ARTIFICIAL HYBRIDIZATION OF RUBBER-BEARING GUAYULE WITH COLD-TOLERANT PARTHENIUM LIGULATUM

ABSTRACT Caespitose and cold-tolerant plants of Parthenium ligulatum (Jones) Barneby (Asteraceae) from a native population in the Uinta Basin, Utah, were uprooted, potted, and transferred to a greenhouse in California. Approximately two years after transfer, the plants flowered and subsequently were crossed to diploid guayule (Parthenium argentatum Gray), the rubber-bearing species, native to the state of Durango, Mexico. Only female guayule × male P. ligulatum crosses produced F₁ hybrids. Only crosses involving guayule as female parent and F₁ plants as male parents produced backcross (BC,) plants. Hybrid plants were variable with respect to their growth habit, inflorescence, and leaf shape. Both parents and F₁ hybrids had 2n = 36 chromosomes. Unlike the parents, however, meiosis was irregular in the hybrids which showed a range of 0–5 and an average of 2.1 univalents at metaphase I. Hybrids averaged 0.87 laggards at anaphase I and 0.83 micronuclei at the tetrad stage. The crossability of guayule and P. ligulatum, the high degree of chromosome pairing of the F₁ hybrids, and the production of BC₁ plants indicate that the two species are related in spite of their distinct morphological and ecological differences. This study suggests that the cold-tolerance trait of P. ligulatum may be transferred to guayule through interspecific hybridization followed by backcrossing. The development of cold-tolerant guayule cultivars is expected to expand the areas of guayule production beyond that of the Chihuahuan desert and similar climates.     

Morphological,chemical, and cytogenetic characters of F1 hybrids betweenParthenium argentatum (Guayule) andP. fruticosum var.fruticosum (Asteraceae) and their potential in rubber improvement

Interspecific hybrids betweenParthenium argentatum, the guayule rubber plant, andP. fruticosum var. fruticosum were evaluated for their potential rubber content and quality. Fifteen-mo-old field-grown plants ofP. fruticosum var.fruticosum measured four times more in height and spread than those ofP. argentatum, but contained less than 0.05% rubber of low mol wt.Parthenium argentatum showed 2% rubber content, with a mol wt of about one million. Resin contents varied little among parents or hybrids. The same age F₁ hybrids were intermediate in height and spread and had low rubber content, but showed presence of high mol wt rubber like the guayule parent. This indicates that high mol wt rubber is expressed over the low mol wt rubber in F₁ hybrids. Despite low rubber content but favorable biomass production, F₁ hybrids revealed irregular meiotic chromosome behavior and low pollen and seed germination. These results suggest that interspecific F₁ hybrids may be used in backcross programs to increase biomass and rubber content in guayule.     

RELATIONSHIP OF WOODY PARTHENIUM ARGENTATUM AND HERBACEOUS P. HISPIDUM VAR. AURICULATUM (ASTERACEAE)

Interspecific hybrids between woody Parthenium argentatum Gray (guayule), native to Mexico and Southwest Texas, and herbaceous perennial P. hispidum var. auriculatum (Britton) Rollins, native to the United States, were obtained successfully. The F₁ hybrids were intermediate for most morphological characters with the exception of the short woody stem, yellow pollen color, and the trichome morphology. Chromosome counts revealed the presence of 2n = 36 A-chromosomes in P. argentatum. The same number of A-chromosomes and four B-chromosomes were found in P. hispidum var. auriculatum. Observations of pollen mother cells showed regular meiosis in both parental species. At diakinesis, chiasmata averaged 1.12 and 1.24 per bivalent for P. argentatum and P. hispidum var. auriculatum, respectively. Meiotic behavior of the F₁ hybrids was irregular. F₁ hybrids averaged 4.43 univalents at metaphase I, 1.95 laggards at anaphase I, and 1.62 micronuclei at the tetrad stage. The low pollen stainability (5.1%) in the F₁ hybrids and the limited number of viable BC₁ seeds (4.07%) may be reflections of the irregular meiosis. Although these primary hybrids are partially fertile, they can be used to introduce desirable characteristics of P. hispidum var. auriculatum, such as herbaceous perennial habit, regrowth ability, and cold tolerance into guayule.     

CHROMOSOME PAIRING AND POLLEN FERTILITY IN INTERSPECIFIC HYBRIDS OF SPECIES OF PARTHENIUM (ASTERACEAE)

Meiotic analyses and pollen viability tests were performed on F, hybrids between diploid guayule (Parthenium argentatum Gray 2n = 36), P. rollinsianum Rzedowski (2n = 36), P. alpinum var. tetraneuris Barneby (2n = 36), and P. alpinum var. alpinum Nutt. (2n = 36). Parthenium chromosomes are small and karyomorphologically similar, and meiotic analysis is difficult because of chromosome clumping. However, cytogenetic studies at metaphase I indicated univalents can be seen in a lateral view of the metaphase plate. Chromosome pairing and the number of univalents varied within and between the interspecific hybrids, with an average univalent number of 1.54 for the P. rollinsianum hybrids, 2.36 for the P. alpinum var. tetraneuris hybrids, and 2.46 for the P. alpinum var. alpinum hybrids. Pollen viability tests for the parental species and the hybrids were conducted by germination of pollen grains on stigmas. The percent of viable pollen recorded for the diploid guayule hybrids with P. rollinsianum, P. alpinum var. tetraneuris, and P. alpinum var. alpinum are 21.94, 13.47, and 11.17, respectively. The degree of chromosome pairing and pollen viability is striking because there are many morphological differences between the parents. The chromosome homology of these species based on their pairing behavior allows for the design of a backcross breeding program that would permit the transfer of the desirable characteristics from these species into diploid guayule.     

STEM ANATOMY OF PARTHENIUM ARGENTATUM,P. INCANUM AND THEIR NATURAL HYBRIDS

Guayule (Parthenium argentatum Gray) contains rubber in the parenchymatous cells of stems and roots. Stem anatomy of P. argentatum is described along with that of P. incanum H.B.K. (mariola). Anatomy of these species differs significantly. Phloem rays in both species increase in width by cell division and expansion; however, the increase observed in mariola is less as compared to that in guayule. Axial xylem parenchyma in guayule is generally a two-cell strand as compared to the fusiform axial xylem parenchyma observed in mariola. Vascular ray cells and cells of the pith region of guayule are parenchymatous, whereas those of mariola are sclerenchymatous. As a result of introgression between guayule and mariola, three forms of guayule exist in the native stands of Mexico. Morphological differences between these guayule plants have been described previously. The stem anatomy of these three groups of plants differ importantly. Group I guayule plants, least introgressed by mariola, have taller rays with the cells of pith region and vascular rays parenchymatous. Group III plants, highly introgressed by mariola, have a few to many cells of vascular rays and pith with lignified secondary walls and shorter rays. Many of the anatomical characteristics of group II plants, somewhat introgressed by mariola, are intermediate between group I and III plants.     

Comparative alkane chemistry of Parthenium argentatum (guayule) and some F1 hybrids

The leaf alkanes of Parthenium argentatum (guayule), P. tomentosum var. stramonium, P. fruticosum var. trilobatum, and the first filial (F₁) generations obtained from crosses with guayule were investigated by GC and mass spectrometry and shown to be useful in chemotaxonomic studies. The identified n-alkanes ranged from C₁₉ to C₄₀ with either n-C₂₉ or n-C₃₁ as the main component. The alkane chemistry of guayule with n-C₃₁ being the main component predominated in most of the F₁ hybrids. The presence of iso-branched alkanes (C₂₇, C₂₉, C₃₁) in P. tomentosum and its hybrids could be detected by GC/MS. These preliminary investigations indicate that epicuticular wax alkanes can be useful in inheritance studies of guayule and its hybrids.     

Comparative analysis of the complete sequence of the plastid genome of Parthenium argentatum and identification of DNA barcodes to differentiate Parthenium species and lines

Background  

Parthenium argentatum (guayule) is an industrial crop that produces latex, which was recently commercialized as a source of latex rubber safe for people with Type I latex allergy. The complete plastid genome of P. argentatum was sequenced. The sequence provides important information useful for genetic engineering strategies. Comparison to the sequences of plastid genomes from three other members of the Asteraceae, Lactuca sativa, Guitozia abyssinica and Helianthus annuus revealed details of the evolution of the four genomes. Chloroplast-specific DNA barcodes were developed for identification of Parthenium species and lines.     

A study of wing morphology and fluctuating asymmetry in interspecific hybrids between Drosophila buzzatii and D. koepferae

In this work we investigate the effect of interspecific hybridization on wing morphology using geometric morphometrics in the cactophilic sibling species D. buzzatii and D. koepferae. Wing morphology in F₁ hybrids exhibited an important degree of phenotypic plasticity and differs significantly from both parental species. However, the pattern of morphological variation between hybrids and the parental strains varied between wing size and wing shape, across rearing media, sexes, and crosses, suggesting a complex genetic architecture underlying divergence in wing morphology. Even though there was significant fluctuating asymmetry for both, wing size and shape in F₁ hybrids and both parental species, there was no evidence of an increased degree of fluctuating asymmetry in hybrids as compared to parental species. These results are interpreted in terms of developmental stability as a function of a balance between levels of heterozygosity and the disruption of coadaptation as an indirect consequence of genomic divergence.     

TRICHOME MORPHOLOGY AS AN INDICATOR OF HIGH RUBBER BEARING GUAYULE (PARTHENIUM ARGENTATUM GRAY) PLANTS IN NATIVE POPULATIONS

Parthenium argentatum Gray (guayule) and P. incanum (mariola) grow in close associations in their native habitat of Mexico. Variation in rubber concentration and morphology of guayule has been known for a long time. Studies of over 75 native guayule plants indicated the presence of at least three prominent forms of guayule plants. Group I guayule plants have oblanceolate leaves and leaf margins are entire to two-toothed. T-shaped leaf trichomes are with centrally attached stalk and cap cell with two blunt ends. Plants in this group are high in rubber, containing on the average 17% rubber. Group II guayule plants have narrow elliptic leaves and leaf margins are entire to four-toothed. T-shaped trichomes have an acentrally attached stalk and a cap cell with short end blunt, long end pointed and straight. Rubber content in these plants averages 10%. Group III guayule plants have ovate leaves and leaf margins are four- to eight-toothed. T-trichomes have an acentrally attached stalk and a cap cell with short end blunt, long end pointed and wavy or curved. These plants on the average contain 6% rubber. Morphological as well as biochemical data indicate the presence of mariola genes in the last two groups of plants and this has resulted in an increase in trichome length and a decrease in the rubber content. Group II guayule plants are of more common occurrence than the others. Based on the data presented here, high rubber bearing guayule plants in native stands can be easily selected by analyzing trichome morphology.     

The interspecific hybrid Petunia parodii × P. inflata and its relevance to somatic hybridization in the genus Petunia

Summary Attempts at the reciprocal cross between Petunia parodii and P. inflata using standard emasculation and pollination techniques failed. Limited pollen tube growth down the style in reciprocal crosses led to reproductive isolation between the self-compatible P. parodii and self-incompatible P. inflata. The interspecific hybrid was successfully produced by bud-pollination of P. parodii with P. inflata as the male parent in 22 percent of attempts, but not in the opposite direction. In vitro pollination of P. parodii ovaries with P. inflata pollen also produced hybrids. The small size of the ovary made it technically impossible to use P. inflata as the female parent for in vitro pollination. The interspecific hybrids were intermediate, as compared to the two parents, for six of the seven plant and flower characters measured. Furthermore, the hybrids had high pollen fertility, set abundant seed upon self-pollination, and readily inter-crossed with the parental species. The results are consistent with a high degree of chromosomal homology in the parental species and with minor genetic divergency leading to reproductive isolation that is pre-zygotic in nature. Overcoming the barriers to cross-incompatibility by practical techniques resulted in fertile interspecific hybrids that segregated for parental characters. The potential value of employing the parental species in somatic hybridization experiments is discussed.Michigan Agricultural Experiment Station Journal Article No. 8404     

Photosynthesis and assimilate partitioning between carbohydrates and isoprenoid products in vegetatively active and dormant guayule: physiological and environmental constraints on rubber accumulation in a semiarid shrub

The stems and roots of the semiarid shrub guayule, Parthenium argentatum, contain a significant amount of natural rubber. Rubber accumulates in guayule when plants are vegetatively and reproductively dormant, complicating the relationship between growth/reproduction and product synthesis. To evaluate the factors regulating the partitioning of carbon to rubber, carbon assimilation and partitioning were measured in guayule plants that were grown under simulated summer‐ and winter‐like conditions and under winter‐like conditions with CO₂ enrichment. These conditions were used to induce vegetatively active and dormant states and to increase the source strength of vegetatively dormant plants, respectively. Rates of CO₂ assimilation, measured under growth temperatures and CO₂, were similar for plants grown under summer‐ and winter‐like conditions, but were higher with elevated CO₂. After 5 months, plants grown under summer‐like conditions had the greatest aboveground biomass, but the lowest levels of non‐structural carbohydrates and rubber. In contrast, the amount of resin in the stems was similar under all growth conditions. Emission of biogenic volatile compounds was more than three‐fold higher in plants grown under summer‐ compared with winter‐like conditions. Taken together, the results show that guayule plants maintain a high rate of photosynthesis and accumulate non‐structural carbohydrates and rubber in the vegetatively dormant state, but emit volatile compounds at a lower rate when compared with more vegetatively active plants. Enrichment with CO₂ in the vegetatively dormant state increased carbohydrate content but not the amount of rubber, suggesting that partitioning of assimilate to rubber is limited by sink strength in guayule.     

DNA Fingerprinting of Individual Species and Intergeneric and Interspecific Hybrids of the Genera Bos and Bison, Subfamily Bovinae

Genome fingerprinting with a hypervariable minisatellite sequence of phage M13 DNA was used to study the genetic variation in individual species of the genera Bosand Bison(subfamily Bovinae) and in their interspecific and intergeneric hybrids. DNA fingerprints were obtained for domestic cow Bos taurus primigenius, vatussy Bos taurus macroceros, banteng Bos javanicus, gaur Bos gaurus, wisent Bison bonasus, bison Bison bison, and for the interspecific and intergeneric hybrids. Compared with the original species, most hybrids showed a greater variation in number and size of hybridization fragments. An association was revealed between the number of hybridization fragments and degree of consanguinity of interspecific hybrids resulting from unique crossing of domestic cow and banteng. Pairwise similarity coefficients were calculated to construct a dendrogram of genetic similarity, which reflected the relationships between the parental species and hybrids varying in degree of consanguinity. The applicability of the method for identifying interspecific and intergeneric hybrids and for studying the consequences of hybridization in the subfamily Bovinae is discussed.     

Detection of hybridization between two loach species (<Emphasis Type="Italic">Paramisgurnus dabryanus</Emphasis> and <Emphasis Type="Italic">Misgurnus anguillicaudatus</Emphasis>) in wild populations

Artificial interspecific hybrids between large scale loach P. dabryanus and tetraploid pond loach M. anguillicaudatus (Cobitidae, Cypriniformes) are viable. To detect the occurrence of possible natural hybridization, genetic analyses by using microsatellite markers were performed for natural populations of large scale loach and pond loach, the reciprocal laboratory hybrids, and “supposed hybrids” with ambiguous morphology. The fertility of the artificial hybrids was also tested. At one diagnostic microsatellite (Mac50), one out of 20 “supposed hybrids” was identified to be F₁ hybrid between the two loach species because it had the same genotype as that of the laboratory hybrids. The triploid hybrids between the two species were confirmed to be female-sterile. The results show that rare hybridization has occurred between diploid large scale loach and tetraploid pond loach in nature although it may have little effect in genetic introgression. This study is helpful for fish conservation and encourages further investigation on natural hybridization and introgression of loaches.     

The effect of micronutrient supply on the growth and seed production of guayule (Parthenium argentatum Gray)

The effect of micronutrient supply on the growth and seed production of guayule (Parthenium argentatum Gray) was investigated. A deficiency of boron, but no other trace element, significantly reduced the growth, seed production and the percentage of seeds that germinated. The optimum concentration of B for seed production was between 20 and 100 μM. Results indicate that guayule may benefit from B fertilisation.     

NATURAL AND ARTIFICIAL HYBRIDS OF HELIANTHUS MAXIMILIANI X H. GROSSESERRATUS

Long , Robert W. (Ohio Wesleyan U., Delaware.) Natural and artificial hybrids of Helianthus Maximiliani × H. grosseserratus. Amer. Jour. Bot. 46(10): 687–692. Illus. 1959.—An investigation of the occurrence of natural hybridization in two perennial sunflowers, Helianthus Maximiliani and H. grosseserratus, was begun in 1950. Subsequently, artificial F₁, F₂, and first and second backcross generations were produced. Fertility and vigor were high in all these plants, but F₁ plants appeared to excel the others in these characteristics. Observations in the experimental garden were supplemented by examination of chromosomes in pollen mother cells, comparisons of herbarium collections, and study of wild populations. Evidence pointed to close genetic relationship of the species and to the occurrence of natural hybridization in areas of distributional overlap. In 1957 and 1958, field work in these areas resulted in the scoring of 18 natural populations, 3 of which consisted of both parental species plus putative F₁ hybrids. Two explanations are offered to account for the seeming absence of introgression. The results support the conclusion that natural hybridization leads to the establishment of F₁ hybrids and that introgression does not occur to any significant extent. Although both species display a high degree of interfertility, they are distinct morphologically. For this reason, it is advisable to maintain them as separate species.     

Homoeologous pairing and recombination in backcross derivatives of tomato somatic hybrids [Lycopersicon esculentum (+) L. peruvianum]

 Genomic in situ hybridization (GISH) was used to examine genome interactions in two allohexa ploid (2n=6x=72) Lycopersicon esculentum (+) L. peruvianum somatic hybrids and their seed progenies originated from subsequent backcrosses to L. esculentum. The ability of GISH to distinguish between chromatin derived from two closely related species, L. esculentum and L. peruvianum (both 2n=2x=24), allowed the precise chromosomal constitution of somatic hybrids and their backcross progenies to be unequivocally established. This enabled the interaction of species genomes to be observed at meiosis, providing clear evidence of strictly regular homoeologous pairing and the high degree of homoeologous recombination in allodiploid plants (2n=2x=24) of the BC₁ generation. In segmental allodiploids of the BC₂ and BC₃ generations, the recombinant chromosomes continued to pair with a homoeologous partner (in the absence of a homologous one), and therefore could be stably incorporated into gametes. Chiasmata were found almost exclusively in more distal, rather subterminal, chromosome segments. A considerable proportion of meiotic recombination was detected in subterminal heterochromatic regions, often involving distal euchromatin, located in close proximity. GISH also supplied information on the extent of the overall sequence homology between the genomes of L. esculentum and L. peruvianum, indicating that despite their different breeding systems, these species may not be differentiated to a high degree genetically. The present study has demonstrated that somatic hybridization between two such closely related, but sexually incompatible or difficult to cross species, provides a way of transferring genes, via homoloeogous crossing-over and recombination, across the incompatibility barriers. Indeed, such hybrids may offer the preferred route for gene transfer, which subsequently results in more stable gene introgression than other methods. Received: 22 July 1996 / Accepted: 23 August 1996     

Natural Hybridization between Two Swallowtail Species Parnassius nomionand Parnassius bremeri(Lepidoptera,Papilionidae) Shown by RAPD–PCR

Genetic evidence for interspecific hybridization between Parnassius nomionand Parnassius bremeriin nature is presented. To demonstrate hybridization between these species, RAPD analysis was used. By testing 25 decamer primers, three and two diagnostic markers were revealed for P. nomionandP. bremeri, respectively. Out of 28 animals examined, 4 were shown to be interspecific hybrids. According to the distribution of diagnostic markers, the interspecific hybrids were intermediate with regard to the parental species. Ecological and biological characteristics of two butterfly species that promote their hybridization in nature are discussed.     

Impact of Asymmetrical Hybridization Followed By Sterile F1 Hybrids on Species Replacement in Pseudorasbora

Pseudorasbora pumila, one of the endangered freshwater minnows in eastern Japan, has been largely replaced by the accidentally introduced species, P. parva, which originated from western Japan. In the contact zone, P. pumila and P. parva have hybridized intensively, producing sterile F₁ hybrids. The present study determined the maternal parent of F₁ hybrids using mtDNA haplotypes to investigate the mating system between P. parva and P. pumila in the hybrid zone. We also pursued the successive changes in the genetic structures of hybridizing populations over a 5-year period using allozymes. A total of 100 natural F₁ hybrids collected from six different populations had P. pumila mtDNA without exception, suggesting that sterile F₁ hybrids resulted from mating only between P. pumila females and P. parva males. Such asymmetrical hybridization implies that P. pumila females waste considerably greater reproductive efforts compared with P. parva males. The data suggest that the rapid replacement of P. pumila by P. parva has been promoted by asymmetrical hybridization, resulting in sterile F₁ hybrids.     

Fertile allotetraploid from the cross betweenPhaseolus coccineus L. andPhaseolus acutifolius A. Gray

Summary As somatic hybridization and genetic transformation are not yet applicable to beans, a programme of hybridization between a male sterile line ofP. coccineus and a wild genotype ofP. acutifolius var.tenuifolius was carried out in order to introduce useful characters from the wild parent into the genome of the cultivated species. This interspecific cross is of particular interest sinceP. acutifolius is a source of resistance to many diseases, drought and high temperature. The difficulties in producing these hybrids were overcome by repeated pollinations and with the help of embryo culture. The F₁ hybrid shows a high sterility which may be explained by the poor pollen quality and the presence of a chromosomic asynapsis at meiosis. Fertile allotetraploids (Co) were successfully produced in progeny from colchicine treated cuttings of F₁ hybrids. Several (C₂) mature seeds were harvested from selfed allotetraploid plants.Abbreviations Co initial allotetraploid plants - C₁, C₂ first and second allotetraploid generation - PMC pollen mother cell