CYTOLOGY OF MAIZE-TRIPSACUM INTROGRESSION

The American Maydinae genera Zea and Tripsacum cross readily when not isolated from each other by gametophytic barriers, and it has been suggested that intergeneric introgression played a role in the evolution of maize. Four Zea chromosomes pair with members of at least one basic genome of tetraploid Tripsacum, and in hybrids involving octaploid Tripsacum all 10 chromosomes of the basic maize genome frequently compete successfully in synapsis with Tripsacum chromosomes. Hybrids that combine 36 Tripsacum and 10 maize chromosomes are female fertile. When they are pollinated by maize their offspring have 36 Tripsacum and 20 maize chromosomes, or again have 36 Tripsacum and 10 maize chromosomes, but the 10 Zea chromosomes are contributed by the new pollen parent. Later backcross generations also include plants with 36 Tripsacum and 12, 14, 16, or 18 maize chromosomes. Individuals with 2n = 56 produce an abundance of offspring with 18 Tripsacum and 20 maize chromosomes when backcrossed with maize. Further backcrossing results in elimination of Tripsacum chromosomes, and eventually plants with 2n = 20 Tripsacum-contaminated maize chromosomes are obtained. Two generations of selfing restore full fertility to these 2n = 20 plants and eliminate all obvious traces of Tripsacum morphology.     

MORPHOLOGY OF TEOSINTOID AND TRIPSACOID MAIZE (ZEA MAYS L.)

Modern races of maize (Zea mays L.) are characterized by indurated glume and rachis tissues. The archaeological record, as well as experimental studies indicate that in North America this induration is associated with hybridization between domesticated maize and its closest wild relative Z. mays subsp. mexicana (Schrad.) Iltis (teosinte). Similar induration can also be introduced into maize through introgression from Tripsacum. North and South American indurated races of maize are not all closely allied morphologically. They evolved independently under domestication. Teosinte is absent from South America, but Tripsacum is widely sympatric with maize from about 42 N to 42 S latitude. For these reasons it has been postulated that induration in South American races may be the result of Tripsacum introgression. However, barriers restricting gene exchange between Zea and Tripsacum are difficult to overcome in nature. It is maintained that indurated South American races of maize were derived from indurated Mexican races, and that the presence or absence of such induration is due to different degrees of expression by intermediate alleles of the tunicate locus.     

Rapid amplification and fixation of new restriction sites in the ribosomal dna repeats in the derivatives of a cross between maize and Tripsacum dactyloides

Some of the derivatives of a cross of maize (Zea mays L.) × Tripsacum dactyloides (L) L (2n = 72) have abnormal development leading to strange and striking morphologies. The Tripsacum chromosomes in these “tripsacoid” maize plants (with Tripsacum-like characteristics) were eliminated and the maize chromosomes were recovered through repeated backcrossing to maize. As an initial attempt to analyze the DNA alterations in tripsacoid maize, we have detected a few restriction site changes in the ribosomal DNA repeat of these plants (Hpa II, Bal I, Sst I, Mbo II, and Sph I) and a new Sph I site was mapped to the spacer region between the 26S and 17S genes. Several possible mechanisms for the generation of a new restriction site are discussed, and we propose that the transient presence of Tripsacum genome during the backcrossing in some way induced a rapid amplification and fixation of new restriction sites in a relatively short period of time.     

Comparative isozyme polymorphisms of north American eastern gamagrass,Tripsacum dactyloides var.dactyloides and maize,Zea mays L.

Random samples, consisting of at least 100 individual seedlings, were taken from the diploid (2n=2x=36) eastern gamagrass (Tripsacum dactyloides var.dactyloides) and assayed to determine which of 12 enzyme marker loci and isozyme systems would be most informative in providing satisfactory resolution of both maize andTripsacum isozyme systems. For comparison, eight maize inbreds were included in the study to aid evaluation and comparison of the various isozyme systems. In addition, evaluations were conducted to identify if the identified optimum isozyme system could be used to detectTripsacum introgression in maize following a maize ×Tripsacum backcrossing scheme. Using the established isozyme techniques for maize (Zea mays L.), theAdh, Pgd, Cat, Est, B-Glu, Got, Idh, Tpi isozyme systems detected no polymorphism among theTripsacum individuals assayed. TheEst andB-Glu systems forTripsacum were unscorable due to poor staining and resolution. TheAcp, Mdh, Pgm, andPhi isozyme systems were found to be satisfactory markers for differentiating between eastern gamagrass individuals as well as detectingTripsacum introgression in maize. The availability of useful isozyme systems which can simultaneously provide significant isozyme resolution of maize,Tripsacum and maize-Tripsacum backcross hybrids, on a single gel system, will be useful for the detection of marker assistedTripsacum introgression into maize. In addition, the identification of a set of variable biochemical markers should also assist breeding, selection and genetic manipulations in eastern gamagrass.The use of company names in this publication does not imply endorsement by the USDA-ARS, or the product names of criticism of similar ones not mentioned. All programs and services of the U.S. Department of Agriculture are offered on a nondiscriminatory basis without regard to race, color, national origin, religion, sex, age, marital status, or handicap.     

Molecular analysis of crosses between Tripsacum dactyloides and Zea diploperennis (Poaceae)

 DNA fingerprinting verified hybrid plants obtained by crossing Eastern gamagrass, Tripsacum dactyloides L., and perennial teosinte, Zea diploperennis Iltis, Doebley & R. Guzmán. Pistillate inflorescences on these hybrids exhibit characteristics intermediate to the key morphological traits that differentiate domesticated maize from its wild relatives: (1) a pair of female spikelets in each cupule; (2) exposed kernels not completely covered by the cupule and outer glumes; (3) a rigid, non-shattering rachis; (4) a polystichous ear. RFLP analysis was employed to investigate the possibility that traits of domesticated maize were derived from hybridization between perennial teosinte and Tripsacum. Southern blots of restriction digested genomic DNA of parent plants, F₁, and F₂ progeny from two different crosses were probed with RFLP markers specifically associated with changes in pistillate inflorescence architecture that signal maize domestication. Pairwise analysis of restriction patterns showed traits considered missing links in the origin of maize correlate with alleles derived from Tripsacum, and the same alleles are stably inherited in second generation progeny from crosses between Tripsacum and perennial teosinte. Received: 11 October 1996/Accepted:8 November 1996     

Composition of Esterase and Peroxidase Isoenzyme Complex, Zein-2 Protein Fraction, and SDS-Protein Complex of Zea Mays L. × Tripsacum Dactyloides L. Hybrids and Parents

The patterns of esterase and peroxidase isoenzymes, subunits of zein-2 fraction and protomers of SDS-protein complex of Zea mays L. × Tripsacum dactyloides L. hybrids and their parents were compared. The study has been made to detect specific to Tripsacum isoesterases and isoperoxidases, zein subunits and SDS-protein protomers which could be used as markers for introgression of gene loci encoding these proteins from Tripsacum into hybrids of Tripsacum with Zea mays. Isoesterases and isoperoxidases as well protomers of SDS-protein complex specific to Tripsacum were detected in all hybrids analyzed. Zein subunits, specific to Tripsacum were detected in some of the analyzed hybrids which i that introgression frequency of the loci encoding proteins studied was different. Chromosome counts taken on the examined hybrids showed the addition of 9 – 13 Tripsacum chromosomes to maize chromosome complement.     

A comparison of apomictic reproduction in eastern gamagrass (Tripsacum dactyloides (L.) L.) and maize-Tripsacum hybrids

The expression of gene(s) governing apomictic reproduction inTripsacum provides the best foundation for comparing the effectiveness of apomictic reproduction in a series of maize-Tripsacum hybrids. Several 38-chromosome, apomictic maize-Tripsacum hybrids are available which possess the gene(s) conferring apomictic reproduction fromTripsacum. Without a base line for comparison, studies directed towards discerning the successful transfer or effectiveness of gene expression in a maize background are hampered. The objectives of this study are to compare the reproductive features found in apomicticTripsacum with those in apomictic maize-Tripsacum hybrids. In addition, this study determined the feasibility of utilizing these maize-Tripsacum hybrid materials to continue an attempt to transfer the genes into a pure maize background. The frequency and occurrence of five unique reproductive features found in apomictic accessions ofTripsacum dactyloides were compared to the reproductive behaviours exhibited in the maize-Tripsacum hybrids. Results indicate the genes controlling apomixis in tetraploidTripsacum are fully functional in maize-Tripsacum hybrids with diploid and triploid maize constitutions. The ability of theTripsacum apomictic genes to retain full expression provides evidence to continue their transfer to a diploid or tetraploid maize background.The use of company names in this publication does not imply endorsement by the USDA-ARS, or the product names or criticism of similar ones not mentioned. All programs and services of the U.S. Department of Agriculture are offered on a nondiscriminatory basis without regard to race, color, national origin, religion, sex, age, marital status, or handicap.     

Crossing maize with sorghum,Tripsacum and millet: the products and their level of development following pollination

Summary Maize was crossed with sorghum, Tripsacum and millet with the aim of introgressing desirable alien characteristics into maize. The products of crosses were analyzed as to their level of differentiation following pollination; their further development on artificial culture medium was compared. In spite of a stimulation rate close to 5%, no evidence of hybridization between maize and sorghum or millet could be obtained. The plants recovered proved to be of maternal origin. However, with an appreciable frequency, stimulation leading to hypertrophic growth of nucellar tissue was observed. This phenomenon is bound to pollination, never occurring in non-pollinated ears. In crosses involving Tripsacum, more than 140 true hybrids were isolated. The influence of the genotypes used as well as factors such as climatic conditions or in vitro techniques are discussed. Except for one haploid maize plant, all the plants recovered proved to be classical hybrids, most of them showing the expected complement of chromosomes from each parent (10 + 36 chromosomes), a few others being slightly hyperploid (2n = 47 to 50 chromosomes). No non-classical hybrids constituted by a nonreduced female gamete and a reduced male gamete were obtained.     

Zeins as indicators of maize-Tripsacum introgression

A survey of zeins in tripsacoid and non-tripsacoid races of maize from Mesoamerica and from South America, annual teosinte, perennial species of Zea and species of Tripsacum revealed at least 33 zein proteins as determined by isoelectric focusing. Zea and Tripsacum and generally also species within these genera are characterized by distinct combinations of zein proteins. Maize is extensively heterogenous, and spans the complete spectrum of zeins present in wild Zea taxa. A comparison of zein proteins failed to distinguish between introgression of maize with Tripsacum or teosinte. The ease with which maize crosses naturally with wild Zea taxa, and the rarity of hybrids with Tripsacum essentially rule out natural Tripsacum introgression as a mode of racial evolution in maize.     

A cross between two maize relatives:Tripsacum dactyloides andZea diploperennis (Poaceae)

Crosses betweenTripsacum dactyloides and teosinte (Zea diploperennis) using standard pollination technique have been successfully attempted and six highly fertile hybrid plants obtained. Previous research had shown other teosintes to be cross-incompatible with Tripsacum and maize to be crossable but highly intersterile withTripsacum. Some investigators believe thatTripsacum played a prominent role in the origin of maize; theTripsacum-diploperennis hybrid provides evidence to support that idea. Ears produced by the hybrid have paired kernel rows, a distinctive characteristic of the oldest archaeological maize that none of the wild relatives have. This unique hybrid is described and discussed in terms of its possible role in the origin and evolution of maize.     

Feature article

Domesticated maize emerged from human selection, exploitation, and cultivation of natural recombinants between two wild grasses that had novel characteristics desired by humans for food. Crossing experiments reconstructing prototypes of ancient archaeological specimens demonstrate how the simple flowering spike of the wild relatives of maize was transformed into the prolific grain-bearing ear within a few generations of intergenomic recombination between teosinte andTripsacum. The high degree of morphological similarities of segregating intercross progeny to archaeological specimens from Tehuacán, Oaxaca, and Tamaulipas provides strong support for this evolutionary scenario. Comparative genomic analysis of maize, teosinte, andTripsacum confirms that maize has inherited unique polymorphisms from aTripsacum ancestor and other unique polymorphisms from a teosinte progenitor. This supports the hypothesis thatTripsacum introgression provided the mutagenic action for the transformation of the teosinte spike into the maize ear. This model for the origin of maize explains its sudden appearance, rapid evolutionary trajectory, and genesis of its spectacular biodiversity.     

Search for evidence of introgression of wheat (Triticum aestivum L.) traits into sea barley (Hordeum marinum s.str. Huds.) and bearded wheatgrass (Elymus caninus L.) in central and northern Europe, using isozymes, RAPD and microsatellite markers

Seeds of English and Austrian populations of bearded wheatgrass (Elymus caninus L.) and sea barley (Hordeum marinum Huds.) growing in the vicinity of wheat (Triticum aestivum L.) fields were collected in order to search for evidence of the introgression of wheat traits into these wild relatives. Seeds were sown and plants grown for subsequent analyses using morphological and genetic (isozymes, RAPD and wheat microsatellites) markers. No F₁ hybrids were found within the individuals of the two species grown, neither with morphological nor with genetic markers. Also, no evidence of introgression of wheat traits into E. caninus was observed. However, in one individual of H. marinum which had the typical morphology of this species, numerous species-specific DNA markers of wheat were amplified, thereby demonstrating previous hybridization. Consequently, the hybridization between wheat and H. marinum under natural conditions and the introgression of wheat traits into this wild relative seems to be possible. Our results contribute to the risk assessment of transgenic wheat cultivation. Received: 20 September 2000 / Accepted: 17 December 2000     

Comparative cytological and molecular analysis of common wheat introgression lines containing genetic material of <Emphasis Type="Italic">Triticum timopheevii</Emphasis> Zhuk

A total of 40 introgression lines of common wheat (2 n = 42) Triticum aestivum L × T. timopheevii Zhuk., resistant to leaf rust and partly to powdery mildew, were examined. Based on cytological analysis of meiosis in pollen mother cells (PMC), hybrid lines were subdivided into two groups characterized by either stable or unstable meiosis. In cytologically stable lines, chromosome configuration at the MI stage of meiosis was mostly bivalent (21II) with small proportion of defect cells (almost 10%), which at most contained two univalents (20II + 2I). Cytologically unstable group was comprised of the lines, containing high proportions of cells with abnormal chromosome pairing in meiotic PMC, as well as the cells with multivalents, and the lines containing aneuploid plants. Localization of the T. timopheevii fragments performed with the use of SSR markers showed that the lines with unstable meiosis were characterized by higher numbers of introgressions compared to stable lines. The influence of certain chromosomes of T. timopheevii on chromosome pairing stability was also demonstrated. In cytologically unstable lines, the increased frequency of 2A substitutions along with the high frequency of introgression of T. timopheevii genetic material into chromosome 7A was observed. Multivalents were scored in all cases of introgression in chromosome 7A. It was suggested that the reason for the genome instability in hybrid forms lied in insufficient compensating ability of certain T. timopheevii chromosomes and/or their parts, involved into recombination processes.     

MORPHOLOGY OF SOME MAIZE-TRIPSACUM HYBRIDS

Diploid (2n = 20) and tetraploid (2n = 40) Zea mays L. were crossed with diploid (2n = 36) and tetraploid (2n = 72) Tripsacum dactyloides (L.) L. to produce a series of hybrids combining different numbers of haploid genomes from each parent. Eight hybrid groups and three parental groups were studied morphologically. Twenty-nine quantitative characters were recorded for each sample. Data were analyzed by univariate analysis of variance, multivariate analysis of variance, and discriminant function analysis, in an attempt to evaluate hybrid differences objectively and determine which morphological characters contribute statistically to group separation. The overall MANOVA F test was significant, establishing the presence of real differences between the hybrids; discriminant function analysis indicated that the percent of paired pistillate spikelets/cupule in the lateral inflorescence was the main variable which differentiated hybrids. Duncan's Multiple Range Tests for significant differences between means were applied to five variables contributing maximally to group discrimination, using the appropriate univariate ANOVAs. Pronounced maize-like attributes of backcross hybrids, as compared with corresponding F₁'s possessing similar genome constitutions, gave possible evidence of gene transfer between Zea mays and Tripsacum during backcrossing to maize.     

ORIGIN OF TRIPSACUM ANDERSONII (GRAMINEAE)

Tripsacum andersonii Gray (Gramineae) is a species with 2n = 64 chromosomes. Chromosome behaviour during meiosis of microsporogenesis suggests that the species combines three homologous haploid Tripsacum genomes of x = 18 (54 chromosomes), and an alien haploid genome of x = 10 chromosomes. Cytogenetic studies indicate that T. andersonii originated as a hybrid between a species of Tripsacum (2n = 36) and a species of Zea (2n = 20). Comparative morphology and flavonoid chemistry fail to identify the Zea species involved in this intergeneric hybrid. Chromosome morphology suggests that it was either Z. mays L. subsp. mays (domesticated maize) or subspecies mexicana (Schrad.) Iltis (annual teosinte). The Tripsacum parent probably was T. latifolium Hitchc. of Central America. It resembles T. andersonii in vegetative morphology. Tripsacum maizar Hernandez et Randolph and T. laxum Nash, which resemble T. andersonii in flavonoid chemistry, are eliminated as possible parents on the basis of growth habit and the morphology of their hybrids with maize.     

SOMATIC EMBRYOGENESIS IN LONG-TERM CALLUS CULTURES OF ZEA MAYS L. (GRAMINEAE)

Long-term (1 yr), soft, embryogenic callus tissue cultures were established from excised immature embryos of a commercial cultivar of hybrid maize (Zea mays L.). Plant regeneration occurred by the formation of somatic embryos, and the regenerated plants were morphologically normal with 2n = 20 chromosomes. Such cultures may be useful for the isolation of mutants and the establishment of embryogenic cell suspension cultures.     

STRUCTURE OF AN ASYMMETRIC HYBRID ZONE BETWEEN TWO WATER STRIDER SPECIES (HEMIPTERA: GERRIDAE: LIMNOPORUS)

The water stricter species Limnoporus dissortis and L. notabilis hybridize across a broad zone in western Canada. Body length and alleles at four allozyme loci show a steep cline along the east slope of the Rocky Mountains in western Alberta, while in central British Columbia the parental phenotypes coexist without merging fully. One sex-linked locus shows little introgression, while there is apparently considerable gene flow at three autosomal loci. Although the hybrid zone has characteristics of a broad tension zone, the spatial distribution of introgression suggests that habitat patchiness and differential habitat associations of the two species also contribute to the pattern of hybridization. Asymmetry in interspecific mating success and incompatibilities of sex chromosomes with each other or with cytoplasmic factors appear to account for the occurrence of L. dissortis genotypes within the range of L. notabilis, and the lack of L. notabilis genotypes within the range of L. dissortis. The genetic structure of this hybrid zone supports the importance of sex-linked traits in maintaining the integrity of species, while its spatial structure suggests that extrinsic habitat features can combine with intrinsic genetic incompatibilities to produce complex hybrid interactions.     

Testcross performance of rye introgression lines developed by marker-assisted backcrossing using an Iranian accession as donor

Introgression libraries facilitate the identification of favorable exotic alleles or genomic regions, which can be exploited for improving elite breeding material. We evaluated the first two introgression libraries in rye (Secale cereale L.) on the phenotypic and molecular level. Our objectives were to detect candidate introgression lines (pre-ILs) with a better testcross performance than the recurrent parent and identify donor chromosome segments (DCS) responsible for the improved performance. We introduced DCS from the self-incompatible heterozygous exotic Iranian primitive rye accession Altevogt 14160 (donor) into the genetic background of the elite inbred line L2053-N (recurrent parent) by marker-assisted backcrossing and developed 40 BC₂S₃ lines in each introgression library. Testcross performance for three agronomic and six quality traits was evaluated in replicated field trials across two testers at five locations over 2 years. The phenotypic effect of the DCS was analyzed for all traits. The pre-ILs had on average a testcross performance comparable to that of the recurrent parent. Significant (P < 0.05) differences between individual pre-ILs and the recurrent parent were detected for all traits except for heading date. For more than 60% of the significant (P < 0.05) differences, the pre-ILs were superior to the recurrent parent. For some pre-ILs, specific DCS were identified containing presumably quantitative trait loci responsible for the superior hybrid performance. Consequently, our study revealed that the development and employment of introgression libraries offers the opportunity for a targeted increase of genetic diversity of elite rye material for hybrid performance of agronomically important traits.     

Malate Dehydrogenase, Alcohol Dehydrogenase, and 6-Phosphogluconate Dehydrogenase Isozymes of Zea mays L. × Tripsacum dactyloides L. Hybrids and Parents

Electrophoretic patterns of malate dehydrogenase (Mdh), alcohol dehydrogenase (Adh), and 6-phosphogluconate dehydrogenase (Pgd) of Zea mays L. × Tripsacum dactyloides L. hybrids and their parents were compared. The components of enzymes specific to T. dactyloides may be used as markers to identify the following T. dactyloides chromosomes in the hybrids: Tr 16 (Mdh 2 and Pdg 1), Tr 7, and/or Tr 13 (Adh 2). The isozymes of Mdh 2 are supposed as a possible biochemical marker to evaluate the introgression of genes, determining an apomictic mode of reproduction from T. dactyloides (localized on Tripsacum 16 chromosome) into Z. mays. The isozymes may be used as markers for the identification of maize chromosomes 1 and 6 in the hybrids as well. Chromosome count taken on the examined hybrids showed the addition of 9 to 13 chromosomes of T. dactyloides to maize chromosome complement.     

Solanum lycopersicoides gene introgression to tomato,Lycopersicon esculentum,through the systematic avoidance and suppression of breeding barriers

Summary While Lycopersicon esculentum and Solanum lycopersicoides have been successfully hybridized, attempts at further direct gene introgression have been unsuccessful due to the presence of incompatibility barriers. A systematic study of the initial hybridization and subsequent backcrosses has identified multiple barriers to introgression. These barriers are expressed as pollen tube inhibition in the upper style and lower pistil, and failures in syngamy, zygote development, and sporogenesis. Upper style cross-incompatibility barriers were successfully avoided by bud pollinations using a stigma complementation procedure to allow pollen germination on otherwise unreceptive stigmas. The inhibition of pollen tube growth was observed in the lower pistil. A combination of environmental, plant, and genetic manipulations facilitated consistent pollen tube growth to the ovule micropyles in all crosses attempted. Failures at syngamy and early zygote formation proved to be the most difficult barriers to overcome — these were particularly severe in crosses to F₁ hybrid plants. Progeny were obtained in all crossing combinations attempted except in the initial hybridization with S. lycopersicoides as the pistillate parent. Although the strong pre-zygotic barriers were overcome in this cross, further progress was restricted by post-zygotic failures. The capability to overcome pre-zygotic barriers and to excise and culture very young embryos has allowed plantlet recovery from male sterile F₁ plants. Partially pollen-fertile F₁ plants were recovered when relatively large F₁ populations were generated from different _{S. lycopersicoides accessions}. In general, barriers to introgression diminished with increased backcrossing, though exceptions were noted. Progeny from the second backcross to L. esculentum possessed adequate fertility to set self-seed under field conditions. Although all backcross progeny were developed from only a few F₁ individuals, considerable genetic variability was recovered for fruit and vegetative characteristics. Potentially useful levels of disease resistance, particularly to Botrytis cinerea, were also recovered.