Herbivory and tree mortality across a pinyon pine hybrid zone

We examined the abundances of three common insect herbivores on pure and hybrid pinyon pines along a 250-km transect in west-central Arizona, United States. Using six morphological traits, we developed a hybrid index to classify trees as pure Pinus californiarum, hybrid, or pure Pinus edulis. The insects (the stem-boring moth, Dioryctria albovittella, the scale insect, Matsucoccus acalyptus, and several species of pitch moths that produce wounds on the trunk and branches) exhibited different distributional patterns across tree types. Stem-boring moths were significantly more abundant on trees at hybrid sites compared to trees at pure sites. In addition, within hybrid sites, hybrids supported significantly more moth larvae than pure trees of either species. These two patterns support the hybrid susceptibility hypothesis in which hybrid breakdown results in increased susceptibility to herbivory. In contrast to stem-borers, there were significantly more pitch moth wounds on trees at pure P. californiarum sites than at hybrid and pure P. edulis sites. Within the hybrid zone, pitch moth abundance was equal on pure P. californiarum and hybrids, and both were significantly greater than on pure P. edulis. These within-site comparisons support the dominance hypothesis where hybrid resistance differs from one tree species, but not the other. Scale insects exhibited the most restricted distribution; over the 250 km transect they were found only in the hybrid zone. This supports the hybrid susceptibility and/or the stress hypothesis (i.e., species at the edge of their range suffer greater stress and are more susceptible to herbivory). We summed the mean numbers of these three common herbivores across sites and found that hybrid sites supported 2.1 and 3.9 times more herbivores than pure P. californiarum and P. edulis sites, respectively. Furthermore, tree mortality was on average, 35 times greater within the hybrid zone compared to pure zones of each species and was associated with the cumulative abundance of herbivores (r ²=0.646). Regardless of whether this mortality is due to insect infestation, stress or a combination of both, these results suggest that hybrid zones are important arenas of natural selection.     

Eucalypt hybrid susceptibility to Gonipterus scutellatus (Coleoptera: Curculionidae)

Abstract Damage caused by the eucalypt snout weevil Gonipterus scutellatus Gyllenhal (Coleoptera: Curculionidae) (=G.gibberus Boisduval) was found on a greater proportion of F₁Eucalyptus amygdalina Labill. X E. risdonii J. D. Hook, hybrid trees than either of the pure species, in a replicated field trial in south‐east Tasmania. A greater proportion of E. risdonii trees was also damaged than E. amygdalina trees. A study of the pattern of oviposition within the trial revealed no difference in oviposition by G. scutellatus between E. risdonii and E. amygdalina. Oviposition by G. scutellatus was significantly higher on the F₁ and F₂interspecific hybrids between these two eucalypt species compared with pure species crosses. There were no apparent differences in damage or oviposition levels between the F₁ hybrids and the single large F₂ progeny included in the trial. This finding provides little evidence that hybrid susceptibility is due to hybrid breakdown after the first generation. Rather, the evidence is consistent with the hypothesis that there may be different mechanisms of host defence operating in each species that are somehow diluted below a threshold level in the hybrids.     

Expansion of the rare Eucalyptus risdonii under climate change through hybridization with a closely related species despite hybrid inferiority

Background and AimsHybridization is increasingly recognized as an integral part of the dynamics of species range expansion and contraction. Thus, it is important to understand the reproductive barriers between co-occurring species. Extending previous studies that argued that the rare Eucalyptus risdonii was expanding into the range of the surrounding E. amygdalina by both seed and pollen dispersal, we here investigate the long-term fitness of both species and their hybrids and whether expansion is continuing.MethodsWe assessed the survival of phenotypes representing a continuum between the two pure species in a natural hybrid swarm after 29 years, along with seedling recruitment. The performance of pure species as well as of artificial and natural hybrids was also assessed over 28 years in a common garden trial.Key ResultsIn the hybrid zone, E. amygdalina adults showed greater mortality than E. risdonii, and the current seedling cohort is still dominated by E. risdonii phenotypes. Morphologically intermediate individuals appeared to be the least fit. Similar results were observed after growing artificial first-generation and natural hybrids alongside pure species families in a common garden trial. Here, the survival, reproduction, health and growth of the intermediate hybrids were significantly less than those of either pure species, consistent with hybrid inferiority, although this did not manifest until later reproductive ages. Among the variable progeny of natural intermediate hybrids, the most E. risdonii-like phenotypes were the most fit.ConclusionsThis study contributes to the increasing number of reports of hybrid inferiority in Eucalyptus, suggesting that post-zygotic barriers contribute to the maintenance of species integrity even between closely related species. However, with fitness rapidly recovered following backcrossing, it is argued that hybridization can still be an important evolutionary process, in the present case appearing to contribute to the range expansion of the rare E. risdonii in response to climate change.     

HYBRIDIZATION AS A DISPERSAL MECHANISM

An example from the genus Eucalyptus is used to argue that hybridization may be of evolutionary significance as a means of gene dispersal where seed dispersal is limited. A previous study of regeneration of E. risdonii and E. amygdalina indicated that the current selective regime was favoring E. risaonii. However, the dispersal of E. risdonii by seeds is shown to be limited (s, = 4.6 m). By comparison, the flow of E. risdonii genes into the range of E. amygdalina by pollen dispersal and F₁ hybridization is widespread (s_p = 82 m). While the actual level of hybridization is low, interspecific hybridization effectively doubles the dispersal of E. risdonii genes into the range of E. amygdalina. This pollen flow can have a significant genetic impact, since isolated hybrids or patches of abnormal phenotypes have been found 200–300 m from the species boundary. Based on lignotuber size, some of these patches appear to have been founded by F₁ hybrids. The frequency of E. risdonii types in the patches appears to increase with patch size suggesting that there is selection for this phenotype in subsequent generations. E. risdonii-like individuals were recovered in the progeny from both intermediate and E. risdonii backcross phenotypes. These results suggest that E. risdonii may invade suitable habitat islands within the E. amygdalina forest, independently of seed migration, by long-distance pollen migration followed by selection for the gene combinations of the pollen parent.     

Allozyme variation and conservation of the Tasmanian endemics, Eucalyptus risdonii, E. tenuiramis and E. coccifera

The rare Tasmanian endemic Eucalyptus risdonii is thought tohave arisen as a result of small, heterochronic changes to the genome ofits more widespread sister species, E. tenuiramis. Previousmorphological studies have shown that genetic differentiation betweenpopulations of E. risdonii and southern E. tenuiramisis continuous and much smaller than the separation between the southernand northern morphotypes of E. tenuiramis. However,morphological traits may be influenced by selection, possibly leading toconvergence, requiring an independent measure of genetic variation. Westudied allozyme frequency variation in E. risdonii, southernE. tenuiramis (parapatric with E. risdonii), northernE. tenuiramis (disjunct from southern populations), and E.coccifera (as an outgroup). Each morphotype had a level of geneticdiversity close to the average reported in ten other eucalypt specieswith similar distributions but the coefficients of populationdifferentiation within morphotypes were lower than in most othereucalypt species. The overall difference between morphotypes wasextremely small, possibly as a result of recent and rapiddifferentiation, but may also be the result of gene flow from otherpeppermint taxa, including E. amygdalina and E.pulchella. Southern E. tenuiramis has greater geneticaffinity with E. risdonii than with northern E.tenuiramis which supports recent evolutionary divergence of E.risdonii. In this study we have shown that taxonomic units are notnecessarily aligned with an equitable partition of the gene pool andthat conservation units should be much broader than single taxa in orderto preserve evolutionary processes.     

The molecular mechanism underlying the “rare allele phenomenon” in a subspecific hybrid zone of the California field mouse,Peromyscus californicus

Natural hybrid zones are known to have unusually high levels of novel or otherwise rare electrophoretic variants (the rare allele phenomenon). These variant alleles are most likely the result either of high levels of unique mutations in hybrids or of intragenic recombination between divergent alleles from the parental populations. This study uses DNA sequence comparisons to determine which process has produced a rare allele of the 6-phosphogluconate dehydrogenase (6PGD) gene in a subspecific hybrid zone of the California field mouse (Peromyscus californicus).About 70% of the coding sequence of 6-PGD was cloned and sequenced from three alleles, including two widespread alleles and one rare allele unique to hybrid populations. Sequence comparisons among the three alleles reveal no patterns that would indicate that the variant was formed by intragenic recombination. Instead, the unique allele of 6-PGD studied seems to have developed by the accumulation of base substitutions, which supports the hypothesis of increased mutation rates in hybrids.     

Production of wide hybrids and backcross progenies between Diplotaxis erucoides and crop brassicas

Intergeneric hybrids were produced between D. erucoides (), a wild species, and four cultivated species of Brassica, B. campestris, B. juncea, B. napus and B. oleracea, through embryo rescue. The hybrid nature of these plants was confirmed through morphological and cytological studies. Backcross pollinations with the pollen of the respective cultivars yielded BC progenies in the hybrids D. erucoides x B. juncea and D. erucoides x B. napus but not in D. erucoides x B. campestris and D. erucoides x B. oleracea. The hybrid D. erucoides x B. campestris was also used as a bridge species and crossed with B. juncea to raise the hybrid and backcross progenies. F₂ progenies were more amenable than f₁ hybrids for raising backcross progenies. Although D. erucoides is considered to be a close relative of B. campestris and B. oleracea, incompatibility barriers of this species with different cultivars do not reflect this relationship.     

Wide hybridization in Brassica

Summary Crosses were made to obtain interspecific hybrids between B. fruticulosa (wild species , 2n = 16) × B. campestris (cultivar , 2n = 20). Although many pollen grains germinated and their tubes entered the style, only about 30% of the ovules received pollen tubes. Fertilized ovules aborted at various stages of development. A few hybrid seeds resulted from hand pollinations in the field, and they showed poor germination and seedling establishment. The in vitro culture of ovaries, ovules, and seeds increased the frequency of obtaining hybrid seeds and plants: the most effective method was ovary culture followed by ovule culture. The hybrid nature of the plants was confirmed through morphological, cytological, and electrophoretic studies. A meiotic analysis of F₁ hybrids (2n = 18) showed that they had 0–5 bivalents and were completely pollen sterile. Electrophoretic analysis of leaf esterases and acid phosphatases of F₁ hybrids revealed bands derived from each parent. Induced amphidiploids of F₁ hybrids contained mostly bivalents, and had about 50% fertile pollen.     

Introgression of the Haynaldia villosa genome into γ-ray-induced asymmetric somatic hybrids of wheat

To study the effect of -ray treatment on donor and derived somatic hybrids, we carried out -ray donor treatment experiments with a wide range of -ray dosages and asymmetric somatic hybridization between protoplasts of wheat (Triticum aestivum L. Jinan 177) and protoplasts of Haynaldia villosa Schur. treated with different dosages of -rays (40, 60 and 80 Gy, respectively). We first screened the putative hybrids by isozyme analysis, followed by characterization of nuclear and organellar genome composition of the hybrids. Genomic in situ hybridization on mitotic metaphases demonstrated that the donor chromosome elimination in the hybrids increased with increased -ray dosage. Intergenomic chromosome recombination/translocations were observed in the hybrids from different dosages of -rays. PCR amplification of 5S rDNA spacer sequences showed that only some of the regenerated hybrid clones inherited donor 5S rDNA sequences, suggesting that the donor DNA was also eliminated randomly. Restriction fragment length polymorphism analysis using mitochondrion (mt) and chloroplast (cp) gene-specific probes showed that the hybrid calli contained mt genomes of both parents and the cp genome of only one of the parents. Recombinations between parental mt as well as cp genes were found in the hybrid clones. Furthermore, development of the hybrid clones was dependent on the -ray dosage used for the donor treatment. Regenerated plants were only obtained from fusion combinations of low (40 Gy) and intermediate (60 Gy) dose irradiation. The possible role and significance of -rays on the introgression of small segments of donor chromosomes to the receptor is discussed.     

Plant genetic differences influence herbivore community structure: evidence from a hybrid willow system

To determine the influence of plant genetic variation on community structure of insect herbivores, we examined the abundances of 14 herbivore species among six genetic classes of willow: Salix eriocephala, S. sericea, their F₁ and F₂ interspecific hybrids, and backcross hybrids to each parental species. We placed 1-year-old plants, grown from seeds generated from controlled crosses, in a common garden. During the growing season, we censused gall-inducing flies and sawflies, leaf-mining insects, and leaf-folding Lepidoptera to determine the community structure of herbivorous insects on the six genetic classes. Our results provided convincing evidence that the community structure of insect herbivores in this hybrid willow system was shaped by genetic differences among the parental species and the hybrid genetic classes. Using MANOVA, we detected significant differences among genetic classes for both absolute and relative abundance of herbivores. Using canonical discriminant analysis, we found that centroid locations describing community structure of the insect herbivores differed for each genetic class. Moreover, the centroids for the four hybrid classes were located well outside of the range between the centroids for the parental species, suggesting that more than additive genetic effects of the two parental species influenced community formation on hybrid classes. Line-cross analysis suggested that plant genetic factors responsible for structuring the herbivore community involved epistatic effects, as well as additive and dominance effects. We discuss the ramifications of these results in regard to the structure of insect herbivore communities on plants and the implications of our findings for the evolution of interspecific interactions.     

Alternative seed defence mechanisms in a palo verde (Fabaceae) hybrid zone: effects on bruchid beetle abundance

Abstract.

• 1 We asked three questions about the patterns of relative abundance of insect herbivores across host plant taxa at a palo verde hybrid zone. (1) What is the morphological structure of the hybrid zone and does this suggest a certain pattern of introgression? (2) How are putative parental seed defence mechanisms expressed in hybrid plants? (3) Do ovipositing females prefer host plant taxa on which their offspring have best survivorship?
• 2 Morphologically, hybrids were either intermediate or tended to resemble one parental species. Previous studies have suggested that unidirectional introgression results in loss of parental defence mechanisms against herbivores. Hybrid plants in general lacked seed coat resistance and early pod abscission which are known to act as plant defence mechanisms against bruchid beetles in the parental palo verde trees.
• 3 All other sources of bruchid mortality that we examined did not vary across parental and hybrid taxa, with the possible exception of egg parasitism which occurred at a lower frequency on one parental palo verde species.
• 4 Thus, survivorship of bruchid offspring should be greater on hybrid palo verdes.
• 5 Patterns of egg densities suggested that females may prefer hybrid hosts in some years but not others. An oviposition choice experiment conducted in the field, however, showed bruchids have no preference for hybrids over one of the parental species.
• 6 These results suggest that some insect herbivores may have higher densities on hybrid host plants because they are less resistant.

     

Hybridisation in the Littorina saxatilis species complex (Prosobranchia : Mollusca)

This paper details attempts to breed members of the L. saxatilis species complex in the laboratory and to construct interspecific hybrids between them. Success in reciprocal crosses between animals from the type localities of the species of Littorina saxatilis (Olivi) and Littorina rudis (Maton) indicates that these two taxa are synonymous. Six of the twelve possible reciprocal hybridisation crosses between the four species (L. saxatilis, L. arcana Hannaford Ellis, L. nigrolineata Gray and L. neglecta Bean) have been attempted, with only one proving successful, that between male L. saxatilis and female L. arcana. This hybrid cross produces viable offspring, although at a lower frequency than either of the parental crosses in the laboratory, while the reciprocal cross has, as yet, proved unsuccessful. Limited work on the F1 hybrids shows them to be inter-fertile and also capable of backcrossing with male L. saxatilis, but not with female L. saxatilis or L. arcana. Details are also given of attempts to find natural hybrids in sympatric populations of the species of the complex, using gel electrophoresis.     

Hybridization as a source of evolutionary novelty: leaf shape in a Hawaiian composite

Hybridization is increasingly recognized as a significant creative force in evolution. Interbreeding among species can lead to the creation of novel genotypes and morphologies that lead to adaptation. On the Hawaiian island of Oahu, populations of two species of plants in the endemic genus Lipochaeta grow at similar elevations in the northern Waianae Mountains. These two species represent extremes of the phenotypic distribution of leaf shape: the leaves of Lipochaeta tenuifolia individuals are compound and highly dissected while leaves of L. tenuis are simple. Based primarily on leaf shape morphology, a putative hybrid population of Lipochaeta located at Puu Kawiwi was identified. Individuals in this population exhibit a range of leaf shapes intermediate in varying degrees between the leaf shapes of the putative parental species. We analyzed individuals from pure populations of L. tenuifolia, L. tenuis and the putative hybrids using 133 AFLP markers. Genetic analysis of these neutral markers provided support for the hybrid origin of this population. The correlation between genetic background and leaf morphology in the hybrids suggested that the genome of the parental species with simple leaves might have significantly contributed to the evolution of a novel, compound leaf morphology.     

Positive and negative effects of herbivory on the population dynamics of Senecio jacobaea L. and Cynoglossum officinale L.

Summary Herbivore effects were studied on populations of the biennial plant species Senecio jacobaea and Cynoglossum officinale. During a three year period (1985–1988) population characteristics (herbivory, number of seedlings, rosettes and flowering plants) were compared in-and outside exclosures, as well as parameters reflecting vegetation cover. In S. jacobaea, a strong negative effect of Tyria jacobaeae was found on seedling establishment, rosette growth and flowering. On the other hand, vertebrate herbivores (mainly rabbits) had an indirect positive effect by limiting the development of the surrounding vegetation (esp. grasses). The increasing vegetation cover in protected populations caused a reduction in germination, seedling- and rosette-growth. Herbivory on C. officinale was low (<10%), no direct effects of herbivores on plant populations were shown. Indirect effects of herbivory through an increasing vegetation were even more pronounced as in S. jacobaea. Therefore, although both plant species may first benefit from herbivore-exclusion, their populations are dependent on rabbits eating other plants (esp. grasses) and reducing competition.Publication of the Meijendel comité, New Series no. 108     

Association of Misexpression with Sterility in Hybrids of <Emphasis Type="Italic">Drosophila simulans</Emphasis>and <Emphasis Type="Italic">D. mauritiana</Emphasis>

Recent studies have identified genes associated with hybrid sterility and other hybrid dysfunctions, but the consequences of introgressions of these speciation genes are often poorly understood. Previously, we identified a panel of genes that are underexpressed in sterile male hybrids of Drosophila simulans and D. mauritiana relative to pure species. Here, we build on this reverse-genetics approach to demonstrate that the underexpression of at least five of these genes in hybrids is associated with hybrid sterility and that these five genes are coordinately regulated. We map one upstream regulator of these genes to a region previously shown to harbor one or more factors causing hybrid sterility. Finally, we show that the genes underexpressed in hybrids are often highly conserved, as might be predicted for downstream targets of the genetic changes that cause hybrid sterility. This approach integrates forward genetics with reverse genetics to show a proximate consequence of the introgression of particular hybrid sterility-conferring regions between species: underexpression of genes necessary for normal spermatogenesis.[Reviewing Editor: Martin Kreitman]     

Risk assessment by crow phenotypes in a hybrid zone

Predation is one of the most selective forces in evolution and, thus, predation may select against hybrids in narrow hybrid zones. It may be possible that parental phenotypes and hybrids differ in their responses towards predators or humans. As predation is difficult to observe I used flight-initiation distance (FID) as a metric of risk assessment. FID is a measurable outcome of the trade-off between fleeing and remaining. Here, I tested whether hybrid and parent crow phenotypes (Corvus corone, Corvus cornix) from the hybrid zone in Eastern Germany differ in their FID. Further, I measured many environmental and social variables to control statistically for their influence on FID. I sampled 154 individuals (53 hooded crows, 54 carrion crows, and 48 hybrids) in the hybrid zone in eastern Germany. I calculated a general linear model using a stepwise backward procedure to establish a minimum model containing only significant variables that explained FID in crows. The variable phenotype (hooded, carrion, hybrid) was then added to the model. There were no differences in FID between hybrids and both parental phenotypes types, suggesting similar risk assessment. This suggests that hybrids may behave similarly in their decision to flee as their parent phenotypes, which, in turn, provides no evidence for a selective disadvantage. An additional analysis focusing on pure phenotypic flocks showed that hybrids in pure hybrid flocks had a lower FID than both parental species in pure flocks. This suggests that hybrids in pure hybrid flocks may be at a disadvantage.     

Embryo development in reciprocal crosses of Phaseolus vulgaris L. and P. coccineus Lam.

Summary Embryo development was examined in reciprocal crosses of Phaseolus vulgaris cv. Great Northern and P. coccineus cv. Scarlet Runner. The formation of abnormal (shrunken and underdeveloped) embryos constituted the primary crossing barrier between the two species when P. coccineus was the female parent. Plants of P. coccineus X P. vulgaris were obtained by embryo culture. Although the P. vulgaris X P. coccineus cross resulted in normal seed development, the fertility of the resulting hybrids was much lower (27%) than that of the reciprocal hybrids (81%). Three classes of F₂ embryos, normal, shrunken, and underdeveloped were formed on reciprocal F₁s and the frequencies did not differ between reciprocal populations. Thus, the interactions between embryo and endosperm and/or maternal parent rather than cytoplasmic-nuclear effects seem to be important in the determination of the extent of embryo growth. The examination of pollen fertility of F₂ plants and the development of F₂ and F₃ embryos suggests that the formation of abnormal embryos and reduced male fertility are independent events. The P. vulgaris — P. coccineus crosses may be useful in studying the possible involvement of interspecific differences in hormonal metabolism in the development of hybrid embryos.     

Somatic embryogenesis and plant regeneration in callus from inflorescences of Hordeum vulgare X Triticum aestivum hybrids

Summary Embryogenic callus cultures were obtained by culturing young inflorescence tissues of Hordeum vulgare cv. PF51811 (2x)XTriticum aestivum cv. Chinese Spring (6x) hybrids on 2,4-D-containing N₆ medium. After subculture for about 10 months the calli retained a high potentiality for somatic embryogenesis and plant regeneration. Of about 300 regenerated plants, approximately 100 were transplanted to potting soil. Eight embryoids and three regenerated plants examined had 28 chromosomes identical to the original hybrid plants, while one regenerated plant was found to be a mixploid composed of cells with 28 and 56 chromosomes. The possibility for obtaining amphiploid hybrids through tissue culture is discussed.     

Naturally occurring interspecific hybridization inSimocephalus (Cladocera,Daphniidae): its potential significance

Among the Cladocera, the occurrence of interspecific hybrids in nature has been reported frequently but rarely adequately documented. During the course of a survey of the genetic variation and population structure of several species ofSimocephalus in ponds in southern Ontario, Canada, readily distinguishable congeners often co-occurred in a pond. Occasionally, clones were recovered that were intermediate in phenotype (as determined from both morphological and electrophoretic analyses) between two species. Two different hybrid clones have been examined in detail to determine the parentals involved in the crosses. Evidence from field and laboratory studies is presented to document the hybrid nature of these clones. Factors favouring formation and long-term survival of hybrids in littoral zone and pelagic species are discussed.     

Morphological and molecular characterization of fertile tetraploid somatic hybrids produced by protoplast electrofusion and PEG-induced fusion between Lycopersicon esculentum Mill. and Lycopersicon peruvianum Mill.

Summary Mesophyl protoplasts of two genotypes of cultivated tomato (Lycopersicon esculentum Mill.) and one of its wild relative species (Lycopersicon peruvianum Mill.) were fused by using electrofusion and polyethyleneglycol-induced fusion. Forty-three fertile tetraploid somatic hybrid plants, each deriving from separate calli, were recovered from both fusion procedures. Electrofusion appeared more efficient than chemical fusion for the production of somatic hybrids. These plants appeared morphologically similar, whatever the fusion procedure and tomato genotype. They had intermediate leaf, inflorescence, and flower morphology. After self-pollination, the hybrids set fruit of intermediate size and color. The hybrid nature of these plants was confirmed by isoelectric focusing of the Rubisco small subunits used as nuclear markers. L. esculentum and L. peruvianum were distinguished by means of two chloroplast markers: CF1-ATPase subunit as analyzed by isoelectro-focusing and ct DNA restriction patterns. All hybrids displayed both ct markers of only one parent with no biased transmission. Mitochondrial (mt) DNAs were prepared from flower buds by using miniaturized CsCl gradients. Preliminary analysis indicated that mt genomes from the hybrids all differed from those of both parents. mt DNA Sall restriction enzyme analysis revealed that all but two hybrids contained one novel fragment of 13.5 kb. Gene mapping experiments showed that the mt apocytochrome b and ATPase subunit 9 homologies in the somatic hybrid mt DNA resembled L. esculentum and L. peruvianum, respectively; the mt nad5 probe distinguished at least four distinct patterns in the hybrids. These results indicated that mt DNA rearrangements involving intergenomic recombinations occurred through protoplast fusion. A greater mt DNA polymorphism was induced with chemical fusion than with electrofusion.