Performance Studies in F6 Lines of Autotetraploid Fenugreek

Hybridization followed by pedigree selection was carried out between two autotetraploid strains of fenugreek (Trigonella foenum-graecum L.). Morphological and cytological investigations were carried out in two autotetraploid parents, their F₁ hybrids and F₆ lines derived from F₁ after selfing. Cytomorphological data of F₆ lines were compared with those of F₁, hybrids and of their parents. The hybrid plants were intermediate with respect to different vegetative traits, however, they showed increased seed set as compared to their autotetraploid parents. The mean pod number per plant in F₆ was one and half times as high as one of the high yielding tetraploid parents and approximately twice as high as that of F₁ hybrid. Meiosis in F₆ was more regular than in F, and the parents. Thus, hybridization followed by pedigree selection is helpful not only in regularizing the meiotic behavior, but vigour and seed fertility have also improved in autotetraploid fenugreek.     

Evaluating the utility of Arabidopsis thaliana as a model for understanding heterosis in hybrid crops

Despite the phenomenon of heterosis having been used to improve crop productivity for almost a century, there is little understanding of the molecular mechanisms involved. Heterosis has been described recently in the widely used plant model Arabidopsis thaliana. To assess the opportunity afforded by this system, we have developed, through pollination to a male sterile female parent (Ler ms1), a range of hybrids of A. thaliana accessions. We tested this method of hybrid production in A. thaliana by comparing seed produced by natural pollination and hand pollination of emasculated plants. Our results demonstrate that using male sterile lines, which more closely represent methods used for hybrid crop production, circumvents the problems associated with the analysis of hybrids produced from emasculated plants. We identified hybrid combinations exhibiting mid-parent heterosis for vegetative fresh weight ranging from −9 to 103% and characterised heterosis in the reciprocal hybrids of one strongly heterotic combination. We found no evidence of a relationship between the extent of genetic relatedness of A. thaliana accessions and the strength of heterosis exhibited by their hybrids. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

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.     

Cytogenetical and molecular investigations on somatic hybrids of Sinapis alba and Brassica juncea and their backcross progeny

Somatic hybrids of Sinapis alba+Brassica juncea (Sal Sal AABB) were synthesized by protoplast electrofusion. They were true genomic allopolyploids since they possessed 60 chromosomes, i.e. the sum of S. alba (2n= 24) and B. juncea (2n= 36) chromosomes. Chromosome pairing was predominantly bivalent with the occasional occurrence of multivalents in the pollen mother cells at diakinesis and metaphase I. Hybrids were completely pollen-sterile, but produced seeds on back-crossing with B. juncea and B. campestris. A total of 37 BC₁ plants were raised from two somatic hybrids (JS-1 and JS-2) and 24 of these were analysed cytologically. The 22 plants originating from the pollinations of somatic hybrids with B. juncea showed a chromosome configuration of 18II+12I and had 42–86% pollen fertility. Two plants from the backcrosses of the somatic hybrid with B. campestris formed 10II +20I, and had 0–4% fertile pollen. Total DNA analysis by probing with pTA71 carrying a full-length 18S–25S rDNA fragment of the wheat nuclear genome revealed that the two somatic hybrids possessed all the characteristic bands of both the species, confirming their hybridity. Probing with the mitochondrial coxI and atp9 genes indicated mitochondrial genome recombination in the hybrids. Hybridization with chloroplast-specific psbD indicated that both the somatic hybrids possessed the cp genome of S. alba origin.     

High embryo sac fertility and diversity of abnormal embryo sacs detected in autotetraploid indica/japonica hybrids in rice by whole‐mount eosin B‐staining confocal laser scanning microscopy

An eosin B staining procedure for use with confocal microscopy (WE‐CLSM) was used to examine the mature embryo sacs in 24 typical autotetraploid (2n = 4x = 48) indica/japonica hybrids and to compare with their original diploid (2n = 2x = 24) indica/japonica hybrids in rice. Four of the 24 hybrids showed >80% embryo sac fertility, and 13 hybrids over 70%. The average embryo sac fertility of the 24 autotetraploid indica/japonica hybrids was 68.08%, which was 17.33% higher than that in diploid hybrids. When compared with the diploid hybrids, 79.17% of the autotetraploid hybrids had higher embryo sac fertility. A diversity of abnormal embryo sacs occurred in autotetraploid indica/japonica hybrids. Embryo sac abnormalities were similar for both autotetraploid and diploid hybrid, but their frequencies were different. Some novel abnormal types were found in autotetraploid hybrids, such as enlarged egg apparatus. The embryo sac fertilities and frequencies of various abnormalities in autotetraploid hybrids varied with parental genotype and environment. The average seed set of the autotetraploid hybrids were higher than their diploid hybrids in different growing seasons.     

Agronomic Traits and Chromosome Behavior of Autotetraploid Sorghums

The agronomic performance and chromosome behavior of autotetraploid sorghum [Sorghum bicolor (L.) Moench] inbreds and F₁ hybrids were investigated. Autotetraploids have possible commercial use and can serve as bridging species in crosses to natural tetraploids. Autotetraploid sorghum inbreds (2n = 40) had higher kernel weight, seed yield, and protein and amino acid content than their diploid counterparts. Stalk height and panicle length of the autotet-raploids, however, were not different from those of the diploids, and seed set of autotetraploids was lower in early generations. Seed set of autotetraploid Ft hybrids was related to and higher than that of the autotetraploid parental inbred lines. Frequency of quadrivalents, anaphase I and II laggards, and quartet micronuclei varied among autotetraploids. Frequency of quadrivalents did not appear to be an important factor in seed set nor was it significantly associated with frequency of laggards. Aneuploids (2n = 39, 41) had higher percentages of laggards and micronuclei than did euploids (2n = 40). Methods of pollination significantly affected seed set in autotetraploid S4 lines. Differences in seed set among autotetraploid lines indicated the feasibility of selecting for lines with high seed set and utilizing those lines for hybrid production.     

Relevance of sexual polyploidization for crop improvement – A review

Colchicine induced polyploids have not directly contributed for crop improvement in the past. On the other hand, the so-called natural polyploids, derived from the functioning of numerically unreduced(2n) gametes have been shown to be more relevant for crop improvement in many cases. Different types of cytological abnormalities during meiosis can give rise to 2n gametes and the genetic composition of these gametes is variable. Depending on the type meiotic abnormalities, various types of 2ngametes, such as first division restitution(FDR), second division restitution (SDR),indeterminate meiotic restitution (IMR) and post meiotic restitution (PMR) gametes,among others, have been described in recent years. For the improvement of autopolyploids such as potato, alfalfa,Vaccinium spp., and some of the fodder grasses, FDR gametes have been proved to be highly useful. However, the use of 2n gametes for the improvement of allopolyploid crops has received much less attention so far. Some of the investigations on allopolyploids, derived from Festuca-Lolium, Alstroemeria and Lilium species hybrids, have revealed that 2ngametes can be most useful for the introgression of alien genes and chromosomes into cultivars. An important feature of using sexual polyploidization in the case of allopolyploids is that introgression can be achieved through recombination due to genetic crossing-over between alien chromosomes as well as addition of alien chromosomes, which is extremely difficult or impossible to achieve in the case of colchicine induced allopolyploids. Because of the recent developments in the field of plant molecular biology, methods have become available for the analysis of 2ngametes and sexual polyploid progenies more accurately and to develop systematic breeding approaches. The methods include DNA in situ hybridization (GISH and FISH)and molecular mapping (AFLP, RFLP, RAPDs).In addition to providing basic information on the genetic and genome composition of the polyploid progenies, these methods can be potentially useful for a more efficient creation of desirable breeding material and cultivars. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cytological analyses of hybrids and derivatives of hybrids between durum wheat and Thinopyrum bessarabicum,using multicolour fluorescent GISH*

The wheat progenitors and other wild relatives continue to be important sources of genes for agronomically desirable traits, which can be transferred into durum wheat (Triticum turgidum; 2n = 4x = 28; AABB genomes) cultivars via hybridization. Chromosome pairing in durum × alien species hybrids provides an understanding of genomic relationships, which is useful in planning alien gene introgression strategies. Two durum cultivars, ‘Lloyd’ and ‘Langdon’, were crossed with diploid wheatgrass, Thinopyrum bessarabicum (2n = 2x = 14; JJ), to synthesize F₁ hybrids (2n = 3x = 21; ABJ) with Ph1. ‘Langdon’ disomic substitution 5D(5B) was used as a female parent to produce F₁ hybrids without Ph1, which resulted in elevation of pairing between durum and grass chromosomes – an important feature from the breeding standpoint. The F₁ hybrids were backcrossed to respective parental cultivars and BC₁ progenies were raised. ‘Langdon’ 5D(5B) substitution × Th. bessarabicum F₁ hybrids were crossed with normal ‘Langdon’ to obtain BC₁ progeny. Chromosome pairing relationships were studied in F₁ hybrids and BC₁ progenies using both conventional staining and fluorescent genomic in situ hybridization (fl‐GISH) techniques. Multicolour fl‐GISH was standardized for characterizing the nature and specificity of chromosome pairing: A–B, A–J and B–J pairing. The A–J and B–J pairing will facilitate gene introgression in durum wheat. Multicolour fl‐GISH will help in characterizing alien chromosome segments captured in the durum complement and in their location in the A and/or B genome, thereby accelerating chromosome engineering research.     

Cytogenetic study of Helianthus laevigatus and its F1 and BC1F1 hybrids with cultivated sunflower, Helianthus annuus

Although the wild sunflower species Helianthus laevigatus has not been extensively studied it may be considered for sunflower breeding as a potential source of desirable genes for Sclerotinia stalk rot resistance and high contents of proteins and linoleic acid in the seed. A set of six H. laevigatus populations was crossed to cultivated sun~ower lines and produced nine F₁ (2-14 plants) and 66 BC₁F₁ hybrid combinations (1-13 plants). Male sterility occurred in F₁ and BC₁F₁ hybrid combinations and pollen viability was lower in the progenies than in the parents (51.6-77.2%in F₁ and in F₁ and 4.8-34.0% in BC₁F₁). Meiosis was normal in the H. laevigatus populations It was found that this tetraploid species also occurred in a hexaploid form Numerous irregularities were observed in the meiosis of the F₁ interspecific hybrids During diakinesis, quadrivalents and hexavalents were recorded in addition to bivalents Dislocated chromosomes and chromosome bridges were present in the other phases The chromosome number in F₁ was 68 (tetraploid). Irregularities in chromosome pairing were observed in the interspecific hybrids at BC₁F₁. There were many univalents, and trivalents quadrivalents and hexavalents were also present The chromosome number in the BC₁F₁ generation ranged from 34 to 60. The occurrence of meiotic irregularities in the F₁ and BC₁F₁ interspecific hybrids indicates that H. laevigatus and the cultivated sunflower differ in genome constitution.     

Rapid Incorporation of D-genome Chromosomes into A and/or B Genomes of Hexaploid Triticale

Two series of progenies were developed from hybrids between octoploid (AABBDDRR) and tetraploid triticale ((AB)(AB)RR). One arose from the successive selfing of the F₁s, while the second was established after one backcross of the F₁ hybrids with the respective 8 × triticale parent. Altogether, 250 F₃ and BC₁F₂ lines were developed, of which 112 were karyotyped in the F₄/F₅ or in BC₁F₃/BC₁F₄ generations using C-banding and SDS-PAGE. The 112 lines represented 61 different karyotypes, of which 39 appeared to be stabilized, having pairs of homologous wheat chromosomes only, while 22 karyotypes exhibited 1—3 heterologous pairs. The frequency of karyotypically stabilized lines originating from the series with one backcrossing was much higher (79.5 %) than those derived from the successive selfing of the F₁ hybrids (51.7%). Six lines had the pure hexaploid triticale chromosome constitution. The frequency of disomic substitutions of D genome chromosomes for their homoeologous A and/or B genome chromosomes ranged from one to six per line with an average of 1.7. Except for 3B and 6B all possible D(A/B) substitutions were obtained. Chromosomes ID and 3D substituted for their homoeologues with the highest frequency, while the substitution of chromosome 4D for 4A or for 4B was the least frequent. D(R) substitutions were found in eight lines only. A complete set of 6x triticale lines was established in which chromosome ID was present in all possible combinations, i.e. single 1D(1A/1B/1R) disomic substitutions as well as disomic ID addition.     

The Diploidised Meiosis of Tetraploid Beta macrocarpa and its Possible Application in Breeding Sugar Beet

Accessions of Beta macrocarpa Guss., collected on the Canary Islands, were tetraploid (2n = 36). Three of these accessions were studied in detail. The plants were rather uniform in their morphological appearance, both within and between accessions, and were annual and fully self-compatible. Meiosis was completely diploidised, suggesting an alloploid nature of the tetraploid species. Crosses with diploid B. vulgaris yielded triploid hybrids which were sterile or nearly so; a few descendants of such hybrids were highly aneuploid. Crosses between tetraploid B. macrocarpa and autotetraploid cytotypes of B. vulgaris showed variable results, only part of these crosses yielded tetraploid hybrids. The tetraploid hybrids exhibited somewhat higher fertility than the triploids. An F₂ generation showed partial hybrid dwarfness, partial fertility and segregation for carliness and coloration of the hypocotyl. All hybrids had multivalents at meiosis (averages: 2.5—5.4 multivalents per pollen mother cell), indicating suppression of the diploidised meiosis. The possibilities of application of the diploidised meiosis in breeding sugar beet are discussed.     

Potato Virus Y Resistance Gene, Ry chc, Mapped to the Distal End of Potato Chromosome 9

Summary A Japanese leading cultivar ‘Konafubuki’ has an extreme resistance gene Ry _chc to Potato virus Y (PVY) in a simplex fashion. To reduce complexity of the highly heterozygous autotetraploid genome and map the Ry _chc gene, haploid plants were parthenogenetically induced from ‘Konafubuki’. The ratio of 7 resistant: 5 susceptible haploids confirmed the single dominant, simplex genotype of ‘Konafubuki’. A selected resistant haploid 98H20-5 was crossed with a susceptible diploid, resulting in 119 F ₁ plants. Using RFLP and RAPD markers, Ry _chc was mapped to the most distal end of the chromosome 9 where the recovery of recombinant genotypes was extremely reduced in this mapping population possibly due to the linked inferiority factor. The location of Ry _chc is different from those of the other extreme resistance genes to PVY, but possibly resides in one of resistance gene clusters.     

Intergeneric hybridization of Diplotaxis erucoides with Brassica napus. I. cytogenetic analysis of F1 and BC1 progeny

Summary Intergeneric hybrids (F₁) Diplotaxis erucoides (DeDe) x Brassica napus (AACC) and the first backcross to B. napus (BC₁) have been obtained through in vitro culture of excised ovaries. The chromosome numbers of F₁ and BC₁ plants proved the occurrence of unreduced gametes. The study of metaphase I chromosome pairing showed that autosyndesis in De genome and allosyndesis between De and A/C genomes might exist. The male fertility of the F₁ plants was low. Some male-sterile plants were found in F₁ and BC₁ progeny. The possibilities of creating addition lines B. napus-D. erucoides and of obtaining a new cytoplasmic male sterility in B. napus are discussed.     

QTL mapping of fiber quality in an elite hybrid derived-RIL population of upland cotton

Xiangzamian 2 (XZM2) was the most widely cultivated cotton hybrid planted as F₁ hybrids and as selfed F₂ seeds in China before the release of transgenic Bt hybrids. By crossing two parents of XZM2, Gossypium hirsutum cv. Zhongmiansuo12 (ZMS12) and G. hirsutum acc. 8891, and through subsequent selfings, we obtained F₈ and F₉ populations of 180 recombinant inbred lines (RILs). A RIL population was cultivated in two cotton-growing regions in China for 2 years. The purpose of the present research was to detect quantitative trait loci (QTL) for fiber quality and provide information applicable to cotton breeding. A genetic map was constructed mainly using SSR markers. QTL controlling fiber quality traits were determined at the single-locus and two-locus levels, and genotype-by-environment interactions were analyzed. Among the main-effect QTL, a fiber length QTL qFL-D2-1 and a reflectance QTL qFR-D2-1 were simultaneously detected at two growing regions in 2 years, which suggested a high degree of stability in different environments, and might be of particular value for a marker-assisted selection (MAS) program. The results suggested that epistatic effects, as well as additive effects, of QTL play important roles in fiber quality in these RILs. In our research, the phenomenon of QTL clusters was detected in the cotton genome.     

Moricandia arvensis cytoplasm based system of cytoplasmic male sterility in Brassica juncea: Reappraisal of fertility restoration and agronomic potential

Cytoplasmic male sterility (CMS) system based on the cytoplasm from Moricandia arvensis (mori) was investigated for fertility restoration and agronomic potential. Fertility restorer gene for mori CMS was introgressed from cytoplasm donor species as all the evaluated Brassica juncea genotypes (155) acted as sterility maintainers. The allosyndetic pairing between M^a and the A/B genome chromosomes in the monosomic addition plants (2n= 18II + 1M^a) facilitated the gene introgression. Partial fertility restoration (43–52% pollen grain stainability) in F₁ hybrids and absence of segregation for male sterility in F₂ progenies suggested gametophytic control of fertility restoration. The pollen fertility in the F₁ hybrids was, however, sufficient to ensure complete seed set upon bag selfing. Introgression from M. arvensis also helped in correction of chlorosis associated with mori cytoplasm in CMS and fertile alloplasmic B. juncea plants. Yield evaluation of thirty F₁ hybrids having the same nuclear genotype but varied male sterilizing cytoplasms (mori, oxy, lyr, refined ogu), in comparison to respective euplasmic hand bred control hybrids, allowed an estimate of yield penalty associated with different CMS systems. It ranged from 1.8% to 61.6%. Hybrids based on cytoplasmically refined ogu were most productive followed by those based on cytoplasmically refined mori CMS. The male sterility systems emanating from somatic hybridization were found superior than those developed from sexual hybridization.     

The Effect of A Genome Substitution on the Resistance of Brassica napus to Infection by Leptosphaeria maculans

Six accessions belonging to four subspecies of Brassica rapa, including three accessions of B. rapa subsp. sylvestris, were crossed with B. oleracea subsp. alboglabra in order to develop a series of synthetic B. napus lines with a common C genome but contrasting A genomes. Different A genomes had significant effects on the efficiency of B. napus resynthesis and the sexual compatibility of the synthetic lines with oilseed rape cultivars. The synthetic lines were used to investigate the effect of A genome substitution on the resistance of B. napus to infection by Leptosphaeria maculans, and to explore the potential for the use of wild forms of B. rapa in oilseed rape breeding programmes. Synthetic lines derived from two wild accessions of B. rapa, and their F₁ hybrids with oilseed rape cultivars, expressed high levels of resistance to L. maculans in glasshouse experiments. One of these lines also expressed high levels of resistance in field experiments in England and Australia when exposed to a genetically diverse pathogen population. All other synthetic lines and cultivars were highly susceptible in both glasshouse and field experiments. F₁ hybrids between oilseed rape cultivars and synthetic lines derived from B. rapa subsp. chinensis were significantly more susceptible than either parent.     

Hybrids of <Emphasis Type="Italic">Avena sativa</Emphasis> with two diploid wild oats (CIav6956) and (CIav7233) resistant to crown rust

Two diploid accessions of wild oat, CIav6956 and CIav7233, were identified as carrying seedling resistance to oat crown rust (caused by Puccinia coronata f. sp. avenae; Pca). Two vigorous interploidy F₁ hybrids were generated from crosses involving the hexaploid oat cultivar Wintaroo and the diploid oat Avena strigosa Schreb. accession CIav6956. An additional interploidy F₁ hybrid, designated “F1-Aa1”, was produced from a cross of Wintaroo and the diploid oat accession CIav7233. All three hybrids were more vigorous and taller than the cultivated parent Wintaroo. The three F₁ hybrids contained full chromosome complements from both parents (2n = 4x = 28), but no seeds were obtained when the three F₁ hybrids were selfed. Meiotic analyses of the hybrids indicated that they exhibited a high degree of inter-genome and intra-genome pairing. Trivalent configurations were detected in 95–96% of meiotic cells and a minimum of three bivalents was present in all cells. An average chiasma frequency of 7.2–7.9 per cell was observed for the three F₁ hybrids. A fourth F₁ hybrid was subsequently generated from a cross between the diploid oat accession CIav7233 and Wintaroo. One octaploid (2n = 8x = 56) was generated from this hybrid and progeny were resistant to two Pca races. The chromosome number of the octaploid progeny varied between 51 and 54 chromosomes. Development of a chromosome addition line(s) with the crown rust resistance should be possible from these partial-octaploids.     

Sexual transfer of Arabidopsis DNA to Brassica napus

Out of 400 reciprocal crosses made between Brassica napus and Arabidopsis thaliana, 200 maternal B. napus flowers produced 31 seeds and 10 seeds developed when A. thaliana was used as the female parent. Of the latter, three seeds successfully germinated but did not develop into mature plants. In contrast, plants from all 32 seeds generated from B. napus were established. DNA from these 31 plants was analysed by 18 probes mapped to the A. thaliana genome, two selectable marker genes and one A. thaliana species-specific repetitive probe. It was found that one plant had identical restriction fragments to A. thaliana with two of the 21 probes used. The gene flow within Brassicaceae is discussed in light of this result.     

Progenies of an interspecific hybrid betweenArachis hypogaea and A.stenosperma — pest resistance and molecular homogeneity

Summary Interspecific triploid hybrids were obtained betweenArachis hypogaea L. andA. stenosperma Krapov. and W.C. Gregory by adopting hybridization coupled with rescue of the developing hybrid embryos. Two hexaploid hybrid populations were generated from triploids, somatically doubled (SD) and sexually polyploidized (SP) hexaploids. Microscopic screening for the occurrence of 2n gametes in triploid hybrids was useful to predict the production of spontaneous hexaploids. In order to facilitate maximum intergenomic recombination, the hexaploids were allowed to self for several generations (F₄) in the greenhouse. Prolific vegetative growth, pollen stainability, and seed set were observed to decline with each selfed generation. Individuals of the F₃ generation from the two hexaploid populations were evaluated for resistance to nematode (Meloidogyne arenaria Chitwood, race 1) and late leafspotCercosporidium personatum (Berk. & Curt.) Deighton under greenhouse conditions. Both SD and SP populations performed significantly better than their cultivated parent, Sunbelt Runner, and a susceptible cultivar, Florunner. At the DNA level, no significant differences were detected among hybrid individuals using cDNA clones and RAPD primers polymorphic for the two parents. Seven cDNA clones were used to probe DNA from 17 F₂ individuals and two parents and 45 RAPD primers were used to amplify DNA from 21 F₂ and F₃ individuals. No significant differences in banding patterns were observed among hybrid individuals which suggested that little or no detectable intergenomic recombination had occurred.     

Crosses between Beta vulgaris L. and Beta lomatogona F. et M.

Summary A population of 2x^ms sugar beets was crossed with 4x Beta lomatogona F. et M. The 3x F₁-plants were male sterile and were backcrossed with 2x and 4x sugar beets and multiplied without pollination as well. After the 1st backcross mainly 3x apomict types arose again and, among others, a small number of successful 4x backcrosses. After pollination by 4x sugar beets this 4x F₁ B₁ produced. besides predominatly apomictically multiplied 4x plants, also about 7% haploid 2x hybrids. The latter probably possess 1 genome from B. vulgaris and 1 genome from B. lomatogona. In the meiosis of the PMC's a certain amount of homeology between a number of chromosomes of both species could be established. The amphihaploid hybrids can be used as breeding parents for the creation of types in which introgession can occur. During hybridization in addition to 2x and 4x B. vulgaris types a number of 2x-, 3x-, 5x- and 6x-hybrids arose. This is presumably caused by the presence of gametes with the somatic number of chromosomes and the occurrence sometimes of haploid apomictic multiplication.The presence of large numbers of bolters in the F₁ and F₁ B₁ suggests that the bolting tendency of both species is based on different genes.