Autogamy and hybridization as evolutionary mechanisms in Panicum subgenus Dicanthelium (Gramineae)

A predominantly autogamous breeding system is described forP. occidentale Scribn.,P. pacificum Hitchc. & Chase, andP. thermale Boland. Five synthetic hybrids and three generations of progeny are also described for crosses among these three species. All parents haven = 9 chromosomes and normal meiosis. Hybrids exhibit various irregularities during meiosis, but nine bivalents are usually formed. One population ofP. occidentale and one ofP. thermale were found to have chromosomes with interchanges larger than those of other populations. The F₂ and F₃ generations had markedly increased fertility, as determined by pollen stainability. It is proposed that autogamy and hybridization are a common means of maintaining and stabilizing genetic variability in these grasses and that these means account for much of the taxonomic difficulty posed by the subgenus. The possibility of a low rate of introgression between populations is also proposed.     

SYNTHETIC HYBRIDS OF AGROPYRON ALBICANS X A. DASYSTACHYUM,SITANION HYSTRIX,AND ELYMUS CANADENSIS

Emasculated crosses of Agropyron albicans Scribn. & Smith with A. dasystachyum (Hook.) Scribn., Sitanion hystrix (Nutt.) J. G. Smith, and Elymus canadensis L. yielded 34, 5, and 9 viable hybrid seeds from 66, 45, and 52 florets, respectively. The hybrids were for the most part morphologically intermediate between their respective parents. The parents behaved cytologically as allotetraploids, 2n = 28; but meiosis in A. albicans was somewhat more irregular than in the other three species. Chromosome pairing was good in all hybrids and indicated that the genomes of the parent species were closely homologous, but only the A. albicans × A. dasystachyum hybrids set seed. Although closely related, A. albicans and A. dasystachyum are not fully conspecific. Agropyron albicans was considered to be a subspecies of A. dasystachyum, as were A. riparium Scribn. & Smith and A. griffithsii Scribn. & Smith ex Piper.     

Somatic hybrids between Brassica juncea (L). Czern. and Diplotaxis harra (Forsk.) Boiss and the generation of backcross progenies

An attempt to transfer genes from droughttolerant Diplotaxis harra, a wild relative of Brassica species, to an elite oil-yielding cultivar, B-85, of mustard (Brassica juncea) was made through protoplast fusion, as the two plant systems are sexually incompatible. By following the standard protocol for PEG-mediated protoplast fusion followed by high pH, high Ca⁺⁺, DMSO treatment and appropriate cell-culture technique, 16 presumptive somatic hybrid plants could be regenerated. Chromosomal analysis of four such somatic hybrids revealed that three of them were asymmetric. Analysis of morphological characters, meiotic chromosomes, and esterase isoenzyme pattern revealed that all the somatic hybrids were different from each other. Furthermore four chromosomes of each genome could undergo homoeologous pairing at meiosis indicating the possibilities for genetic recombination and chromosomal rearrangements. Irregular distribution of chromosomes at anaphase-II at meiosis has been a consistent feature of these plants. Eventually, pollen of all the somatic hybrids showed complete infertility preventing the recovery of any selfed seed. Nevertheless, ovule fertility of one somatic hybrid was not totally impaired as it had set some seeds upon backcrossing with the B. juncea parent. The esterase isoenzyme banding pattern of 24 individual progeny plants of this backcross provided evidence for their recombinant nature. It was thus confirmed that a transfer of genetic traits from Diplotaxis harra to B. juncea had indeed taken place. Furthermore, it was conceptualised that a transfer of alien genes through the protoplast-fusion technique is primarily possible in situations where meiotic pairing of the chromosomes of the two participating genomes generates recombinant gametocytes which can pass through subsequent filial generations.     

Intergeneric Somatic Hybridization Between Brassica napus L. And Sinapis alba L.

Electrically induced protoplast fusion was used to produce somatic hybrids between Brassica napus L. and Sinapis alba L. Seven hybrids were obtained and verified by the simple sequence repeat and cleaved amplified polymorphic sequence analysis of the genefael, indicating that the characteristic bands from S. alba were present in the hybrids. The hybridity was also confirmed by chromosome number counting because the hybrids possessed 62 chromosomes, corresponding to the sum of fusion-parent chromosomes. Chromosome pairing at meiosis was predominantly normal, which led to high pollen fertility,ranging from 66% to 77%. All hybrids were grown to full maturity and could be fertilized and set seed after self-pollination or back-crosses with B. napus. The morphology of the hybrids resembled characteristics from both parental species. An analysis of the fatty acid composition in the seeds of F1 plants was conducted and the seeds were found to contain different amounts of erucic acid, ranging from 11.0% to 52.1%.     

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.     

APOMIXIS IN THE POA SECUNDA COMPLEX

Observation of cleared ovules of Poa secunda Presl showed that plants produce aposporous embryo sacs, and emasculation studies showed that they do not set seed without pollen. The ratio of sexual to asexual embryo sacs on a given plant does not correlate with percent stainable pollen, nor does pollen stainability correlate with meiotic regularity or with chromosome number. Percent apomictic ovules, percent pollen stainability, and regularity of meiosis all vary within a given population; they also vary if the same plant is moved from one site to another. Although there is morphological variation in the species, none of the reproductive features correlates with morphology. Pollen from any morph will stimulate seed set in any other morph, and hybrids have been produced in low frequency. Plants will also set seed using their own pollen. In its reproductive biology, P. secunda exhibits many similarities with P. pratensis L.     

Impaired Male Meiosis due to Irregular Synapsis Coupled with Cytomixis in a New Diploid Cytotype of Dianthus angulatus (Caryophyllaceae) from Indian Cold Deserts

Dianthus angulatus (Caryophyllaceae) is cytologically examined here for the first time for the area of India. The diploid chromosome count of 2n?=?30, ascertained here, represents a new cytotype, supplementing the earlier report of a hexaploid cytotype with 2n?=?90 from outside of India. We report here the occurrence of two plants showing impaired meiosis due to irregular synapsis and cytomixis collected from Kinnaur district, Himachal Pradesh (India). The other plants of this species collected in Lahaul-Spiti region showed normal male meiosis (n?=?15) with a high (95%?100%) pollen fertility and normal seed set, and they reproduce sexually. Irregular synapsis in two plants from the Kinnaur region is characterized by the complete absence of chromosome pairing and the presence of 30 univalents at diakinesis and meta-anaphase. In addition, other meiotic irregularities were found, such as unoriented chromosomes, laggards, precocious movements of univalents at anaphase-I and micronuclei at telophase. Microsporogenesis was also abnormal, resulting in the formation of monads, dyads, triads, polyads, and tetrads with micronuclei. The occurrence of intra- and intermicrosporal chromatin material transfer during microsporogenesis was observed, which is a rather rarely observed phenomenon. The synaptic irregularities coupled with chromatin transfer in these plants seem to be responsible for the high pollen sterility (38%?42%) and heterogeneously sized pollen grains. In these plants no seeds were set, and plants reproduced vegetatively through root suckers.     

Fertile,intermediate hybrids between Phaseolus vulgaris and P. acutifolius from congruity backcrossing

Summary Recurrent or introgressive backcrossing of Phaseolus vulgaris — P. acutifolius hybrids with either P. vulgaris or P. acutifolius increased fertility, as measured by pollen stainability and seed per pod from non-manipulated flowers, while invariably resulting in loss of traits from the non-recurrent parent. When hybrids were backcrossed with each of the parent species in alternate generations (congruity backcrossing), fertility decreased in early generations but gradually increased in later generations. By the fourth or fifth generation, congruity-backcross hybrids produced numbers of seeds per pod from non-manipulated flowers comparable to those of parent species, although the percent of stainable pollen tended to be lower. Congruity-backcross hybrids were intermediate to parent species when pedigrees contained equal representation of parents but favored the majority parent when pedigrees were unbalanced. Individuals exhibiting symptoms of developmental incongruity, such as abnormal leaflet number, foliar variagation, or unusual growth patterns, occurred in each generation. However, completely new characteristics appeared after the second backcross generation, presumably due to recombination. Backcrossing to both parent species in alternate generation reversed incongruity, allowing selection for fertility without loss of traits from one of the parent species.Scientific Journal Series Paper Number 15, 671 of the Minnesota Agricultural Experimental Station     

A CYTOGENETIC ANALYSIS OF CERTAIN POLYPLOIDS IN GRINDELIA (COMPOSITAE)

Two diploid taxa, Grindelia procera and G. camporum, and 3 tetraploid ones, G. camporum, G. hirsutula, and G. stricta, have been studied to ascertain their interrelationships. Meiosis in diploid parental strains was regular, the common chromosome configuration being 5 rod bivalents and 1 ring bivalent. The average chiasmata frequency per chromosome was 0.60. Pollen fertility was about 90% in all strains examined. Diploid interspecific hybrids had normal meiosis with an average chiasmata frequency of 0.56 per chromosome. No heterozygosity for inversions or interchanges was detected, and pollen fertility was above 85%. Meiosis in parental tetraploid strains was characterized by the presence of quadrivalents in addition to a complementary number of bivalents. The average chiasmata frequency per chromosome was 0.59 and pollen fertility was generally about 80%. Tetraploid interspecific hybrids also had quadrivalents, normal meiosis, and high pollen fertility. Close genetic relationships between the diploids and between the tetraploids are indicated, and geographical, ecological, and seasonal barriers to gene exchange exist. Attempts to obtain hybrids between diploids and tetraploids were successful in a few cases. The hybrids were tetraploid and had normal meiosis and fertility similar to parental and F₁ tetraploids. Their origin was by the union of unreduced gametes of the diploid female parent and normal pollen from the tetraploid parent. On the basis of chromosome homology, normal meiosis, plus high fertility exhibited in the diploid, tetraploid, and diploid X tetraploid interspecific hybrids, these species of Grindelia are considered to be a part of an autopolyploid complex. Gene exchange between diploids and diploids, tetraploids and tetraploids, and diploids and tetraploids is possible. Tetraploid G. camporum may have originated by hybridization between G. procera and diploid G. camporum with subsequent doubling of chromosomes and selection for the combined characteristics of the diploids.     

Cytogenetic characterization and fae1 gene variation in progenies from asymmetric somatic hybrids between Brassica napus and Crambe abyssinica.

Sexual progenies of asymmetric somatic hybrids between Brassica napus and Crambe abyssinica were analyzed with respect to chromosomal behavior, fae1 gene introgression, fertility, and fatty-acid composition of the seed. Among 24 progeny plants investigated, 11 plants had 38 chromosomes and were characterized by the occurrence of normal meiosis with 19 bivalents. The other 13 plants had more than 38 chromosomes, constituting a complete chromosomal set from B. napus plus different numbers of additional chromosomes from C. abyssinica. The chromosomes of B. napus and C. abyssinica origin could be clearly discriminated by genomic in situ hybridization (GISH) in mitotic and meiotic cells. Furthermore, meiotic GISH enabled identification of intergenomic chromatin bridges and of asynchrony between the B. napus and C. abyssinca meiotic cycles. Lagging, bridging and late disjunction of univalents derived from C. abyssinica were observed. Analysis of cleaved amplified polymorphic sequence (CAPS) markers derived from the fae1 gene showed novel patterns different from the B. napus recipient in some hybrid offspring. Most of the progeny plants had a high pollen fertility and seed set, and some contained significantly greater amounts of seed erucic acid than the B. napus parent. This study demonstrates that a part of the C. abyssinica genome can be transferred into B. napus via asymmetric hybridization and maintained in sexual progenies of the hybrids. Furthermore, it confirms that UV irradiation improves the fertility of the hybrid and of its sexual progeny via chromosomal elimination and facilitates the introgression of exotic genetic material into crop species.     

甘蔗-滇蔗茅杂交F_1花粉母细胞减数分裂过程GISH分析

研究甘蔗属与滇蔗茅属间远缘杂交F1染色体遗传行为具有重要科学意义,可为发掘利用滇蔗茅野生优异基因资源提供细胞学依据。本研究采用常规压片技术对可育父本及不育杂交F1花粉母细胞减数分裂过程进行比较观察,结果显示可育父本云南95-19减数分裂正常,而不育杂交F1分裂异常;进一步对F1花粉母细胞减数分裂过程进行基因组荧光原位杂交(GISH,genome in situ hybridization)分析,结果表明:滇蔗茅与甘蔗属热带种的亲缘关系较远,双亲染色体在F1细胞中不能进行同源配对,终变期,15条滇蔗茅染色体以单价体形式存在,花粉母细胞减数分裂细胞中存在滞后染色体、染色体丢失和不均衡分离现象。甘蔗-滇蔗茅属间远缘杂交F1花粉母细胞减数分裂异常因而杂交F1花粉完全败育。     

Asymmetric hybridization between Nicotiana tabacum and N. repanda by donor recipient protoplast fusion: transfer of TMV resistance

Summary Genetically asymmetric hybrids were recovered by fusion of Nicotiana tabacum protoplasts with irradiated protoplasts of kanamycin-resistant, nopalineproducing plants of N. repanda. Hybrid calli were selected by culture on media containing kanamycin and were regenerated. These plants were morphologically similar to N. tabacum but produced nopaline, indicating they retained genes from N. repanda. Esterase isozyme profiles also indicated that the plants are somatic hybrids, but are more similar to N. tabacum than N. repanda. Chromosome counts showed most of the hybrids had 55–62 chromosomes, which is consistent with extensive, although incomplete elimination of N. repanda chromosomes. The hybrids were largely male sterile, but about half of them set seed when crossed with N. tabacum. Chromosome numbers of the progeny and the pattern of inheritance of kanamycin resistance indicated the continued elimination of N. repanda genetic material in these backcrosses. The N. repanda parent used in these fusions gave a hypersensitive response to TMV, whereas the N. tabacum parent was TMV sensitive. When inoculated with TMV, plants from two hybrid clones gave a hypersensitive response. Plants from the other clones became systemically infected with the virus.     

THE GENUS COLLINSIA. XVI. SUPERNUMERARY CHROMOSOMES

Dhillon , T. S., and E. D. Garber . (U. Chicago, Chicago, Ill.) The genus Collinsia. XVI. Supernumerary chromosomes. Amer. Jour. Bot. 49 (2) : 168–170. 1962.—The pairing and transmission of supernumerary chromosomes in Collinsia solitaria and in hybrids between C. sparsiflora subsp. arvensis and C. bruceae were studied. Two supernumerary chromosomes usually formed a bivalent with 1 chiasma at metaphase I; 4 supernumerary chromosomes occasionally yielded a trivalent and univalent at this stage. Plants with 2–4 supernumerary chromosomes were fertile and plants with 5–8, sterile. Plants with a given number of supernumerary chromosomes when used as seed or pollen parent gave gametes with a higher number of such chromosomes.     

Intergeneric Somatic Hybridization Between Brassica napus L. and Sinapis alba L.

Electrically induced protoplast fusion was used to produce somatic hybrids between Brassica napus L. and Sinapis alba L. Seven hybrids were obtained and verified by the simple sequence repeat and cleaved amplified polymorphic sequence analysis of the genefael, indicating that the characteristic bands from S. alba were present in the hybrids. The hybridity was also confirmed by chromosome number counting because the hybrids possessed 62 chromosomes, corresponding to the sum of fusion-parent chromosomes. Chromosome pairing at meiosis was predominantly normal, which led to high pollen fertility,ranging from 66% to 77%. All hybrids were grown to full maturity and could be fertilized and set seed after self-pollination or back-crosses with B. napus. The morphology of the hybrids resembled characteristics from both parental species. An analysis of the fatty acid composition in the seeds of F1 plants was conducted and the seeds were found to contain different amounts of erucic acid, ranging from 11.0% to 52.1%.     

GISH confirmation of somatic hybrids between Solanum melongena and S. torvum: assessment of resistance to both fungal and bacterial wilts

Interspecific somatic hybrids between Solanum melongena L. (2n = 2x = 24) and two accessions of Solanum torvum Sw. (2n = 2x = 24) were produced in view of transferring resistance to two soil-born pathogens, Ralstonia solanacearum and Verticillium dahliae, from the wild species into the cultivated eggplant. All somatic hybrids were phenotypically homogenous and intermediate between the parents. Their hybrid nature was confirmed by analysis of isozymes and RAPDs. They showed reduced pollen viability, and all but one possessed the chloroplasts from either one or the other parent. As S. melongena and S. torvum chromosomes were morphologically indistinguishable, genomic in situ hybridisation (GISH) was applied to recognise the chromosomes from each parent in the hybrids. As expected, the selected tetraploid plants contained one complete set of chromosomes from each fusion partner. On spread preparations, the two parental genomes were not spatially separated at any time of the cell cycle. Translocation or recombinant chromosomes could not be demonstrated in the mitotic metaphase. Tests for resistance performed in vitro by using suspensions of two strains of R. solanacearum (race 1 and 3) and filtrate of culture medium of one strain of V. dahliae, revealed that S. melongena was susceptible, whereas both accessions of S. torvum had high levels of resistance. Except for two hybrid clones, which were found susceptible to race 3, as was S. melongena, all somatic hybrids tested showed good levels of bacterial and fungal resistance, either intermediate or as high as that of the wild parent.     

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.     

Cytogenetics of double cross hybrids between Pennisetum americanum — P. purpureum amphiploids and P. americanum x Pennisetum squamulatum interspecific hybrids

Summary Pearl millet, Pennisetum americanum L. Leeke-napiergrass, Pennisetum purpureum Schum. amphiploids (2n=42) were crossed with pearl millet X Pennisetum squamulatum Fresen. interspecific hybrids (2n=41) to study the potential of germplasm transfer from wild Pennisetum species to pearl millet. These two interspecific hybrids were highly cross-compatible and more than two thousand trispecific progenies were produced from 17 double crosses. All doublecross hybrids were perennial and showed a wide range of morphological variations intermediate to both parents in vegetative and inflorescence characteristics. Some crosses resulted in sublethal progenies. Chromosomes paired mainly as bivalents (¯x15.88) or remained as univalents. At metaphase I, trivalents, quadrivalents, an occasional hexavalent and a high frequency of bivalents indicated some homeology among the genomes of the three species. Delayed separation of bivalents, unequal segregation of multivalents, lagging chromosomes, and chromatin bridges were observed at anaphase I. Although approximately 93% of the double-cross hybrids were male-sterile, pollen stainability in male-fertile plants ranged up to 94%. Seed set ranged from 0 to 37 seed per inflorescence in 71 plants under open-pollinated conditions. Apomictic embryo sac development was observed in double-cross progenies when crosses involved a pearl millet x P. squamulatum apomictic hybrid as pollen parent. These new double-cross hybrids may serve as bridging hybrids to transfer genes controlling apomixis and other plant characteristics from the wild Pennisetum species to pearl millet.     

Crossability of Brassica tournefortii and B. rapa, and morphology and cytology of their F1 hybrids

The crossability between Brassica tournefortii (TT, 2n = 20) and Brassica rapa (AA, 2n = 20) and the cytomorphology of their F₁ hybrids were studied. Hybrids between these two species were obtained only when B. tournefortii was involved as a female parent. The hybrid plants were intermediate for most of the morphological attributes and were found to be free from white rust under field conditions. The F₁ plants showed poor pollen fertility, although occasional seed set was achieved from open pollination. Self-pollination or backcrosses did not yield any seeds in these plants. The occurrence of chromosome association ranging from bivalents (0–7), trivalents (0–2) to a rare quadrivalent (0–1) in the dihaploid hybrids indicates pairing between the T and A genomes. The homoeologous pairing coupled with seed set in the F₁ plants offer an opportunity for interspecific gene transfers from B. tournefortii to B. rapa and vice-versa through interspecific hybridization. Received: 3 July 2000 / Accepted: 22 September 2000     

Aneuploidy and isolation in two Hypochoeris species

The annual species Hypochoeris glabra (2n = 10) and the perennial H. radicata (2n = 8) hybridise readily in nature and in experiment. During meiosis in F₁ hybrids the maximum association is a chain of seven and a bivalent indicating that at least three interchanges differentiate the two genomes. The nucleolar chromosomes in the two species are homologous and form a ring bivalent. They are, however, differentiated since in the F₁ hybrid only one nucleolar-organiser region is expressed. Although chromosomal differentiation reduces the egg fertility of F₁ hybrids to about 1%, viable backcross hybrids to H. radicata as pollen parent have been experimentally produced and occur in natural populations. Backcrosses with 8, 9 and rarely 13 chromosomes are found and those with 2n = 8 are fully interfertile with H. radicata. Gene flow may therefore take place in natural populations across an aneuploid barrier. The direction of gene flow in Hypochoeris is probably unidirectional from the annual to the perennial.     

Airborne‐pollen pool and mating pattern in a hybrid zone between Pinus pumila and P. parviflora var. pentaphylla

The reproductive isolation barriers and the mating patterns among Pinus pumila, P. parviflora var. pentaphylla and their hybrids were examined by flowering phenology and genetic assays of three life stages: airborne‐pollen grains, adults and seeds, in a hybrid zone on Mount Apoi, Hokkaido, Japan. Chloroplast DNA composition of the airborne‐pollen was determined by single‐pollen polymerase chain reaction. Mating patterns were analysed by estimating the molecular hybrid index of the seed parent, their seed embryos and pollen parents. The observation of flowering phenology showed that the flowering of P. pumila precedes that of P. parviflora var. pentaphylla by about 6 to 10 days within the same altitudinal ranges. Although this prezygotic isolation barrier is effective, the genetic assay of airborne‐pollen showed that the two pine species, particularly P. pumila, still have chances to form F₁ hybrid seeds. Both parental species showed a strong assortative mating pattern; F₁ seeds were found in only 1.4% of seeds from P. pumila mother trees and not at all in P. parviflora var. pentaphylla. The assortative mating was concluded as the combined result of flowering time differentiation and cross‐incompatibility. In contrast to the parental species, hybrids were fertilized evenly by the two parental species and themselves. The breakdown of prezygotic barriers (intermediate flowering phenology) and cross‐incompatibility may account for the unselective mating. It is suggested that introgression is ongoing on Mount Apoi through backcrossing between hybrids and parental species, despite strong isolation barriers between the parental species.