MEIOTIC IRREGULARITY IN SPECIES AND INTERSPECIFIC HYBRIDS OF PINUS

This paper describes a comparative analysis of meiotic conditions in 61 individual trees representing 21 species and 22 interspecific hybrid combinations of the genus Pinus. Material was collected during three successive growing seasons at the Eddy Arboretum of the Institute of Forest Genetics at Placerville, California. Meiotic irregularity occurred in all species and hybrids examined; mean irregularity frequencies of individual trees ranged from 0 to 47.2 percent. Abnormalities in chromosome movement and their consequences, (1) precocious disjunction associated with the occurrence of univalents and (2) the failure of chiasma terminalization leading to lagging chromosomes and to chromosome breakage and fragments, account for most of the observed irregularity. The same kinds of irregularity occur both in the species and in the hybrids, but they were considerably more frequent in certain of the hybrids than in the related species. These abnormalities in chromosome movement seem to be characteristic of Pinus and are attributed primarily to rrechanical difficulties which attend the large pine chromosomes in meiosis. The most spectacular meiotic irregularities were the characteristic bridge-fragment configurations considered to be the result of crossing-over in heterozygous paracentric inversions. Inversion bridges were observed in 59 of the 61 trees and were as frequent in the species as in the hybrids. They apparently do not result from interspecific differentiation in chromosome structure but from spontaneous intra-specific rearrangements. The literature and work now in progress provide increasing evidence that introgression has been an important factor in the evolution of pine populations. The cytological study of pine chromosomes has failed to produce qualitative evidence of introgression, but the quantitative measurement of meiotic irregularity may serve as a useful criterion for recognizing the results of past hybridization.     

Gametes with somatic chromosome number and their significance in interspecific hybridization in Fuchsia

Sexual polyploidization has both a theoretical as well as an applied significance. Morphological screening for large pollen grains and shape of pollen produced by the individual, cytological investigation of hybrid progeny, and unbalanced separation of chromosomes at anaphase I in pollen mother cells were used to detect the gametes with somatic chromosome number in Fuchsia. The interspecific hybrids of F. fulgens (sect. Ellobium) × F. magellanica (sect. Quelusia), F. fulgens (sect. Ellobium) × F. splendens (sect. Ellobium), and F. triphylla (sect. Fuchsia) × F. splendens (sect. Ellobium) produced at the University of Auckland, New Zealand, showed both large and normal pollen grains in the same anther indicating the presence of unreduced gametes. Cytological investigation carried out on the hybrid progeny of F. fulgens (diploid, 2n=22, sect. Ellobium) × F. magellanica (tetraploid, 2n=44, sect. Quelusia) and F. triphylla (diploid, sect. Fuchsia) × F. arborescens (diploid, sect. Schufia) revealed unexpected chromosome numbers of 2n=44 and 2n=33, respectively. In general, the hybrids showed low fertility caused by genetically unbalanced gametes resulted from random disjunction of chromosomes at anaphase I. Studies on meiosis together with the presence of different shapes and sizes of pollen grains in Fuchsia proved indirectly that unreduced gametes are the products of first division meiotic nuclear restitution. These unreduced gametes were viable irrespective of pollen shape, their predominance in the hybrids, nuclear DNA amount and species phylogenetic position.     

Using DNA‐based techniques to identify hybrids among linear‐leaved Potamogeton plants collected in China

Abstract It is well known that interspecific hybrids occur in the genus Potamogeton. The linear‐leaved Potamogeton species commonly have highly variable morphological characteristics. Their hybrids often show similar vegetative characters to their parental species and their identification based solely on morphology is not always conclusive. In order to clarify whether there are any hybrids from the linear‐leaved Potamogeton plants collected in China, we used internal transcribed spacers (ITS) of nuclear ribosomal DNA and chloroplast rbcL gene sequences to identify the hybrids. Using ITS sequence additivity, we identified four hybrids, namely P. orientalis (P. pusillus×P. oxyphyllus), P. pusillus×P. berchtoldii, P. foliosus×P. octandrus, and P. cristatus×P. octandrus. The latter three hybrids should be considered as new hybrids in Potamogeton. The maternal parents of the four hybrids were confirmed using chloroplast rbcL gene sequences.     

不同松树种间杂交类型的可育性分析

探究不同松树种间杂交可育性,为松树种间杂交育种亲本选配提供依据,进而为松树的杂种优势研究积累材料.该研究利用随机区组设计,通过92个松树种间杂交组合与20个半同胞对照,分析9种松树种间杂交类型子代的平均球果产种数、平均球果产种量、百粒重、发芽率与成苗率5项育性指标,综合球果期、种期与苗期3个阶段的表现,利用模糊数学隶属...     

Artificial and natural hybrids in pycnanthemum (Labiatae)

Further chromosome counts, principally from the southeastern United States, are reported for 38 accessions of 11 species, together with new or corrected counts forP. nudum (2n = 40),P. montanum (2n = 40),P. curvipes (2n = 40), andP. torrei (2n = 80). Four species have now been shown to have intraspecific polyploidy, five are known only as diploids, and nine occur only at the tetraploid level. Artificially produced interspecific hybrids range from vigorous to dwarfed or nonflowering, and from semifertile to sterile as measured by anther development and stainable pollen. Naturally occurring hybrids are described for the following species combinations:P. clinopodioides ×P. incanum,P. pilosum ×P. virginianum, P. flexuosum ×P. nudum, andP. albescens ×P. loomisii.

     

Paternal chloroplast inheritance patterns in pine hybrids detected with trnL-trnF intergenic region polymorphism

The inheritance patterns of the chloroplast genomes of shortleaf pine (Pinus echinata Mill.), loblolly pine (Pinus taeda L.) and slash pine (Pinus elliottii Engelm.) were investigated through the trnL-trnF intergenic spacer polymorphism analysis. The DNA sequences of this spacer differ among these three closely related Pinus species. A modified 'cold' PCR-SSCP (single-strand conformation polymorphism) analysis of this spacer shows that the artificial hybrids (F1) from the shortleaf pine (seed parent) 2 loblolly pine (pollen parent) cross, exhibit the loblolly pine profile. Additionally, nine putative hybrids between shortleaf pine and loblolly pine, previously identified by the IDH (Isocitrate dehydrogenase) allozyme marker, presented the shortleaf pine profile indicating that shortleaf pine, not loblolly pine, sired all of the putative hybrids. Nondenatured polyacrylamide-gel electrophoresis of the trnL-trnF intergenic spacer demonstrated that the artificial hybrids (F1) from the cross, slash pine (seed parent) 2 shortleaf pine (pollen parent), present the shortleaf pine profile. Those results confirmed that the chloroplast genome is paternally inherited in these three species of the genus Pinus. The significance of the trnL-trnF intergenic region polymorphism and our modified 'cold' SSCP protocol for population genetic studies is discussed.     

Phylogenetic relationships amongPinus species (Pinaceae) inferred from different numbers of 6PGDH loci

Electrophoretic examination of variousPinus species from both subgenera revealed that several taxa differ in the number of loci that control the enzyme system 6-phosphogluconate dehydrogenase (6PGDH). Based on inheritance analyses and published data, it was established that all species of subg.Pinus possess only two 6PGDH loci, whereas all stone pines of subg.Strobus exhibit four controlling loci. In order to trace the phylogenetic links at which one or two gene duplications occurred during pine evolution, several species of subsect.Strobi (sectionStrobus) and two species of sect.Parrya were additionally investigated. Based on conclusions about the uniqueness of gene duplications and the different numbers of 6PGDH loci, a phylogenetic tree of the pine taxa was constructed. This tree shows some new features not recognized in earlier studies and supports several novel assignments postulated in very recent pine classifications.     

Difference in cuticular transpiration and sclerophylly in juvenile and adult pine needles relates to the species-specific rates of development

Pinus species show remarkable ontogenetic differences in needle morphology (heterophylly) between juvenile and adult vegetative phases. This developmental shift may play an adaptative role in their success under diverse habitats. As a first step to know the functional differences between each vegetative phase, we compared water loss through the cuticles of juvenile and adult needles of 21-month-old nursery-grown seedlings of nine hard pine species. Cuticular transpiration (CT), calculated after complete stomatal closure, was obtained by leaf-drying curves, and was related to leaf, ontogenetic and climatic parameters. The rate of cuticular transpiration (RCT) between juvenile and adult needles differed across pine species, and in particular segregated the Mediterranean species Pinus canariensis and P. halepensis, from the Eurasian P. uncinata and introduced species P. radiata. For these species, RCT was always higher in juvenile needles. The different leaf and ontogenic parameters studied were correlated with the variation in RCT among the nine pine species. We discuss this relationship in the light of the species ecology. Besides their possible adaptive interpretation, these results suggest an underlying need to consider the ontogenetic heterophylly when assessing functional traits in hard pine seedlings, in particular those traits that govern water relations.     

Phylogenetic Relationships and Species Delimitation in Pinus Section Trifoliae Inferrred from Plastid DNA

Recent diversification followed by secondary contact and hybridization may explain complex patterns of intra- and interspecific morphological and genetic variation in the North American hard pines (Pinus section Trifoliae), a group of approximately 49 tree species distributed in North and Central America and the Caribbean islands. We concatenated five plastid DNA markers for an average of 3.9 individuals per putative species and assessed the suitability of the five regions as DNA bar codes for species identification, species delimitation, and phylogenetic reconstruction. The ycf1 gene accounted for the greatest proportion of the alignment (46.9%), the greatest proportion of variable sites (74.9%), and the most unique sequences (75 haplotypes). Phylogenetic analysis recovered clades corresponding to subsections Australes, Contortae, and Ponderosae. Sequences for 23 of the 49 species were monophyletic and sequences for another 9 species were paraphyletic. Morphologically similar species within subsections usually grouped together, but there were exceptions consistent with incomplete lineage sorting or introgression. Bayesian relaxed molecular clock analyses indicated that all three subsections diversified relatively recently during the Miocene. The general mixed Yule-coalescent method gave a mixed model estimate of only 22 or 23 evolutionary entities for the plastid sequences, which corresponds to less than half the 49 species recognized based on morphological species assignments. Including more unique haplotypes per species may result in higher estimates, but low mutation rates, recent diversification, and large effective population sizes may limit the effectiveness of this method to detect evolutionary entities.     

Field-based artificial crossings indicate partial compatibility of reciprocal crosses between <Emphasis Type="Italic">Pinus sylvestris</Emphasis> and <Emphasis Type="Italic">Pinus mugo</Emphasis> and unexpected chloroplast DNA inheritance

Crossability relationships between Scots pine (Pinus sylvestris L.) and mountain dwarf pine (Pinus mugo Turra) was studied, using artificial pollination approach. Partial compatibility of the reciprocal crossings of these species was proved experimentally, validating the idea of a spontaneous formation of their hybrid swarms under natural conditions. The hybrids were validated using organellar DNA markers and nuclear DNA microsatellites. Based on the percentage of filled seeds, the interspecific crossings were less efficient than the intraspecific cross-pollinations of P. sylvestris and P. mugo individuals. Both species were found to intercross readily with individuals of their putative hybrid swarm, P. mugo exhibiting a higher hybridological affinity towards putatively hybrid individuals than P. sylvestris. Validation of the hybrids confirmed the paternal inheritance of chloroplast DNA (cpDNA) in the combination P. sylvestris × P. mugo only. Surprisingly, in the reciprocal crossing P. mugo × P. sylvestris, maternal inheritance of cpDNA was revealed. Obtained results offer a new insight into the direction and intensity of gene flow within the hybrid swarms of Scots pine and mountain dwarf pine.     

ARTIFICIAL INTERSPECIFIC HYBRIDIZATION IN RUELLIA (ACANTHACEAE)

Observations were made on the morphology, seed fertility, pollen viability and meiotic configurations of 22 different interspecific hybrids of 12 North American taxa representative of 3 sections of Ruellia. The hybrids were classified as (1) fully vigorous and fertile; (2) vigorous but with reduced fertility; (3) reduced vigor and fertility; (4) weak and sterile. Hybrids showed meiotic irregularities in microsporogenesis: univalents, chains, precocious separation of bivalents, nondisjunction, and micronuclei were commonly observed. Interspecific pollinations within sect. Dipteracanthus resulted in fertile or partially sterile F₁ hybrids. Intersectional crosses produced partially or completely sterile hybrids or failed. Assuming that crossability is correlated with degree of evolutionary relationship, the parental species are interpreted in genecological terms to be grouped in polytypic ecospecies. Hybridization experiments doubtless would be of value in clarifying relationships in tropical and subtropical species of Ruellia.     

Hybridization experiments in Geranium

Hybridization experiments were carried out with species of Geranium sect. Anemonifolia, sect. Lucida, sect. Pyrenaica and sect. Ruberta. All 9 species tested proved to be self-compatible. Of the 13 interspecific combinations tested only two were successful (G. brutium X G. molle and G. purpureum X G. robertianum) but both hybrids were completely sterile. It is concluded that at least the European species are reproductively isolated and that hybridization experiments are of little value for an infrageneric classification of Geranium.     

Band composition of electrophoretic spectra of storage proteins in interspecific pea hybrids

Electrophoretic spectra of storage proteins in parental plants and interspecific F₁ and F₂ hybrids Pisum sativum × Pisum fulvum have been studied. Correspondence between the polymorphism levels of protein components among the species and within the species P. sativum was established. Accessions of P. fulvum I609881 and I609885 manifested low polymorphism. Storage proteins of both parents were observed in spectra of F₁ hybrids. F₂ hybrids segregated at a limited set of bands. Accession I609881 of P. fulvum is characterized by unique band 7, which was inherited in F₁.     

HYBRIDIZATION BETWEEN ASTER AND MACHAERANTHERA AND ITS TAXONOMIC SIGNIFICANCE

A controlled crossing program resulted in the production of four interspecific and three intergeneric hybrids among species of Aster sect. Oxytripolium and Machaeranthera. Meiotic analysis revealed stronger degrees of genome homology in some of the intergeneric hybrids than in some of the interspecific hybrids. The significance of these results to the taxonomy of Aster and Machaeranthera was considered and the result was the transfer of Aster sonorae to Machaeranthera.     

Transpecific microsatellites for hard pines

Microsatellites are difficult to recover from large plant genomes so cross-specific utilisation is an important source of markers. Fifty microsatellites were tested for cross-specific amplification and polymorphism to two New World hard pine species, slash pine (Pinus elliottii var. elliottii) and Caribbean pine (P. caribaea var. hondurensis). Twenty-nine (58%) markers amplified in both hard pine species, and 23 of these 29 were polymorphic. Soft pine (subgenus Strobus) microsatellite markers did amplify, but none were polymorphic. Pinus elliottii var. elliottii and P. caribaea var. hondurensis showed mutational changes in the flanking regions and the repeat motif that were informative for Pinus spp. phylogenetic relationships. Most allele length variation could be attributed to variability in repeat unit number. There was no evidence for ascertainment bias.     

Mapped DNA probes from loblolly pine can be used for restriction fragment length polymorphism mapping in other conifers

A high-density genetic map based on restriction fragment length polymorphisms (RFLPs) is being constructed for loblolly pine (Pinus taeda L.). Consequently, a large number of DNA probes from loblolly pine are potentially available for use in other species. We have used some of these DNA probes to detect RFLPs in 12 conifers and an angiosperm. Thirty complementary DNA and two genomic DNA probes from loblolly pine were hybridized to Southern blots containing DNA from five species of Pinus (P. elliottii, P. lambertiana, P. radiata, P. sylvestris, and P. taeda), one species from each of four other genera of Pinaceae (Abies concolor, Larix laricina, Picea abies, and Pseudotsuga menziesii), one species from each of three other families of Coniferales [Sequoia sempervirens (Taxodiaceae), Torreya californica (Taxaceae) and Calocedrus decurrens (Cupressaceae)], and to one angiosperm species (Populus nigra). Results showed that mapped DNA probes from lobolly pine will cross-hybridize to genomic DNA of other species of Pinus and some other genera of the Pinaceae. Only a small proportion of the probes hybridized to genomic DNA from three other families of the Coniferales and the one angiosperm examined. This study demonstrates that mapped DNA probes from loblolly pine can be used to construct RFLP maps for related species, thus enabling the opportunity for comparative genome mapping in conifers.     

Production and characterization of interspecific hybrids between Brassica maurorum and crop brassicas

 Seven interspecific hybrids were produced between Brassica maurorum (♀), a wild species resistant to Alternaria blight and white rust, and all the monogenomic (B. campestris, B. nigra and B. oleracea) and digenomic (B. juncea, B. napus and B. oleracea) crop brassicas (♂) through embryo rescue. The hybrids were confirmed by means of morphological and cytological studies. All the hybrids were pollen-sterile. Amphidiploids were induced in three of the hybrids: B. maurorum×B. napus, B. maurorum×B. carinata, B. maurorum×B. nigra. The hybrids were also confirmed through DNA analyses for nuclear and organelle genomes using RAPD and RFLP techniques. Received: 31 July 1998 / Accepted: 14 August 1998     

Phylogenetic analysis of the hard pines (Pinus subgenus Pinus,Pinaceae) from chloroplast DNA restriction site analysis

Phylogenetic analysis of plastid DNA restriction site and rearrangement mutations suggests a number of major revisions to taxonomy and phylogenetic concepts in the hard pines. Total genomic DNA from 18 species that sampled all nine subsections was digested with 19 restriction enzymes, blotted, and probed with 70% of the Douglas-fir (Pseudotsuga menziesii) chloroplast genome, or, with clones encompassing the entire chloroplast genome of Pinus contorta. A total of 204 site mutations and five rearrangement mutations were generated, of which 126 were phylogenetically informative. Wagner parsimony analyses revealed 11 clades that were strongly supported by bootstrap and decay index analyses. All North American species except P. resinosa formed a distinct monophyletic group that was strongly separated from the Eurasian species. Within the Eurasian clade subsect. Sylvestres was polyphyletic; its Mediterranean species were closely allied with members of sect. Pinea. Sect. Pinea appeared polyphyletic as well; both species of its subsect. Leiophyllae showed a close affinity to Mesoamerican pines of subsect. Oocarpae in sect. Pinus. Within the North American pines subsects. Ponderosae and Oocarpae were polyphyletic. Despite its shallow fossil record, subsect. Contortae emerged as a sister group to all of the North American pines apart from P. resinosa, which was allied with Eurasian species of subsect. Sylvestres. The remaining North American subsections formed two groups: a poorly resolved clade with subsects. Ponderosae and Sabinianae, and sequentially nested clades represented by: P. radiata; P. taeda; representatives of subsects. Oocarpae and Ponderosae from Mesoamerica, and subsect. Leiophyllae. We present estimates of divergence times for each of these major clades based on molecular clocks calibrated using two hard pine fossil observations.     

Ribosomal DNA evidence for the diversification of Tropaeolum sect. Chilensia (Tropaeolaceae)

Maximum parsimony and Bayesian likelihood analysis of ribosomal DNA internal transcribed spacer (ITS) sequences from 55 samples representing 25 species, subspecies, and/or hybrids of Tropaeolum sect. Chilensia yielded a tree with a large proportion of well-supported nodes at the interspecific and intersubspecific level. However, the tree shows samples from two species, T.azureum and T. brachyceras, arising in two different positions. Samples representing subspecies of T. hookerianum and T. leptophyllum suggest that these species are polyphyletic. The data corroborate evidence for hybridization between T. brachyceras and T. tricolor. Consideration of interfertility data, past and present Chilean ecology, and empirical evidence for the behavior of phenotypic and genotypic characters in known hybrids suggest a high likelihood that reticulate evolution has played a role in the diversification of T. sect. Chilensia. This reticulate evolution may explain the discordance between the ITS tree and the conventional taxonomy. High divergence in ITS sequences between T. sect. Chilensia and other members of Tropaeolum prohibits reliable outgroup-rooting, but midpoint rooting places the root at a partition comprising taxa whose distribution conforms to a relictual eastern-western South American disjunction described for other taxa. Within the Chilean taxa, the analysis suggests that biogeographic diversification has been from the mesophytic southern habitats northward to central mediterranean and northern arid zones.