东亚七筋姑多倍体ITS区的序列测定与分析

通过对13个东亚七筋姑二倍体居群和5个四倍体居群的ITS区(包括ITS1、5.8SrDNA和ITS2)进行克隆测序。ITS序列长度显示出比较高的多态性,ITS1长度为171~270bp,ITS2长度为205~238bp,5.8S序列的长度在162~164bp之间,序列中G C含量为45.83%~54.13%,当空位(gap)作缺失处理时,东亚七筋姑ITS区全序列排序后的长度为682bp,其中简约性位点289个;最大简约法分析获得一致性指数(CI)为0.630,保留系数(RI)为0.684,四倍体居群没有单独聚为一支,而是和相邻地理单元的二倍体居群聚为一支,本研究结果支持东亚七筋姑异地多起源的推断。     

东亚七筋姑的传粉生物学研究

以陕西镇坪化龙山地区的东亚七筋姑自然居群为对象,通过重力玻片法、套袋法和人工授粉对其进行连续的传粉生物学观察和研究,以明确其散粉规律和繁育系统.结果表明,东亚七筋姑的花期在五月中下旬,单花花期2～3 d,整个花序的花期随花序大小的不同而异,一般为3～7 d;东亚七筋姑为虫媒传粉,其访花昆虫大多为小型昆虫,主要种类有小型甲虫、蚁类和蝇类等;东亚七筋姑自花传粉是可育的,存在异花传粉,不存在无融合生殖,自然条件下结籽率为58%,人工异株异花授粉结籽率为82%.可见,东亚七筋姑的繁育系统为兼性异交类型,需要传粉者.     

New Steroidal Glycosides from Rhizomes of Clintonia udensis

A total of 10 steroidal glycosides, together with three new spirostanol glycosides (6–8), a new furostanol glycoside (9), and a new cholestane glycoside (10), were isolated from the rhizomes of Clintonia udensis (Liliaceae). The structures of the new compounds were determined on the basis of extensive spectroscopic analyses, including 2-D nuclear magnetic resonance (NMR) data, and of hydrolytic cleavage followed by chromatographic or spectroscopic analyses. The isolated glycosides were evaluated for their cytotoxic activity against HL-60 leukemia cells. Spirostanol glycosides 1 and 2, and furostanol glycoside 4 showed cytotoxic activity with IC₅₀ values of 3.2±0.02, 2.2±0.12, and 2.2±0.06 μg/ml, respectively. Neither the spirostanol and furostanol saponins with a hydroxy group at C-1 (6 and 9) and C-12 (7 and 8) nor cholestane glycosides (5 and 10) exhibited apparent cytotoxic activity at a sample concentration of 10 μg/ml.     

Clonal diversity of Clintonia udensis Trautv. et Mey. populations and its correlation with ecological factors

The clonal diversity of Clintonia udensis Trautv.et Mey.was detected by ISSR markers among 16 populations,and its correlation with ecological factors was analyzed as well in this work.Results showed that individuals(clonal ramets)per genotype were 1.12 and 1.149 at population and species levels,respectively,and that the 16 populations were all multiclonal.The detected genotypes were localized,without exception,within populations but demonstrated relatively high clonal differentiation among populations.The clonal diversity of the studied populations was high,with the average Simpson's index of 0.975,while the genets showed a clonal architecture of"guerilla".The population genetic diversities revealed by genet were consistent with those by ramet,further confirming their genetic differentiation among populations.And its genotype diversity within populations probably resulted largely from the frequent seedling regeneration and self-compatibility.In addition,the correlation analysis further revealed that,among the ecological factors,Simpson's index of C.udensis had a significant positive correlation(P<0.05)with pH values in the soil but not others.     

A Cytogeographical Study on Clintonia udensis (Liliaceae)

The genus Clintonia has four species in North America and one in eastern Asia (Fig．1)．In this paper,the karyotypes and the intraspecific morphological and cytogeographic differentiations of ten populations of C．udensis from China are analysed,and the probable origin area of the genus is also discussed．The conclusions are as follows： (1)Based On the chromosome numbers 2n=28 from the North American species,the Japanese and Himalayan groups,the earlier investigators established x=14 as the basic chromosome number of Clintonia,and they thought that there was polyploid in this genus except for aneuploid only in C．borealis (Utech,1975;Utech and Suda,1975),but a few authors(Sen,1975;Wang et al,1993)pointed out the x=7 basic number of this genus based on 2n=14 in C．udensis from Primorskiy Kray of Russia(Skolovskaya,1966)and Yunlong,Yunnan Province of China(Wang et al,1993)respectively．Our result along with there reports by previous authors(see table 1)shows that at least two ploid levels exist in C．udensis,i.e．2n＝14 and 2n＝28．The diploids are widely distributed from northwest Yunnan of China to Primorskiy Kray of Russia,while the tetraploids are located in northwest Yunnan,Himalayas,Japan,and a narrow area in Mt．Hualongshan of southern Shaanxi(07 population)(Fig．2)．Therefore,the basic chromosome number of Clintonia certainly is x=7 ratherthan x=14,while 2n=28 in North America,Japan and Himalayan area are tetraploid, not diploid, According to the previous data,so far no diploid taxa of Clintonia has been found in North America and Himalayas．So we consider that the primitive type of Clintonia is in eastern Asia,and the secondary diversity center and the present distribution center of this genus are in North America． (2)C．udensis is widely distributed in eastern Asia(Fig．2);it has two cytotypes．The karyotypes for all the diploid populations are remarkablely similar. Taken together, they can be roughly repersented by the formula:K(2n)=14=2V+6J+2v+4j(2NOR+2j). The chromosomes range in length 25.55～12.78 μm, with the ratio of the longest to the shortest 2.0. The karyotype belongs to Stebbins (1971)2A(→2B). For the tetraploid taxa, except for 07 population, karyotypes are also identical and may be roughly symbolized as: K (2n)=28=4V + 12J + 4v+8j ( 2NOR + 6j ). The length of chomosomes is from 27.87 to 13.93 μm, with the ratio of the longest to the shortest 2.0, and thus the karyotype belongs to 2A(→2B). The karyotype of 07 population is similar to those of above tetraploid taxa but also has some differences, especially in the position of satellites and the morphology of 10th and 14th pairs of chromosomes. Its formula is K(2n)=28=4V+12J(2NOR+10J)+4v+ 8j(2n=28=10m+16sm(2SAT)+2st). The ratio of the longest (23.72μm)to the shortest(12.97μm) chromosomes is 1.83. The Karyotype belongs to 2A. And the distribution range of this population is very narrow. We think that it is probably a recent evolutionary event in C. udensis. (3)Wether Clintonia in eastern Asia has 1 sp. or 2 spp. or 1 spp. and 1 var. has been debated for a long period. According to our observation, within C. udensis, only the size of seeds is related to its ploidy level, i.e. diploid individuals have smaller seeds and tetraploid ones have larger ones; the colour of seeds is related to its geographic distribution, i.e. the materials from the Himalayas through Yunnan, Sichuan to eastern Qinling Range have pale brown seeds, while those from Mt. Lüliang, Shanxi Province via Hebei, Liaoning, Jilin to Siberia and Japan have dark brown seeds. Some other morphological characters, such as the size of leaves and fruits, inflorescence type and flower numbers between individuals in one locality, even within one populaion have evident variation. Therefore, we consider that evidence(see Table 4 )for separating C. alpina or C. udensis var. alpina from C. udensis is notsufficient.     

基于叶绿体DNA序列分析东亚七筋姑的遗传进化

通过对东亚七筋姑(Clintoniaudensis Trautv.etMey.)19个居群的cpDNA基因间隔区(cpDNA intergenic regions)rpl20-rps12和trnL-F进行序列分析,结果表明,当空位(Gap)作缺失处理时,trnL-F全序列排序后的长度为754个位点,G C含量为35.2%～35.8%,rpl20-rps12序列排序后长度为814bp,G C含量为34.1%～34.3%.聚类结果显示四倍体居群没有单独聚为一支,其多倍体可能是同源多倍体,起源方式属于异地多起源.     

Seed germination time course and seedling development of Clintonia udensis Trautv. et Mey. (Uvulariaceae): a typical cool temperate woodland perennial in Japan

Seed germination time course and seedling development mechanisms of Clintonia udensis Trautv. et Mey. (Uvulariaceae) were investigated under experimental condition. Seed germination tests were carried out under four thermal regimes, i. e. 10, 15, 20, and 25°C, after seeds were harvested, and stored at 5°C in wet conditions for 6 months under light‐exposed or shaded conditions. Approximately 63% of all seeds produced had the potential to germinate beyond 4 years and 6 months. The developmental process after germination continued for over 2 years. Phase I: the radicle first breaks through the seed coat 2 years after fructification. Phase II: the radicle becomes much larger with a hypocotyle. Phase III: part of the cotyledon elongates over 20 mm. Phase IV: the plumule further develops in two steps, i. e. the plumule is first formed, while cotyledon is disappearing, and then the plumule appears with second and third radicles, growing with cotyledon.     

Genetic differentiation and spatiotemporal history of diploidy and tetraploidy of Clintonia udensis

Polyploidy is an important factor shaping the geographic range of a species. Clintonia udensis (Clintonia) is a primary perennial herb widely distributed in China with two karyotypic characteristics—diploid and tetraploid and thereby used to understand the ploidy and distribution. This study unraveled the patterns of genetic variation and spatiotemporal history among the cytotypes of C. udensis using simple sequence repeat or microsatellites. The results showed that the diploids and tetraploids showed the medium level of genetic differentiation; tetraploid was slightly lower than diploid in genetic diversity; recurrent polyploidization seems to have opened new possibilities for the local genotype; the spatiotemporal history of C. udensis allows tracing the interplay of polyploidy evolution; isolated and different ecological surroundings could act as evolutionary capacitors, preserve distinct karyological, and genetic diversity. The approaches of integrating genetic differentiation and spatiotemporal history of diploidy and tetraploidy of Clintonia udens would possibly provide a powerful way to understand the ploidy and plant distribution and undertaken in similar studies in other plant species simultaneously contained the diploid and tetraploid.     

七筋菇自然居群的遗传结构分析

采用ISSR分子标记,对七筋菇(Clintonia udensis)17个居群的遗传多样性与遗传结构进行了研究。结果表明:七筋菇不同居群的多态位点百分率PPB为11.90%~59.52%,总的多态位点百分率PPB为98.8%,具有高的遗传多样性。Shannon多样性指数(0.6903)和基因分化系数(GST=0.6944)均揭示出七筋菇居群间存在明显的遗传差异,AMOVA分析结果也显示遗传变异主要发生在居群之间(81.47%),而居群内部的遗传变异仅为18.53%。七筋菇居群间的遗传距离从0.1871~0.6632,平均为0.3838,大于同一物种居群间的平均遗传距离值(0.05),同样表明七筋菇居群间的遗传多样性存在较大差异。七筋菇居群间的基因流Nm=0.2200,远远低于一般广布种植物的基因流(Nm=1.881)。Mantel检测显示居群间的遗传距离与地理距离之间没有显著相关性(r=0.029,P=0.3196)。七筋菇分布范围广以及其进化历史是其具有高遗传多样性的原因;居群间存在较高遗传变异可能是由于七筋菇本身的生物学特性、有限的基因流以及遗传漂变等原因造成的。     

Asexual and sexual reproductive strategies in clonal plants

Most plants can reproduce both sexually and asexually (or vegetatively),and the balance between the two reproductive modes may vary widely between and within species.Extensive clonal growth may affect the evolution of life history traits in many ways.First,in some clonal species,sexual reproduction and sex ratio vary largely among populations.Variation in sexual reproduction may strongly affect plant's adaptation to local environments and the evolution of the geographic range.Second,clonal growth can increase floral display,and thus pollinator attraction,while it may impose serious constraints and evolutionary challenges on plants through geitonogamy that may strongly influence pollen dispersal.Geitonogamous pollination can bring a cost to plant fitness through both female and male functions.Some co-evolutionary interactions,therefore,may exist between the spatial structure and the mating behavior of clonal plants.Finally,a trade-off may exist between sexual reproduction and clonal growth.Resource allocation to the two reproductive modes may depend on environmental conditions,competitive dominance,life span,and genetic factors.If different reproductive modes represent adaptive strategies for plants in different environments,we expect that most of the resources should be allocated to sexual reproduction in habitats with fluctuating environmental conditions and strong competition,while clonal growth should be dominant in stable habitats.Yet we know little about the consequence of natural selection on the two reproductive modes and factors which control the balance of the two reproductive modes.Future studies should investigate the reproductive strategies of clonal plants simultaneously from both sexual and asexual perspectives.     

Asexual and sexual reproductive strategies in clonal plants

Most plants can reproduce both sexually and asexually (or vegetatively), and the balance between the two reproductive modes may vary widely between and within species. Extensive clonal growth may affect the evolution of life history traits in many ways. First, in some clonal species, sexual reproduction and sex ratio vary largely among populations. Variation in sexual reproduction may strongly affect plant’s adaptation to local environments and the evolution of the geographic range. Second, clonal growth can increase floral display, and thus pollinator attraction, while it may impose serious constraints and evolutionary challenges on plants through geitonogamy that may strongly influence pollen dispersal. Geitonogamous pollination can bring a cost to plant fitness through both female and male functions. Some co-evolutionary interactions, therefore, may exist between the spatial structure and the mating behavior of clonal plants. Finally, a trade-off may exist between sexual reproduction and clonal growth. Resource allocation to the two reproductive modes may depend on environmental conditions, competitive dominance, life span, and genetic factors. If different reproductive modes represent adaptive strategies for plants in different environments, we expect that most of the resources should be allocated to sexual reproduction in habitats with fluctuating environmental conditions and strong competition, while clonal growth should be dominant in stable habitats. Yet we know little about the consequence of natural selection on the two reproductive modes and factors which control the balance of the two reproductive modes. Future studies should investigate the reproductive strategies of clonal plants simultaneously from both sexual and asexual perspectives. Translated from Acta Phytoecologica Sinica, 2006, 20(1): 174–183 [译自: 植物生态学报]     

Reproductive biology of the 38 extant felid species: a review

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Reproductive mode and genetic variation suggest a North American origin of European Letharia vulpina

Our data on the intercontinental population biology of Letharia vulpina show an unexpected shift from a recombining North American population with unique haplotypes to genetically depauperate Swedish and Italian populations, each with many representatives of a single repeated haplotype. Analysis of eight loci in 47 individuals supported recombination in North American populations and showed almost no variation among European populations. We infer that a genetic bottleneck caused by limited long-distance dispersal accounts for the lack of genetic variation found in marginal populations. This lack of variation in the European populations makes it impossible to use population genetic means to distinguish clonal reproduction from self-fertilization or even outcrossing, but phenotype indicates that reproduction in the marginal populations is by clonal spread, via soredia and isidioid soredia.     

Reproductive biology of Sykes and blue monkeys (Cercopithecus mitis)

Thirty-five live births were recorded over a 5-year period from three subspecies of the mitis (Sykes and blue) monkey (Cercopithecus mitis) maintained outdoors in single-male harem breeding groups. There was no indication of birth seasonality. Females of one subspecies, C. m. kolbi, were individually caged, and basic reproductive parameters were monitored. A menstrual cycle length of 31.9 ± 6.0 days was recorded, and there were a large number of irregular cycles. Within individuals, mean progesterone and estrogen concentrations during the luteal phase of the cycle ranged from 5.5 to 10.7 nmol/liter and from 542 to 829 pmol/liter, respectively. Attempts at timed matings were unsuccessful. It is concluded that the mitis monkey is slow to adapt reproductively to captivity and is, therefore, not especially well-suited for routine reproductive studies. However, it may prove valuable to elucidate some of the more complex factors controlling guenon reproduction.     

Reproductive biology of Philodryas patagoniensis (Snakes: Dipsadidae) in south Brazil: Female reproductive cycle

In this study, we describe the female reproductive cycle of Philodryas patagoniensis in south Brazil, which was described through morpho‐anatomical and histological analyses. The peak of secondary vitellogenesis occurred during winter–spring (July–December), ovulation in spring (October–December), mating and fertilization in spring–summer (October–February), oviposition in spring–autumn (October–May) and births from late spring to autumn (December–July). The diameter of vitellogenic follicles/eggs was larger in winter–spring than in other seasons. The diameter of the shell glands was also larger in winter–spring. In spite of the clear reproductive peak, gonads only showed reduced activity in the autumn. Therefore, at the individual level, females have a discontinuous cyclical reproduction; in the populational level, the reproductive cycle is seasonal semisynchronous. We support the hypothesis that P. patagoniensis have the ability to produce multiple clutches with long‐term stored sperm. Sexual dimorphism in body size was evident, and females are significantly larger and heavier than males. Larger females were able to produce follicles and eggs in larger amount and size. The maternal body size was positively related to the reproductive effort and fecundity. To conclude, we deliberated about the proximal and distal causes that influence the reproductive traits and patterns of P. patagoniensis.     

Reproductive biology in an invasive plant Solidago canadensis

Solidago canadensis,a perennial Compositae plant originating from North America,was introduced into China as a horticultural plant in 1935.Under natural conditions,S.canadensis allocates large amounts of energy to sexual reproduction and produces many seeds,which reflects an r-strategy with high seed number and small seed size.In addition,naturalized populations have a great capacity to grow clonally with underground stems.S.canadensis has become an invasive weed in eastern China,and has caused serious damages to agricultural production and ecosystems in several provinces in China.In order to understand the reproductive characteristics of S.canadensis and effectively control its spread,we examined soil conditions,seed characteristics,seed germination and the capacity for asexual reproduction in different plant parts.We investigated the population dispersion of S.canadensis in fixed sites for three years,and analyzed the seasonal dynamics of the morphological parameters of the underground parts and the caloric values of different organs of S.canadensis.We also compared differences in the root systems of S.canadensis and composite exotic weeds.The following results were obtained:1)Under natural conditions,the germination season of S.canadensis lasts from March to October,with a peak from April to May.Vegetative growth and asexual reproduction are especially vigorous during summer due to high temperatures and soil drought stress.On the other hand,the rainy season proves suitable for seed germination.Most S.canadensis flower between September and January,and fruit in late October.A mature plant can produce about 20000 seeds.The mean weight of 1000 seeds ranges from 0.045 g to 0.050 g,and the mean seed moisture content ranges from 60% to 80%.The light-winged seeds disperse readily by air,water,vehicles,human activity or through livestock.2)S.canadensis seeds have a wide tolerance for different values of pH,salinity and soil moisture.The mean percent germination of seeds is 30% under suitable conditions.The results of seed germination under various environmental stresses and investigation of soil conditions indicate that well-aerated,slightly acidic soils with low salinity are suitable for the growth of S.canadensis.Additionally,S.canadensis has a high tolerance for contamination by heavy metal elements including Zn,Cu and Pb,but has low accumulation coefficients for these elements.3)S.canadensis reproduces asexually via underground rhizomes and nodes on the stem base to recruit new individuals,and in plants that experience mechanical damage,this reproductive strategy is used to produce clonal shoots.The capacity for asexual reproduction among different plant parts rank as follows:underground parts＞stem-base (20 cm)＞stem-base (30 cm)＞stem-base (45 cm)＞stem.Further,with increasing mechanical damage,the quantity of shoots produced by the plant decreases.4)The morphological parameters of the root system of S.canadensis including length,surface area,volume,and average diameter are greater than for composite exotic weeds.These parameters indicate that S.canadensis has the physiological potential to widely invade China.5)The aboveground growth rate and most of the underground morphological parameters vary remarkably among the seasons,with a peak normally occurring in September.In August,a fraction of the energy in leaves and stems is allocated underground to increase fine root growth and water uptake during hot weather.Additionally,the seasonal dynamics of the underground morphological parameters and the caloric values of different organs of S.canadensis enhance its reproductive ability.Based on the results above,we conclude that S.canadensis has great invasive potential in China.We suggest that urgent measures should be taken to control its further spread,and to minimize its impact on local plant diversity.     

Reproductive biology of Psammobatis extenta

Psammobatis extenta collected from the commercial bottom trawl fishery operating in Puerto Quequén, Argentina, between 2000 and 2001, had a sex ratio of 1:1 (535 specimens). Both the largest male and female measured 313 mm total length ( L _T). The relationship between total body mass and L _T was significantly different between sexes; mature females weighed more than mature males for a given L _T. The size at which 50% of males were sexually mature was 262 mm L _T (83·6% of the maximum size), whereas 50% of females were mature at 249 mm L _T (80·0% of the maximum size). The gonadosomatic index in mature males and females was slightly higher during summer, but did not show significant variation over the sampling period. The hepatosomatic index of males and females was higher during the autumn and winter and followed a distinct seasonal pattern. Evidence suggested a continuous reproductive cycle during the year, with a maximum number of females carrying egg-cases in summer.     

Reproductive biology in an invasive plant Solidago canadensis

Solidago canadensis, a perennial Compositae plant originating from North America, was introduced into China as a horticultural plant in 1935. Under natural conditions, S. canadensis allocates large amounts of energy to sexual reproduction and produces many seeds, which reflects an r-strategy with high seed number and small seed size. In addition, naturalized populations have a great capacity to grow clonally with underground stems. S. canadensis has become an invasive weed in eastern China, and has caused serious damages to agricultural production and ecosystems in several provinces in China. In order to understand the reproductive characteristics of S. canadensis and effectively control its spread, we examined soil conditions, seed characteristics, seed germination and the capacity for asexual reproduction in different plant parts. We investigated the population dispersion of S. canadensis in fixed sites for three years, and analyzed the seasonal dynamics of the morphological parameters of the underground parts and the caloric values of different organs of S. canadensis. We also compared differences in the root systems of S. canadensis and composite exotic weeds. The following results were obtained:

Reproductive biology of the Caribbean brooding octocoral Antillogorgia hystrix

The reproductive biology of the Caribbean gorgonian Antillogorgia hystrix was studied in the shallow‐water reefs of Cross Harbour, Great Abaco (The Bahamas) from 2009 to 2010. Antillogorgia hystrix is an internal brooder that reproduces annually. The population at Cross Harbour was gonochoric and the sex ratio was skewed toward females (~3:1). Oogenesis precedes spermatogenesis by several months, and lasts at least 9 months, with oocytes >100 μm in diameter first becoming visible in dissections of samples from February; mature oocytes are present in late October–November. The size of mature oocytes (400–900 μm in diameter) was greater than that of the spermaries, which were rarely larger than 400 μm. Brooded planulae were observed in polyps from early November to mid‐December, and planula release was observed in aquaria in December 2009, which suggests that planulation occurs continuously over this period. Planulae of A. hystrix contained dinoflagellate symbionts, presumably acquired during embryogenesis and/or by mature planulae while they were in the gastrovascular cavity of the polyp. Brooding is an uncommon reproductive strategy among Caribbean gorgonians and this is the first report of internal brooding in the genus Antillogorgia. The genus contains a number of sympatric species with different modes of reproduction, and knowledge of their reproductive biology is critical to understand their ecology and evolution.