海桑属植物分子鉴定的ISSR分析

为准确鉴别海桑属植物，采用ISSR（inter—simple sequence repeat）分子标记技术，对6种海桑属植物（海桑、拟海桑、杯萼海桑、海南海桑、卵叶海桑及无瓣海桑）基因组DNA进行PCR扩增。建立了海桑属植物ISSR标记的标准程序和海桑属各物种的DNA指纹数据库。采用的11个引物共产生71个物种特征性标记，其中无瓣海桑23个，海南海桑16个，海桑15个，杯萼海桑6个，拟海桑6个，卵叶海桑5个。运用这些特征性标记，可迅速区分海桑属植物。研究表明ISSR技术是在分子水平上鉴定海桑属植物的一种行之有效的方法。     

国产海桑属植物的两个杂交种

作者通过对红树林海桑属植物进行形态学、花粉学、细胞学以及其它方面的比较研究,证实了拟海桑和海南海桑为两个杂交种,它们的嫌疑亲本分别是杯萼海桑和海桑、杯萼海桑和卵叶海桑·ｓｏｎｎｅｒａｔｉａ×ｈａｉｎａｎｅｎｗｓｉｓｋｏ,ＥＹ．ＣｈｅｎｅｔＷ．Y　Ｃｈｅｎ（ｐｒｏｓｐ）为海南海桑的新名称,而拟海桑S．ｐａｒａｃｓｅｏｌａｒｉｓＫｏ,Ｅ.Ｙ.Ｃｈｅｎ,ｅｔＷ,ＣｈｅｎＹ.Ｃｈｅｎ则被归并入ｓｏｎｎｅｒａｔｉａ×ｇｕｌｎｇａｉＮ.C．Ｄｕｋｅ．     

国产海桑属（Sonneratia Linn．f．）植物的形态解剖

本文描述我国海南岛产的卵叶海桑、海南海桑、杯萼海桑、拟海桑、海桑等５种海桑属植物的叶、花、果实、种子、木材等的形态结构;分析它们的主要特征在各种植物之间的异同．观察结果表明,海南海桑和拟海桑两新种的形态结构特征与其它各种植物有明显的不同,并讨论它们之间的分类关系。     

海桑属(Sonneratia)植物的木材结构及其系统演化意义

研究了海桑科海桑属(Sonnerotia)6种植物的木材结构特征,并与同科的八宝树属(Duabanga)、千屈菜科的紫薇属(Lagerstroemia)植物的木材结构进行比较。结果表明：1.射线高度和射线宽度可作为卵叶海桑区别于其它种类的鉴定特征或辅助特征;2．海桑属形成一单系的类群,并与紫薇属有更近的亲缘关系,而与同科的八宝树属的亲缘关系更远;3．导管数量特征的聚类分析可以推测海桑属植物沿两支进化,一支进化为水分输导效率高的种类(即导管直径宽和输导面积大,管孔密度小,如海桑和拟海桑),另一支进化为水分输导安全性高的种类(即导管直径窄、输导面积小,管孔密度大,如杯萼海桑、卵叶海桑、无瓣海桑、海南海桑)。     

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。     

海桑属红树植物次生木质部解剖特征极其对潮间带生境的适应

通过光学显微镜和扫描电子显微镜详细观察了相似生境条件下生长的海桑属(Sonneratia)植物以及低、高潮位生长的海桑(S. caseolaris)和杯萼海桑(S. alba)次生木质部的形态特征,应用Lasersharp软件测量了其次生木质部的数量特征.结果表明:海桑属植物次生木质部形态特征的特化是与潮间带生境相适应的,能在水分胁迫的生境中,有效地协调水分输导的有效性和安全性.其特化结构主要包括:1)宽、窄导管并存,2)管孔密度较大,复孔率高,3)存在纤维状导管、形状不规则的导管和少量环管管胞,4)螺旋雕纹、附物纹孔、管壁具疣等许多导管壁的微观结构,有利于水分输导的安全性,5)射线细胞和分隔木纤维内的淀粉粒是渗透调节的物质基础,有利于促进水分上升,6)薄的纤维壁厚和宽的纤维腔径有利于水分的贮存,7)具胶质纤维.相似生境条件下生长的海桑属植物次生木质部数量特征的测量结果表明海桑和拟海桑水分输导效率高,但水分输导安全性差,而杯萼海桑水分输导效率低,但其输导安全性高.与高潮位生长的海桑和杯萼海桑相比,低潮位生长的海桑和杯萼海桑次生木质部导管分子更加"小型化"(有更小的导管分子长度、管孔弦向直径、管孔面积),有更高的导管聚合度和管孔密度,数量特征随生境不同的种内变动有利于海桑和杯萼海桑提高水分输导的安全性,但海桑和杯萼海桑输导安全性的获得是以牺牲水分输导的有效性为前提的.与生长在高潮位生境的海桑和杯萼海桑相比,低潮位生境生长的海桑和杯萼海桑的木材结构有更小的管孔面积,更厚的纤维壁,更低的导管比率和射线比率,更高的纤维比率,胶质纤维分布均匀且数量增加,这些特征有利于低潮位生境生长的海桑和杯萼海桑提高木材的韧性和强度,增强抗风浪冲击的能力.     

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?     

无瓣海桑、海桑、秋茄红树人工林群落动态及物种多样性研究

对红树植物无瓣海桑、海桑、秋茄3种人工林群落动态及物种多样性特征的系统研究结果表明,无瓣海桑群落和海桑群落的乔木层明显分为上层和中层两个亚层,上层木为无瓣海桑或海桑,中层木主要为秋茄和桐花树;秋茄群落的乔木层为单一层次,基本由秋茄组成．在无瓣海桑群落和海桑群落中,优势种群无瓣海桑或海桑仅有高龄级个体存在,未出现自然更新现象;秋茄和桐花树为旺盛增长种群,有可能成为优势种群,表明无瓣海桑和海桑为先锋造林树种,在裸滩种植可以促进其它红树植物的天然定居生长;在秋茄群落中,秋茄为旺盛增长种群,能够自然：更新演替,桐花树和海莲属初生增长种群．无瓣海桑群落和海桑群落的物种组成和物种多样性指标较接近,基本包含秋茄群落中的主要物种秋茄、桐花和海莲。表明无瓣海桑和海桑能与这些物种协调共生,同时种植无瓣海桑或海桑可以形成多样化的红树林群落;无瓣海桑群落和海桑群落在形成初期,种植密度较大时,物种多样性较高;密度相近时。形成初期随林龄的增加。其物种多样性略有提高．     

中国海桑属小志

本文描述中国海桑属一新种:拟海桑(Sonneratia paracaseolaris Ko, E. Y. Chen et W. Y.Chen).重新发表海桑属7种检索表,并将原隶属海桑组的海南海桑S.hainanensis Ko, E. Y.Chenet W.Y.Chen改隶无瓣海桑组,因其过去被认为是花瓣的部分实为退化雄蕊.     

中国海桑属小志

本文描述中国海桑属一新种:拟海桑(Sonneratia paracaseolaris Ko, E. Y. Chen et W. Y.Chen).重新发表海桑属7种检索表,并将原隶属海桑组的海南海桑S.hainanensis Ko, E. Y.Chenet W.Y.Chen改隶无瓣海桑组,因其过去被认为是花瓣的部分实为退化雄蕊.     

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.     

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.     

红皮云杉与嫩江云杉RAPD和ISSR分子标记反应体系优化和特异性检测

以红皮云杉和嫩江云杉为材料，采用现代分子标记技术-RAPD和ISSR标记研究红皮云杉与嫩江云杉间在基因组水平上的遗传差异。通过正交试验设计筛选RAPD和ISSR反应程序和反应体系。从近100个引物中筛选出3个RAPD引物和一个ISSR引物用来区分红皮云杉和嫩江云杉，它们分别是OPN07、OPA17、S25和ISSR67。用这4个引物扩增两种云杉基因组DNA，分别在1 000，950，1 500，2 000 bp处嫩江云杉有特异性谱带而红皮云杉没有。研究表明，采用RAPD和ISSR分子标记可以将红皮云杉和嫩江云杉在分子水平上加以区分，为今后进一步阐明云杉的系统进化，物种鉴定和种间杂交育种提供了理论依据。     

Chemical constituents from the fruits of Sonneratia caseolaris and Sonneratia ovata (Sonneratiaceae)

Nine (1–9) and seven (1–6, 10) compounds were isolated from the fruits of Sonneratia caseolaris and Sonneratia ovata, respectively. Their structures were identified by comparing their MS and NMR data as well as the physical properties with the literature. All the isolated compounds were screened against a rat glioma C-6 cell line using the MTT assay method; only compounds (-)-(R)-nyasol (1), (-)-(R)-4′-O-methylnyasol (2) and maslinic acid (6) were found to show moderate cytotoxic activity. Our findings from these two kinds of fruits can be used as a foundation for further chemotaxonomic studies on Sonneratia species. The nor-lignans (1, 2) and 6H-benzo[b,d]pyran-6-one derivatives (3, 4) were isolated from this genus for the first time, indicating that these two classes of compounds may tentatively be considered as taxonomic markers for Sonneratia genus.     

Phylogenetic Analysis of the Sonneratiaceae and its Relationship to Lythraceae Based on ITS Sequences of nrDNA

Lagerstroemia nested within the Sonneratiaceae. The Sonneratiaceae occurred within the Lythraceae with high bootstrap value support (96%). The two traditional genera constituting Sonneratiaceae were in different well-supported clades. Duabanga (Sonneratiaceae) is sister to the clade of Lagerstroemia (Lythraceae) (82%). The mangrove genus Sonneratia (100%) formed the other monophyletic group. It was located terminally within the Lythraceae clade and comprised two clades: one consisting of S. apetala, S. alba, S. ovata, and S. hainanensis; the other including S. caseolaris and S. paracaseolaris. The results indicated that species previously placed in two different sections (Sect. Sonneratia and Sect. Pseudosonneratia) of Sonneratia occurred within the same clade, and the taxonomic classification was not supported by the molecular analysis of the ITS region sequences. Based on the phylogenetic analyses of the ITS regions, the Sonneratiaceae were shown to be nested within the family Lythraceae. Therefore, the sequence data presented here do not support the recognition of the Sonneratiaceae as a distinct family, but instead support the inclusion of Sonneratiaceae in the Lythraceae proposed by other authors. Received 27 December 1999/ Accepted in revised form 25 June 2000     

落叶松种间及无性系间ISSR鉴别技术的研究

采用ISSR分子标记技术对兴安、长白和日本落叶松种间以及不同无性系进行了鉴别。从49条引物中筛选出13条ISSR引物可以对落叶松种间和无性系间进行鉴别，特异条带个体的百分率为100%，该项技术为落叶松新品种以及良种的准确鉴别提供了新的途径和手段：其中5条引物在日本落叶松、兴安落叶松和长白落叶松不同位置扩增出特异谱带，作为种的鉴定的标准，有9条引物可以对落叶松种内不同无性系分别扩增出特异片段，进行无性系的鉴别。     

A comparison of genetic variation among wild and cultivated Morus Species (Moraceae: Morus) as revealed by ISSR and SSR markers

In this study, inter-simple sequence repeats (ISSR) ans simple sequence repeat (SSR) markers were used to investigate genetic diversity of 27 mulberry accessions including 19 cultivated accessions (six M. multicaulis, three M. alba, two M. atropurpurea, two M. bombycis, one M. australis, two M. rotundiloba, one M. alba var. pendula, one M. alba var. macrophylla, and one M. alba var. venose) and 8 wild accessions (two M. cathayana, two M. laevigata, two M. wittiorum, one M. nigra and one M. mongolica). ISSRs and SSRs were compared in terms of their informativeness and efficiency in a study of genetic diversity and relationships among 27 mulberry genotypes. SSRs presented a higher level of polymorphism and greater information content. All index values of genetic diversity both markers analyzed using Popgene 32 software indicated that within wild species had higher genetic diversity than within cultivated species. Cultivation may caused the lose of genetic diversity of mulberry compared with wild species revealed by ISSR and SSR markers. The mean genetic similarity coefficients among all mulberry genotypes ascribed by ISSR and SSR matrices were 0.7677 and 0.6131, respectively. For all markers a high similarity in dendrogram topologies was obtained although some differences were observed. Cluster analysis of ISSR and SSR using UPGMA method revealed that the wild species are genetically distant from the domesticated species studied here. The correlation coefficients of similarity were statistically significant for both marker systems used. Principal coordinates analysis (PCA) for ISSR and SSR data also supports their UPGMA clustering. These results have an important implication for mulberry germplasm characterization, improvement, molecular systematics and conservation.     

Hyperspectral leaf signature as an added dimension for species discrimination: case study of four tropical mangroves

Hyperspectral leaf reflectance of a plant provides unique information that is characteristic of that plant. The present investigation is a preliminary attempt to assess whether spectra of leaves of mangrove species recorded under field conditions contain adequate spectral information for discerning mangroves at species rank. The paper highlights the hyperspectral characteristics of leaf surfaces of four prominent tropical mangrove species, viz., Avicennia alba, Avicennia marina, Rhizophora mucronata and Sonneratia caseolaris, found in the tidal forests of India. Hyperspectral observations were recorded using a field spectroradiometer, and pre-processed and averaged reflectance values of samples for three types of arrangements, viz., (1) randomly arranged leaves, (2) dorsal leaf surfaces and (3) ventral leaf surfaces of the species were statistically tested using one-way ANOVA to see whether the values significantly differed at every spectral location. All the four species were statistically different at all the spectral locations with majority of the bands exhibiting 99% confidence level. Finally, discriminant analysis was performed to identify the bands for maximum separability for the three types of arrangement of the leaves of the species taken separately and in different combinations. The optimal Wilks’ Lambda (L) were achieved with: six, three, eleven, six, five, thirteen and eleven wavelengths for discriminating random leaves of the four species, dorsal and ventral surfaces of A. alba, A. marina, R. mucronata, S. caseolaris, upper leaf surfaces of all the species, lower leaf surfaces, respectively. Factor analysis was used as an added tool to identify the wavelengths that were uncorrelated and contained maximum information in the combination of selected wavelengths. The results confirmed the unique spectral signatures of the four species, which could be explained in terms of leaf properties unique to the species. Cellular structure and pigmentation of the isolateral leaves of S. caseolaris are very different from the dorsiventral ones of the other three, which significantly changed the reflectance characteristics of the species.     

Sonneratia apetala Buch.Ham in the mangrove ecosystems of China: An invasive species or restoration species?

By the end of 1990s when China initiated a 10-year mangrove reforestation project, the mangrove forest area had decreased from 250,000 to 15,000 ha. Over 80% of current Chinese mangroves are degraded secondary forests or plantations. As an initial restoration and reforestation effort, Sonneratia apetala, a native of India, Bengal and Sri Lanka, was introduced in 1985 to Dong Zhaigang Mangrove Nature Reserve in Hainan Island from Bengal. It has then been introduced into other places since 1991. However, the further use of the species is becoming increasingly controversial as there are emerging signs that it may become invasive in certain locations. A comprehensive evaluation of the species’ condition in China regarding benefits and risks is critically needed. Here, we map the introduction and dispersal routes and monitor the growth of S. apetala in China from 1985 to 2006. S. apetala grows fast and performs well in the introduced 2300 ha muddy beaches area. It greatly improves the soil fertility and shows a suite of suitable characteristics as a pioneer restoration species. Currently, no natural invasion of S. apetala has been observed in the northern mangrove area. However, invasion into natural forests does occur in southerly locations such as Shenzhen, Zhanjiang and Dong Zhaigang. In these locations, S. apetala exhibits invasive characteristics such as overgrowth and high spreading ability that evidently affects local mangrove ecosystem structure and function. While the species clearly offers some benefits at some locations where it cannot naturally invade, it appears harmful to other native mangrove species, posing a major practical problem to both ecologists and land managers. This situation will be similar to previously imported non-native and invasive intertidal wetland species, Spartina alterniflora (smooth cordgrass), with similar results and problems.     

22种木莲属植物亲缘关系的ISSR分析

利用ISSR标记分析技术探讨了22种木莲属植物的遗传亲缘关系。选用10对ISSR引物组合对木莲属22个种的基因组进行分析。结果表明：22个种共扩增出595条DNA带,其中多态性条带占94.45%;根据Nei Li遗传相似性系数在0.68处,UPGMA聚类结果将22个种划分为6个类群,在0.70处进一步可分为12个亚类群;22种木莲属植物的Nei-Li遗传相似性系数变化范围为0.605(乳源木莲与球果木莲之间)～0.956(中缅木莲与滇南木莲之间),平均遗传相似性系数为0.698。说明木莲属植物种间差异性大。其中中缅木莲、滇南木莲之间遗传相似性系数为0.956,两者亲缘关系最近。该结果同时支持将巴东木莲、乳源木莲、滇桂木莲分别作为独立的种。