CHROMOSOME NUMBERS OF NORTH AMERICAN SPECIES OF ANTENNARIA GAERTNER (ASTERACEAE: INULEAE)

Chromosome numbers are presented for 99 populations of 13 species of Antennaria, including A. plantaginifolia, A. neglecta, A. virginica, A. solitaria, A. racemosa, A. corymbosa. A. rosea, A. media, A. Parlinii, A. fallax, A. neodioica, A. canadensis, and A. petaloidea. Four species from the eastern United States (A. plantaginifolia, A. neglecta, A. solitaria, and A. virginica) were determined as diploid (n = 14), and these are all sexual. Diploid counts were also obtained for two sexual species (A. racemosa and A. corymbosa) from the western United States. Chromosome counts are presented for two heteroploid agamic complexes occurring in the eastern United States; these include what have traditionally been referred to as A. Parlinii, A. fallax, A. neodioica, A. canadensis, and A. petaloidea. Determinations of 2n = 56, 70, 84, and 112 were obtained for the A. Parlinii and A. fallax groups, where 2n = 84 had been the only number previously reported. Numbers of 2n = 84 were confirmed for A. petaloidea and A. canadensis and 2n = 56 for A. neodioica. The western United States polyploid species (A. rosea and A. media) are reported as 2n = 56. The presence of apomixis is correlated with polyploidy. The distribution of chromosome numbers in eastern United States Antennaria demonstrates that two diploids and many polyploids occur above the glacial margin, and thus there is an increase in the frequency of polyploidy with latitude. Colonization of the glaciated region by Antennaria following the recession of the Wisconsin ice sheet is also discussed. Many of the polyploids occur only in the glaciated region, thus suggesting a recent origin for these cytotypes. There is evidence indicating that the original base number in Antennaria may be x = 7.     

ALLOZYME DIVERGENCE AMONG FIVE DIPLOID SPECIES OF ANTENNARIA (ASTERACEAE: INULEAE) AND THEIR ALLOPOLYPLOID DERIVATIVES

The Antennaria parlinii and A. neodioica agamic complexes are widely distributed across North America. Morphological data have suggested that these dioecious, perennial, entire-leaved herbs are of multiple hybrid origin from among five sexual diploid species. Antennaria neglecta, A. plantaginifolia, A. racemosa, and A. virginica are hypothesized to be the diploid progenitors of the A. neodioica complex, whereas A. parlinii sensu lato is thought to include the genomes of A. plantaginifolia, A. racemosa, and A. solitaria. An electrophoretic study was initiated to assess the degree of divergence among the five diploid species and to test the hypotheses of the hybrid (allopolyploid) origins of A. parlinii and A. neodioica. Twenty genetic loci were surveyed in 76 populations of the diploid and polyploid taxa. The diploid species are well defined morphologically, although the genetic basis of differences distinguishing them have not been determined. The species exhibit little divergence at genes specifying soluble enzymes, however each species has unique alleles in highest frequency at one or two genes. Allozymes indicate that gene diversity in the obligately outcrossing diploids occurs primarily within rather than among populations. The shale barren endemic, A. virginica, is as genetically diverse as the more edaphically diverse and widespread species. Tetraploid cytotypes of diploid (2n = 28) A. virginica possess the same allozymes as the diploids and these cytotypes appear to be of autopolyploid (non-hybrid) origin. Enzyme electrophoresis is concordant with morphological data in suggesting that A. neodioica contains the genomes of A. neglecta, A. virginica, A. plantaginifolia, A. racemosa and perhaps A. solitaria whereas the latter three species are the progenitors of A. parlinii.     

用ISSR标记技术分析山药品种遗传多样性

利用ISSR标记技术对28个山药品种的遗传多样性进行分析。结果表明,从44条ISSR引物中可筛选出7条能够扩增出清晰、稳定条带的引物;这7条ISSR引物对28个山药品种扩增条带间存在较大差异,多态性条带比率为83.01%,Shannon多样性指数为0.3191;构建的分子树状图将28个山药品种划分为4组:第一组含有日本白、花山药和日本园3个品种;第二组为小叶山药;第三组为嵩野1号;其余23个品种归入第四组。而且主成分分析结果支持上述的聚类分析结果。这为利用ISSR标记技术鉴定山药品种,为有效地利用山药种质资源提供了依据。     

PATTERNS OF ISOZYME VARIATION IN WESTERN NORTH AMERICAN ANTENNARIA (ASTERACEAE: INULEAE) II. DIPLOID AND POLYPLOID SPECIES OF SECTION ALPINAE

Antennaria is a genus of dioecious, perennial, herbaceous Composites that are especially diverse in the cordillera of western North America. Section Alpinae consists of about nine taxa, among them A. aromatica, A. densifolia, A. media, A. pulchella, and A. umbrinella. Although diploids are morphologically distinct, the polyploid derivatives of the diploids obscure the morphological distinctness of the groups. A survey of 19 putative isozyme loci indicates that the diploids have diverged only moderately from one another with respect to biochemical genetics (I = 0.838 to 0.961). Additionally, only moderate amounts of genetic diversity were detected. Isozyme data are supportive of a hypothesis of a rapid mode of speciation in Antennaria, where morphological differentiation has been accompanied by small amounts of allozyme divergence. Polyploids have significantly higher amounts of heterozygosity than diploids and tetrasomic inheritance is inferred. Evidence from morphology and biochemical genetics suggests that the polyploids represent a continuum between interracial autopolyploids and segmental allopolyploids. In light of the relatively low degree of genetic and morphological divergence among many Antennaria species, taxonomic conservatism is advocated.     

PATTERNS OF CLONAL DIVERSITY IN THE ANTENNARIA ROSEA (ASTERACEAE) POLYPLOID AGAMIC COMPLEX

The perennial herbaceous species, Antennaria rosea, is a large, morphologically diverse, polyploid agamic complex that is widespread in the cordillera of western North America. The species consists of triploid and tetraploid, nonpseudogamous, gametophytic apomicts. Populations of A. rosea are gynoecious, consisting almost entirely of pistillate clones. Clonal diversity among 63 populations of A. rosea was studied over a large portion of its range. Isozyme electrophoresis utilizing four polymorphic enzyme systems detected 192 multilocus genotypes among the populations. Populations of A. rosea tend to be composed of one or a few genotypes (range 1–11; mean 3.5), and these genotypes usually occur in only one or a few localized populations. Geographic patterns of clonal diversity may be a result of frequent genesis of new clones in populations that occur in areas where sexual relatives of A. rosea donate compatible pollen to facultatively sexual apomicts. Populations from previously glaciated regions tend to have fewer clones per population than those from unglaciated portions of the range.     

GENETIC DIFFERENTIATION IN THE FLORIDA SUBSPECIES OF HELIANTHUS DEBILIS (ASTERACEAE)

Techniques of horizontal starch gel electrophoresis were employed to estimate levels of genetic variation within and genetic differentiation among populations of the three Florida subspecies of Helianthus debilis. The subspecies are H. d. debilis, H. d. vestitus, and H. d. tardiflorus. These taxa are very similar with respect to levels of genetic variability. The average values across all populations for proportion of polymorphic loci (25.5%) and number of alleles per polymorphic locus (2.35) are comparable to those found in other outcrossing plants. The average frequency of heterozygotes per locus (0.05) is lower than that found in other outcrossers. Local populations within each subspecies are genetically very similar. The average genetic distance between local populations is D = 0.010 ± 0.001. Subspecies vestitus and subspecies tardiflorus are also genetically very similar (D = 0.015 ± 0.001). However, subspecies debilis is differentiated from both subspecies vestitus (D = 0.121 ± 0.006) and subspecies tardiflorus (D = 0.103 ± 0.005). This genetic differentiation parallels morphological differentiation. The average genetic distance among all three subspecies is D = 0.080 ± 0.003. A moderate amount of genetic differentiation accompanies the process of subspeciation in Helianthus debilis.     

EVIDENCE FOR A MOSAIC HYBRID ZONE IN THE GRASS SHRIMP PALAEMONETES KADIAKENSIS (PALAEMONIDAE) AS REVEALED BY MULTIPLE GENETIC MARKERS

Molecular techniques provide powerful tools for studying the geographic structure of hybrid zones and the dynamics of gene exchange between incipient species. We examined allozyme variation at five loci (PGM, GPI, MDH-1, MDH-2, and LDH) for 27 populations of Palaemonetes kadiakensis from the central, coastal, and eastern regions of Texas. Central Texas populations of P. kadiakensis exhibited highly significant linkage disequilibrium and departures from Hardy-Weinberg genotype proportions. In populations with linkage disequilibrium, allelic differences at GPI defined two types of P. kadiakensis, designated A and B. Both types existed in central Texas with little or no evidence of interbreeding, whereas the populations from all other localities showed complete introgression of type B alleles into the type A gene pool. We also examined ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) variation in a subset of populations, chosen to cover a range of geographic locations and levels of linkage disequilibrium. Two groups of mtDNA haplotypes and two restriction fragment patterns for the rDNA corresponded to allozyme type A and B individuals in populations exhibiting linkage disequilibrium. In populations with ongoing hybridization, all hybrid animals (N= 15) exhibited type A mtDNA. Exhibition of type A mtDNA indicated that type A females had mated successfully with type B males, but type B females had not mated successfully with type A males. Genotype distributions suggest reduced reproduction by hybrid offspring in central Texas populations. These patterns are consistent with a mosaic model of hybrid zone dynamics.     

RADIATION-INDUCED GENETIC MARKERS IN THE STUDY OF LEAF GROWTH IN ZEA

Stein , Otto L. (Montana State U., Missoula.), and Dale Steffensen . Radiation-induced genetic markers in the study of leaf growth in Zea. Amer. Jour. Bot. 46(7): 485–489. Illus. 1959.—Corn embryos heterozygous for the plant color mutant yellow-green were treated with moderate doses of radiation. The sectors resulting from the loss of the dominant gene Yg₂ furnish a graphic illustration of the growth of an organ in space and time. Wet embryos were irradiated and leaves 3 through 6 were analyzed. The total number of sectors per leaf increases in relation to the amount of leaf tissue present at time of irradiation and their distribution varies, indicating a change in mitotic activity during early ontogeny of the leaf. As represented by leaf 6 in the mature embryo, cell division is rather uniform throughout the young primordium. Soon thereafter (leaf 5) cell divisions in the apical region increase, though cell lineages remain short. In the basal region of the leaf, the proportion of cell division decreases but those cells which do divide continue their activity over a longer period than the apical cells. Longer files of mutant cells are formed in such a way. The mid regions of the leaf maintain their status quo in terms of distribution of sectors. The results indicate that differentiation of the intercalary meristem as a thin band occurs early in the development of the leaf primordium. The use of genetic techniques in conjunction with radiation, in this case as a “timed” release of genetic markers, may be a useful tool in the precise analysis of the dynamic aspects of organogenesis and histogenesis.     

中国胭脂鱼种群的遗传分析

采用RAPD和PCR—RFLP技术,分析了长江中游两个中国胭脂鱼群体的遗传结构。50个随机引物进行RAPD分析,有3个引物显示了多态,宜昌、金口群体内个体之间的遗传相似度分别为0．9274、0．9313,群体之间遗传相似度为0．9000。12个限制性内切酶分析了两群体线粒体DNAND-5／6基因的限制性片段长度多态性,仅内切酶Ncil的酶切图谱显示了多态,基因型间的核苷酸序列歧化距离为0．235%,核苷酸多样性为0．004。分析表明,长江中游两个中国胭脂鱼群体遗传结构较为单一,群体之间表现了较为明显的遗传分化。     

EVIDENCE FOR A LACK OF PHOTOSYSTEM SEGREGATION IN CHLAMYDOMONAS REINHARDTII (CHLOROPHYCEAE)

The distribution of photosystems I and II (PSI and PSII) in cells of Chlamydomonas reinhardtii Dangeard was studied by immunogold electron microscopy using cultures grown autotrophically at moderate irradiance and harvested in the middle of the light period. Sections of Lowicryl-embedded cells were labeled with monospecific heterologous antisera raised against the reaction center proteins of PSI (CP1-e) or the core antenna proteins of PSII (CP40 and CP47). All three antisera labeled both the appressed and the nonappressed thylakoid membranes at essentially similar densities. Labeling with both PSI and PSII antisera was slightly more concentrated over the outer nonappressed membranes of the thylakoid bands (1.7- to 2.4-fold with anti-CP1- e and 1.5- to 1.8-fold with anti-CP47 and anti-CP40). However, since appressed membranes comprised 73% of the total thylakoid membranes, 50%–62% of the PSI and 58%–65% of the PSII labeling were localized on appressed membranes. We conclude that photosystem distribution in C. reinhardtii is similar to that reported for other algae and different from the lateral heterogeneity observed in higher plants.     

HOMOPLOID RETICULATE EVOLUTION IN HELIANTHUS (ASTERACEAE): EVIDENCE FROM RIBOSOMAL GENES

Phylogenetic relationships among the 21 taxa comprising Helianthus sect. Helianthus and three outgroup species were assessed by restriction site mapping of the 18S-25S nuclear ribosomal RNA gene family. Wagner parsimony analysis of the 41 restriction site or length mutations observed produced a single 59-step most parsimonious tree. This tree was then compared to a cytoplasmic-based plastid phylogeny for this group. Several major discrepancies were observed between the two phylogenies suggesting both recent and ancient introgression. Furthermore, three cases of diploid hybrid speciation are unambiguously documented and a fourth case is suggested. These data are interpreted to suggest that evolution in Heliathus is reticulate rather than exclusively dichotomous and branching.     

GENETIC EVIDENCE FOR MULTIPLE ORIGINS OF DIOECY IN THE HAWAIIAN SHRUB WIKSTROEMIA (THYMELAEACEAE)

Dioecy is unusually common in the Hawaiian Islands, yet little is known about the evolutionary biology of this breeding system. A native shrub, Wikstroemia, has an unusually diverse array of breeding systems: two forms of dioecy, cryptic and morphological dioecy, as well as hermaphroditism (perfect flowers). The existence of two forms of dioecy is significant for three reasons: 1) the presence of cryptic unisexuals that are functionally unisexual, but retain the appearance of hermaphroditism in both sexes, is strong evidence for the ancestral status of hermaphroditism; 2) the production of nonfunctional pollen, by female cryptic unisexuals, is a new instance of a phenomenon which has previously been reported for a few other species; 3) the two forms of dioecy are morphological markers which are useful in hybridization studies for tracing the genetic basis of their inheritance. Crosses were made between cryptically unisexual individuals (C), between morphologically unisexual individuals (M), and between the two types of unisexuality. The offspring of crosses between individuals with the same sex type usually resulted in offspring with that sex type, but most of the progeny of between-sex type crosses were, unexpectedly, perfect-flowered hermaphrodites. These results show that genetic control of sex determination is not homologous in all populations, suggesting that dioecy has evolved at least twice in Hawaiian Wikstroemia. The genetic data further suggest that males are the heterozygous sex.     

北京地区野生大豆种群SSR标记的遗传多样性评价

 使用40对SSR引物分析了北京地区野生大豆(Glycine soja)天然种群的遗传结构与遗传多样性。10个种群共检测到526个等位变异, 平均每对引物等位基因数为13.15个, 种群平均Shannon指数(I)为0.658, 群体平均位点预期杂合度(H_e)为0.369, 群体平均位点杂合度(H_o)为1.29 %。平均种群内遗传多样度(H_s)为0.362, 平均种群间遗传多样度(D_ST)为0.446, 基因分化程度(G_ST)为0.544。该研究显示, 中-西部生态区种群比北部和东部山区种群有较高的遗传多样性。在地理上, 环绕北京地区的太行山和燕山两大余脉区域野生大豆种群遗传分化表现出地理差异。可能是经过干旱选择而形成的有抗旱潜力的种群在遗传上表现单一化。期待该种群提供耐旱基因。     

GENETIC DIVERGENCE AMONG MEDITERRANEAN AND MACARONESIAN GENERA OF THE SUBTRIBE CHRYSANTHEMINAE (ASTERACEAE)

Genetic variation at 17 isozyme loci was used to assess divergence among the four genera comprising subtribe Chrysantheminae (Anthemideae: Asteraceae). The Macaronesian endemic genus Argyranthemum is supported as monophyletic and is about equally divergent at isozyme loci from the other three genera of the Chrysantheminae, Chrysanthemum, Heteranthemis, and Ismelia. Chrysanthemum is native to the Mediterranean whereas Heteranthemis occurs in southern Iberia and Morocco, and Ismelia is endemic to Morocco. The genera Chrysanthemum and Ismelia have a genetic identity of 0.9283, which is comparable to values often seen for congeneric species and indicates that they should be treated as one genus. The isozyme data indicate that three lines consisting of Argyranthemum, Chrysanthemum-Ismelia, and Heteranthemis radiated rapidly from a common ancestor. Divergence times estimated from isozyme data suggest that the initial radiation of the three lines occurred 2.5–3.0 mya. If this is so, then Argyranthemum or its ancestor arrived in Macaronesia after all the islands except La Palma and El Hierro were formed. The evolutionary history of the subtribe is discussed in relation to the climatic and geological events that took place in the Western Mediterranean between the Tertiary and Quaternary, i.e., the first Northern Hemisphere glaciation and desertification of the Sahara region. The high mean genetic identities between species of Argyranthemum suggest that it might have subsequently undergone a second more recent radiation in the Macaronesian Islands. Also, the high mean identity (0.860) between populations in Chrysanthemum and Ismelia suggest that these continental genera might be in the early stages of secondary speciation.     

大豆灰斑病菌生理小种的RAPD标记*

运用随机扩增多态性DNA（RandomamplifiedpolymorphicDNA,RAPD）技术对发生于中国东北的大豆发斑病菌（Cercosporidiumsojinum）的10个生理小种进行基因组DNA多态性分析。用13个10-核苷酸随机引物共计获得了111个RAPD标记,其中86.5％具有多态性,通过聚类分析确定了供试小种间的亲缘关系。试验证明,RAPD技术分析大豆灰斑病菌遗传变异可提供大量分子标记,综合分析13个随机引物的扩增谱带可将供试菌株清楚分开。RAPD技术是一项操作简单、快速和灵敏的方法,极具对病菌群体遗传分析的潜力。     

INDUCTION OF APOMIXIS BY OUTCROSSING BETWEEN GENETICALLY DIVERGENT ENTITIES OF CALOGLOSSA LEPRIEURII (CERAMIALES,RHODOPHYTA) AND EVIDENCE OF HYBRID APOMICTS IN NATURE1

Our previous study revealed that apomixis, recycling of tetrasporophytes, can be generated through outcrossing between genetically divergent entities of Caloglossa monosticha M. Kamiya, though such apomicts have never been found in nature. In the case of C. leprieurii (Mont.) G. Martens, the most widespread species in this genus, many apomictic strains have been isolated worldwide, but it is unknown whether these apomicts evolved through an outcrossing process similar to that in C. monosticha. In this study, heterogeneity of the apomicts and their sexual relatives as well as their evolutionary relationships was examined using the nuclear‐encoded actin gene and plastid‐encoded RUBISCO spacer region. Thirteen out of 18 apomictic strains were heterogeneous and contained divergent actin alleles, whereas only two out of 23 sexual strains were heterogeneous. The five homogeneous apomicts were genetically identical, or quite similar, to the sexual strains isolated from adjacent sites. Furthermore, three of the five homogeneous apomicts frequently produced tetraspores that grew into gametophytes, while all the heterogeneous apomicts never generated gametophytes. Apomictic strains from Florida were allotriploid, and each of the three actin sequences was closely related to those of sexual strains from Florida, Peru, and Mexico/Guatemala. In crossing tests, obligate apomixis was generated through the outcrossing between the male from Madagascar and the female from the northwestern Atlantic. These results suggest that outcrossing between genetically divergent sexual entities is one factor that induces apomixis in C. leprieurii.     

大豆灰斑病菌生理小种的RAPD标记

运用随机扩增多态性DNA（RandomamplifiedpolymorphicDNA,RAPD）技术对发生于中国东北的大豆发斑病菌（Cercosporidiumsojinum）的10个生理小种进行基因组DNA多态性分析。用13个10-核苷酸随机引物共计获得了111个RAPD标记,其中86.5％具有多态性,通过聚类分析确定了供试小种间的亲缘关系。试验证明,RAPD技术分析大豆灰斑病菌遗传变异可提供大量分子标记,综合分析13个随机引物的扩增谱带可将供试菌株清楚分开。RAPD技术是一项操作简单、快速和灵敏的方法,极具对病菌群体遗传分析的潜力。     

长竹蛏不同地理居群的遗传多样性

研究以大连、烟台、莱州、青岛和赣榆近海5个不同地理居群的长竹蛏(Solen strictus Gould)为实验材料,利用ISSR分子标记进行了遗传多样性的研究。结果表明,13个ISSR引物在5个居群中共扩增出200个位点,平均每个引物记录15.4个位点,5个居群的多态位点比例为43.56%—60.43%。长竹蛏在物种水平上的Nei’s基因多样性指数和Shannon’s信息指数分别为0.2854和0.4390,在居群水平上分别为0.1674和0.2530。NJ聚类分析显示,青岛居群与赣榆居群的亲缘关系最近,而烟台居群与其他4个居群的亲缘关系较远。经Mantel检测,长竹蛏5个居群间的遗传距离与地理距离并无相关性(r=-0.0834,P>0.1)。AMOVA分子变异分析表明,长竹蛏的遗传变异有47.71%发生在居群间,52.29%发生在居群内,居群内的遗传变异大于居群间的遗传变异。长竹蛏5个居群间的遗传分化系数(Gst)为0.2889,基因流(Nm)为1.3194。结果表明,长竹蛏具有较高的遗传多样性,但居群间已发生了一定程度的遗传分化。       

VERY LOW GENETIC HETEROZYGOSITIES IN SEXUAL AND AGAMOSPERMOUS POPULATIONS OF EUPATORIUM ALTISSIMUM (ASTERACEAE)

An electrophoretic study revealed very low genetic heterozygosities for both sexual and agamospermous populations of Eupatorium altissimum in eastern North America. Low heterozygosity or gene diversity (H_T = 0.03) can be explained by the small sizes of the extant sexual populations, estimated to be on the order of 10⁴. There was no substantial difference in observed heterozygosity between sexual and agamospermous races. This result shows that a higher level of heterozygosity is not a prerequisite for the evolution of agamospermous races from the sexual ancestor.