Polymorphic microsatellite DNA markers in the mangrove tree Avicennia alba

Like most species of mangrove trees of the genus Avicennia, A. alba is widely distributed among tropical and subtropical coasts around the world. Mangroves play an essential role in ecosystem dynamics but are reported to be regressing as human pressure increases on coastal zones. Hypervariable genetic markers are useful for population genetics studies, to estimate the level of impact and the populations potential for recovery. Microsatellite markers for A. alba were obtained by screening a partial genomic library enriched for microsatellite dinucleotide repeats. Among 20 primer pairs defined, six amplified polymorphic microsatellites with a satisfying level of variability.     

Generation and maintenance of suspension cultures from cotyledons and their organogenic potential of two mangrove species, Sonneratia alba and S. caseolaris

Liquid cultures were successfully generated from cotyledons of two Sonneratia species, S. alba and S. caseolaris in Murashige and Skoog (MS) medium containing 0.1 μmol L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D). Adventitious roots differentiated from cotyledons of S. alba. Proliferated cells were subcultured and a large volume of suspension cells was subsequently established in 100-mL flasks. All the cytokinins tested inhibited cell proliferation. After three years of culture, the potential to differentiate was tested as indicated by greening of the cells. Greening occurred when suspension cells were transferred to solid MS medium with and without 0.1 μmol L⁻¹ 2,4-D. Greening was stimulated by low concentrations of the weak auxins indolebutyric acid (IBA) and naphthaleneacetic acid (NAA) while 2,4-D stimulated late-stage greening. Abscisic acid (ABA) inhibited greening. Gibberellic acid (GA₃) at 1.0 μmol L⁻¹ stimulated callus greening and was not inhibitory even when tested at high concentrations. Cytokinins were inhibitory in combination with 0.1 μmol L⁻¹ of either IBA or NAA. The cause of different effects of plant hormones on growth and differentiation was discussed. Small-scale liquid media and 24-well culture plates of solid media methods developed in this paper are suitable for the optimization of hormonal conditions for cell proliferation and differentiation.     

Root system architecture and gravity perception of a mangrove plant,Sonneratia alba J. Smith

We describe the features of the root system and the gravitropism of roots produced bySonneratia alba. The root system consists of four root types with different growth directions: (a) Pneumatophores, which are negatively orthogravitropic and their statocytes are very large (922 μm²) and the statolith is located near the proximal wall, (b) Cable roots and (c) Feeding roots which are both diagravitropic and their statoliths are settled along the longitudinal wall, and (d) Anchor roots which are positively orthogravitropic. The statocyte cells are the smallest (420 μm²) and statoliths settled at the distal wall. We found that all roots with marked gravitropism have statoliths that settle along different walls of the statocyte. This implies that the statoliths sensing of gravity is done by gravity on mass, and that they are denser than surrounding cytoplasm and this position is related to root growth direction. This finding matches the statoliths sediment under the effect of gravity. Irrespective of statolith, position and direction of growth may be stable.     

三种海桑属濒危红树植物的资源分布与濒危现状

通过文献查阅和访问咨询,结合野外调查,对3种海桑属(Sonneratia)濒危红树植物的资源分布和濒危现状进行了初步研究。结果显示：卵叶海桑(Sonneratia ovata Backer)有2个自然居群和3个人工居群,个体数量分别为231和38棵;海南海桑(S. × hainanensis W. C. Ko, E.Y. Chen & W.Y. Chen)有4个自然居群和2个人工居群,个体数量分别为139和11棵;拟海桑(S. × gulngai N. C. Duke & B. R. Jackes)有4个自然居群和3个人工居群,个体数量分别为345和12棵;它们的自然居群更新能力都极差,人工种群也均未见新生个体。气候变化和人为干扰是种群规模急剧减小的直接原因,自身更新困难是种群规模难以恢复的制约因素。因此,建议在天然种群就地保护的基础上,优化迁地保护策略,加强人工辅助育种研究,加快突破育种难题,并科学抚育野外回归种群,以保证其种群的正常更新。     

An inverse relationship between allelopathic activity and salt tolerance in suspension cultures of three mangrove species,Sonneratia alba,S. caseolaris and S. ovata: development of a bioassay method for allelopathy,the protoplast co-culture method

A bioassay method for allelopathy, the ‘protoplast co-culture method’ was developed to study the relationship between salt tolerance and allelopathy of three mangrove species, Sonneratia alba, S. caseolaris, and S. ovata. Plants of S. alba grow in the seaward-side high salinity region and plants of the latter two species grow in upstream-side regions of a mangrove forest, respectively. Effects of five sea salts (NaCl, KCl, MgCl₂, MgSO₄ and CaCl₂) on the growth of the suspension cells of the latter two species were first investigated by a small-scale method using 24-well culture plates. S. ovata cells showed higher tolerance than S. caseolaris cells to NaCl and other salts, but were not as halophilic as S. alba cells. Protoplasts isolated from suspension cells were co-cultured with lettuce protoplasts in Murashige and Skoog’s (MS) basal medium containing 1 μM 2,4-dichlorophenoxyacetic acid, 0.1 μM benzyladenine, 3 % sucrose and 0.6–0.8 M osmoticum. S. caseolaris protoplasts had a higher inhibitory effect on lettuce protoplast cell divisions than S. alba protoplasts at any lettuce protoplast density, and the effect of S. ovata was intermediate between the two. These results were similar to those obtained from a different in vitro bioassay method for allelopathy, the ‘sandwich method’ with dried leaves. The inverse relationship between allelopathic activity and salt tolerance in suspension cells of Sonneratia mangroves is discussed.     

海桑属红树植物离子积累、光合和抗氧化能力及相关性分析

以海桑属（SonneratiaLinn.f.）红树植物无瓣海桑（S.apetalaBuch.-Ham.）、海桑也S.caseolaris（Linn.）Engl.页、杯萼海桑（S.albaSmith）、卵叶海桑（S.ovataBacker）、拟海桑（S.×gulngaiN.C.DukeetB.R.Jackes）和海南海桑（S.×hainanensisW.C.Koetal.）为研究对象,比较了根际土壤和叶片中离子含量以及叶片光合和叶绿素荧光参数、叶绿素含量、抗氧化酶活性和O-·2产生速率的差异,并分析了叶片中Na＋和Cl-含量与部分生理生化指标的相关性。比较结果表明：海桑和拟海桑叶片中K＋含量最高、Na＋和Cl-含量均显著低于其他种类,但它们的根际土壤中Na＋和Cl-含量却较高。供试6种植物中仅海桑叶片对Cl-的富集系数小于1,各供试种类对K＋、Na＋、Cl-、Ca2＋和Mg2＋的富集系数均大于1;无瓣海桑对离子的富集系数由大至小依次为Mg2＋、K＋、Ca2＋、Na＋、Cl-,其他种类对离子的富集系数由大至小均依次为K＋、Ca2＋、Mg2＋、Na＋、Cl-。无瓣海桑和海桑的chla、chlb和总叶绿素含量差异不显著,但均高于其他种类;供试种类的chla/chlb比值均约为3,可能与海桑属植物为阳生植物有关。无瓣海桑的Pn、Tr和Gs均最高,而杯萼海桑的Pn、Tr和Gs均最低,但6种植物的Ci无明显差异。供试种类的Fv/Fm、qP、ETR和ΦPSⅡ均无显著差异,仅部分种类间的NPQ差异显著。无瓣海桑叶片中SOD、CAT和POD活性均显著高于其他种类,但O-·2产生速率最低;而卵叶海桑叶片中O-·2产生速率最高,其APX活性也均显著高于其他种类。相关性分析结果表明：供试6种植物叶片中Na＋和Cl-含量与Pn、qP、ΦPSⅡ和ETR负相关,与NPQ及SOD、CAT、APX和POD活性正相关。其中,Na＋含量与qP、ΦPSⅡ、ETR和SOD活性极显著相关,与NPQ和CAT活性显著相关;Cl-含量与SOD活性极显著相关,与qP、ΦPSⅡ和ETR显著相关。研究结果表明：供试海桑属植物对高盐生境有不同的耐性机?     

光合和抗氧化能力及相关性分析海桑属红树植物离子积累

以海桑属(Sonneratia Linn. f.)红树植物无瓣海桑(S. apetala Buch.-Ham.)、海桑也S. caseolaris (Linn.) Engl.页、杯萼海桑(S. alba Smith)、卵叶海桑(S. ovata Backer)、拟海桑(S.× gulngai N. C. Duke et B. R. Jackes)和海南海桑(S.× hainanensis W. C. Ko et al.)为研究对象,比较了根际土壤和叶片中离子含量以及叶片光合和叶绿素荧光参数、叶绿素含量、抗氧化酶活性和 O-·2产生速率的差异,并分析了叶片中 Na+和 Cl-含量与部分生理生化指标的相关性。比较结果表明：海桑和拟海桑叶片中 K+含量最高、Na+和 Cl-含量均显著低于其他种类,但它们的根际土壤中Na+和 Cl-含量却较高。供试6种植物中仅海桑叶片对 Cl-的富集系数小于1,各供试种类对 K+、Na+、Cl-、Ca2+和Mg2+的富集系数均大于1;无瓣海桑对离子的富集系数由大至小依次为 Mg2+、K+、Ca2+、Na+、Cl-,其他种类对离子的富集系数由大至小均依次为 K+、Ca2+、Mg2+、Na+、Cl-。无瓣海桑和海桑的 chla、chlb 和总叶绿素含量差异不显著,但均高于其他种类;供试种类的 chla/ chlb 比值均约为3,可能与海桑属植物为阳生植物有关。无瓣海桑的 Pn、Tr 和Gs 均最高,而杯萼海桑的 Pn、Tr 和 Gs 均最低,但6种植物的 Ci 无明显差异。供试种类的 Fv / Fm、qP、ETR 和ΦPSⅡ均无显著差异,仅部分种类间的 NPQ 差异显著。无瓣海桑叶片中 SOD、CAT 和 POD 活性均显著高于其他种类,但O-·2产生速率最低;而卵叶海桑叶片中 O-·2产生速率最高,其 APX 活性也均显著高于其他种类。相关性分析结果表明：供试6种植物叶片中 Na+和 Cl-含量与 Pn、qP、ΦPSⅡ和 ETR 负相关,与 NPQ 及 SOD、CAT、APX 和 POD 活性正相关。其中,Na+含量与 qP、ΦPSⅡ、ETR 和 SOD 活性极显著相关,与 NPQ 和 CAT 活性显著相关;Cl-含量与 SOD 活性极显著相关,与 qP、ΦPSⅡ和 ETR 显著相关。研究结果表明：供试海桑属植物对高盐生境有不同的耐性机制,其中,海桑和拟海桑通过拒吸 Na+和 Cl-抵御盐胁迫的伤害;供试6种植物对海岸潮间带生境的适应性有明显差异,无瓣海桑最适宜在此生境中生长。     

Cell differentiation in protoplast cultures from embryogenic callus ofAbies alba L.

Protoplasts were isolated from embryogenic callus ofAbies alba L. which originated from immature seeds. The protoplasts were immobilized in alginate beads in order to follow the development of single protoplasts.Surrounding culture medium was modified from Kao and Michayluk (1975). After cell wall regeneration subsequent cell divisions lead to the formation of colonies showing an early differentiation of small meristematic cells and large vacuolated suspensor-type cells.Abbreviations 6-BA N⁶-benzyladenine - IAA 3-indoleacetic acid (potassium salt) - KM Kao and Michayluk (1975) - MES 2-(N-morpholino)ethanesulfonic acid - NAA 1-naphthalene acetic acid (sodium salt) - PVP Polyvinylpyrrolidone - SH Schenk and Hildebrandt (1972) - Tween 80 Polyoxyethylene- sorbitan - monooleate - WPM (woody plant medium), Lloyd and McCown (1981)     

Micropropagation of grey mangrove Avicennia marina

This study was conducted to develop a micropropagation protocol for grey mangrove, Avicennia marina (Forsk.)     

Salt tolerant sugarbeet progeny from tissue cultures challenged with multiple salts

Lines of sugarbeet (Beta vulgaris L.) tolerant of multiple salts was accomplished by an in vitro multiple salt challenge. Petioles were placed on RV medium amended with 5 different salts along with Murashige and Skoog base salts for one month. Surviving shoots were cultured on RV medium to obtain petioles for subsequent challenges. During the first, second and third challenges, organogenically regenerated shoots developed from 5%, 46%, and 80% of the petioles, respectively. After the third multiple salt challenge, tolerant shoots were rooted and transplanted in soil. Salt was added to this soil at 1.0% by weight and plants were observed for 2 months. The ten most salt tolerant plants were vernalized to obtain seed. The R₁ seed and controls were planted in soil containing 0%, 0.61% or 0.77% multiple salts per dry soil weight. Emergence of R₁ seedlings was significantly greater than the controls under salt stress. Multiple salt tolerant R₁ plants were maintained in salt amended soil to the 8–10 leaf stage and appeared as healthy and vigorous as the control growing in salt free soil.Contribution from Missouri Agricultural Experiment Station Journal Series No. 10948. University of Missouri, Columbia, MO, USA Mention of trade names does not constitute a guarantee or warranty of the product by University of Missouri of Holly Sugar Corporation and does not imply their approval to the exclusion of other products.     

无瓣海桑-海桑红树林的防风效能

为了解热带风暴(台风)期间红树林的防风功能,在2007年热带风暴"范斯高"影响期间(2007年9月24日0:00—15:00)采用自动气象站对海南东寨港无瓣海桑-海桑人工红树林的防风效应进行观测,结果表明:(1)各位置风速平均值大小为林后1H(H为平均树高)<林前1H,林后风速大幅度消减;(2)热带风暴期间防风效能为28%～68%,小于非热带风暴情况下的65%～77%,并随着风速的增大而减小;(3)热带风暴期间,其主导风向的变化幅度为45°,当风向与红树林接近垂直时,风速消减值越大,防风效果越显著。综上,热带风暴期间无瓣海桑-海桑人工红树林的防风效果显著。     

Genetic and chromosomal variation inPetunia hybrida plants regenerated from protoplast and callus cultures

Plants regenerated from callus cultures derived from leaf discs and mesophyll protoplasts ofPetunia hybrida cv. Rose of Heaven exhibit a high frequency of genetic and chromosomal variation. Of twelve leaf disc-derived plants examined, only three had the normal diploid chromosome number (2n=14) while seven were tetraploid and two were aneuploid (16 and 27 chromosomes). Of seventeen plants derived from two protoplasts, none had the diploid chromosome number. Most had 28 chromosomes, one 29, two 27, one 26 and one had variable numbers (14–28) in different root tip cells. In all cases aneuploidy was associated with developmental abnormality. In addition, heritable differences in growth, morphology and flower pigmentation were observed in callus-derived tetraploids and diploids, including one diploid which differed from parent plants in at least four characters. These results are discussed in terms of the importance ofPetunia in genetics research and for studies of somaclonal variation.     

Cytogenetics of protoplast cultures of Brachycome dichromosomatica and Crepis capillaris and regeneration of plants

Summary Callus derived protoplasts of Brachycome dichromosomatica (2n=2x=4) and Crepis capillaris (2n=2x=6) have been regenerated into karyologically normal plants, i.e. plants without visible alterations of the diploid chromosome set. However, metaphase analysis of protoplast cultures derived from both callus as well as mesophyll cells showed karyological changes in the overwhelming majority of cells in both species leading to multinucleated, polyploid and aneuploid cells. Furthermore, callus derived protoplasts sometimes exhibited changes at the chromosome level as indicated by translocations. The vast majority of aberrant karyotypes arose from failures during mitosis and cytokinesis, pointing to inadequate microtubules as a possible underlying cause. Karyological events of the kind described herein greatly affect the plating efficiency of isolated protoplasts and the viability of protoplast derived calli. Plant regeneration, although demonstrated in this study for the first time in both species, seems to be limited to rarely occurring, protoplast-derived colonies with a relatively stable genome. Our experiments, performed with chromosomal model species, emphasize the need for controlled, non-mutagenic culture conditions.     

Salinity, growth and root respiration in the grey mangrove, Avicennia marina

Relationships between growth parameters and root respiration under various conditions of salinity were investigated in seedlings of the grey mangrove Avicennia marina (Forsk.) Vierh. Growth, root/shoot ratios, leaf succulence and osmotic potential of leaves were measured for seedlings grown for 6–8 weeks in 100, 50, 25 and 0% seawater. Oxygen uptake of root segments, from distal to proximal ends of roots, was measured for all treatments. Total growth was maximal in 25% seawater, highest leaf succulence was obtained in 50% seawater, and highest leaf osmotic potential in 100% seawater. Oxygen uptake in distal root segments, as measured both by Clark oxygen electrode and Warburg manometry, showed a stimulation in the presence of salt that closely paralleled growth stimulation. The rates of respiration were highest in 25% seawater. The oxygen uptake was not stimulated by salt per se, since concentrations higher than 25% were associated with a decline in rate of oxygen uptake from the maximum. Values for the respiratory quotient approximated to one in all treatments. Avicennia marina has been reported to exclude from its roots about 90% of the salt in the surrounding medium. It might have been expected that increased concentrations of salt in the growth medium would be associated with a standard salt respiration response in the roots; however, this was not obtained.     

无瓣海桑与三种乡土红树植物混交的生态效应

为了解人工种植红树林的生态效应,对2001年种植于广州市南沙区的三种人工混交红树林进行了群落结构和土壤理化性状的调查分析。结果表明:由于红海榄的不耐荫导致样地3无法形成复层结构,但也不排除是红海榄化感作用的影响;无瓣海桑+木榄群落、无瓣海桑+秋茄群落的群落结构和对土壤中重金属的吸收效果优于无瓣海桑+红海榄群落,因此在该实验的立地条件下营造红树林混交林时可以考虑无瓣海桑与秋茄、木榄混交,不应将无瓣海桑与红海榄混交;随着演替的进行,该实验地在无重大干扰的情况下将向着近天然红树林生态系统演化。     

Salt balance of leaves of the mangrove Avicennia marina

The contribution of each of the salt-transporting processes to the NaCl balance of the leaves of the salt-recreting mangrove Avicennia marina (Forssk.) Vierh. was quantitatively investigated. Transpiration rates, xylem sap concentration, leaf salt content, recretion rates and rates of salt retranslocation out of the leaves were continuously monitored during three day periods and the salt fluxes in and out of the leaves were calculated. The results indicated that salt filtration by the roots is by far the most important salt-rejecting mechanism, preventing some 80% of the salt which is carried towards the root surface by the transpiration stream, from entering the shoot. Out of the remaining quantity of salts which enter the root xylem and reach the leaves, only 40% is removed by the salt-recreting glands.     

Low salinities adversely affect photosynthetic performance of the mangrove,Avicennia marina

Photosynthetic performance of the highly salt tolerant mangrove, Avicennia marina, was compared at two sites differing insubstrate soil salinities. Carbon dioxide exchange and chlorophyll fluorescence weremonitored at a high salinity site in Durban Bay (35) and at a low salinitysite in Beachwood (< 12). Mean CO₂ exchange, conductanceand transpiration were consistently higher at the high salinity site. Carbondioxide response curves indicated that carboxylation efficiency was higherand stomatal limitation lower at the Durban Bay site. PSII quantum yield,electron transport rates (ETR) and intrinsic PSII efficiency(F_v/F_m) were significantly higher at the high salinity site.Quenching analysis indicated a higher degree ofphotoinhibition/photoprotection in leaves at the low salinity site. Predawnand midday leaf water potentials were –1.6 and –3.1 MPa at Beachwood,compared to –2.6 and –3.8 MPa, respectively, at Durban Bay. Leafconcentrations of Na⁺, K⁺, Ca²⁺, Mg²⁺,Cl^- and N were significantly higher at Durban Bay. Photosyntheticperformance is apparently impaired at the low salinity site in Beachwood asa result of K⁺ and N deficiencies in the leaves.     

外来红树植物无瓣海桑生物学特性与生态环境适应性分析

根据近年来的调查观测数据和收集国内外有关资料较全面地论述外来红树植物无瓣海桑的形态特征、地理分布、生物学特性 (包括物候、种子品质与苗木生长、苗木抗逆性、种苗扩散性、种内竞争特点 )和生态环境适应性 (包括气温、光照、盐度、潮滩与土壤 ) ;据此分析了无瓣海桑生态入侵的可能性 ,并提出相关的建议 ,为我国进一步研究该外来种引进后的生态效应以及该树种的合理开发利用提供科学依据