Conservation genetics of Amorpha georgiana (Fabaceae), an endangered legume of the Southeastern United States

Amorpha georgiana (Fabaceae) is an endangered legume species found in longleaf pine savannas in the Southeastern United States. Approximately 900 individuals and 14 populations remain, most of which are concentrated in North Carolina. Eleven microsatellite loci were used to explore genetic diversity, population structure and recent population bottlenecks using genotypic data from 132 individuals collected at ten different localities. Although A. georgiana is quite rare, it exhibited high levels of genetic diversity (17.7 alleles/locus; H _o = 0.65, H _E = 0.75). Most of the genetic variation was found within rather than between populations of this species. The single remaining Georgia population was well differentiated from populations of the Carolinas ( F _ST > 0.1), which had weaker structure among them ( F _ST < 0.1). Only a geographically disjunct population showed strong evidence of a recent population bottleneck, perhaps due to a recent founder event. Hybridization with A. herbacea was also detected. For conservation management plans, A. georgiana populations in each geographic region (North Carolina, South Carolina and Georgia) plus a disjunct population in North Carolina (Holly Shelter) should be treated as separate management units for which in situ conservation, including habitat restoration and use of prescribed burns, should ensure persistence of this species and preservation of its evolutionary potential.     

Fitness consequences of variable maternal provisioning in quacking frogs (Crinia georgiana)

Variable maternal provisioning may evolve when there is variation in the quality of offspring environments. The frog Crinia georgiana has high variability in egg size both within and between clutches, independent of female phenotype. It breeds in ponds with high spatial and temporal variation in habitat quality. Egg size strongly affected offspring fitness in good and poor quality offspring environments, whether the egg size difference was from between or within clutches. Since there is a trade-off in egg size and number, these fitness consequences translate to strong effects on maternal fitness. In the variable and unpredictable offspring environment of C. georgiana, the maintenance of variable maternal provisioning both within and between clutches is likely to be an evolved response to the offspring environment.     

紫穗槐果实和叶子的紫外光谱指纹图谱研究

本文通过研究紫穗槐的紫外光谱指纹图谱,鉴别紫穗槐及其混淆品。分别取紫穗槐果实和叶子各19个样品以及6个混淆品,应用紫外光谱法对其共有峰和特征峰进行检测。结果显示:紫穗槐果实有2个共有峰,其峰值比为1.330:1;紫穗槐叶有4个共有峰,其峰值比1.45:1:0.71:0.69。     

紫穗槐种子提取物的抑菌活性研究

目的研究紫穗槐种子提取物的抑菌活性。方法将紫穗槐种子乙醇提取物分别通过石油醚、乙酸乙酯和正丁醇萃取,选择金黄色葡萄球菌和肺炎克雷伯杆菌为供试菌,采用试管二倍稀释法测定紫穗槐种子提取物的最小抑菌浓度（MIC）和最小杀菌浓度（MBC）,涂布平板法绘制杀菌曲线,电镜下观察药物对细菌超微结构的影响。结果紫穗槐种子提取物经乙酸乙酯萃取后对供试菌抑制作用较强,其中对金黄色葡萄球菌的MIC和MBC分别为2．5、5．0mg／mL;对肺炎克雷伯杆菌的MIC和MBC分别为5．0、10．0mg／mL;杀菌曲线结果表明,药物对供试菌的抑制作用存在浓度和时间依赖性;电镜结果说明,药物的作用可能与破坏菌体细胞壁、改变细胞膜通透性有关。结论紫穗槐种子提取物具有显著的抗菌活性。     

Fitness consequences of parental compatibility in the frog Crinia georgiana

Theory suggests that multiple mating by females can evolve as a mechanism for acquiring compatible genes that promote offspring fitness. Genetic compatibility models predict that differences in fitness among offspring arise from interactions between male and female haplotypes. Using a cross-classified breeding design and in vitro fertilization, we raised families of maternal and paternal half-siblings of the frog Crinia georgiana, a species with a polyandrous breeding system and external fertilization. After controlling for variation in maternal provisioning, we found significant effects of interacting parental haplotypes on fertilization success, and nonadditive genetic effects on measures of offspring fitness such as embryo survival, and survival to, size at, and time to metamorphosis. Additive genetic variation due to males and females was negligible, and not statistically significant for any of the fitness traits measured. Combinations of parental haplotypes that resulted in high rates of fertilization produced offspring with higher embryo survival and rapid juvenile development. We suggest that a gamete recognition mechanism for selective fertilization by compatible sperm may promote offspring fitness in this system.     

Amorinin,A prenylated chromenoflavanone from Amorpha fruticosa

A diprenylated chromenoflavanone has been characterized from the root bark of Amorpha fruticosa.     

紫穗槐的离体快速繁殖

以子叶节为外植体,建立起了紫穗槐的快速离体再生系统.经过四周的培养,在附加8mg·L-16-BA的MS培养基上能够获得再生频率为100%,平均每个外植体5.21个芽点的高效再生植株.以再生植株的茎节为外植体所进行的继代能够在相同的培养基上连续的产生新的不定芽,但芽点数要少于起始培养.经过3周的培养,有82.53%切下的再生茎段能够在含2.0mg·L-1IAA的MS培养基上生根.在所有进行分析过的再生植株中,它们的染色体数目都没有发生变异(2n=40).经过练苗以后,再生植株成功地定植于土壤当中并展示了一致的外部形态和生长特性.     

Karyotype analysis and physical mapping of 45S rDNA in eight species of Sophora,Robinia, and Amorpha

The karyotype analysis and physical locations of 45S rDNA were carried out by means of fluorescence in situ hybridization in three species,and two forms of Sophora,two species of Robina,and one species of Amorpha.S.japonica L.,S.japonica L.f.oligophylla Franch.,S.japonica L.f.pendula Loud.,and S.xanthantha C.Y.Ma.are all tetraploids with 2n=28.There were four 45S rDNA sites in pericentromeric regions of two Pairs of chromosomes in each of them.S.rubriflora Tsoong.is a triploid with 2n=21,and three sites were located in each satellite of group 5 chromosomes.In R.pseudoacacia L.(2n=2x=22),we examined four intensive signals in telomeric regions of two pairs of satellite chromosomes.In R.hispida L.(2n=2x=30),there were four other signals in centromeric regions besides those like in R.pseudoacacia.Amorpha fruticosa L.has most chromosomes(2n=40)among the eight materials,however,there were only six 45S rDNA loci and they laid in centromeric regions,and satellites of three pairs of chromosomes.45S rDNA is a valuable chromosomal landmark in karyotype analysis.The distribution and genomic organization Of rDNA in the three genera were also discussed.     

Synchronous polyandry and multiple paternity in the frog Crinia georgiana (Anura: Myobatrachidae)

Multiple paternity has rarely been reported in anuran amphibians, with only three previous documented examples. For the Australian frog Crinia georgiana, we observed synchronous polyandry in an average of 44% of matings observed at four field sites. This suggests matings involving more than one male are common in this species. One to eight males were observed in amplectant groups with second males amplexed ventrally. Genetic analyses, using allozyme electrophoresis, of offspring from two matings indicated that at least two of three possible males fathered offspring. Third males were unlikely to have shared paternity, explained by their inappropriate position during amplexus. Multiple paternity may be more common in frogs than has been reported. Copyright 1999 The Association for the Study of Animal Behaviour.     

Amoradin,amoradicin and amoradinin,three prenylflavanones from Amorpha fruticosa

Three new prenylflavanones, amoradin, amoradicin and amoradinin, were isolated from the root bark of Amorpha fruticosa. Their structures were deduced from chemical and spectral evidence.     

基于rbcL序列试论鸢尾属部分物种间的系统发育关系

选用叶绿体rbcL基因对鸢尾属植物的分类和系统发育进行了分析。以雄黄兰为外类群,对21份鸢尾属材料和GenBank下载的共49条序列采用最大简约法构建系统进化树,探讨了鸢尾属的分类系统。针对有争议的物种,聚类结果表明：马蔺与白花马蔺关系较远;扁竹兰和蝴蝶花亲缘关系较扇形鸢尾近;细锐果鸢尾和锐果鸢尾亲缘关系近,大锐果鸢尾较之远;四川鸢尾和薄叶鸢尾聚类近;西伯利亚鸢尾系内物种间聚类复杂,亲缘关系不清楚。     

Diverse rhizobia associated with woody legumes Wisteria sinensis, Cercis racemosa and Amorpha fruticosa grown in the temperate zone of China

Fifty-nine bacterial isolates from root nodules of the woody legumes Wisteria sinensis, Cercis racemosa and Amorpha fruticosa grown in the central and eastern regions of China were characterized with phenotypic analysis, PCR-based 16S and 23S rRNA gene RFLP, Box PCR and 16S rRNA gene sequencing. Seven main phena were defined in numerical taxonomy, which corresponded to distinct groups within the genera Agrobacterium, Bradyrhizobium, Mesorhizobium and Rhizobium in 16S and 23S rRNA gene PCR-RFLP. The phylogenetic relationships of the 16S rRNA genes supported the grouping results of PCR-RFLP. Most of the isolates from Amorpha fruticosa were classified into two groups closely related to Mesorhizobium amorphae. Seventeen of the 21 isolates from Wisteria sinensis were identified as two groups related to Rhizobium and Agrobacterium. Six out of 10 isolates from Cercis racemosa were identified as a group related to Bradyrhizobium. Our results indicated that each of the investigated legumes nodulated mainly with one or two rhizobial groups, although isolates from different plants intermingled in some small bacterial groups. In addition, correlation between geographic origin and grouping results was found in the isolates from Amorpha fruticosa. These results revealed that the symbiotic bacteria might have been selected by both the legume hosts and the geographic factors.     

Testing appropriate habitat outside of historic range: The case of Amorpha herbacea var. crenulata (Fabaceae)

In the next century, safeguarding plant species against extinction from complete land conversion may require introducing species to novel locations. Although regulatory agencies caution against translocation outside of known historic ranges, when most wild populations and their habitats have been severely altered few viable options may be available for conserving rare plants. We introduced 345 endangered Amorpha herbacea var. crenulata along a pine rockland/transverse glade gradient with similar attributes to historically known occurrences for south Florida, USA, and monitored their survival and growth for five years. The experimental phase addressed: (1) Is the recipient site suitable for colonisation of this species despite hydrological manipulation in the region? (2) Can translocated plants grow equally well in four microhabitats along a gradient within the recipient site? We characterised soil water content, soil nutrient, and vegetation cover to assess the microhabitats at the recipient site. From 2006 to 2008 plants survived in all four microhabitats, but had highest survival in pineland. Translocated plants grew best in microhabitats with less grass cover and higher P content – the pineland and the restoration glade. Through 2008 we observed consistently higher soil water content with less total vegetation cover in pineland and significantly higher P content in the restoration glade.Using 2006–2008 data, we implemented the adaptive management phase, moving 20 plants from the lowest survival microhabitat to the highest survival microhabitat. This tactic improved the survival of plants by 2011, though growth rates of moved plants did not improve. Short-distance translocation, assessing environmental attributes related to plant survival and growth, quantifying similarity of soil, temperature, precipitation, and community as in this study are recommended to evaluate prospective introduction sites for translocations within or outside of range.     

Rescue and Restoration: Experimental Translocation of Amorpha herbacea Walter var. crenulata (Rybd.) Isley into a Novel Urban Habitat

Though translocations of rare populations should be considered only as the last resort for species’ conservation, when habitat destruction is imminent, it may be the only means to preserve a species. With over half the known, wild federally endangered Crenulate leadplant (Amorpha herbacea var. crenulata), Fabaceae, growing on unprotected land slated for development, preserving this unprotected population was critical. We rescued whole plants, cuttings, and seeds for an experimental translocation. Into a restored pine rockland, once dominated by the invasive exotic tree Brazilian pepper (Schinus terebinthifolius), we transplanted plants from different sources and of different sizes. Plants used were rescued from an unprotected site, seedlings, and 1‐, 2‐, and 7‐year‐old plants from Fairchild Tropical Botanic Garden’s ex situ collection, creating a novel population in a new habitat. We also evaluated which propagule type and source had the best survival, growth, and reproduction. After 40 months, overall transplant survival was 71%. Large whole plants, rescued and nursery grown, had the best survival rates (86 and 78%), whereas cuttings had 67% survival and seedlings had only 26% survival. The restored site, once nearly a monoculture of S. terebinthifolius, is now dominated by 104 native plant species, including 17 naturally recruited state listed, plus the one translocated federally endangered plant species. In addition, one federally threatened snake species was observed on the site. These studies demonstrate that botanic garden collections not only play a vital role in the conservation of species’ genetic diversity but also can be used as source material for habitat restoration.     

干旱区人工防护林带不同林分凋落叶分解及养分释放

2007年10月下旬至2008年11月,采用原位模拟分解网袋法,对新疆克拉玛依市区北郊人工防护林新疆杨、紫穗槐及二者混合凋落叶进行为期365 d的分解及养分释放动态试验.结果表明：树种不同,凋落叶质量损失率的动态变化不同;凋落叶组成对质量损失率有显著影响,与单优林凋落叶相比,紫穗槐与新疆杨凋落叶混合后更易于分解.经修正Olson负指数衰减模型分析,新疆杨凋落叶分解系数最低(k=0.167),混合凋落叶分解系数最高(k=0.275),估测3种凋落叶半分解和95％分解所需时间为2.41～4.19 a和10.79～17.98 a.不同的分解时期3种凋落叶中N、P和K的残留率不同,分解1年后,K为净释放,N和P为固持或从周围环境中吸收而富集.分解过程中,除紫穗槐凋落叶在分解中期有机碳分解率下降外,其他处理凋落叶有机碳分解率均不断上升,1年后分解率在35.5%～44.2%之间.C/N值基本呈下降趋势,分解前期和中期下降幅度较小,后期下降较快.     

Analysis of cutaneous and internal gill gas exchange morphology in early larval amphibians, Pseudophryne bibronii and Crinia georgiana

This study uses stereological techniques to examine body, internal gill and cardiovascular morphology of two larval amphibians, Pseudophryne bibronii and Crinia georgiana, to evaluate the roles of diffusive and convective gas exchange. Gosner stage 27 specimens were prepared for light microscopy and six parallel sections of equal distance taken through the body as well as a further six through the heart and internal gills. Body, internal gill and heart volume as well as body and internal gill surface areas were determined. The harmonic mean distance across the internal gills was also measured and used to estimate oxygen diffusive conductance, DO?. The species were of similar body size and surface area, but the heart and internal gills were larger in P. bibronii, which may represent precursors for greater growth of the species beyond stage 27. The much larger surface area of the skin compared to the internal gills in both species suggests it is the main site for gas exchange, with the gills supplementing oxygen uptake. The sparse cutaneous capillary network suggests diffusion is the main oxygen transport mechanism across the skin and directly into deeper tissues. A numerical model that simplifies larval shape, and has an internal (axial vessels) and external oxygen source, confirms that diffusion is able to maintain tissue oxygen with limited convective input.     

多效唑对紫穗槐生长及生理特性的影响

采用盆栽试验的方法研究了土施不同浓度多效唑(1、5、10、15、20 mg/L)对紫穗槐(Amorpha fruticosa Linn.)生长及生理特性的影响,以探明多效唑对紫穗槐的作用机制和最佳处理方式。结果显示:随着多效唑处理浓度(1~20 mg/L)的增加,紫穗槐幼苗株高、单叶面积和主根长呈下降趋势,基径、叶片长宽比、根鲜重和根冠比呈上升趋势;多效唑各处理均使紫穗槐幼苗叶片的相对含水量、叶绿素、可溶性糖和可溶性蛋白含量显著升高,POD活性显著增加,MDA含量和相对电导率显著下降。采用隶属函数法对各项生长、生理指标进行综合评价,结果发现20 mg/L多效唑处理下紫穗槐幼苗的抗性最强。说明多效唑可通过调节紫穗槐幼苗的生物量分配、水分状况、细胞渗透性和抗氧化性等,有效改善其生长、生理特性及提高抗逆性。本研究结果为多效唑在边坡植被建成和恢复中的应用提供了理论依据。     

Influence of environmental oxygen on development and hatching of aquatic eggs of the australian frog, Crinia georgiana

The effect of oxygen partial pressure (Po(2)) on development and hatching was investigated in aquatic embryos of the myobatrachid frog, Crinia georgiana, in the field and in the laboratory. Eggs from 29 field nests experienced widely variable Po(2) but similar temperatures. Mean Po(2) in different nests ranged between 2.9 and 19.3 kPa (grand mean 12.9 kPa), and mean temperature ranged between 11.9 degrees and 16.8 degrees C (grand mean 13.7 degrees C). There was no detectable effect of Po(2) or temperature on development rate or hatching time in the field, except in one nest at 2.9 kPa where the embryos died, presumably in association with hypoxia. Laboratory eggs were incubated at 15 degrees C at a range of Po(2) between 2 and 25 kPa. Between 5 and 25 kPa, there was almost no effect of Po(2) on development rate to stage 26, but the embryos hatched progressively earlier-at earlier stages and lower gut-free body mass-at lower Po(2). At 2 kPa, development was severely delayed, growth of the embryo slowed, and morphological anomalies appeared. A high tolerance to low Po(2) may be an adaptation to embryonic development in the potentially hypoxic, aquatic environment.     

紫穗槐根瘤菌的分离与回接鉴定

采用YMA培养基对新鲜的紫穗槐(Amorpha fruticosa L.)根瘤提取物进行根瘤菌的分离培养,获得的菌落为圆形乳白色,表面凸起,直径0.4~0.5 cm,革兰反应阴性,显微观察显示为短杆状,分离菌株回接原宿主植物,同时接种于经60℃15 min水浴加热处理的紫穗槐种子,蛭石盆栽实验显示,接种此菌的紫穗槐植株都有结瘤,并与对照在结瘤数、瘤重、植株鲜重与干重间都有显著差异,实验证明所分离的菌株均为紫穗槐根瘤菌。