Production of an interspecific hybrid between Texas and Argentine bluegrass

Texas and Argentine bluegrass are both dioecious Poa species with distinct morphological characteristics. Argentine bluegrass has a bunch-type growth habit, fine upright leaves and does not produce rhizomes. Texas bluegrass spreads by rhizomes and has wider, longer leaves. In an effort to produce novel germplasm with forage or turf potential, experiments were conducted to determine whether hybrid seed could be produced. Crosses were made using pollen from an Argentine male to fertilize two Texas female plants. Parents were induced to flower in the greenhouse by extending the photoperiod to 18 h with 400 W high-pressure sodium lamps from late December to April. The two crosses produced >200 seeds each. Hybrid seedlings segregated for traits from both parents. A sorghum-derived SSR marker confirmed the hybrid nature of a group of seedlings. Seeds harvested from F1 plants using Texas or Texas × Argentine pollen lost the undesirable cottony characteristic of Texas bluegrass. Evaluations for forage and turf potential of hybrid and later generation seed are planned.     

Hybridization between Camelina sativa (L.) Crantz (false flax) and North American Camelina species

The potential for gene flow between Camelina sativa, a promising edible and industrial oil crop in Canada, and its wild North American relatives C. alyssum, C. microcarpa and C. rumelica subsp. rumelica, was investigated. Sequence analysis of the nuclear ribosomal internal transcribed spacer region (ITS) was used to differentiate Camelina species and/or accession‐specific nucleotide markers and identify interspecific F₁ hybrids. ITS analysis identified hybrids in progeny of C. alyssum × C. sativa, C. microcarpa accession 36010 × C. sativa and C. sativa × C. rumelica subsp. rumelica. Seed set on C. alyssum × C. sativa F₁ and F₂ progeny was similar to the parents; few seeds were produced on hybrid progeny of C. microcarpa accession 36010 × C. sativa and C. sativa × C. rumelica subsp. rumelica. The study provided evidence that should the species have sympatric distributions and overlapping flowering periods, gene flow between C. sativa and its wild North American relatives is possible and that it would most likely occur with C. alyssum given the high fertility of the F₁ hybrids recovered.     

Pollen fertility among BC2 offspring of Impatiens interspecific hybrids of New Guinea and Indonesian ancestry

BC_T2 seedlings, derived from the cross Tangeglow (I. hawkeri Bull. × I. aurantiaca Teysm.) × 7851-1 (I. hawkeri Bull. × I. platypetala Lindl.) with 7851-1 as recurrent parent, were tested for presence of pollen fertility. Of 59 BC₂ seedlings, 11 were capable of in vitro pollen germination and also of effecting fertilization and subsequent seed set as pollen parent when crossed with Tangeglow as the seed-parent tester. The Spearman's rank correlation coefficient [r_s] of + 0.63 indicated a moderately good correlation of seedling ranking, based on pollen germination percent in vitro and seed set in vivo. Pollen germination of the 11 pollen-fertile BC₂ seedlings varied from a mean low of 4% to a mean high of 41%. The 3 highest pollen-germinating BC₂ seedlings also had the highest seed sets, but siblings showed a wide range in pollen-germinating ability. The best pollen germination for a seedling was 8% in the BC₁ and 41% in the BC₂. Pollen fertility in the BC₂ is discussed in relation to using interspecific hybridization in an Impatiens breeding program. This revised version was published online in August 2006 with corrections to the Cover Date.     

Production of interspecific hybrids in the genus Delphinium via ovule culture

The melon (Cucumis melo) plant introduction PI 414723 was observed to segregate for resistance after inoculation with a highly virulent non-aphid-transmissible strain of zucchini yellow mosaic potyvirus (ZYMV-NAT). By self-pollination of selected resistant plants, the percentage of resistant plants increased in successive generations until nearly 100% in the S₇ progeny. A reciprocal cross was made between a resistant S₅ plant of PI 414723 and a plant of the susceptible cultivar Dulce. Parental-, filial-, and backcross- generation plants were inoculated with ZYMV-NAT. One-half of the F₁ plants, regardless of the direction of the cross, were resistant, indicating that the PI 414723 individual used for crossing was heterozygous. The F₂ generation obtained by self-pollination of resistant F₁ plants segregated to resistant and susceptible in accordance with a 27:37 ratio, indicating that, in this cross, three complementary dominant genes are needed for resistance to be expressed. The resistance to ZYMV carried by PI 414723 is oligogenic with the number of genes observed to segregate in crossing depending on the genotype of the susceptible parent. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cytogenetics of interspecific hybrids in the genus Capsicum L.

Summary Crosses between a wild species C. chacoense and three cultivated species of chili pepper viz. C. annuum, C. frutescens and C. chinense yielded hybrids when C. chacoense was the seed parent but the reciprocal crosses were unsuccessful. C. chacoense × C. annuum F₁ hybrids were partly fertile and therefore an F₂ population could be raised; the other two F₁ hybrids were totally sterile. Chromosome pairing in the F₁ plants resulted largely in bivalents and a few multivalents and univalents. The genomes of the four species share large homologies and the role of chromosome structural changes in genome differentiation is suggested. Hybrid sterility is the major reproductive isolation mechanism.     

Successful backcrosses of somatic hybrids between Sinapis alba and Brassica oleracea with the Brassica oleracea parent

Somatic hybrids between Sinapis alba (2n= 24) and Brassica oleracea (2n= 18) have been backcrossed with the B. oleracea parent. Whereas backcrosses with the diploid B. oleracea parent were unsuccessful, 344 BC₁ seeds could be obtained from inter-valence crosses with tetraploid B. oleracea (2n= 4x= 36). The investigated 96 BC₁ plants segregated for morphological traits and for fertility. They were backcrossed with diploid B. oleracea or self-pollinated, depending on their male fertility. The BC₁F₂ and BC₂ progenies segregated well for the morphological traits. Disturbances were observed especially in the generative phase (flower development and pollen fertility). Both male fertile and male sterile BC₁F₂ and BC₂ plants were obtained and backcrossed or self-pollinated with the B. oleracea parent. The presence of either one of the parental or the cybrid organelle genomes was detected. In the progenies, a stable maternal inheritance of the organelle genome patterns was observed. Isozyme analyses revealed polymorphism for the leucine aminopeptidase (LAP) which was used for the identification of S. alba genes in the progenies. Cytological investigations showed a clear differentiation between the BC₁F₂ and BC₂ plants. Whereas the BC₁F₂ plants possess large numbers of chromosomes ranging from 34 to 40, the BC₂ material was strongly reduced to chromosome numbers ranging from 20 to 22. Preliminary investigation of the meiosis suggests the possibility of introgressions of S. alba-DNA into the B. oleracea genome.     

Reproductive behaviour and broomrape resistance in interspecific hybrids of sunflower

Interspecific hybrids and backcross generations between the wild perennial species Helianthus resinosus, Helianthus paucifiorus, Helianthus laevigatus, Helianthus nuttallii ssp. nuttallii T. & G. and Helianthus giganteus, resistant to broomrape (Orobanche cernua) and susceptible inbred lines were obtained to study crossability to cultivated sunflower and the transmission and expression of resistance to this parasitic weed. Conventional crosses with all the species tested were successful except for the crosses with diploid H. giganteus, for which embryo rescue techniques were needed to overcome hybrid incompatibility. Pollen viability and seed set were highest for F₁ hybrids with hexaploid species and lowest for those with the diploid H. giganteus. We evaluated F₁, BC₁F₁, some BC₂F₁ plants and the wild and cultivated parents. The wild species and interspecific hybrids were resistant to broomrape infection except for H. nuttallii, which showed segregation, indicating that the resistance is dominant. The crossability and resistance of F₁, and back-cross generations of species with different ploidy levels indicate that the transfer of broomrape resistance to cultivated sunflower is feasible.     

Relationships between African species of the genus Cucumis L. Estimated by the production,vigour and fertility of F1 hybrids

Summary The production, vigour, and fertility of F₁ hybrids between nine African species of the genus Cucumis L. were studied as a measure of the relationships between the species. Hybrid plants were obtained from 29 out of the 72 possible cross combinations. Two F₁ hybrids died as seedlings, and 27 hybrids were raised to maturity. Pollen production and stainability varied greatly amongst these hybrids, as did fruit and seed set following self pollination and backcrossing with either parental species. The fruit shape of the hybrids was always intermediate between that of both parental species.Two species appeared to be closely related: C. prophetarum L. and C. anguria var. longipes A. Meeuse. Most other species produced highly to moderately fertile F₁ hybrids with at least one other species. C. metuliferus Naud. produced only sterile hybrids with C. zeyheri Sond. 2x. The results sustain the recent taxonomic classification of the genus (Jeffrey, 1980).     

DNA fingerprint of F1 interspecific hybrids from the Triticeae tribe using ISSRs

Inter-simple-sequence repeat (ISSR) were used to fingerprint three kinds of F1 interspecific hybrids: the Portuguese durum wheat cultivar ‘Candial’ × tritordeum, the Portuguese bread wheat cultivar ‘Barbela’ × tritordeum and the Portuguese triticale cultivar ‘Douro’ × tritordeum. Among the 30 primers tested, the number of discriminative primers varies from 13 to 20 in each hybrid combination, the total number of ISSR loci analysed ranged from 101 to 183, and the number of discriminative ISSR loci varied from 50 to 71. ISSR-PCR technique revealed to be a valuable fingerprint system for all F1 hybrid combinations and respective parents analysed here. The complementary use of molecular cytogenetic techniques and molecular markers could allow a more accurately evaluation and characterisation of different plant species.     

Production and cytogenetics of tetraploid–octoploid Avena hybrids

The objectives of this research were to gather hybridization information about crosses between tetraploid and synthetically derived octoploid lines of Avena and their reciprocals, and to study the cytology of the hybrids obtained. When the octoploid plants were used as females, only 2.6% of the pollinated florets set seed, while seed set increased to 39.0% in reciprocal crosses. However, germination was 95.7% for the first group of hybrids and only 18.2% for the reciprocals. These diverse results can be explained by the paternal: maternal endosperm genomic ratios and the hypothesis of polar nuclei activation. The tetraploid-octoploid hybrids did not always contain 42 chromosomes. Octoploid lines derived indirectly from tetraploid (AABB) × hexaploid (AACCDD) crosses were found to retain their A and B genome chromosomes. Univalents in the above hybrids averaged 15.44 in 1990 and 15.15 in 1991, indicating the presence of C and D genome chromosomes inherited from the octoploid lines. Directly synthesized octoploids from diploid (A_cA_c) × hexaploid (AACCDD) crosses were found to form 20.44 univalents, 5.30 bivalents, and 2.69 trivalents when crossed with AABB tetraploid species. The high number of univalents indicates that chromosomes from at least three genomes, i.e. B, C, and D, remain unpaired, while trivalents are the result of homology between the A and A_c genomes. Hybrids between tetraploid and octoploid lines can be obtained at low frequencies but, regardless of the direction of the cross, these hybrids are highly sterile.     

Production and characterization of hybrids between Cajanus cajan × C. reticulatus var. grandifolius

Cajanus reticulatus var. grandifolius, endemic to Australia and a wild relative of the cultivated species, C. cajan, was successfully crossed with the latter as the female parent. The major wild species characters such as persistent stipules, long pod hairs, pod shattering, brown seeds with grey speckles, and presence of seed strophiole were dominant in the hybrid. For growth and branching habit, and leaflet, flower, pod, and seed size, the hybrid was intermediate between the parents. The meiotic cells of the hybrid were found to have quadrivalents, trivalents, univalents,and showed reduced chromosome pairing as revealed by the increased number of rod bivalents per cell at metaphase-I, and stickiness and precocious movement of chromosomes to poles in the second division. In comparison to the parents, the hybrid had fewer pods and seeds. However,these anomalies in the interspecific hybrid are not significant enough to preclude the gene transfer from C. reticulatus to the cultivated species through a sexual route. This revised version was published online in August 2006 with corrections to the Cover Date.     

Production of an interspecific hybrid between Brassica fruticulosa and B. rapa

An interspecific hybrid between a wild species, Brassica fruticulosa (2n = 16, FF) and a crop Brassica species, B. rapa (2n = 20, AA) was synthesized using sequential ovary‐ovule culture on MS medium supplemented with casein hydrolysate. Morphological, molecular and cytogenetic analysis confirmed the true hybrid nature of the offspring. The F1 plants (2n = 18) were intermediate in morphology, highly pollen‐sterile as well as self‐sterile. A maximum of three bivalents per PMC was recorded, but 14 I + 2 II was the most common meiotic configuration. Normal pollen fertility and regular bivalent (18 II) formation was observed in the amphiploid sectors of the hybrid plants. The F₁ hybrid harboured significantly lower aphid populations than the crop Brassica parent.     

Production of hybrids between Cajanus acutifolius and C. cajan

There are many wild species of pigeonpea which are endemic to Australia. These wild species are cross incompatible with cultivated species of Indian origin. Cajanus acutifolius is one such species which does not easily cross with cultivated pigeonpea. Interspecific pollinations lead to hybrid seeds which were semi-shrivelled. Very few seeds germinated to give rise to F1 plants. Backcrossing the hybrid plants to C. cajan, the male parent, gave rise to aborting seeds which did not germinate in vivo hence BC1 plants are obtained after saving the aborting embryos in vitro. BC₁ plants showed normal meiotic pairing, but had low pollen fertility. The reasons for embryo abortion and low pollen fertility in spite of normal meiosis could be due to the effect of wild species cytoplasm. This revised version was published online in August 2006 with corrections to the Cover Date.     

Rapid estimation of ploidy levels in in vitro-regenerated interspecific Arachis hybrids and fertile triploids

Summary A significant correlation among chromosome number, chloroplast number and pollen grain size was observed using interspecific hybrids (2n=30,60), Arachis hypogaea (2n=40), A. stenosperma (PI 338280) (2n=20), A. batizocoi (PI 468329) (2n=20) and A. villosulicarpa Hoehne (PI 336984) (2n=20), representing four ploidy groups. Both chloroplast number in guard cells and pollen grain size were found to be positively correlated with ploidy, and provided a reliable method to distinguish diploid, triploid, tetraploid and hexaploid plants from each other regardless of their taxonomic backgrounds. These methods in combination with root-tip chromosome counts enabled us to confirm ploidy determinations on all three dissue layers, L₁, L₂ and L₃ (guard cells, microsporocytes and root meristematic cells, respectively), and to detect the chimeric nature of some colchicine-treated tissue culture-derived plants. Screening pollen grains by size and the detection of highly stainable and viable, 30-chromosome pollen grains have enhanced the use of triploid hybrids in peanut breeding programs.     

Physiological and anatomical traits associated with tolerance to long‐term partial submergence stress in the Lotus genus: responses of forage species,a model and an interspecific hybrid

Cattle production based on natural pastures is often subject to flooding periods, which affect plant performance and as a result, forage production. Although most forage legumes are not tolerant to flooding, Lotus spp. are outstanding alternatives, since species, such as L. tenuis (Lt) and L. corniculatus (LcT), have high forage quality and are adaptable to different environments. We recently obtained a L. tenuis × L. corniculatus hybrid (LtxLc) with potential new cultivar traits, although its tolerance to flooding stress has not yet been evaluated. In the present study, the performance of LtxLc, its parental diploid accessions, the model legume L. japonicus and tetraploid LcT were evaluated under 55 days of partial submergence stress and a 35‐day recovery period. Physiological, morphological and anatomical traits were analysed, showing that tolerance to partial submergence was positively associated with aerenchyma and adventitious root formation and relative growth rates. Overall, Lt and LtxLc showed the best responses under stress and during the recovery period. Nevertheless, the higher forage value of LtxLc makes it recommendable for use in environments affected by flooding. Our results could be used as breeding criteria for the generation of new cultivars tolerant to partial submergence stress.     

Identifying the Pathogen of Cotton Leaf Spot Based on the Analysis of rDNA-ITS and Histone 3 Gene Sequence in Xinjiang

[Objective] The objective of this experiment is to identify pathogens causing cotton leaf spot in Xinjiang. [Method] Two hundred and six leaf samples displaying typical leaf spot symptoms were collected from 30 loci in 7 different regions of Xinjiang in 2016 and 2017. The fungal isolates were purified using conventional methods. Ten representative fungal strains were selected for further tests based on different sampling times, locations, and colony characteristics. The strains were identified based on their morphological characteristics and the sequence analysis of both rDNA internal transcribed spacer and histone 3 gene. [Result] In north of Xinjiang, Alternaria alternata and A. tenuissima were the dominant pathogens at the seedling stage (from May to June), whereas in August and September, A. tenuissima was the main pathogen and few were A. alternata and A. macrospora. In south of Xinjiang, A. tenuissima was the main pathogen at the seedling stage, whereas in August and September, A. tenuissima and A. macrospora dominated. Further, Gossypium hirsutum and G. barbadense was mainly infected with A. tenuissima and A. macrospora, respectively. [Conclusion] The main pathogens causing cotton leaf spot in Xinjiang are A. alternata, A. tenuissima and A. macrospora.     

Production of high yield short duration Brassica napus by interspecific hybridization between B. oleracea and B. rapa

Brassica napus is a leading oilseed crop throughout many parts of the world. It is well adapted to long day photoperiods, however, it does not adapt well to short day subtropical regions. Short duration B. napus plants were resynthesized through ovary culture from interspecific crosses in which B. rapa cultivars were reciprocally crossed with B. oleracea. From five different combinations, 17 hybrid plants were obtained in both directions. By self-pollinating the F₁ hybrids or introgressing them with cultivated B. napus, resynthesized (RS) F₃ and semi-resynthesized (SRS) F₂ generations were produced, respectively. In field trial in Bangladesh, the RS B. napus plants demonstrated variation in days to first flowering ranging from 29 to 73 days; some of which were similar to cultivated short duration B. napus, but not cultivated short duration B. rapa. The RS and SRS B. napus lines produced 2–4.6 and 1.6–3.7 times higher yields, respectively, as compared to cultivated short duration B. napus. Our developed RS lines may be useful for rapeseed breeding not only for subtropical regions, but also for areas such as Canada and Europe where spring rapeseed production can suffer from late spring frosts. Yield and earliness in RS lines are discussed.     

Production of interspecific hybrids between Alstroemeria pelegrina L. var. rosea and A. magenta Bayer by ovule culture

Interspecific hybrids were efficiently produced in the cross-incompatible combination between Alstroemeria pelegrina L. var. rosea and A. magenta Bayer by culturing immature ovules with placenta 7–14 days after pollination on 2 g/l Gelrite-solidified MS medium containing 3% (w/v)sucrose. The plants showed intermediate characteristics between the parents and their hybridity was confirmed by karyotype and DNA analyses. The mean number of chromosome association per PMC at metaphase I was 2.60I+6.70II, pollen stainability was20.8%, and they produced viable seeds after self-pollination. Furthermore, mature plants were obtained when the hybrids were backcrossed as male parents with both the parents. The backcross-progeny from A. pelegrina var. rosea × hybrids exhibited 3.8 to 79.7% pollen stainability and that from A. magenta × hybrids 78.8 to 98.3%. Almost all of these plants produced viable seeds after self-pollination, which implies that they can beutilized for breeding of novel cultivars of Alstroemeria. This revised version was published online in July 2006 with corrections to the Cover Date.     

Production of interspecific F1 hybrids, BC1, BC2 and BC3 populations between Lycopersicon esculentum and two accessions of Lycopersicon peruvianum carrying new root-knot nematode resistance genes

Two accessions of Lycopersicon peruvianum Mill. (PI270435, PI126443) carrying Mi-2 and Mi-3, respectively, new root-knot nematode resistance genes, were selected as the male parents for crosses with L. esculentum Mill. in order to produce interspecific hybrids. Crossability barriers between these two distantly related species were circumvented by ovule culture. A total of ten interspecific F₁ hybrid plants were produced. The hybrid nature of the putative F₁ plants was confirmed by a comparison of several morphological characteristics and a PCR-based assay. Eight of ten hybrid plants were backcrossed with L. esculentum to generate a total of 98 BC₁ progeny. Two lines were advanced to the BC₂ and BC₃ levels. Based on these results, ovule culture was found to be an effective method for the production of novel interspecific F₁ and BC₁ plants. This revised version was published online in July 2006 with corrections to the Cover Date.     

Characterization of morphological variation and cold resistance in interspecific somatic hybrids between potato (Solanum tuberosum L.) and S. brevidens Phil.

Summary Somatic hybrids between Solanum tuberosum L. cv. Gracia (2n=4x=48) and Solanum brevidens Phil. (2n=2x=24) were produced via fusion of mesophyll protoplasts. Selection of the protoplast derived putative hybrid calli was based on their vigorous growth. Additive isozyme patterns and chromosome numbers as well as the expression of parental morphological characters have proved the hybrid origin of the selected regenerants. Extensive chromosome loss during the regeneration process resulted in aneuploid hybrids with high frequency. Genomic instability could not be detected in these plants during the period of vegetative propagation. Regenerants from hybrid tissues exhibited wide morphological variation especially in tuber formation. The detailed morphological analysis based on the use of multivariate method (principal component analysis, PCA) enabled to identify morphological groups among the hybrid clones. The positioning of hybrid clones in the PCA space could not be correlated with chromosome numbers. The genomic ratio represented by the tetraploid and diploid parents influenced the morphology of somatic hybrid population according to the applied analytical system. Two selected hybrid clones have exhibited an intermediate degree of frost tolerance compared to the parents, based on the recovery of plants from lower buds after freezing of potted plants.