorrhiza, Kandelia candel, and Rhizophora stylosa. Noncoding regions of chloroplast DNA (cpDNA) from each species were separately amplified using universal chloroplast primers. Six, two, and three polymorphic cpSSR loci in B. gymnorrhiza, K. candel, and R. stylosa, respectively, were developed from amplified noncoding cpDNA regions. Characterization of 216, 156, and 253 individuals of B. gymnorrhiza, K. candel, and R. stylosa, respectively, collected from different natural mangrove populations (B. gymnorrhiza, 9; K. candel, 7; R. stylosa, 9) on Iriomote Island in Japan showed that these loci provide cpSSR markers with polymorphisms ranging from two to four alleles per locus and gene diversity between 0.027 and 0.480. These cpSSR markers will be useful for analyzing the maternal lineage distributions and population genetic structures of the three species. Several of these markers may also be useful in similar studies of other mangrove species.     

Dispersal and Settlement Properties of Kandelia candel (Rhizophoraceae) Propagules

Abstract Three different types were found in the floating pattern of Kandelia candel (L.) Druce propagules: horizontal-floating type, vertical-floating type and sinking type. These correspond to the specific gravity, which increases with flooding in sea water and the repeated exposure to air. The floating types gradually change to the sinking type. In nature, such a condition is found only under the intertidal zone of shallow lagoons where floating propagules can increase their specific gravity, lie on the mud deposits and develop their roots in contact with the muddy soil. The J-shaped seedlings frequently found in K. canel populations in mangrove swamps might be recognized as evidence of growing up in the intertidal zone.
It is speculated that the horizontal-floating propagule has a role as a drifter in the ocean and long-distance disperser, the vertical-floating one as a colonizer of nearby populations and the sinking one as a successor in the original habitat.     

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.     

Accumulation and Dynamics of Cu,Pb, Zn and Mn Elements in Kandelia candel (L.) Druce Mangrove Community of Jiulong River Estuary of Fujian

The accumulation and biocycle of Cu, Pb, Zn and Mn elements were studied in Kandelia candel (L.) Druce mangrove community of Jiulong River Estuary of Fujian. The pool amounts of Cu, Pb, Zn and Mn elements in the forest soil (0～30 cm in depth) were 6.86, 4.23, 25.64 and 134. 67 g · m-2, respectively. The respective element contents ranged from in different parts of the plant, 1.85～6.97, 0.37～3.74, 16.0～25.2 and 140～1405 μg· g-1 of Cu, Pb, Zn and Mn. The pool amounts of elements in standing crop of the community were 87.98, 40. 34, 335.34 and 8006.99 mg · m-2 for Cu, Pb, Zn and Mn respectively. The pool amounts of Cu, Pb, Zn and Mn elements in residues were: 593.06, 49.27, 2450.79 and 43486.70 μg · m-2 respectively. The biocycle of the elements in the community were described as follows: Annual uptake of Cu was 10. 17, Pb 4.32, Zn 49. 14 and Mn 2268. 16 mg · m-2; Annual return of Cu was 2.21, Pb 0.70, Zn 18.63 and Mn 1574.98 mg· m-2; Annual retention of Cu was 7.96, Pb 3.62, Zn 30. 51 and Mn 693.18 mg · m-2. The turnover periods of Cu, Pb, Zn and Mn were 40, 58, 18 and 5 year, respectively. The absorption, utilatizion and cycle coefficients were in the order of Mn〉Zn〉Cu〉Pb.     

Isolation and characterization of microsatellite markers for Carolina hemlock (Tsuga caroliniana)

We describe the isolation and characterization of 31 polymorphic di- and trinucleotide microsatellite marker loci for Carolina hemlock (Tsuga caroliniana Englem.). In addition, primer pairs for 16 loci amplified scoreable alleles in six other Tsuga species. In eastern North America, both Carolina hemlock and eastern hemlock (Tsuga canadensis [L.] Carr.) populations are declining due to infestation by hemlock woolly adelgid, Adelges tsugae. The markers described here should enhance population genetic studies of hemlocks, providing valuable information for conserving and restoring these important forest tree species.     

Isolation and characterization of eight microsatellite loci from Phaedranassa tunguraguae (Amaryllidaceae)

Phaedranassa tunguraguae is an endangered species endemic to Ecuador. Eight highly polymorphic microsatellite loci were isolated from an enriched genomic library for this species. Levels of polymorphism were evaluated using a total of 31 individuals from a single natural population. An average of 14.1 alleles per locus was detected, and observed heterozygosity ranged from 0.387 to 0.903. All but one locus depart significantly from Hardy–Weinberg equilibrium. These loci are the first microsatellite primers isolated for Amaryllidaceae and will be utilized to investigate patterns of genetic variation of P. tunguraguae, which will contribute data relevant to the conservation of the species.     

中国红树林北缘区秋茄树种群空间分布格局和结构特征分析

对位于中国红树林北缘区福鼎市的秋茄树〔Kandelia candel(Linn.)Druce〕天然林和人工林的高度结构和径级结构进行了分析,并以"空间替代时间"的方法研究了秋茄树天然林和人工林的空间分布格局以及天然林的种群结构特征。结果表明:不同滩位秋茄树天然林和人工林的高度结构及径级结构明显不一致。天然林中株高0.0~0.5和1.5~2.0 m的个体占多数;而人工林以株高1.5~2.0 m的个体为主,无株高0.0~0.5 m的幼苗,株高超过2.0 m的植株也极少,表明天然林的高度结构均匀而人工林的高度结构极不合理。根据基径(D)可将秋茄树种群分为13个径级,其中天然林不同径级个体数量随径级增大依次减少,而人工林中径级Ⅲ(6 cm≤D<10 cm)的植株数量最多,且没有径级Ⅸ(30 cm≤D<34 cm)以上的个体,表明天然林的径级结构良好。从分布格局看,天然林幼苗群呈现聚集分布,而其小树群、大树群和老树群均为随机分布;人工林无幼苗群和老树群,小树群呈均匀分布,大树群呈聚集分布。种群静态生命表分析结果表明:秋茄树天然林种群的期间死亡率和消失率在龄级1(D<2 cm)和龄级7(22 cm≤D<26 cm)达到最大,在其他龄级均较小且处于稳定状态;其个体生存率单调下降、累计死亡率单调上升,仅在龄级1时个体生存率高于累计死亡率,从龄级2(2 cm≤D<6 cm)开始均表现为累计死亡率高于生存率;其存活曲线属于典型的Deevey-Ⅲ型,表明秋茄树天然林种群中幼苗丰富但期间死亡率较高,成树的期间死亡率相对较低且个体数量稳定。     

Development and polymorphism of simple sequence repeat DNA markers for Bruguiera gymnorrhiza (L.) Lamk

Microsatellite markers of Bruguiera gymnorrhiza were developed. Fifty‐four of 161 clones were found to contain microsatellite repeats. Primer pairs were designed for 20 of these clones according to their sequence data. Of these, seven primers showed polymorphism for 32 individuals from Iriomote Island, Japan. Two to five alleles per locus were detected, and the observed heterozygosities ranged from 0.031 to 0.500. Cross‐species amplification using five of the seven primers also worked well for B. cylindrica and B. parviflora. Because our previous study reported very low levels of genetic diversity for allozymes in the same Iriomote population, these microsatellite markers should be a powerful tool for various kinds of genetic analysis.     

福建漳江口红树林区秋茄幼苗生长动态

通过福建漳江口红树林自然保护区内8个样方24个小样方人工种植600个秋茄胚轴,在3a时间内对秋茄胚轴建立、幼苗生长以及环境因子进行定期观测。研究结果表明：林缘空地的秋茄生长状况良好,白骨壤林内最不利于秋茄幼苗的生长。潮位、盐度、底质土壤理化因子不是造成该样地各样方间秋茄幼苗生长差异的主要原因。动物取食、光照状况以及种间竞争是限制秋茄生长的主要环境因子。秋茄胚轴在长根前易于随潮水漂走,底质土壤中自骨壤致密的根系抑制了秋茄胚轴的定植,导致白骨壤林内秋茄幼苗漂走的数量最多。昆虫和螃蟹等动物的取食是导致林内已经固着生长的秋茄幼苗大量死亡的最主要原因,而林外被取食的幼苗个体极少。此后秋茄幼苗能否继续成长,主要取决于幼苗所接受到的光照条件。3a后,在荫蔽的树冠下,秋茄幼苗无法存活;而在林外,秋茄幼苗已经长成幼树。在林外滩涂上迅速生长的互花米草,也将影响秋茄幼苗的更新和生长。     

Genetic characterization of 12 heterologous microsatellite markers for the giant tropical tree Cariniana legalis (Lecythidaceae)

Twelve microsatellite loci previously developed in the tropical tree Cariniana estrellensis were genetically characterized in Cariniana legalis. Polymorphisms were assessed in 28 C. legalis individuals found between the Pardo and Mogi-Guaçu River basins in the state of São Paulo, Brazil. Of the 12 loci, 10 were polymorphic and exhibited Mendelian inheritance. The allelic richness at each locus ranged from 2-11, with an average of 7 alleles per locus, and the expected heterozygosity ranged from 0.07-0.88. These loci showed a high probability of paternity exclusion. The characteristics of these heterologous microsatellite markers indicate that they are suitable tools for investigating questions concerning population genetics in C. legalis.