Congruity backcrossing as a means of creating genetic variability in self pollinated crops: seed morphology of Phaseolus vulgaris L. and P. acutifolius A. Gray hybrids

Summary Seeds of early generations of three reciprocal congruity-backcross (CBC) pedigrees, developed by backcrossing Phaseolus vulgaris-P. acutifolius hybrids to each of the parent species in alternate generations, exhibited a preponderance of traits (size, shape, color, and pattern) of the cytoplasmic parent. The large size of Red Cloud (V₁), the P. vulgaris parent common to all of the pedigrees, dominated pedigrees with V₁ as the cytoplasmic parent, while the small size and rounded or square shapes of the tepary parents, wild P. acutifolius var. acutifolius PI 263590 or G400445 (A₁₉), wild P. acutifolius var. latifolius PI 406622 (A₁₀), or cultivated P. acutifolius var. latifolius Serowi PI 319443 (A₉), were the majority phenotypes when P. acutifolius was the cytoplasmic parent. Continuing through the second cycle of CBC, that is the second backcross with each of the parent species or the fourth backcross, began an amelioration of the apparent cytoplasmic effect on gene expression, as reciprocal pedigrees became more alike, usually with intermediate expression of parental traits or the appearance of new traits. The large seed size of V₁ was recovered in hybrids with P. acutifolius cytoplasm and the kidney shape of V₁ became rare in hybrids with P. vulgaris cytoplasm. Although the tepary-bean parents represented two subspecies and both cultivated and wild P. acutifolius, the three sets of reciprocal-hybrid pedigrees with P. vulgaris Red Cloud are surprisingly similar. It may be that the exotic parent used to develop a CBC pedigree should be selected more for combining ability in the interspecific cross than for specific economic traits. While the number of generations (six or more) required to produce fertile, intermediate CBC hybrids (that did not require embryo rescue) may preclude routine use of this method by practical plant breeders, the crossability of advanced hybrids with both parental species and the amount of variability apparent in advanced-hybrids progenies suggests that CBC would be valuable for maintaining exotic germplasm in immediately useful forms.     

Genetic Control of Abnormal Seedling Development in Phaseolus vulgaris L. and P. vulgaris×P. acutifolius A. Gray Hybrids

Abnormal hybrid seedling development hinders attempts to introgress diverse germplasm in common bean (Phaseolus vulgaris L.) breeding programs. In this study, the hypothesis that DL₁ and DL₂ loci control abnormal seedling development both in Phaseolus vulgaris L. intraspecific hybrids and in P. vulgaris×P. acutifolius A. Gray interspecific hybrids was tested. Hybrids from crosses between six P. vulgaris lines (two Andean dl₁ d₁, DL₂DL₂, two Mesoamerican DL₁DL₁ dl₂ dl₂, and two facilitators dl₁dl₁, dl₂dl₂) and P. vulgaris cultivar Ex Rico 23 developed normally, indicating that Ex Rico 23 has the facilitator genotype dl₁, dl₁, dl₂dl₂. Previous studies have shown that Ex Rico 23 ×P. acutifolius results in abnormal seedlings, but ‘ICA Pijao’, also dl₁, dl₁ dl₂dl₂, ×P. acutifolius results in normal seedlings. Neither the Andean nor the Mesoamerican lines crossed successfully with P. acutifolius, but crosses between one of the two facilitators (G3807) and P. acutifolius resulted in normal seedlings. These results support the hypothesis that different loci control intra- and interspecific hybrid seedling development.     

Genotypic diversity enhances recovery of hybrids and fertile backcrosses of Phaseolus vulgaris L. × P. acutifolius A. Gray

Summary Homozygous and heterozygous Phaseolus vulgaris and P. acutifolius parental genotypes were hybridized to determine if genotypic diversity would aid gene transfer between these species. Certain P. vulgaris female parents resulting from diverse intraspecific crosses increased the frequency of species hybrids obtained. From a total of 19 self-sterile hybrids, 20 backcross-1 (BC₁) lines (P. vulgaris recurrent parent) were produced from 4 partially female-fertile hybrids, each of which had a heterozygous P. vulgaris female parent. Heterozygous P. acutifolius male parents did not influence the frequency at which interspecific hybrids could be produced but apparently improved female-fertility. Fertility of the F₁BC₁ generation was highly variable. The frequency of fertile individuals increased in each subsequent backcross generation (F₁BC₂ and F₁BC₃).Purdue University Agricultural Experiment Station Publication 9946.     

Simple Genetic Control of Hybrid Plant Development in Interspecific Crosses between Phaseolus vulgaris L. and P. acutifolius A. Gray

Crosses were performed between nine Phaseolus vulgaris lines (as females) and seven P. acutifolius lines (as- male to examine parental compatibility for the production of vigorous hybrid And backcross plants, in vitro embryo rescue techniques were required to secure hybrid and backcross proseny following interspecific crossing. Seedling development appeared to be dependent on which allele the P. vulgaris parent carried at an interspecific incompatibility locus. Seven of the nine P. vulgaris lines tested carried an allele at this locus which interacted with a nuclear factor in the P. acutifolius genome resulting in stunted, sub-lethal hybrids. The lines, ICA pijao' and ‘Sacramento Light Red Kidney’ did not carry this allele and produced vigorous hybrid progeny in combination with all P. acutifolius parents. Intensive backcrossing produced progeny which also segregated for sub-lethal and viable plant development. The observed segregation patterns suggest that a bridge crossing scheme would facilitate the introgression of P. acutifolius germplasm into incompatible P. vulgaris lines. Similarities, with an intraspecific incompatibility system are discussed.     

Attempts to overcome postfertilization barrier in interspecific crosses of the genus Lens

Wild relatives are a potential source of genetic diversity to lentil (Lens culinaris Medik). The objective of this research was to obtain viable interspecific hybrids between the domesticated lentil and its wild relatives. The paper details the results of a number of interspecific crosses among L. culinaris, L. orientalis, L. odemensis, L. ervoides and L. nigricans. Viable hybrids were produced between L. culinaris × L. orientalis, L. culinaris × L. nigricans, L. culinaris × L. ervoides and between L. culinaris × L. odemensis. Further viable hybrids were obtained between L. culinaris and L. ervoides, which have the potential to be a ‘bridge’ in hybridization to L. culinaris for specific L. nicrigans lines which proved recalcitrant in L. culinaris × L. nigricans crosses. This is the first time that four wild species of lentils have been used successfully in hybridization with cultivated lentils, and viable hybrids produced. This paper also suggests that the artificial supplement of GA₃, hormone is needed after fertilization for the normal growth of the hybrid embryo, possibly as the natural GA₃ production is restricted with alien pollinations in cultivated lentils in both F₁ and backcross hybrids.     

Introgression of Phaseolus acutifolius A. Gray Genes into the Phaseolus vulgaris L. Genome

The tepary bean (Phaseolus acutifolius A. Gray) is a desirable genetic resource for incorporation of improved disease, pest, and stress resistance into common bean (P. vulgaris L.). Reproductive barriers separate the two species and the degree to which tepary genes may be introgressed into the common bean genome has not been well described. Greenhouse studies of gene introgression through recurrent back-crossing to common bean were performed using two first backcross (BC₁) and nine second backcross (BC₂) populations. The truncate primary leaf characteristic of tepary bean was readily observed but the short primary leaf petiole and narrow bract tepary traits were infrequently observed in both BC₁ and BC₂ populations. In one BC, population high frequencies of adaxial stomata (characteristic of P. acutifolius) and the presence of a 30 kD cotyledon polypeptide from P. acutifolius were also observed; however, a diapho-rase isozyme from P. acutifolius appeared to be eliminated from BC₁ progeny at a high rate whereas 6-phosphogluconate dehydrogenase alleles appeared to be transmitted normally. The expression of tepary primary leaf truncate morphology and bract width were correlated with decreased fertility in one of the two BC₁ populations. Given suitable parental genotypes and population sizes it should be possible to transfer genes from P. acutifolius to P. vulgaris, especially in regions of the genome which are not associated with inviability or sterility. It will be difficult to transfer factors from certain regions of the tepary genome which are preferentially eliminated during introgression.     

Transfer of powdery mildew resistance from Brassica carinata to Brassica oleracea through embryo rescue

Interspecific hybrid plants and backcross 1 (BC₁) progeny were produced through sexual crosses and embryo rescue between Brassica carinata accession PI 360883 and B. oleracea cvs Titleist’and‘Cecile’to transfer resistance to powdery mildew to B. oleracea. Four interspecific hybrids were obtained through application of embryo rescue from crosses with B. carinata as the maternal parent, and their interspecific nature confirmed through plant morphology and random amplified polymorphic DNA (RAPD) analysis. Twenty‐one BC₁ plants were obtained through sexual crosses and embryo rescue although embryo rescue was not necessary to produce first backcross generation plants between interspecific hybrids and B. oleracea. All interspecific hybrids and eight of the BC₁ plants were resistant to powdery mildew.     

Overcoming unilateral breeding barriers between Lycopersicon peruvianum and cultivated tomato,Lycopersicon esculentum

Summary Treatment of Lycopersicon peruvianum stigmas with an artificial medium analogous to stigmatic exudate allowed pollen germination and growth on immature pistils. Growth of Lycopersicon esculentum pollen tubes to L. peruvianum ovules, an otherwise incompatible cross, was achieved following such treated bud-pollinations. No plantlets were recovered, although a few embryos from this cross at the globularity heart stage of development were excised at 22 days after pollination, indicating the presence of crossing failures as severe as in the reciprocal cross. Hybrid plants were obtained from the reciprocal cross, using as pollen parent an L. peruvianum line selected for congruity with L. esculentum. Bud pollinations to L. peruvianum, using these interspecific F₁ hybrids as the pollen parent, allowed viable embryo development and plantlet recovery. Resulting backcross plants, which possess approximately 1/4 L. esculentum genome in a L. peruvianum cytoplasm, may facilitate further introgression of the L. esculentum nuclear genes into in foreign cytoplasm.     

Interspecific hybridization between cultivated american species of the genusphaseolus

Summary Attempted interspecific hybridization between the speciesPhaseolus vulgaris, P. coccineus, P. acutifolius andP. lunatus has indicated that a close genetic relationship exists betweenP. vulgaris andP. coccineus. Viable hybrids of varying fertility can be made reciprocally between these species. Affinity is indicated withP. acutifolius by the formation of viable but sterile hybrids. Less affinity is apparent between these three species andP. lunatus with which they do not produce viable interspecific hybrids. Nevertheless morphological resemblances between the four species justify their retention within the same section of the genus.Effects of cytoplasmic constitution on both fertility and viability were shown in the F₂ and backcross progenies ofP. vulgaris () ×P. coccineus () hybrids and on viability in theP. vulgaris () ×P. acutifolius () and reciprocal hybrids. In slightly fertileP. vulgaris () ×P. coccineus () hybrids it was concluded that distorted F₂ segregations were due to differential loss ofP. coccineus genes in sporogenesis and embryogenesis occurring in aP. vulgaris plasmon.Formerly at The School of Agriculture, University of Cambridge.     

The transfer of triazine resistance from Brassica napus L. to B. oleracea L. II. Morphology,fertility and cytology of the F1 hybrid

Summary F₁ hybrids of triazine resistant Brassica napus and triazine susceptible B. oleracea were morphologically intermediate to the parent species. Of 49 hybrids examined, 44 had 28 chromosomes, two had 37, one had 38 and two had 56. The 38-chromosome plant was thought to be a matromorph, the others, A₁C₁C (28), A₁C₁CC (37) or A₁A₁C₁C₁CC (56) type hybrids. Pollen stainability averaged 9.0% in the sesquidiploid, 32.0% in the tetraploids and 89.5% in the hexaploids. All the interspecific hybrids were resistant to 1.0×10^-4 mol L^-1 atrazine. The sesquidiploid hybrids produced gametes with chromosome numbers ranging from 9 to 17 and the tetraploid hybrid gametes had chromosome numbers from 15 to 22. Most hybrids produced self-seed. The partial fertility of these hybrids may permit their backcrossing to one or both parents.     

Pre- and post-fertilization barriers to backcrossing the interspecific hybrid between Allium fistulosum L. and A. cepa L. with A. cepa

Summary To facilitate the introgression of desirable traits of Allium fistulosum into the genome of A. cepa, several accessions of the hybrid between these species were pollinated with A. cepa as the recurrent parent, and in vitro ovary and ovule culture were performed to obtain an increase in the recovery of backcross progeny. Compared to the results obtained from seed development in planta, the increase in the number of backcross progeny was generally very limited, and in some cases even a decrease was found. Raising the sucrose concentration in the ovary culture medium resulted in a higher frequency of ovules developing back seed coats but this was not followed by an increase in the number of backcross progeny obtained. Pollen tube growth of A. cepa was disturbed in the styles of the interspecific hybrids. Per ovule, frequencies of micropylar penetration exceeded frequencies of backcross progeny only to a limited extent. Hence, it was concluded that in the tested interspecific hybrid accessions the attainable gain in viable backcross progeny by the application of in vitro culture techniques is limited by strong pre-fertilization barriers acting at the level of stylar incongruity.     

Results of crossing Brassica pekinensis (Lour.) Rupr. with B. Oleracea L. Var. Acephala Dc.

Success in obtaining an interspecific hybrid between B. pekinensis and B. oleracea var. acephala was possible through the use of the Chinese cabbage as the seed parent and the use of the growth regulator, N-m-tolylphthalamic acid. The hybrids appeared vegetatively intermediate between the parents except for the inflorescence which appeared more like the kale parent. The F₁ was backcrossed with ease to the Chinese cabbage parent using the hybrid either as the pollen or seed parent. No backcross was made to the kale parent. Segregation in the F₂ was continuous for most characters studied.Michigan Agricultural Experiment Station, Journal Article Number 2588.     

Hybridization of Sinapis alba L. and Brassica napus L. via Embryo Rescue

Embryo rescue techniques were used to obtain hybrids between Sinapis alba L. (white mustard) and Brassica napus L. (oilseed rape) with the goal of improving the disease tolerance of oilseed rape. Hybrid plants with 31 or 43 chromosomes were only recovered, when S. alba, was used as the female parent. One hybrid was obtained from the cross S. alba L. cv. ‘Kirby’×B. napus L. cv. ‘Topas’, while 26 hybrids were obtained, when various S. alba L. cultivars were pollinated with the rapid cycling B. napus line CrGC 5006. All F₁, hybrid plants were male sterile; however, the first generation backcross to B. napus L., also obtained by embryo rescue, produced plants with 50 chromosomes and 61–84 % pollen viability. Second backcross generation seed was produced by normal sexual crossing. Preliminary cytological analyses of pollen mother cells of hybrid plants suggests the possibility of genetic exchange between the two species.     

Heterosis for symbiotic nitrogen fixation in Phaseolus interspecific hybrids

Summary Interspecific hybrids of Phaseolus vulgaris and Phaseolus acutifolius or Phaseolus filiformis have been evaluated for evidence of heterosis for symbiotic nitrogen fixation activity. Hybrids showing composite leghemoglobin profiles were selected for comparison. Rooted cuttings from hybrids and their respective parents were used to estimate nitrogen fixation (acetylene reduction) rates and to determine nitrogen accumulation during the growth period. Some hybrids of Phasolus vulgaris × Phaseolus acutifolius and Phaseolus vulgaris × Phaseolus filiformis had significantly (P<0.01) higher nitrogen fixation rates and in the latter case, accumulated more total nitrogen, than either parent. The interspecific hybrids also showed more complex leghemoglobin profiles than either parent.     

Different Behavior of Brassica juncea and B. carinata as Sources of Phoma lingam Resistance in Experiments of Interspecific Transfer to B. napus

With the aim to transfer Phoma lingam resistance into rape, successful interspecific crosses were made between three oilseed rape varieties (Brassica napus) and the resistant species B. carinata and B. carinata. Although both hybrid types B. napus×B. juncea and B. napus×B. carinata showed the same high level of resistance as the respective resistant parent, the resistance could be only transferred from juncea crosses. After three backcross generations, lines morphologically undistinguishable from rape, fertile, and with a high degree of resistance were obtained. The resistance of B. carinata was practically lost in the first backcross. A possible explanation of this different behavior could be a higher recombination between the genomes B and C (juncea crosses) than between B and A (carinata crosses). The: applied embryo culture increased the yield of hybrids and first backcross plants and reduced considerably the generation time.     

Interactions of BA, GA3, NAA, and surfactant on interspecific hybridization of Lycopersicon esculentum × L. chilense

Time consuming and expensive tissue culture type techniques are currently used for achieving Lycopersicon esculentum × L. chilense interspecific hybrids. The objective of this study was to determine if the number of viable hybrid seed produced directly from this wide cross could be improved by optimizing female/male parent selection when treated with various hormones in the presence or absence of a surfactant (Triton X-100). Individual or combination treatments of BA(6-benzylaminopurine), GA₃ (gibberellic acid), and/or NAA(α-naphthaleneacetic acid) were applied (both with and without surfactant)to the ovaries of two L. esculentum female parents (Fla 7613 and89S) one day following pollination by nineL. chilense accessions. Hormone applications over 1-, 3-, and 5-dayperiods were made and all treatments were compared to a distilled water control. Across all experiments 21 viable hybrids from 1920 fruits (1.1%) were obtained from2128 pollinations. There were five consistent trends observed across three experiments. First, treatments utilizing NAA produced more hybrids than any other treatment hormone. Second, eight of the total 21 hybrids resulted from five consecutive days of hormone treatments (P-value ≤0.01), and six of those eight were realized when NAA was involved. Third, the use of Triton X-100 in combination with the hormone treatments significantly reduced the number of fruits/pollination (P-value ≤ 0.01), but did not significantly reduce the number of overall viable hybrids (P-value > 0.4). Fourth, the L. chilense male parent LA2759 significantly produced the most interspecific hybrids (P-value ≤ 0.04),seven hybrids from 295 fruits, compared to the male parent LA130 which did not produce any hybrids (from 293 fruits) -other accessions were intermediate. Fifth, nearly ten times more hybrids were produced from crosses with Fla 7613 than with 89S. These experiments provide a foundation for future studies to discern specific concentrations, timing and frequency of application of NAA and possibly, GA₃, to efficiently produce interspecific hybrids of L. esculentum × L. chilense. This revised version was published online in August 2006 with corrections to the Cover Date.     

Interspecific hybridization in sexual diploid Allium senescens var. minor× apomictic tetraploid A. nutans and sexual diploid A. senescens var. minor× apomictic hexaploid A. senescens

Fourteen interspecific hybrids in sexual diploid Allium senescens var. minor× apomictic tetraploid Allium nutans L. crosses, and eight interspecific hybrids in sexual diploid A. senescens var. minor× apomictic hexaploid A. senescens L. crosses were produced. The number of chromosomes was 2n= 24 in interspecific hybrids of diploid × tetraploid, and 2n= 32 in diploid × hexaploid crosses. Triploid and tetraploid interspecific hybrids showed intermediate parental morphological characteristics. Tetraploid interspecific hybrids of A. senescens var. minor×A. senescens crosses formed two groups based on leaf colour and leaf width. Seeds were formed in 11 out of 14 triploid interspecific hybrids under natural conditions. In cytological observations of parthenogenesis, three out of 12 triploid interspecific hybrids and five out of eight interspecific tetraploid hybrids were observed. Parthenogenesis ranged from 26.0% to 86.0% in five tetraploid interspecific hybrids. Non-parthenogenesis to parthenogenesis segregated in a 3:5 ratio in A. senescens var. minor×A. senescens crosses.     

Relationship between molecular marker heterozygosity and hybrid performance in intra- and interspecific hybrids of cotton

The present study was conducted to investigate the relationship between parental molecular marker diversity and hybrid performance in both intra‐ and interspecific hybrids of cotton to evaluate the feasibility of predicting hybrid performance using molecular markers. Three cytoplasmic male sterile (CMS) lines were crossed with 10 restorer lines to produce 22 F₁ hybrids during 2003. Of 22 F₁s, 14 hybrids were intraspecific (Gossypium hirsutum × G. hirsutum) and eight interspecific (G. hirsutum × G. barbadense). These 22 F₁ hybrids and their parents were evaluated for yield and fibre quality traits at Zhejiang University, Hangzhou, China during 2004 and 2005. Genetic distances (GD) among the parents were calculated from 56 random‐amplified polymorphic DNAs (RAPD) and 66 simple sequence repeat (SSR) marker data, and their correlation with hybrid performance and heterosis were analysed. The parents could be discriminated into G. hirsutum and G. barbadense clusters by cluster analysis based on both RAPD and SSR markers data. The correlation (r = 0.503, P ≤ 0.05) was calculated between GD_rapd (GD based on RAPD markers) and GD_ssr (GD based on SSR markers). Correlation of GD with hybrid performance and heterosis differed considerably between intra‐ and interspecific hybrids. The correlation between GD and hybrid performance was non‐significant for most of traits within the hybrids of G. hirsutum species. However, it was significantly and positively correlated for fibre length, fibre strength and elongation in interspecific hybrids. The relationship between GD and heterosis was observed to be positively significant for boll weight within hybrids of G. hirsutum with significant and negative correlations for fibre length and elongation. In conclusion, the power of predicting hybrid performance using molecular markers in cotton is low. But, the relationship between SSR marker heterozygosity and hybrid performance can be used to predict fibre length during interspecific hybrid cotton breeding.     

Development of Yellow Seeded Brassica napus Through Interspecific Crosses

Yellow seeded Brassica napus was developed through interspecific crosses with the two mustard species, B. juncea and B. carinata. The objective of these two interspecific crosses was the introgression of genes for yellow seed colour from the A genome of B. juncea and C genome of B. carinata into the A and C genomes of B. napus, respectively. The interspecific F₁ generations were backcrossed to B. napus in an attempt to eliminate B genome chromosomes and to improve fertility. Backcross F₂ plants of the (B. napus×B. juncea) ×B. napus cross were then crossed with backcross F₂ plants of the (B. napus×B. carinata) ×B. napus cross. The objective of this intercrossing was to combine the A and C genome yellow seeded characteristics of the two backcross populations into one genotype. The F₂ generation of the backcross F₂ intercrosses was grown in the field, plants were individually harvested and visually rated for seed colour. Ninety-one yellow seeded plants were identified among the 4858 plants inspected. This result indicated that the interspecific crossing scheme was successful in developing yellow seeded B. napus.     

The use of ISSR markers to identify Texas bluegrass interspecific hybrids

Seventeen ISSR primers were screened on Texas bluegrass (Poa arachnifera), Kentucky bluegrass (P. pratensis), Canadian bluegrass (P. compressa), Argentine bluegrass (P. ligularis), cv. ‘Sherman’ (P. secunda), putative Texas × Kentucky (TK) (P. arachnifera ×P. pratensis) hybrids and hybrids involving Texas, Canadian, and Sherman bluegrass [(P. arachnifera × P. compressa) × (P. arachnifera ×P. secunda)], to determine whether they could be used to produce robust and reproducible DNA fingerprints and identify interspecific hybrids. Nine of the 17 primers consistently produced robust fingerprints and nine 2‐way primer combinations were also selected. DNA fingerprints were highly reproducible and the majority of the selected primers (16/18) amplified hybrid profiles using two putative TK full‐sib hybrids. Combined with a rapid DNA extraction protocol, the ISSR technique enabled a fast and practical way to detect F₁ interspecific hybrids early in the breeding programme and could also be useful for other applications that require DNA‐based markers.