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.     

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.     

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。     

Micropropagation of grey mangrove Avicennia marina

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

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)     

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。     

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.     

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.     

Allometric growth and carbon storage in the mangrove Sonneratia apetala

Allometric growth reflects different allocation patterns and relationships of different components or traits of a plant and is closely related to ecosystem carbon storage. As an introduced species, the growth and carbon storage of Sonneratia apetala are still unclear. To derive allometric relationships of the mangrove S. apetala and to estimate carbon storage in mangrove ecosystems, we harvested 12 individual Sonneratia apetala trees from four different diameter classes in the Futian National Nature Reserve, Guangdong, China. Allometric growth models were fitted. The results showed that diameter at breast height (DBH) and wood density were better variables for predicting plant biomass (including above- and below-ground biomass) than plant height. There were significant power function relationships between biomass and DBH, with a mean allometric exponent of 2.22, and stem biomass accounted for 97% of the variation in S. apetala total biomass. Nearly isometric scaling relationships were developed between stem biomass and other biomass components. To better understand the carbon stocks of the S. apetala ecosystem, we categorized all trees into five age classes and quantified vegetation carbon storage. The S. apetala vegetation carbon storage ranged from 96.48 to 215.35 Mg C ha^?1, and the carbon storage significantly increased with stand age. The allometric equations developed in this study are useful to estimate biomass and carbon storage of S. apetala ecosystems.

     

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.     

Drought effects on photosynthesis of the mangrove, Avicennia germinans, under contrasting salinities

 Drought effects on leaf photosynthesis of A. germinans growing under two contrasting salinities were studied in a Venezuelan fringe mangrove. During both wet and dry seasons, severe chronic-photoinhibition at predawn was not observed but strong down regulation occurred at midday during both seasons. Carbon assimilation rates (A, μmol CO₂ m⁻² s⁻¹) declined during the dry season from 11.9±1.8 to 7.0±1.5 and from 9.6±2.0 to 4.7±2.5 in plants from low and high salinity sites, respectively. Changes in carbon assimilation per unit of chlorophyll (A/Chl, mmol CO₂ mol⁻¹ Chl) were from 31.6±4.7 to 20.5±4.3 and from 21.9±4.7 to 15.2±8.2 in the low and high salinity plants, respectively. Therefore, neither changes in Chl nor seasonal differences in photoprotective down regulation could account fully for the decrease in leaf photosynthesis during drought. A reduction in CO₂ diffusion due to lowered stomatal conductance was not large enough to explain such a dramatic effect of drought on leaf photosynthesis. Stomatal response could be mitigated by the capability of A. germinans for osmotic adjustment under high salinity and/or drought. However, this intracellular salt accumulation may reduce carbon assimilation capacity further by decreasing the metabolism of leaf cells, increasing dark respiration and/or photorespiration. Received: 10 June 1998 / Accepted: 5 October 1998     

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.     

杯萼海桑和格氏海桑自然杂交的分子证据

Interspecific hybridization has been frequently observed in the mangrove genus Sonneratia. However, no natural hybridization has been reported between Sonneratia alba and S. griffithii to date, despite their overlapping distribution in the coast of Andaman Sea. In this study, cysteine proteinase inhibitor gene (cpi) from the nuclear genome, and two intergenic spacers (trnL-trnF and trnV-trnM) from the chloroplast genome, were sequenced to determine whether natural hybridization took place between the two species. Our results revealed two distinct types of cpi sequences from the putative hybrid matching those acquired from S. griffithii and S. alba, respectively. Sequencing of the chloroplast trnL-trnF and trnV-trnM regions showed that S. alba differed from S. griffithii by one nucleotide in each region, and the putative hybrid had the identical sequences with S. griffithii. Molecular data demonstrated clearly that there indeed existed natural hybridization between S. alba and S. griffithii, and that S. griffithii was the maternal parent in this hybridization event.     

Allometric models for aboveground biomass estimation of the mangrove Avicennia schaueriana

As mangroves become recognized as important carbon storages, the need for reducing the uncertainty of carbon inventories becomes increasingly emphasized. Accordingly, the objective of this study was to develop allometric models to estimate the total aboveground biomass (AGB) and the biomass per compartment of Avicennia schaueriana and to compare them with other models previously published for the genus Avicennia. Fifty three A. schaueriana trees, with different diameters at breast height (DBH) and height, were felled in a mangrove from Southeastern Brazil and their dry weight determined. Simple linear regression analysis was used to develop the equations after log-transformation, using the following independent variables: DBH and DBH² * height. All the equations were significant and presented high R _a ² (adjusted coefficient of determination). DBH provided the lowest SEE (standard error of estimation) in the regressions associated to leaves and total AGB, while DBH² * height generated the most precise regressions for trunk, branches, and twigs. In comparison with other 11 equations previously developed for the genus Avicennia, the equation developed in the present study for total AGB showed the lowest mean deviation in relation to trees with known biomass, underscoring the importance of developing species- and site-specific equations.