The evolution and maintenance of monoecy and dioecy in Sagittaria latifolia (Alismataceae)

Plant species rarely exhibit both monoecious and dioecious sexual systems. This limits opportunities to investigate the consequences of combined versus separate sex function on mating patterns and genetic variation and the analysis of factors responsible for the evolution and maintenance of the two sexual systems. Populations of the North American clonal aquatic Sagittaria latifolia are usually either monoecious or dioecious and often grow in close geographic proximity. We investigated mating patterns, genetic structure, and relationships between the two sexual systems using allozyme variation in populations from southern Ontario, Canada. As predicted, selfing rates in monoecious populations (n = 6, mean = 0.41) were significantly higher than in dioecious populations (n = 6, mean = 0.11). Moreover, marker-based estimates of inbreeding depression (delta) indicated strong selection against inbred offspring in both monoecious (mean delta = 0.83) and dioecious (mean delta = 0.84) populations. However, the difference in selfing rate between the sexual systems was not reflected in contrasting levels of genetic variation. Our surveys of 12 loci in 15 monoecious and 11 dioecious populations revealed no significant differences in the proportion of polymorphic loci (P), number of alleles per locus (A), or observed and expected heterozygosity (H(o) and H(e), respectively). Strong inbreeding depression favoring survival of outcrossed offspring may act to maintain similar levels of diversity between monoecious and dioecious populations. Despite geographical overlap between the two sexual systems in southern Ontario, a dendrogram of genetic relationships indicated two distinct clusters of populations largely corresponding to monoecious and dioecious populations. Reproductive isolation between monoecious and dioecious populations appears to be governed, in part, by observed differences in habitat and life history. We suggest that selfing and inbreeding depression in monoecious populations are important in the transition from monoecy to dioecy and that the maintenance of distinct sexual systems in S. latifolia is governed by interactions between ecology, life history, and mating.     

Phenotypic plasticity of hermaphrodite sex allocation promotes the evolution of separate sexes: an experimental test of the sex-differential plasticity hypothesis using Sagittaria latifolia (Alismataceae)

Separate sexes can evolve under nuclear inheritance when unisexuals have more than twice the reproductive fitness of hermaphrodites through one sex function (e.g., when females have more than twice the seed fertility of hermaphrodites). Because separate sexes are thought to evolve most commonly via a gynodioecious intermediate (i.e., populations in which females and hermaphrodites cooccur), the conditions under which females can become established in populations of hermaphrodites are of considerable interest. It has been proposed that resource-poor conditions could promote the establishment of females if hermaphrodites are plastic in their sex allocation and allocate fewer resources to seed production under these conditions. If this occurs, the seed fertility of females could exceed the doubling required for the evolution of unisexuality under low-, but not high-resource conditions (the sex-differential plasticity hypothesis). We tested this hypothesis using replicate experimental arrays of the aquatic herb Sagittaria latifolia grown under two fertilizer treatments. The results supported the sex-differential plasticity hypothesis, with females having more than twice the seed fertility of hermaphrodites under low-, but not high-fertilizer conditions. Our findings are consistent with the idea that separate sexes are more likely to evolve under unfavorable conditions.     

Chloroplast haplotype variation among monoecious and dioecious populations of Sagittaria latifolia (Alismataceae) in eastern North America

Aquatic plants commonly have extensive geographical distributions, implying few restrictions to dispersal. Here we investigate the postglacial history of an aquatic plant with contrasting sexual systems (monoecy and dioecy), which are predicted to affect dispersal ability. We examined the distribution of cpDNA haplotypes using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) among 76 populations (32 monoecious, 38 dioecious, two mixed and four undetermined populations) of Sagittaria latifolia sampled throughout eastern North America. We also use these data to investigate the polarity of the evolutionary transition between monoecy and dioecy. Using PCR-RFLP, we identified eight cpDNA haplotypes. All haplotypes were found in unglaciated areas of the species' range, clustered primarily in the southeastern United States, providing evidence that glacial refugia probably occurred in this area. Genetic diversity (hT) was more than six times greater among monoecious compared to dioecious populations. All seven of the haplotypes for which the sexual system could be determined were represented among monoecious populations. In contrast, only four haplotypes were detected in dioecious populations and 94% of individuals from dioecious populations possessed a single haplotype. Monoecious populations possessing this widespread haplotype were restricted to the southern portion of the range, indicating that dioecy probably originated in this region and then spread northwards. The distribution of cpDNA haplotypes in dioecious populations represents a subset of the variation found in monoecious populations, a pattern expected if dioecy has evolved from monoecy in S. latifolia.     

Pollinator responses to variation in floral display and flower size in dioecious Sagittaria latifolia (Alismataceae)

In animal-pollinated plants with unisexual flowers, sexual dimorphism in floral traits may be the consequence of pollinator-mediated selection. Experimental investigations of the effects of variation in flower size and floral display on pollinator visitation can provide insights into the evolution of floral dimorphism in dioecious plants. Here, we investigated pollinator responses to experimental arrays of dioecious Sagittaria latifolia in which we manipulated floral display and flower size. We also examined whether there were changes in pollinator visitation with increasing dimorphism in flower size. In S. latifolia, males have larger flowers and smaller floral displays than females. Visitation by pollinators, mainly flies and bees, was more frequent for male than for female inflorescences and increased with increasing flower size, regardless of sex. The number of insect visits per flower decreased with increasing floral display in males but remained constant in females. Greater sexual dimorphism in flower size increased visits to male inflorescences but had no influence on the number of visits to female inflorescences. These results suggest that larger flower sizes would be advantageous to both females and males, and no evidence was found that females suffer from increased flower-size dimorphism. Small daily floral displays may benefit males by allowing extended flowering periods and greater opportunities for effective pollen dispersal.     

Gender variation of sequential inflorescences in a monoecious plant Sagittaria trifolia (Alismataceae)

In protogynous plants, female flowers of early blooming plants are at a reproductive disadvantage because they cannot set fruit due to the lack of available pollen. To study this phenomenon, gender expression of the monoecious herb Sagittaria trifolia was investigated over the entire flowering season in two field and two cultivated populations in Hubei and Hunan Provinces, China. In racemes of S. trifolia, flowers open sequentially from bottom to top, with female flowers opening first followed by male flowers. This creates a temporal separation of sexes in the species. Under field conditions small plants are often male, with production of both male and female flowers increasing with plant size. Femaleness increased among sequential inflorescences since female flower production increased whereas male flower production did not. Seed production was greater in large inflorescences because they contain more female flowers, and the number of ovules increased in female flowers at basal positions within the raceme. A consistent pattern of high seed set was observed in flowers from both field and cultivated populations. About 1 % of unfertilized ovules resulted from no pollination and 2 % of the seeds produced were only partly developed due to resource limitation. In the first inflorescence of the six experimental populations, 6.7-40.0 % of individuals produced only male flowers, and female flowers of 1.9-6.5 % individuals were aborted. The occurrence of male flowers in early blooming inflorescences could be an adaptive strategy to conserve resources and enhance pollination of female flowers in protogynous S. trifolia.     

Gender plasticity in Sagittaria sagittifolia (Alismataceae), a monoecious aquatic species

 Aquatic plants are well known for their high degree of phenotypic plasticity in vegetative structures, particularly leaves. Less well understood is the extent to which their sexuality can be modified by environmental conditions. Here we investigate gender plasticity in the European clonal monoecious aquatic Sagittaria sagittifolia (Alismataceae) to determine how floral sex ratios may vary with plant size and inflorescence order. We sampled two populations from aquatic habitats in East Anglia, U.K. and measured a range of plant attributes including ramet size and the number of female and male flowers per inflorescence. The two populations exhibited similar patterns of phenotypic gender, despite contrasting patterns of total allocation to female and male flower number. Plants produced male-biased floral sex ratios but female flower number increased from the first to the second inflorescence whereas male flower number decreased. Size-dependent gender modification occurred in both populations, but the patterns of allocation to female flower production differed between the two populations. Our results are consistent with the view that monoecy is a sexual strategy that enables plants to adjust female and male allocation in response to changing environmental conditions. Received September 16, 2002; accepted October 23, 2002 Published online: March 20, 2003     

Sixteen polymorphic microsatellite markers from Zizania latifolia Turcz. (Poaceae)

Sixteen polymorphic microsatellite markers were isolated and identified in Zizania latifolia Turcz. (Poaceae), a perennial aquatic plant widespread in Eastern Asia. The microsatellite-enriched library was constructed using the fast isolation by AFLP of sequences containing repeats method. These markers revealed two to 14 alleles, with an average of 5.6 alleles per locus. The observed and expected heterozygosities varied from 0.071 to 0.690 and from 0.174 to 0.812, respectively. These markers will be useful for studying of gene flow and evaluating the genetic diversity of the Zizania latifolia population.     

Population genetics of two rare perennials in isolated wetlands: Sagittaria isoetiformis and S. teres (Alismataceae)

We investigated genetic structure in two closely related perennial plants that occur in isolated wetlands: Sagittaria isoetiformis, restricted to the southeastern Coastal Plain of North America, and S. teres, endemic to the northeastern Coastal Plain. Using horizontal starch-gel electrophoresis, we screened 527 individuals from 11 populations of S. isoetiformis and 367 individuals from seven populations of S. teres. A high proportion of the 16 loci were polymorphic (%P(S) = 93.8% in S. isoetiformis and %P(S) = 75.0% in S. teres), with higher mean numbers of alleles per polymorphic locus and effective alleles per locus in S. isoetiformis (AP = 3.27, A(E) = 1.90) than in S. teres (AP = 2.58, A(E) = 1.30). Species- and population-level expected heterozygosities were higher in S. isoetiformis (H(ES) = 0.399, H(EP) = 0.218) than in S. teres (H(ES) = 0.177, H(EP) = 0.101). Jackknife estimates of F statistics indicated moderate levels of inbreeding in S. teres (F(IS) = 23.1%). Strong differentiation characterized these geographically isolated populations (G(ST) = 39.9% in S. isoetiformis, and G(ST) = 26.1% in S. teres). Genetic identities varied substantially within (ī = 75%, range = 0.558-0.963 in S. isoetiformis; ī = 89%, range = 0.776-0.963 in S. teres) and among species (ī = 81%, range = 0.506-0.882), leading to the discrimination of four regional population clusters using nonmetric multidimensional scaling (NMDS). It appears that S. isoetiformis and S. teres are a progenitor-derivative species pair.     

Subandrodioecy and male fitness in Sagittaria lancifolia subsp. lancifolia (Alismataceae)

Sagittaria lancifolia subsp. lancifolia is described as cosexual (monoecious), but the study population consisted of 84% cosexuals that typically had 35% pistillate buds and 16% predominant males that typically had 0-2% pistillate buds. Hand-pollinations showed that pistillate flowers required pollination to set seed, and pollen from both male and cosexual plants was potent. No gender switching was seen in the field or greenhouse. From 24 experimental crosses, 890 offspring were grown to maturity. Among these, all offspring of cosexual sires were cosexual, but approximately half the offspring of male sires were male, implying that maleness was inherited as a single, dominant allele. These results indicate that S. lancifolia is subandrodioecious, a very rare breeding system. It is rare, in part because its maintenance requires a large male-fitness differential between male and cosexual plants. In the study population, this condition was met by the differential survival of staminate buds on male racemes. Larvae of the weevil Listronotus appendiculatus killed many staminate buds. They did so in a vertical gradient, with buds lower on racemes safer. Male plants have replaced pistillate with staminate buds at these safer positions and thereby enjoy disproportionally higher male fitness.     

Aerenchyma develops by cell lysis in roots AND CELL SEPARATION IN LEAF PETIOLES IN Sagittaria lancifolia (Alismataceae)

Aerenchyma gas spaces are important for plants that survive flooding because these spaces provide an internal pathway for oxygen transport to the root zone. The objective of this study was to characterize the development of aerenchyma gas spaces in Sagittaria lancifolia L., a dominant species in freshwater wetlands adjacent to the Gulf of Mexico. Tissue at different developmental stages was collected from hydroponically grown plants, embedded in plastic, and sections were observed with a light microscope. In S. lancifolia roots, lysigeny (cell lysis) produced gas spaces that increased in volume from the root meristem to the most mature root tissue. Shoot aerenchyma occurred in the large petioles of S. lancifolia and through the blade midrib, but not in the laminar portion of the blade. In contrast to the roots, gas spaces in the petiole were formed by schizogeny (cell separation during development). Shoot initials produced cells that formed interlocking cylinders in the cortex and diaphragm cells that bridged the central portion of the cylinders. Division and expansion of both these cell types increased the diameter of the cylinders and created schizogenous gaps between diaphragm layers that produced large gas spaces in mature tissue. Therefore, aerenchyma development occurs by two different processes in S. lancifolia.     

Isolation and characterization of polymorphic microsatellite markers from European pear (Pyrus communis L.)

This study reports the development and characterization of 19 microsatellite primer pairs developed from genomic DNA of European pear (Pyrus communis) and their transferability to other Pyrus and Malus material. The primers were designed from two different genomic libraries enriched for di‐ and trinucleotide repeats. When tested in six P. communis cultivars and 15 other Pyrus species, 13 primers revealed single‐locus polymorphism and six showed more complex patterns that suggest multiple loci. Two to 18 alleles were detected per locus and two primer pairs were sufficient to discriminate all accessions. Transferability of nine primer pairs to Malus was demonstrated through amplification of discrete products in two accessions.     

Isolation and characterization of 10 microsatellite loci from Iris hexagona (Iridaceae)

Ten microsatellite loci isolated from Iris hexagona are described. All these loci are polymorphic, with three to 13 alleles across 24 individuals from a single natural population. Heterozygosity ranged from 0.125 to 0.870. Three loci depart significantly from Hardy–Weinberg equilibrium in our test population. The test population shows significant heterozygote deficiency in these and two other loci. Three loci exhibit significant linkage disequilibrium. These loci will be utilized to investigate patterns of genetic variation in the species throughout the Florida peninsula.     

Isolation and characterization of eight microsatellite loci from Phaedranassa tunguraguae (Amaryllidaceae)

Phaedranassa tunguraguae is an endangered species endemic to Ecuador. Eight highly polymorphic microsatellite loci were isolated from an enriched genomic library for this species. Levels of polymorphism were evaluated using a total of 31 individuals from a single natural population. An average of 14.1 alleles per locus was detected, and observed heterozygosity ranged from 0.387 to 0.903. All but one locus depart significantly from Hardy–Weinberg equilibrium. These loci are the first microsatellite primers isolated for Amaryllidaceae and will be utilized to investigate patterns of genetic variation of P. tunguraguae, which will contribute data relevant to the conservation of the species.     

Isolation and characterization of microsatellite markers in an endemic Hawaiian planthopper (Nesosydne chambersi: Delphacidae)

We have isolated and characterized 17 microsatellite loci for the endemic Hawaiian planthopper Nesosydne chambersi (Delphacidae), a member of a large Hawaiian Nesosydne radiation. Thirty individuals from one population and 10 individuals from two populations across the species' range were tested to investigate polymorphism. The observed loci contained two to nine alleles per locus. Expected heterozygosity within this species ranged from 0.2 to 0.85. These markers will be used to assess intraspecific differentiation and population structure within N. chambersi.     

Isolation and characterization of polymorphic microsatellite markers from Primula nutans (Primulaceae)

Seven polymorphic microsatellite loci were developed for a perennial seashore plant, Primula nutans. Degenerate oligonucleotide‐primed (DOP)–polymerase chain reaction (PCR)‐amplified DNA was ligated to TOPO TA vector and screened with radioactively labelled dinucleotide repeat probes. A sample of 378 individuals from Finland, Norway and Russia were used to characterize those loci, which exhibited two to four alleles per locus with observed heterozygosity of 0.003–0.229 and expected heterozygosity of 0.016–0.527. No linkage disequilibrium was found between these seven loci. These are the first microsatellite markers reported for P. nutans.     

慈姑属植物的生态及进化生物学研究进展

慈姑属(Sagittaria L.)隶属于泽泻科,是世界广布的水生植物,其生境多样、叶形和繁殖表型复杂,进化地位较特殊,是生态和进化生物学研究较典型的材料。笔者在查阅相关慈姑属研究的大量文献的基础上,对前人研究的物种、探讨的问题及研究结果进行了归纳与总结:慈姑属物种存在广布种和濒危种,且个别物种的濒危现状可能与其生境条件相关;慈姑属的个别种类成为稻田入侵杂草,其竞争能力因慈姑种类及水稻栽培品种的不同而各异;环境对慈姑属植物有很强的塑造作用,不同环境还会造成慈姑所在的水生植物群落结构发生变化;慈姑属植物具有较高的遗传多样性;在繁殖方面,慈姑属植物体现出不同繁殖方式和两性功能的权衡,以及在繁育系统上从雌雄同株到雌雄异株的进化途径。本文还提出了相关研究存在的不足、研究中应注意的科学问题,并对慈姑属植物未来的研究方向提供了新思路。     

矮慈姑的传粉机制与交配系统

对矮慈姑（ＳａｇｉｔａｒｉａｐｙｇｍａｅａＭｉｑ）的传粉过程与花粉流作了观察,用同工酶遗传标记法对其一个自然居群的异交率作出定量估计。在自然及人工居群中均观察到虫媒传粉,其中有较大比例的近距离传粉。未检出风媒花粉流。自交可育,同时存在雌雄同株和花序内雌雄花异熟等异交机制。异交率估计结果（ｔ＝４９９％）表明其交配系统为异交／自交兼性系统。作者认为,其较大比例的自交应归因于同株之不同克隆分株间交配的频繁发生