Unreduced gamete formation in wheat × <Emphasis Type="Italic">Aegilops</Emphasis> spp. hybrids is genotype specific and prevented by shared homologous subgenomes

Key message

The presence of homologous subgenomes inhibited unreduced gamete formation in wheat × Aegilops interspecific hybrids. Unreduced gamete rates were under the control of the wheat nuclear genome.

Abstract

Production of unreduced gametes is common among interspecific hybrids, and may be affected by parental genotypes and genomic similarity. In the present study, five cultivars of Triticum aestivum and two tetraploid Aegilops species (i.e. Ae. triuncialis and Ae. cylindrica) were reciprocally crossed to produce 20 interspecific hybrid combinations. These hybrids comprised two different types: T. aestivum × Aegilops triuncialis; 2n = ABDU^tC^t (which lack a common subgenome) and T. aestivum × Ae. cylindrica; 2n = ABDD^cC^c (which share a common subgenome). The frequency of unreduced gametes in F₁ hybrids was estimated in sporads from the frequency of dyads, and the frequency of viable pollen, germinated pollen and seed set were recorded. Different meiotic abnormalities recorded in the hybrids included precocious chromosome migration to the poles at metaphase I and II, laggards in anaphase I and II, micronuclei and chromosome stickiness, failure in cell wall formation, premature cytokinesis and microspore fusion. The mean frequency of restitution meiosis was 10.1 %, and the mean frequency of unreduced viable pollen was 4.84 % in T. aestivum × Ae. triuncialis hybrids. By contrast, in T. aestivum × Ae. cylindrica hybrids no meiotic restitution was observed, and a low rate of viable gametes (0.3 %) was recorded. This study present evidence that high levels of homologous pairing between the D and D^c subgenomes may interfere with meiotic restitution and the formation of unreduced gametes. Variation in unreduced gamete production was also observed between T. aestivum × Ae. triuncialis hybrid plants, suggesting genetic control of this trait.

     

Meiotic chromosome pairing behaviour of natural tetraploids and induced autotetraploids of Actinidia chinensis

Key message

Non-preferential chromosome pairing was identified in tetraploid Actinidia chinensis and a higher mean multivalent frequency in pollen mother cells was found in colchine-induced tetraploids of A. chinensis compared with naturally occurring tetraploids.

Abstract

Diploid and tetraploid Actinidia chinensis are used for the development of kiwifruit cultivars. Diploid germplasm can be exploited in a tetraploid breeding programme via unreduced (2n) gametes and chemical-induced chromosome doubling of diploid cultivars and selections. Meiotic chromosome behaviour in diploid A. chinensis ‘Hort16A’ and colchicine-induced tetraploids from ‘Hort16A’ was analysed and compared with that in a diploid male and tetraploid males of A. chinensis raised from seeds sourced from the wild in China. Both naturally occurring and induced tetraploids formed multivalents, but colchicine-induced tetraploids showed a higher mean multivalent frequency in the pollen mother cells. Lagging chromosomes at anaphase I and II were observed at low frequencies in the colchicine-induced tetraploids. To investigate whether preferential or non-preferential chromosome pairing occurs in tetraploid A. chinensis, the inheritance of microsatellite alleles was analysed in the tetraploid progeny of crosses between A. chinensis (4x) and A. arguta (4x). The frequencies of inherited microsatellite allelic combinations in the hybrids suggested that non-preferential chromosome pairing had occurred in the tetraploid A. chinensis parent.     

Pathways to polyploidy: indications of a female triploid bridge in the alpine species <Emphasis Type="Italic">Ranunculus kuepferi</Emphasis> (Ranunculaceae)

Polyploidy is one of the most important evolutionary processes in plants. In natural populations, polyploids usually emerge from unreduced gametes which either fuse with reduced ones, resulting in triploid offspring (triploid bridge), or with other unreduced gametes, resulting in tetraploid embryos. The frequencies of these two pathways, and male versus female gamete contributions, however, are largely unexplored. Ranunculus kuepferi occurs with diploid, triploid and autotetraploid cytotypes in the Alps, whereby diploids are mostly sexual, while tetraploids are facultative apomicts. To test for the occurrence of polyploidization events by triploid bridge, we investigated 551 plants of natural populations via flow cytometric seed screening. We assessed ploidy shifts in the embryo to reconstruct female versus male gamete contributions to polyploid embryo and/or endosperm formation. Seed formation via unreduced egg cells (B_III hybrids) occurred in all three cytotypes, while only in one case both gametes were unreduced. Polyploids further formed seeds with reduced, unfertilized egg cells (polyhaploids and aneuploids). Pollen was highly variable in diameter, but only pollen >27 μm was viable, whereby diploids produced higher proportions of well-developed pollen. Pollen size was not informative for the formation of unreduced pollen. These results suggest that a female triploid bridge via unreduced egg cells is the major pathway toward polyploidization in R. kuepferi, maybe as a consequence of constraints of endosperm development. Triploids resulting from unreduced male gametes were not observed, which explains the lack of obligate sexual tetraploid individuals and populations. Unreduced egg cell formation in diploids represents the first step toward apomixis.     

Micronucleation by mitosis inhibitors in developing microspores of Spathiphyllum wallisii Regel

Key message

We developed an efficient protocol for chromosome scattering in Spathiphyllum microspores. The effects of plant material, developmental age, genotype and antimicrotubular toxin type, exposure and concentration were evaluated.

Abstract

Asymmetric hybridization through microprotoplast-mediated chromosome transfer (MMCT) is a known method for overcoming sexual breeding barriers between distantly related plant species. To obtain microprotoplasts, it is necessary to induce mass micronucleation either in somatic or gametic cells. We have tested the efficiency for micronuclei induction of five mitosis inhibitors, amiprophos-methyl (APM), butamiphos (BUT), chlorpropham (CIPC), oryzalin (ORY) and propyzamide (PRO), on developing microspores of diploid Spathiphyllum wallisii Regel. Besides the used toxins, also the effect of their concentrations and incubation period as well as plant genotypes and material was tested. We observed micronuclei (MNi) in pollen mother cells, dyads and tetrads as well as other abnormalities such as ball metaphases and chromosome bridges. The flower position on the spadix and the type of starting material (dissected anthers vs. complete spadices) did not significantly influence micronucleation frequencies. The highest micronucleation index of 86 % was obtained in microspores treated with 10 μM ORY during 72 h. All six genotypes tested formed micronuclei after this particular treatment, although the efficiency varied between cultivars. Next to ORY, CIPC was also a very efficient MNi inducer. The average number of MNi found in micronucleated cells varied between 1.67–6.44 for CIPC and 0.83–5.50 for ORY. The maximal number of MNi observed was 12 for CIPC and 9 for ORY. Our results demonstrate that CIPC and ORY can be applied for mass micronucleation on developing microspores of S. wallisii as a first step of MMCT in aroid interspecific or intergeneric breeding.     

Inheritance in doubled-diploid clementine and comparative study with SDR unreduced gametes of diploid clementine

Key message

Tetraploid clementine displays mainly tetrasomic inheritance. Genetic structures of 2n SDR and 2 × gametes from DD clementine are complementary and will guides triploids citrus breeding strategies.

Abstract

Triploid breeding is developed worldwide to create new seedless cultivars. Citrus triploid hybrids can be recovered from 2x × 2x sexual hybridizations as a consequence of the formation of unreduced gametes (2n), or from 4x × 2x interploid hybridizations in which tetraploid parents used are most often doubled-diploid (DD). Here we have analyzed the inheritance in doubled-diploid clementine and compared the genetic structures of gametes of DD clementine with SDR unreduced gametes of diploid clementine. Parental heterozygosity restitution (PHR) with DD parents depends on the rate of preferential chromosome pairing and thus the proportion of disomic versus tetrasomic segregations. Doubled-diploid clementine largely exhibited tetrasomic segregation. However, three linkage groups had intermediate segregation and one had a tendency for disomy. Significant doubled reduction rates (DR) rates were observed in six of the nine LGs. Differences of PHR between 2n SDR and 2x DD gametes were highest in the centromeric region and progressively decreased toward the distal regions where they were not significant. Over all markers, PHR was lower (two-thirds) in SDR 2n gametes than in DD-derived diploid gametes. The two strategies appear complementary in terms of genotypic variability. Interploid 4x × 2x hybridization is potentially more efficient for developing new cultivars that are phenotypically closer to the diploid parent of the DD than sexual hybridization through SDR 2n gametes. Conversely, 2x × 2x triploidisation has the potential to produce novel products with characteristics for market segmentation strategies.

     

Microsatellite and flow cytometry analysis to help understand the origin of Dioscorea alata polyploids

Background and Aims Dioscorea alata

is a polyploid species with a ploidy level ranging from diploid (2n = 2x = 40) to tetraploid (2n = 4x = 80). Ploidy increase is correlated with better agronomic performance. The lack of knowledge about the origin of D. alata spontaneous polyploids (triploids and tetraploids) limits the efficiency of polyploid breeding. The objective of the present study was to use flow cytometry and microsatellite markers to understand the origin of D. alata polyploids.

Methods

Different progeny generated by intracytotype crosses (2x × 2x) and intercytotype crosses (2x × 4x and 3x × 2x) were analysed in order to understand endosperm incompatibility phenomena and gamete origins via the heterozygosity rate transmitted to progeny.

Results

This work shows that in a 2x × 2x cross, triploids with viable seeds are obtained only via a phenomenon of diploid female non-gametic reduction. The study of the transmission of heterozygosity made it possible to exclude polyspermy and polyembryony as the mechanisms at the origin of triploids. The fact that no seedlings were obtained by a 3x × 2x cross made it possible to confirm the sterility of triploid females. Flow cytometry analyses carried out on the endosperm of seeds resulting from 2x × 4x crosses revealed endosperm incompatibility phenomena.

Conclusions

The major conclusion is that the polyploids of D. alata would have appeared through the formation of unreduced gametes. The triploid pool would have been built and diversified through the formation of 2n gametes in diploid females as the result of the non-viability of seeds resulting from the formation of 2n sperm and of the non-viability of intercytotype crosses. The tetraploids would have appeared through bilateral sexual polyploidization via the union of two unreduced gametes due to the sterility of triploids.     

Chromosomal structural changes and microsatellite variations in newly synthesized hexaploid wheat mediated by unreduced gametes

Allohexaploid wheat was derived from interspecific hybridization, followed by spontaneous chromosome doubling. Newly synthesized hexaploid wheat by crossing Triticum turgidum and Aegilops tauschii provides a classical model to understand the mechanisms of allohexaploidization in wheat. However, immediate chromosome level variation and microsatellite level variation of newly synthesized hexaploid wheat have been rarely reported. Here, unreduced gametes were applied to develop synthesized hexaploid wheat, NA0928, population by crossing T. turgidum ssp. dicoccum MY3478 and Ae. tauschii SY41, and further S0–S3 generations of NA0928 were assayed by sequential cytological and microsatellite techniques. We demonstrated that plentiful chromosomal structural changes and microsatellite variations emerged in the early generations of newly synthesized hexaploid wheat population NA0928, including aneuploidy with whole-chromosome loss or gain, aneuploidy with telosome formation, chromosome-specific repeated sequence elimination (indicated by fluorescence in situ hybridization) and microsatellite sequence elimination (indicated by sequencing), and many kinds of variations have not been previously reported. Additionally, we reported a new germplasm, T. turgidum accession MY3478 with excellent unreduced gametes trait, and then succeeded to transfer powdery mildew resistance from Ae. tauschii SY41 to synthesized allohexaploid wheat population NA0928, which would be valuable resistance resources for wheat improvement.     

Chromosome segregation in an allotetraploid banana hybrid (AAAB) suggests a translocation between the A and B genomes and results in eBSV-free offsprings

Many banana cultivars (including the Plantain type) are triploid interspecific hybrids between Musa acuminata (A genome) and Musa balbisiana (B genome). M. balbisiana contains endogeneous Banana streak virus sequences (eBSVs) that can, in interspecific genome context, spontaneously release infectious viral genomes. We analyzed, a triploid progeny of 184 individuals from a cross between a tetraploid AAAB breeding accession (CRBP39) and the diploid AA accession (Pahang) with 38 SSR and eBSV-specific PCR markers. The results showed that (1) most of the alleles are found/transmitted in the expected frequency to the progeny with only 10 % biased; (2) 70 % of the loci displayed a tetrasomic allele segregation and (3) interspecific intrachromosomal recombinations occurred for all the chromosome segments surveyed. However, half of the offspring obtained resulted from maternal unbalanced gametes transmission. Analysis of gamete composition and marker association suggested the presence of a large translocation between A and B genome involving chromosome 1 and 3. The two infectious eBSVs present in the maternal parent CRBP39 are located on chromosome 1B and appeared in a higher proportion than expected in the progeny. Interestingly, we showed that both eBSVs were absent from 24 offspring that represent promising material for breeding.     

Focus Issue on the Plant Physiology of Global Change: Production of Diploid Male Gametes in Arabidopsis by Cold-Induced Destabilization of Postmeiotic Radial Microtubule Arrays

Whole-genome duplication through the formation of diploid gametes is a major route for polyploidization, speciation, and diversification in plants. The prevalence of polyploids in adverse climates led us to hypothesize that abiotic stress conditions can induce or stimulate diploid gamete production. In this study, we show that short periods of cold stress induce the production of diploid and polyploid pollen in Arabidopsis (Arabidopsis thaliana). Using a combination of cytological and genetic analyses, we demonstrate that cold stress alters the formation of radial microtubule arrays at telophase II and consequently leads to defects in postmeiotic cytokinesis and cell wall formation. As a result, cold-stressed male meiosis generates triads, dyads, and monads that contain binuclear and polynuclear microspores. Fusion of nuclei in binuclear and polynuclear microspores occurs spontaneously before pollen mitosis I and eventually leads to the formation of diploid and polyploid pollen grains. Using segregation analyses, we also found that the majority of cold-induced dyads and triads are genetically equivalent to a second division restitution and produce diploid gametes that are highly homozygous. In a broader perspective, these findings offer insights into the fundamental mechanisms that regulate male gametogenesis in plants and demonstrate that their sensitivity to environmental stress has evolutionary significance and agronomic relevance in terms of polyploidization.The spontaneous formation of polyploid species through whole-genome duplication is a major force driving diversification and speciation in plant evolution (Wang et al., 2004). The redundant genomic material produced by polyploidization provides genotypic plasticity that facilitates adaptation and confers enhanced competitiveness compared with diploid progenitors (Adams and Wendel, 2005a, 2005b; Leitch and Leitch, 2008). Molecular analyses suggest that the genomes of most angiosperms (more than 90%) retain evidence of one or more ancient genome-wide duplication events (Cui et al., 2006). Moreover, recently, Wood et al. (2009) established that up to 15% of angiosperm and 31% of gymnosperm speciation events were accompanied by polyploidization. Polyploidization in plants is also commercially beneficial. Many important crop species including wheat (Triticum aestivum), potato (Solanum tuberosum), tobacco (Nicotiana tabacum), coffee (Coffea arabica), and numerous fruit varieties are polyploid (Bretagnolle and Thompson, 1995). Although several mechanisms can yield polyploids, it is thought that most polyploid plants are formed by the spontaneous production and fusion of diploid (2n) gametes (Bretagnolle and Thompson, 1995; Ramsey and Schemske, 1998). However, despite the evolutionary and agricultural significance of sexual polyploidization in plants (Ramanna and Jacobsen, 2003), the molecular mechanism underlying 2n gamete formation in natural populations is poorly understood.Several cytological defects lead to diploid gamete formation in both male and female reproductive lineages. In some species, premeiotic and postmeiotic genome doubling events are reported, but diploid gametes typically result from a defect in one of the two meiotic divisions, a phenomenon referred to as “restitution” (Bretagnolle and Thompson, 1995; Ramsey and Schemske, 1998). Meiotic restitution mechanisms are categorized into three classes: (1) omission of one of the meiotic cell divisions; (2) alterations in meiosis I (MI) or meiosis II (MII) spindle morphology; or (3) defects in meiotic cytokinesis (Ramanna and Jacobsen, 2003). Additionally, depending on the genetic makeup of the resulting 2n gametes, meiotic restitution mechanisms can be further subdivided into two classes: first division restitution (FDR) and second division restitution (SDR). In FDR, the sister chromatids disjoin and segregate to opposite poles, yielding 2n gametes that largely retain the heterozygosity of the parental plant. In SDR, sister chromatids do not disjoin in MII and segregate to the same pole, generating highly homozygous 2n gametes (Köhler et al., 2010).Several genes governing 2n gamete formation have been identified and characterized in potato, maize (Zea mays), and Arabidopsis (Arabidopsis thaliana; Consiglio et al., 2004; Brownfield and Köhler, 2011). Mutations in Arabidopsis DYAD/SWITCH1 and maize ARGONAUTE104 (AGO104) and AM1 induce a complete loss of MI and, consequently, convert the meiotic cell cycle into a mitotic one (Ravi et al., 2008; Pawlowski et al., 2009; Singh et al., 2011). Lesions in Arabidopsis OSD1/GIG1 and TAM/CYCA2;1, two proteins involved in progression of the meiotic cell cycle, cause a complete loss of MII, generating highly homozygous 2n gametes in both male and female meiosis (d’Erfurth et al., 2009, 2010). Spindle-based meiotic restitution mechanisms have been reported in both Arabidopsis jason and atps1 mutants and in the potato ps mutant, in which parallel, fused, and tripolar spindles in male MII lead to the formation of FDR 2n spores (Mok and Peloquin, 1975; d’Erfurth et al., 2008; De Storme and Geelen, 2011). Disruption of postmeiotic male cytokinesis, which is regulated by a mitogen-activated protein kinase (MAPK) kinase signaling pathway, also results in polyploid gametes. Mutations in TES/STUD/AtNACK2, MKK6/ANQ1, and MPK4, three main components of the cytokinetic MAPK signaling cascade, induce a complete loss of cytokinesis following male meiosis, generating fully restituted tetraploid pollen grains (Hulskamp et al., 1997; Spielman et al., 1997; Soyano et al., 2003; Zeng et al., 2011).Despite progress on understanding cytological mechanisms and genetic factors governing the formation of 2n gametes in natural populations, less is known about the environmental factors involved. There is evidence that 2n gamete production can be stimulated by both biotic and abiotic stresses, such as nutritional deprivation, wounding, disease, herbivory, and temperature stress (Ramsey and Schemske, 1998). In Lotus tenuis, temperature stresses, and in particular high temperatures, increase the level of parallel spindle-driven 2n gamete production (Negri and Lemmi, 1998). Similarly, in rose (Rosa spp.), short periods of high temperature (48 h at 30°C–36°C) can induce cytomixis and parallel and tripolar spindles at male metaphase II, generating dyads and triads at the end of male sporogenesis (Pécrix et al., 2011). Low-temperature environments can also stimulate 2n gamete formation. For example, Solanum phureja grown in cool field environments produces more restituted spores compared with lines grown under normal conditions (McHale, 1983). Similarly, in Datura spp. and Achillea borealis, unreduced pollen formation is higher at low temperatures (Ramsey and Schemske, 1998; Ramsey, 2007). Recently, Mason et al. (2011) demonstrated that cold stress significantly stimulates 2n pollen production in some interspecific Brassica spp. hybrids. Temperature-induced diploid gamete formation is not restricted to plants. Low temperatures have also been shown to stimulate the formation of 2n spores in some animal species, particularly among fish and amphibians (Bogart et al., 1989; Mable et al., 2011). Moreover, ecological population studies have demonstrated that polyploid plant and animal species occur more frequently at higher altitudes and at latitudes closer to the poles (Beaton and Hebert, 1988; Barata et al., 1996; Dufresne and Hebert, 1998), leading to the suggestion that cold climates stimulate the production of polyploid gametes.In this study, we demonstrate that short periods of cold stress induce a development-specific production of meiotically restituted spores in Arabidopsis, which thereby constitutes an ideal model system to identify potential cytological and molecular factors involved in stress-induced sexual polyploidization. Using a combination of cytological and genetic approaches, we reveal the cytological basis for cold-induced meiotic restitution and additionally demonstrate that restituted binuclear and polynuclear spores spontaneously develop into diploid and polyploid pollen grains. We also use pollen tetrad-based segregation analysis to monitor the genetic makeup of cold-induced 2n gametes and Arabidopsis mutants to examine the potential role of some candidate regulators (e.g. TAM/CYCA1;2 and MKK2) in the sensitivity of male meiosis to low-temperature stress.     

Integrated response of intercropped maize and potatoes to heterogeneous nutrients and crop neighbours

Background and Aims

In communities, plants often simultaneously interact with intra- and inter-specific neighbours and heterogeneous nutrients. How plants respond under these conditions and then affect the structure and function of communities remain important questions.

Methods

Maize (Zea mays L.) was intercropped with potatoes (Solanum tuberosum L.). In the field experiment, we applied fertilizer both homogeneously and heterogeneously under monocropping and intercropping conditions. The heterogeneous nutrient treatment in intercropping was designed with different fertilizer placements, at intraspecific and interspecific rows, respectively. In the pot experiment, crops were grown under both homogeneous and heterogeneous nitrogen conditions with single plant, intraspecific and interspecific competition. Shoot and root biomass and yield were measured to analyse crop performance.

Results

In the field experiment, the heterogeneous nitrogen, compared with the homogenous one, enhanced the performance of the intercropped crop. Importantly, this effect of heterogeneous nitrogen was greater when fertilizer was applied at interspecific rows, rather than at intraspecific rows. Moreover, in pot experiments, the root foraging precision of the two crops was increased by interspecific neighbours, but only that of potatoes was increased by intraspecific neighbours.

Conclusions

The integrated responses of plants to heterogeneous neighbours and nutrients depend on the position of nutrient-rich patches, which deepen our understanding of the function of plant diversity, and show that fertilizer placement within multi-cropping systems merits more attention. Moreover, the enhanced utilization of heterogeneous nitrogen could drive overyielding in multi-cropping systems.     

Low interspecific pollen transfer between invasive aquatic <Emphasis Type="Italic">Ludwigia grandiflora</Emphasis> and native co-flowering plants

Showy invasive alien plants are often integrated in the diet of generalist pollinators and because of the lack of co-evolvement with the native plant community, a high amount of interspecific pollen transfer (IPT) can be expected. We investigated pollinator switching and magnitude plus distance of IPT between the alien aquatic Ludwigia grandiflora and the native Lythrum salicaria in both directions in uninvaded and invaded sites with a different relative abundance of L. grandiflora (% cover of the alien plant: no cover; low cover: <5%; high cover: 50–75%). A field experiment was conducted to include both pollinator interspecific movements and tracking of IPT, using fluorescent dye as a pollen analogue. Despite a substantial overlap in pollinators between L. grandiflora and the native L. salicaria, less than 10% of the observed flights were interspecific. Similar results were found in dye transfer patterns. The proportions of stigmas with conspecific dye were always higher than the proportions of stigmas with heterospecific dye for L grandiflora and L. salicaria. There were no differences in conspecific dye loads for L. salicaria between uninvaded and invaded sites. Conspecific pollen loss (native CPL) and heterospecific pollen deposition (alien HPD) were in general low and species-specific. The distance of HPD ranged respectively from 1.7 to 39 m and from 0.3 to 54.8 m in the low cover and high cover sites while CPL ranged respectively from 6.40 to 68.02 m and from 0.60 to 40.18 m in the low cover and high cover sites. We can conclude that, in this system, CPL and HPD will play a minor role in pollinator-mediated interaction. Furthermore, interspecific competition for pollinators will cover a larger distance than just neighboring individuals. Our results suggest the necessity to consider the combined effect of insect visitation, pollen deposition, relative alien abundance, distance and seed set when investigating pollinator-mediated interactions of invasive plants.     

Capturing flavors from Capsicum baccatum by introgression in sweet pepper

Key message

Biochemical characterization in combination with genetic analyses in BC ₂ S ₁ plants and near-isogenic lines led to the detection and validation of C. baccatum loci affecting flavor, terpenoid content and Brix level.

Abstract

The species Capsicum baccatum includes the most common hot peppers of the Andean cuisine, known for their rich variation in flavors and aromas. So far the C. baccatum genetic variation remained merely concealed for Capsicum annuum breeding, due to post-fertilization genetic barriers encountered in interspecific hybridization. However, to exploit the potential flavor wealth of C. baccatum we combined interspecific crossing with embryo rescue, resulting in a multi-parent BC₂S₁ population. Volatile and non-volatile compounds plus some physical characters were measured in mature fruits, in combination with taste evaluation by a sensory panel. An enormous variation in biochemical composition and sensory attributes was found, with almost all traits showing transgression. A population-specific genetic linkage map was developed for QTL mapping. BC₂S₁ QTLs were validated in an experiment with near-isogenic lines, resulting in confirmed genetic effects for physical, biochemical and sensory traits. Three findings are described in more detail: (1) A small C. baccatum LG3 introgression caused an extraordinary effect on flavor, resulting in significantly higher scores for the attributes aroma, flowers, spices, celery and chives. In an attempt to identify the responsible biochemical compounds few consistently up- and down-regulated metabolites were detected. (2) Two introgressions (LG10.1 and LG1) had major effects on terpenoid content of mature fruits, affecting at least 15 different monoterpenes. (3) A second LG3 fragment resulted in a strong increase in Brix without negative effects on fruit size. The mapping strategy, the potential application of studied traits and perspectives for breeding are discussed.     

Timing of invasive pollen deposition influences pollen tube growth and seed set in a native plant

Invasive plants may threaten the reproductive success of native sympatric plants by modifying the pollination process. One potential mechanism takes place through the deposition of invasive pollen onto native stigmas when pollinators are shared among species. We explore how pollen from the invasive plant Brassica nigra influences pre- and post-fertilization stages in the native plant Phacelia parryi, through a series of hand pollination experiments. These two species share pollinators to a high degree. P. parryi flowers were hand-pollinated with either pure conspecific pollen (the control) or with B. nigra pollen applied prior to, simultaneously with, or following conspecific pollen. Application of B. nigra pollen lowered seed set, with the simultaneous application resulting in the highest reduction. Pollen tube growth was also influenced by the presence of invasive pollen, with fewer conspecific pollen tubes reaching the base of P. parryi styles in treatments where B. nigra pollen was applied prior to or simultaneously with conspecific pollen. The deleterious effects of invasive pollen on native seed set in this study are likely not due to loss of stigmatic receptivity since seed set was less affected when heterospecific pollen was applied prior to conspecific pollen, but may instead involve interactions between interspecific pollen grains on the stigma or within the style. Our study highlights the importance of timing of foreign pollen deposition on native stigmas and suggests that interspecific pollen transfer between native and exotic plants may be an important mechanism of competition for pollination in invaded plant communities.     

Gamete attachment process revealed in flowering plant fertilization

Sex-possessing organisms perform sexual reproduction, in which gametes from different sexes fuse to produce offspring. In most eukaryotes, one or both sex gametes are motile, and gametes actively approach each other to fuse. However, in flowering plants, the gametes of both sexes lack motility. Two sperm cells (male gametes) that are contained in a pollen grain are recessively delivered via pollen tube elongation. After the pollen tube bursts, sperm cells are released toward the egg and central cells (female gametes) within an ovule (Fig. 1). The precise mechanism of sperm cell movement after the pollen tube bursts remains unknown. Ultimately, one sperm cell fuses with the egg cell and the other one fuses with the central cell, producing an embryo and an endosperm, respectively. Fertilization in which 2 sets of gamete fusion events occur, called double fertilization, has been known for over 100 y. The fact that each morphologically identical sperm cell precisely recognizes its fusion partner strongly suggests that an accurate gamete interaction system(s) exists in flowering plants.Open in a separate windowFigure 1.Illustration of the fertilization process in flowering plants. First, each pollen tube accesses an ovule containing egg and central cells. Next, the 2 sperm cells face the female gametes in the ovule after the pollen tube bursts. Finally, each sperm cell simultaneously fuses with either egg or central cell.     

BcMF11, a novel non-coding RNA gene from Brassica campestris, is required for pollen development and male fertility

Key message

BcMF11 as a non-coding RNA gene has an essential role in pollen development, and might be useful for regulating the pollen fertility of crops by antisense RNA technology.

Abstract

We previously identified a 828-bp full-length cDNA of BcMF11, a novel pollen-specific non-coding mRNA-like gene from Chinese cabbage (Brassica campestris L. ssp. chinensis Makino). However, little information is known about the function of BcMF11 in pollen development. To investigate its exact biological roles in pollen development, the BcMF11 cDNA was antisense inhibited in transgenic Chinese cabbage under the control of a tapetum-specific promoter BcA9 and a constitutive promoter CaMV 35S. Antisense RNA transgenic plants displayed decreasing expression of BcMF11 and showed distinct morphological defects. Pollen germination test in vitro and in vivo of the transgenic plants suggested that inhibition of BcMF11 decreased pollen germination efficiency and delayed the pollen tubes’ extension in the style. Under scanning electron microscopy, many shrunken and collapsed pollen grains were detected in the antisense BcMF11 transgenic Chinese cabbage. Further cytological observation revealed abnormal pollen development process in transgenic plants, including delayed degradation of tapetum, asynchronous separation of microspore, and aborted development of pollen grain. These results suggest that BcMF11, as a non-coding RNA, plays an essential role in pollen development and male fertility.     

Morphology of the unusual polyad in Amazonian Parkia legume trees

Key message

An unusual polyad occurs in three Parkia species, named cavitate polyad. It has an internal central space full of lipoprotein substances, contacting all pollen grains, probably aiding pollen germination.

Abstract

This study details the unusual morphology of polyads of three species of Parkia (P. multijuga Benth., P. ulei (Harms) Kuhlm., and P. pendula (Willd.) Benth. ex Walp.) and suggests functions for polyad adaptive traits that are linked to the reproductive success of the species. Polyads within the anthers of the three Parkia species were analysed by surface (scanning electron microscopy) and anatomical (light microscopy) studies. Ultrastructure and development studies were carried out for P. pendula polyads. Polyads are globose and cavitated, i.e., exhibit an internal cavity that varies in size, being more conspicuous in P. ulei and P. pendula. Other differences among species are related to the polyad size, exine ornamentation and the type of substances stored in the pollen grain. The polyad internal cavity is filled with an exudate that may be related to the pollen germination through the internal pores and/or translocation of substances from the anther loculus to the inside or vice versa. This inference is supported by the following observations: the spaces between the pollen grains in a polyad are also filled with the exudate, and the exudate inside the polyad is similar to the anther locular fluid. The morphology and substances stored within the pollen grains of Parkia polyads seem to be more related to polyad functionality and physiology than to the selective pressures exerted by different pollinators on the group.     

Ploidy manipulation of the gametophyte, endosperm and sporophyte in nature and for crop improvement: a tribute to Professor Stanley J. Peloquin (1921�C2008)

Background

Emeritus Campbell-Bascom Professor Stanley J. Peloquin was an internationally renowned plant geneticist and breeder who made exceptional contributions to the quantity, quality and sustainable supply of food for the world from his innovative and extensive scientific contributions. For five decades, Dr Peloquin merged basic research in plant reproduction, cytology, cytogenetics, genetics, potato (Solanum tuberosum) improvement and education at the University of Wisconsin-Madison. Successive advances across these five decades redefined scientific comprehension of reproductive variation, its genetic control, genetic effects, evolutionary impact and utility for breeding. In concert with the International Potato Center (CIP), he and others translated the advances into application, resulting in large benefits on food production worldwide, exemplifying the importance of integrated innovative university research and graduate education to meet domestic and international needs.

Scope

Dr Peloquin is known to plant breeders, geneticists, international agricultural economists and potato researchers for his enthusiastic and incisive contributions to genetic enhancement of potato using haploids, 2n gametes and wild Solanum species; for his pioneering work on potato cultivation through true seed; and as mentor of a new generation of plant breeders worldwide. The genetic enhancement of potato, the fourth most important food crop worldwide, benefited significantly from expanded germplasm utilization and advanced reproductive genetic knowledge, which he and co-workers, including many former students, systematically transformed into applied breeding methods. His research on plant sexual reproduction included subjects such as haploidization and polyploidization, self- and cross-incompatibility, cytoplasmic male sterility and restorer genes, gametophytic/sporophytic heterozygosity and male fertility, as well as endosperm dosages and seed development. By defining methods of half-tetrad analysis and new cytological techniques, he elucidated modes, mechanisms and genetic controls and effects of 2n gametes in Solanum. Ramifications extend to many other crops and plants, in both basic and applied sciences.

Achievements

Based upon a foundation of genetics, cytogenetics and plant reproductive biology, Dr Peloquin and co-workers developed methods to use 2n gametes and haploids for breeding, and used them to move genes for important horticultural traits from wild tuber-bearing Solanum species to cultivated potato for the betterment of agriculture. The resulting potato germplasm included combinations of yield, adaptation, quality and disease resistance traits that were previously unavailable. This elite plant germplasm was utilized and distributed to 85 countries by the CIP, because it not only increased potato yields and quality, it also broadened the adaptation of potato to lowland tropical regions, where humanity has benefited from this addition to their food supply.Key words: 2n gametes, endosperm balance number, haploid, Solanum, true potato seed     

Development of a complete set of monosomic alien addition lines between Brassica napus and Isatis indigotica (Chinese woad)

Key message

A complete set of monosomic alien addition lines of Brassica napus with one of the seven chromosomes of Isatis indigotica and the recombinant mitochondria was developed and characterized.

Abstract

Monosomic alien addition lines (MAALs) are valuable for elucidating the genome structure and transferring the useful genes and traits in plant breeding. Isatis indigotica (Chinese woad, 2n = 14, II) in Isatideae tribe of Brassicaceae family has been widely cultivated as a medicinal and dye plant in China. Herein, the intertribal somatic hybrid (2n = 52, AACCII) between B. napus cv. Huashuang 3 (2n = 38, AACC) and I. indigotica produced previously was backcrossed recurrently to parental B. napus, and 32 MAAL plants were isolated. Based on their phenotype, 5S and 45S rDNA loci and chromosome-specific SSR markers, these MAALs were classified into seven groups corresponding to potential seven types of MAALs carrying one of the seven I. indigotica chromosomes. One of the MAALs could be distinguishable by expressing the brown anthers of I. indigotica, other two hosted the chromosome with 5S or 45S rDNA locus, but the remaining four were identifiable by SSR markers. The simultaneous detection of the same SSR maker and gene locus in different MAALs revealed the paralogs on the chromosomes involved. The recombinant mitochondrial genome in MAALs was likely related with their male sterility with carpellody stamens, while the MAAL with normal brown anthers probably carried the restoring gene for the male sterility. The complete set of MAALs should be useful for exploiting the I. indigotica genome and for promoting the introgression of valuable genes to B. napus.