Male gametophytic selection in maize

Summary There is evidence that male gametophyte selection is a widespread phenomenon in higher plants. The pollen tube growth rate is one of the main components of gametophyte selective value; genetic variability for this trait, due to the effect of single genes or to quantitative variation, has been described in maize. However, indication of gametophytic selection has been indirectly obtained; its effect was revealed by the positive relation observed between gametophyte competitive ability and sporophyte metrical traits.This paper considers the results of selection applied to gametophyte populations produced from single plants. The competitive ability of the lines was evaluated in comparison with that of a standard line by means of the pollen mixture technique. Sporophytic traits were measured in the hybrid progeny obtained by crossing selected S₃ and S₄ families with an unrelated single cross and an inbred line. Gametophyte selection produced inbred lines with high gametophyte competitive ability. In view of the selection procedure adopted, this result was interpreted as an indication of haploid expression of genes involved in the control of pollen tube growth. Moreover, this gametophytic trait was positively correlated with sporophytic traits (seedling weight, kernel weight and root tip growth in vitro), indicating that both groups of characters have a common genetic basis.     

Gene expression during the male gametophytic phase

Abstract

In recent years a number of experimental findings have indicated that in higher plants the gametophytic phase is able to express its own genetic information, a large part of which it shares with the sporophytic generation. Quantitative estimates of haploid and haplodiploid gene expression have been obtained by mRNA and isozyme analysis in several plant species: 60-70% of the genes are expressed in both pollen and plant, about 10% are pollen-specific, and 20% represent the sporophytic domain. Moreover, it has been demonstrated that stage-specific genes are expressed in the gametophytic generation: at least two sets of genes are activated during pollen development, others are expressed only in the postshedding period, during germination and tube growth. Studies have been made to ascertain the role played by gametophyte-expressed genes in pollen development; the in vivo and in vitro pollen tube growth rate has been revealed to be controlled by the gametophyte genome itself. Differential effects of specific chromosomal deficiencies on the development of maize pollen grains have indicated that components of normal microspore development are controlled by genes located in specific parts of the genome. For single gene analysis, gene transfer can be used; on the contrary, for traits with a multifactorial genetic control, direct proof of gene expression both in the gametophytic and the sporophytic generation can be obtained when selection is applied to the pollen population of a hybrid plant, and response to selection is observed in the resulting sporophytic progeny. Response to selection, applied at different stages of the gametophytic phase, has been described in the sporophytic progeny and this with regard to many adaptive traits; thus the phenomenon can have an important bearing on the genetic structure of natural populations and on higher plant evolution, it can also be used as a breeding tool to increase the efficiency of conventional selection methods.     

GENETIC VARIABILITY OF GELIDIUM CANARIENSIS (RHODOPHYTA) DETERMINED BY ISOZYME ELECTROPHORESIS1

The populations of Gelidium canariensis (Grunow) Seoane-Camba from the Canary Islands were analyzed for genetic variability by isozyme electrophoresis in 1989 and 1990. Each population was divided into sporophytic and gametophytic subpopulations. Twenty-three to 27 putative alleles corresponding to 22 gene loci were analyzed. Sev-enteen loci were monomorphic in all six subpopulations, and five were polymorphic in at least one subpopulation. Significant deviations from Hardy-Weinberg equilibrium were found. The amount of genetic variability (percentage of polymorphic loci, mean number of alleles per locus, and average gene diversity) of haploid subpopulations was lower than that of diploid subpopulations. No correlation between genetic distance and geographical distance was found. Low genetic differentiation between sporophytic and gametophytic subpopulations of the same locality was obsewed in two populations. The low genetic diversity and genetic differentiation suggest that the genetic structure of the populations of G. canariensis from the Canary Islands is due to a combination of founder effects and the predominance of asexual reproduction. Initial differences in gene frequencies may have persisted because of insufficient time to reach a higher level of differentiation.     

POLLINATION DYNAMICS IN EPILOBIUM CANUM (ONAGRACEAE): CONSEQUENCES FOR GAMETOPHYTIC SELECTION

Several hand-pollination experiments have shown that pollen tube competition leads to gametophytic selection, and thus affects genotypic and phenotypic characteristics of the next generation. This study is one of the first to quantify natural levels of pollen tube competition. In a population of Epilobium canum, I measured both the amount of pollen deposited on stigmas and the timing of deposition. Approximately 20 tetrads were required for full seed set within fruits. Hummingbirds deposited >20 tetrads at 50–70% of the flowers examined, often in a single load. When pollen arrived in 2 loads, a portion of the ovules within each ovary was probably sired by competing pollen from the second load. Competition may be relatively weak unless at least 30 tetrads compete for ovules. About 20% of the flowers received >30 tetrads in the first load, and 13% acquired > 30 tetrads in 2 loads. The frequency and intensity of pollen tube competition varied among plants. In some styles, 80% of the pollen tubes were excluded from access to ovules, but in others no competition occurred. Further studies of pollination rates and progeny fitness are needed before we can assess the role of pollen tube competition in natural populations. Potential effects of gametophytic selection are discussed.     

Isolation and Early in Vitro Development of Young Pollen Protoplasts in Nicotiana tabacum

For isolating young pollen protoplasts in Nicotiana tabacum. The authors had established two efficient enzymatic methods via anther preculture or pollen starvation pretreatment. Procedure of the first method included the following steps: 1. Cold pretreatment of flower buds with pollen at late unicellular to early bicellular stage; 2. Anther floating culture for pollen shedding into the culture medium followed by dehiscence of exine; 3. Enzymatic maceration of exine-dehisced pollen resulting in degradation of intine and release of pollen protoplasts in large quantity. Procedure of the second method involved the following steps: 1. Culture of pollen at middle bicellular in Kyo and Harada' B medium for starvation: 2. Enzymatic maceration of starvated pollen resulting in release of pollen protoplasts and subprotoplasts. Factors affecting the results of both methods as well as early in vitro developmental events of young pollen protoplasts were studied. The protoplasts could be induced either to trigger the first sporophytic division or to continue the gametophytic pathway leading germinatation of pollen tubes !ndicating their potentiality of inducing both sporophytic and gametophytic development of pathway. In rare instance a quite interesting phenomenon was observed that a pollen protoplast first divided into two daughter cells and one of which then germinated a pollen tube. It may insinuate that such pollen protoplasts initially induced a sporophytic pathway could reverse induce a gametophytic pathway.     

花粉管生长和极性引导的孢子体和配子体控制研究进展

双受精是被子植物特有的生殖方式,精细胞只有通过花粉管穿过花柱才能到达子房、胚珠受精。花粉管在母本组织中的生长和引导包括孢子体控制(sporophytic control)和配子体控制(gametophytic control)两个连续的过程,现已克隆出不同阶段花粉管生长和引导的基因,通过分析其表达调控揭示出花粉管生长和引导的分子机制。该文就近年来国内外有关花粉管生长和极性引导的调控机制研究进展进行综述,并对禾本科(Poaceae)和十字花科(Brassicaceae)植物花粉管引导的异同点进行了比较分析。     

Sporophytic response to pollen selection for Alachlor tolerance in maize

In order to assess the efficiency of male gametophytic selection (MGS) for crop improvement, pollen selection for tolerance to herbicide was applied in maize. The experiment was designed to test the parallel reactivity to Alachlor of pollen and plants grown in controlled conditions or in the field, the response to pollen selection in the sporophytic progeny, the response to a second cycle of MGS, and the transmission of the selected trait to the following generations. The results demonstrated that pollen assay can be used to predict Alachlor tolerance under field conditions and to monitor the response to selection. A positive response to selection applied to pollen in the sporophytic progeny was obtained in diverse genetic backgrounds, indicating that the technique can be generally included in standard breeding programs; the analysis of the data produced in a second selection cycle indicated that the selected trait is maintained in the next generation.     

THE GENETIC STRUCTURE OF SPOROPHYTIC AND GAMETOPHYTIC POPULATIONS OF THE MOSS,FUNARIA HYGROMETRICA HEDW

Patterns of phenotypic and genotypic variability in two populations of the moss, Funaria hygrometrica, were investigated using measurements of gametophytic and sporophytic morphology, sporophytic reproductive output, spore germination, gametophytic growth rates and tolerances of copper, cadmium, and low nutrient conditions, and electrophoretically detectable enzyme variation. The two populations differed in all traits measured, but complete monomorphism within populations at 14 enzyme loci suggested that each represented a single clone. Variability in gametophytic growth rates and responses to different experimental media, however, occurred among haploid sib families (families of meiotic progeny derived from the same sporophyte) and among sibs within families within both populations, suggesting high levels of genetic variability. Low mean reproductive output and a high level of variability among sporophytes in a mine site population probably reflected heavy metal toxicity. Based on this study, in combination with previous work on F. hygrometrica (Shaw, 19906), somatic mutation and/or nongenetic effects appear to contribute significantly to phenotypic variability in natural populations.     

花粉管引导机制的研究进展

花粉管引导是指显花植物在受精过程中,雌蕊组织与花粉管相互作用使花粉管定向生长并最终到达胚囊的过程,其机制颇为复杂。该文基于调控花粉管生长的孢子体引导和配子体细胞引导两个主要过程,阐述雌蕊中不同蛋白分子和其它小分子物质的浓度梯度在花粉管的孢子体组织引导中的作用,以及胚囊中不同类型的细胞及其相关基因与蛋白在花粉管的配子体细胞引导中的作用。同时,该文也对精细胞在花粉管引导中的作用进行了阐述。     

Variation in sporophytic and gametophytic vigor in wild and cultivated varieties of Cucurbita pepo and their F1 and F2 generations

 This study examines sporophytic and gametophytic vigor in wild and cultivated varieties of Cucurbita pepo L. and their hybrids in order to determine whether hybrid vigor extends to the microgametophyte generation. It also examines the variation in sporophytic and gametophytic vigor to discern the non-genetic influences of pollen provisioning by the sporophyte on pollen performance from the genetic influences of the microgametophyte’s own genotype on pollen performance. A cultivated and a wild C. pepo and their F1 and the F2 generations were grown under field conditions and flower and fruit production were monitored over one summer. We found that the four types of plants differed significantly in the number of male and female flowers and the number of fruits they produced. The F1 plants produced significantly more male flowers and marginally more female flowers and fruits than the parental lines. To estimate gametophytic vigor pollen was germinated in vitro and pollen tube length measured after 30 min. We found that pollen tubes from the F1 plants had significantly greater growth than tubes from the parental lines or the F2 generation, indicating that hybrid vigor extends to the microgametophytic generation. By partitioning the variance of pollen tube growth into ’within’ and ’among’ plant components of variation, we were able to show that the genotype of the microgametophyte influences pollen performance in vitro, but that expression of hybrid vigor in the microgametophyte is likely to be due to an environmental effect related to provisioning of the pollen grains during development. Received: 16 April 1998 / Revision accepted: 18 August 1998     

A new look at incompatibility relationships in higher plants

Summary Pollen tube growth was evaluated using an 18-step scale after both intra- and interspecific pollinations of genotypically widely differing diploid potato species and potato dihaploids expressing monofactorial gametophytic incompatibility. The results obtained account for a wide array of types of pollen tube growth resulting from crossing partners with distinct incompatibility behavior. Based on the assumption that inhibition of pollen tubes is the rule and solely prevented by the pollen itself, a model proposing one common cause underlying different mechanisms for both intraspecific selfincompatibility and interspecific incompatibility in diploids is put forward. Data supporting the model are presented from the experimental results of this study and from the literature. The strength of pollen-style interaction depends on particular S-alleles in combination with recognition and activity properties of the basic S-genotypes. The model is suitable to explain all former observations on incompatibility in diploids with a gametophytic system of incompatibility and, with modifications of the manner of phenotypic expression, also in plants with sporophytic incompatibility. The proposed scheme of pollen tube growth phenotypes permits prediction of pollen tube growth behavior in an intended cross combination. The model is based on both classical Mendelian genetics and recent molecular genetic insight.     

Quantitatively determined self-incompatibility

Summary It is shown by simulation that a hypothetical multilocus, quantitatively determined self-incompatibility system, whether gametophytic or sporophytic, should maintain variability in small populations at a higher level than would panmixia. Studies of more than 20 isozyme loci show that borage has almost no variability.     

Pollen selection — past,present and future

 A series of studies, recently reviewed, has established that approximately 60% of the structural genes which are expressed in the sporophytic portion of the angiosperm life cycle are also expressed and exposed to selection in the pollen. Given the haploidy and large population sizes of pollen grains, a substantial portion of the sporophytic genome could thus be periodically exposed to a bacterial type of mass screening. This extraordinary possibility is often subject to some skepticism which may, of course, be justified. However, recent attempts to apply models appear to be inappropriate in this context, in part because these attempts overlook an important source of genetic variation, and also because they assume fixed values for selection and fitness. More recently, studies of pollen/pollen interactions have suggested that what Linskens termed the ”programic phase” may represent an arena for important, and largely unexplored phenomena, some of which are discussed here. Received: 3 June 1996 / Revision accepted: 26 July 1996     

Evolutionary Dynamics of Sporophytic Self-Incompatibility Alleles in Plants

The stationary frequency distribution and allelic dynamics in finite populations are analyzed through stochastic simulations in three models of single-locus, multi-allelic sporophytic self-incompatibility. The models differ in the dominance relationships among alleles. In one model, alleles act codominantly in both pollen and style (SSIcod), in the second, alleles form a dominance hierarchy in pollen and style (SSIdom). In the third model, alleles interact codominantly in the style and form a dominance hierarchy in the pollen (SSIdomcod). The SSIcod model behaves similarly to the model of gametophytic self-incompatibility, but the selection intensity is stronger. With dominance, dominant alleles invade the population more easily than recessive alleles and have a lower frequency at equilibrium. In the SSIdom model, recessive alleles have both a higher allele frequency and higher expected life span. In the SSIdomcod model, however, loss due to drift occurs more easily for pollen-recessive than for pollen-dominant alleles, and therefore, dominant alleles have a higher expected life span than the more recessive alleles. The process of allelic turnover in the SSIdomcod and SSIdom models is closely approximated by a random walk on a dominance ladder. Implications of the results for experimental studies of sporophytic self-incompatibility in natural populations are discussed.     

Pollen-tube growth rates in Collinsia heterophylla (Plantaginaceae): one-donor crosses reveal heritability but no effect on sporophytic-offspring fitness

Background and Aims

Evolutionary change in response to natural selection will occur only if a trait confers a selective advantage and there is heritable variation. Positive connections between pollen traits and fitness have been found, but few studies of heritability have been conducted, and they have yielded conflicting results. To understand better the evolutionary significance of pollen competition and its potential role in sexual selection, the heritability of pollen tube-growth rate and the relationship between this trait and sporophytic offspring fitness were investigated in Collinsia heterophylla.

Methods

Because the question being asked was if female function benefited from obtaining genetically superior fathers by enhancing pollen competition, one-donor (per flower) crosses were used in order to exclude confounding effects of post-fertilization competition/allocation caused by multiple paternity. Each recipient plant was crossed with an average of five pollen donors. Pollen-tube growth rate and sporophytic traits were measured in both generations.

Key Results

Pollen-tube growth rate in vitro differed among donors, and the differences were correlated with in vivo growth rate averaged over two to four maternal plants. Pollen-tube growth rate showed significant narrow-sense heritability and evolvability in a father–offspring regression. However, this pollen trait did not correlate significantly with sporophytic-offspring fitness.

Conclusions

These results suggest that pollen-tube growth rate can respond to selection via male function. The data presented here do not provide any support for the hypothesis that intense pollen competition enhances maternal plant fitness through increased paternity by higher-quality sporophytic fathers, although this advantage cannot be ruled out. These data are, however, consistent with the hypothesis that pollen competition is itself selectively advantageous, through both male and female function, by reducing the genetic load among successful gametophytic fathers (pollen), and reducing inbreeding depression associated with self–pollination in plants with mix-mating systems.Key words: Collinsia heterophylla, evolvability, female fitness, good genes, heritability, male fitness, mixed-mating system, Plantaginaceae, pollen competition, sexual selection     

Intra- and extracellular lipid composition and associated gene expression patterns during pollen development in Brassica napus

Pollen development in angiosperms is regulated by the interaction of products contributed by both the gametophytic (haploid) and sporophytic (diploid) genomes. In entomophilous species, lipids are major products of both sporophytic and gametophytic metabolism during pollen development. Mature pollen grains of Brassica napus are shown to contain three major acyl lipid pools as follows: (i) the extracellular tryphine mainly consisting of medium-chain neutral esters; (ii) the intracellular membranes, particularly endoplasmic reticulum, mainly containing phospholipids; and (iii) the intracellular storage lipids, which are mostly triacylglycerols. This paper reports on the kinetics of accumulation of these lipid classes during pollen maturation and the expression patterns of several lipid biosynthetic genes and their protein products that are differentially regulated in developing microspores/ pollen grains (gametophyte) and tapetal cells (sporophyte) of B. napus. Detailed analysis of three members of the stearoyl-ACP desaturase (sad) gene family by Northern blotting, in situ hybridization and RT-PCR showed that the same individual genes were expressed both in gametophytic and sporophytic tissues, although under different temporal regulation. In the tapetum, maximal expression of two marker genes for lipid biosynthesis (sad and ear) occurred at a bud length of 2–3 mm, and the corresponding gene products SAD and EAR were detected by Western blotting in 3–4 mm buds, coinciding with the maximal rates of tapetal lipid accumulation. These lipids are released following tapetal cell disintegration and are relocated to form the major structural component of the extracellular tryphine layer that coats the mature pollen grain. In contrast, in developing microspores/pollen grains, maximal expression of the lipid marker genes sad, ear, acp and cyb5 was at the 3–5 mm bud stages, with the SAD and EAR gene products detected in 4–7 mm buds. This pattern of expression coincided with accumulation of the intracellular storage and membrane lipid components of pollen. These results suggest that, although the same genes may be expressed in the sporophytic tapetal cells and in gametophytic tissues, they are regulated differentially leading to the production of the various contrasting lipidic structures that are assembled together to give rise to a viable, fertile pollen grain.     

Overlap of gametophytic and sporophytic gene expression in barley

Summary The overlap of gametophytic and sporophytic gene expression in barley was studied by means of enzyme electrophoresis. Of the isozymes found, 60% were expressed in both gametophyte and sporophyte, 30% were sporophyte specific, and 10% were gametophyte specific. A considerable amount of the barley genome is thus potentially amenable to gametophytic selection. The estimated sizes of the common, sporophytic and gametophytic domains in barley gene expression correspond with the estimates obtained in other plant species.     

The joint evolution of mating system and pollen performance: Predictions regarding male gametophytic evolution in selfers vs. outcrossers

Studies of sexual selection in plants historically have focused on pollinator attraction, pollen transfer, gametophytic competition, and post-fertilization discrimination by maternal plants. Pollen performance (the speeds of germination and pollen tube growth) in particular is thought to be strongly subject to intrasexual selection, but the effect of mating system on this process has not been rigorously evaluated. Here we propose four predictions derived from the logic that pollen performance should evolve with mating system as an adaptive response to: (1) the competitive environment among pollen genotypes and (2) variation among female genotypes regularly encountered by a given pollen genotype. First, as previously predicted, due to the higher potential for intense selection among diverse pollen genotypes in outcrossing relative to selfing taxa, pollen should evolve to germinate and/or to grow more rapidly in outcrossers than in selfers. Second, due to stronger selection on pollen performance in outcrossing than in selfing taxa, heritable variation in pollen tube growth rate is more likely to be purged in outcrossers. In selfers, by contrast, genetic variation in pollen tube growth rates may readily accumulate because selfing reduces the number of genetically distinct male gametophytes likely to be deposited on any given stigma, thereby relaxing selection on male gametophytic traits. A summary of published studies presented here provides preliminary support for this prediction. Third, due to the high probability that the pollen of outcrossing individuals will be exposed to multiple pistil genotypes, we predict that the pollen of habitually outcrossing taxa will evolve to perform more consistently across female genotypes than the pollen of selfing taxa. Fourth, we predict that epistatic interactions between pollen and pistil genotypes are more likely to evolve in selfers than in outcrossers. We suggest several empirical approaches that may be used to test these predictions.