Investigations on the breeding behaviour of aneuploids of guava

Summary The breeding behaviour of aneuploids of guava (Psidium guajava L.) such as trisomic, tetrasomic, and higher aneuploids has been studied. Reciprocal crosses between aneuploids and diploids indicated less than 100% crossability. The aneuploids when used as male parents crossed less frequently than as female parents and certain aneuploids crossed more readily than others. Differences were observed in fruit size, fruit weight, and seed number in the reciprocal crosses.The extra chromosome was found to be transmitted through both the egg cell and the pollen. However, the frequency of transmission was greater through the egg cell than the pollen. As high as 26% transmission of extra chromosomes were obtained through the egg cell. There was no clear cut difference between trisomics and higher aneuploids with regard to the frequency of transmission of extra chromosomes.     

The identification of faba bean (Vicia faba L.) trisomics by translocation tester set

Summary Five subtelocentric primary trisomics of Vicia faba L. were isolated from crosses between asynaptic mutants and normal plants or translocation tester stocks. The extra chromosome of each trisomic was identified cytologically by critical multivalent association at metaphase I of meiosis.     

Cytogenetical studies on African rice,Oryza glaberrima Steud. 3. Primary trisomics produced by pollinating autotriploid with diploid

Summary In order to promote cytogenetical studies on Oryza glaberrima (2n=2x=24), I produced primary trisomics in the progeny of the autotriploid. When the autotriploid was pollinated with the diploid, 136F1 seeds (5.8% of the number of spikelets pollinated) were obtained. The numbers of chromosomes of 31 surviving progeny varied from 24 to 28, with plants having 2n=25 occurring at the highest frequency (34.4%). The primary trisomics differed from one another as well as from the normal disomics in many aspects. The differences were attributed to a single extra chromosome in each trisomic type. Nine primary trisomics were classified into 8 types on the basis of morphological and reproductive features. Seeds were obtained from all primary trisomics except for Type D by self pollination, open pollination or pollination with the disomics. I was able to maintain 7 types of primary trisomics by seed propagation. The primary trisomics which are capable of expressing subtle genetic dissimilarities should be useful for studying the relationship between the two cultivated species of rice at the level of individual chromosomes.     

Basic information for the use of primary trisomics in genetic and breeding research

Summary An attempt is made to arrange the basic information needed for using primary trisomics in localizing single and complementary dominant and recessive genes, accounting for random complete chromatid and random chromosome association, for heterozygosity of the trisomics and for female and male transmission rates. The expected ratios are given separately for the di-, tri- and tetrasomic group in each population. Also the ratios in the total populations have been calculated for different rates of female and male transmission of the extra chromosome. Both the backcross procedure and that of selfing trisomic F₁ plants are considered. Finally the use of trisomics for studying quantitative characters is discussed in connection with the breeding and incompatibility system of the crops and in relation to the frequent morphological differences among trisomics as well as between trisomics and their disomic counterpart.     

Identification of primary trisomics and other aneuploids in foxtail millet

A complete set of nine primary trisomics (2n+ 1) for cv.‘Yugu No. 1’of foxtail millet, Setaria italica (L.) Beauv. (2n= 2x= 18), was identified cytologically from progenies derived from crosses between autotriploids(2n= 2x= 27) and their diploid counterparts. Five autotriploid plants were identified from 2100 seedlings derived from 4x-2x crosses; the reciprocal crosses (2x-4x) failed to produce autotriploids. Autotriploids grew vigorously and were morphologically very similar to diploids. Theprimary trisomics (2n= 2x= 19) constituted ?32.5% of the total progeny from the 3x-2x crosses, whereas 59.8% of the descendants were aneuploids with chromosome numbers ranging from 20 (double trisomics and tetrasomics) to 37 (2n= 4x+ 1; or autotetraploid with one additional chromosome). The nine primary trisomics identified were self-fertile; seven had characteristic morphology, whereas trisomics VIII and IX resembled the disomics. The seed set for trisomic V was the lowest (20%), and trisomic VIII the highest (74%). Other aneuploids with 20 or more somatic chromosomes were either self-sterile or partially fertile with various, but low, levels of seed set. Each of the primary trisomics showed its unique transmission rate when self-pollinated; trisomic IX had the highest (45.8%), whereas trisomic V had the lowest (19.6%) transmission rate.     

Chromosomal location of fertility restoring genes for ‘wild abortive’ cytoplasmic male sterility using primary trisomics in rice

Summary Identification and location of fertility restoring genes facilitates their deployment in a hybrid breeding program involving cytoplasmic male sterility (CMS) system. The study aimed to locate fertility restorer genes of CMSWA system on specific chromosomes of rice using primary trisomics of IR36 (restorer), CMS (IR58025A) and maintainer (IR58025B) lines. Primary trisomic series (Triplo 1 to 12) was crossed as maternal parent with the maintainer line IR58025B. The selected trisomic and disomic F₁ plants were testcrossed as male parents with the CMS line IR58025A. Plants in testcross families derived from disomic F₁ plants (Group I crosses) were all diploid; however, in the testcross families derived from trisomic F₁ plants (Group II crosses), some trisomic plants were observed. Diploid plants in all testcross families were analyzed for pollen fertility using 1% IKI stain. All testeross families from Group I crosses segregated in the ratio of 2 fertile: 1 partially fertile+partially sterile: 1 sterile plants indicating that fertility restoration was controlled by two independent dominant genes: one of the genes was stronger than the other. Testcross families from Group II crosses segregated in 2 fertile: 1 partially fertile+ partially sterile: 1 sterile plants in crosses involving Triplo 1, 4, 5, 6, 8, 9, 11 and 12, but families involving triplo 7 and triplo 10 showed significantly higher X² values, indicating that the two fertility restorer genes were located on chromosome 7 and 10. Stronger restorer gene (Rf-WA-1) was located on chromosome 7 and weaker restorer gene (Rf-WA-2) was located on chromosome 10. These findings should facilitate tagging of these genes with molecular markers with the ultimate aim to practice marker-aided selection for fertility restoration ability.     

Trisomic genetic analysis of aroma in three Japanese native rice varieties (Oryza sativa L.)

Information on the genetics of aroma in rice facilitates breeding and selection of new aromatic varieties with high yield and good quality. Objective of the present study was to make clear the number of genes controlling aroma, and the allelism of aroma genes and the location of aroma gene(s) on the chromosome in three Japanese native aromatic rice varieties (Kabashiko, Shiroikichi and Henroyori). Lack of leaf aroma in all F₁ plants of non-aromatic/aromatic crosses indicated the recessive nature of aroma, and the segregation ratios (3:1) of non-aromatic to aromatic plants in its F₂ populations from Nipponbare/aromatic varieties crosses revealed that each of the three aromatic varieties contains a single recessive gene for aroma. Through trisomic analysis, the segregation of non-aromatic and aromatic plants in all F₂ populations from the crosses between trisomics lines NT8, with an extra chromosome 8, and aromatic varieties deviated significantly from disomic segregation of 3:1 ratios, and fitted to trisomic segregation, however, in other F₂ populations derived from other 7 types of trisomic F₁ plants, the segregation ratios of non-aromatic to aromatic were 3:1, indicating that the single recessive aroma gene was located on chromosome 8 in three aromatic varieties. This revised version was published online in July 2006 with corrections to the Cover Date.     

The first record of tetrasomy in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.)

Tetrasomic plants with two additional small chromosomes were identified, with a frequency about 2.5%, in the trisomic pea line TRUST-R, which normally contains one extra chromosome covering a sporophyte lethal in the regular chromosome set. As compared to trisomics, tetrasomics exhibited an enhanced expression of the traits resulting from extra chromosome addition: slow growth, enlarged bracts, shortened peduncles, wavy leaflets and stipulae. They were almost sterile, their pollen contained a variable proportion of empty grains and some anomalously large, small or deformed grains. In metaphase I, two extra chromosomes did not form a stable bivalent and only in some cases were situated close to each other. In anaphase I, the extra chromosomes migrated independently to either pole or retarded in the equatorial plain, the same was observed for chromatids in anaphase II. This retardation resulted in anomalous cytokinesis, so that triads, dyads and half-divided or non-divided monads appeared. The retarded extra chromosomes may form small extra nuclei either included into one of the microspores or forming a separate miniature cell; in this way tetrasomics may eliminate extra chromosomes. One of the tetrasomics analysed formed an exceptionally high proportion of microspore pentads. In the regluar TRUST-R trisomics, the sole extra chromosome retarded in the equatorial plain in anaphases I and II. The retardation in anaphase II often makes cytokinesis in trisomics (in general more regular than in tetrasomic) to proceed in two steps: at first cell wall formation separates a pollen mother cell into two dyads and then each of them into two microspores.     

Tertiary trisomics in lentil

Summary Tertiary trisomics have been selected among derivatives of hybrids between the cultivated lentil and its wild progenitor Lens orientalis, and between the cultivated species and L. ervoides. Hybrids heterozygotes to a single translocation were more useful as a source of trisomic plants then those heterozygotes to a number of translocations. The trisomic plants were weeker compared to the euploid plants and many have not reached maturity. The meiosis, fertility and transmission rate of the extra chromosome were studied in a single trisomic line.     

Interspecific hybridization between rice bean (Vigna umbellata) and its wild relative (V. minima): Fertility-sterility relationships

Summary The pre-and post-fertilization barriers in the interspecific crosses between Vigna umbellata and V. minima were investigated. In the reciprocal crosses (V. minima as the parent) the entry of pollen tubes into the ovary was delayed by about 4 h, and no seed set was observed. However, no pre-fertilization barriers were encountered in crosses involving V. minima as the parent and V. umbellata as the parent (normal cross). The delay/absence of divisions in the endosperm and the failure of embryo to divide were the post-fertilization barriers responsible for somatoplastic sterility in normal crosses which yielded a few hybrid seeds. The hybrid seeds showed poor germinability. The F₁ hybrids were intermediate between the parents in most morphological characters, and are completely sterile for pollen. Backcrossing of F₁ hybrid with both the parents did not restore fertility in the progenies. V. minima is considered as the tertiary gene pool of the rice bean.     

An intersubgeneric hybrid between Glycine tomentella Hayata and the soybean,G. max (L.) Merr.

Summary The objective of the present paper is to provide information on the morphology and cytology of an intersub-generic hybrid (2n=59) between Glycine tomentella Hayata (2n=78) and G. max (L.) Merr. (2n=40) obtained through in vitro immature seed culture. The hybrid plant was slow in vegetative growth and twinning like the female parent but morphologically was intermediate between both parents for several traits. At metaphase I, the average chromosome associations and ranges for 25 cells were 44.0 I (37–51)+7.5 II (3–11). The plant was completely pollen and seed sterile. The present investigation suggests that wild perennial Glycine species can be exploited as either the male or female parent in wide hybridization programs with the soybean, G. max.     

Identification of seven telotrisomics of rye (Secale cereale L.)

Summary Seven monotelotrisomics emerged spontaneously from primary trisomics of rye cv. Esto. Analyses of karyotype, of chromosome pairing in meiosis and of Giemsa banding pattern were used to identify monotelotrisomics in rye as IRS, 1RL, 3RS, 4RS, 4RL, 6RS and 7RL. Vitality and fertility were better than in primary trisomics. The extra telocentrics were well transmitted through the female gamete. Transmission through the male gamete was in all cases less than trasmission through the female gamete.     

Evidence of a partial reproductive barrier between wild and cultivated pearl millets (Pennisetum glaucum)

Summary The occurrence of seed malformation in association with reduced thousand grain weight and germination ability has been observed in crosses between cultivated female plants and wild male plants. A survey of 16 cultivated accessions (P. glaucum subsp. glaucum) and 11 wild accessions (P. glaucum subsp. monodii) ranging over the whole species diversity showed this postzygotic incompatibility was general, but its intensity varied greatly with the cultivated female accession used and very little with the wild male parent origin. About 15% of the 123 cultivated x wild crosses observed gave normal seeds. Seed malformation has never been observed in crosses between cultivated accessions and appeared independent of genetic distances between the parents. The reciprocal crosses between wild female plants and cultivated male plants gave normal-looking seeds with good germination but consistently reduced thousand grain weight. Both seed malformation and seed small size are an expression of a genetic imbalance. These slight reproductive barriers seem to have been built during the domestication process.Abbreviation ICRISAT International Crop Research Institute for the Semi-Arid Tropics     

A new source of cytoplasmic male sterility in sunflower

Summary Interspecific substitutions of the nucleus of Helianthus annuus (2n=34) cv. Saturn into the cytoplasm of H. petiolaris (2n=34) by successive backcrossing, resulted in progenies with indehiscent anthers containing white, rather than normal yellow, pollen. Further backcrossing by cv. Saturn failed to restore pollen shed, suggesting that the male sterility was cytoplasmic. In vivo germination tests of pollen from 23 such plants from eight BC₅ lines, indicated complete pollen sterility for 14 plants, but normal seed set, suggesting that female fertility was not affected. Meiosis in all plants examined was normal.Crosses between seven sources of pollen-fertility restorer, one collection of wild H. annuus, and an existing source of cytoplasmic male sterility, resulted in a high frequency of plants with normal pollen shed in all F₁ progenies. However, no normal pollen shed was evident in F₁ progenies for similar crosses between BC₅ male-steriles and three of the seven restorer sources, nor for the single wild H. annuus evaluated. The foregoing suggests that the backcross substitution lines are a new source of cytoplasmic male sterility. The inheritance of restoration of pollen shed was complex and not fully elucidated. Some data suggested that two independent, complementary, dominant genes were required, but others indicated two to three independent, dominant genes.     

Cross compatibility between Abelmoschus esculentus and A. moschatus

Interspecific cross compatibility between cultivated and wild okra (Abelmoschus esculentus and A. moschatus) and pollen tube growth behaviour in the crosses among a local cultivar of A. esculentus, A. moschatus and their F₁s were studied. Fruit set was observed in all the crosses except one and seed setting was absent in two of the crosses which set fruit. All seed produced were shrivelled but F₁ plants were obtained from two crosses where cultivated okra was used as the seed parent. The F₁ plants were perennial in nature with very low pollen viability and seed set. A high percentage of pollen germination and profuse pollen tube penetration in the style were observed in the cross A. esculentus × A. moschatus but low pollen tube penetration with abnormal pollen tubes was observed in the reciprocal cross. The number of pollen tubes was very low but they appeared to be normal in the backcross A. esculentus × F₁, but were generally abnormal in the reciprocal cross. Both pre- and postzygotic barriers seemed to occur in crosses between the two species. The present studies indicate that these barriers can be overcome and desirable characters from A. moschatus transferred to cultivated okra using conventional hybridisation techniques. This revised version was published online in July 2006 with corrections to the Cover Date.     

Parthenogenetic monohaploids (2n=x=12) from Solanum tuberosum L. and S. verrucosum Schlechtd. and the production of homozygous potato diploids

Summary Dihaploid and dihaploid derived clones of Solanum tuberosum and diploid genotypes of S. verrucosum produced 85 viable monohaploids by female parthenogenesis. All were induced using diploid S. phureja clones, homozygous for embryo spot, as pollinator. Frequency of S. tuberosum monohaploids per 100 berries was rather constant in three successive years (14, 17 and 17 respectively). No male and female fertility was found in flowering monohaploids.Colchicine-induced chromosome doubling yielded homozygous s. tuberosum diploids with low pollen quality but good seed fertility.Two diploid self-incompatible species (S. multidissectum and S. berthaultii) produced no monohaploids. The presence of genes for female parthenogenesis in some dihaploids is discussed.     

Inheritance of seed colour in Brassica campestris L. and breeding for yellow-seeded B. napus L.

Summary Seed colour inheritance was studied in five yellow-seeded and one black-seeded B. campestris accessions. Diallel crosses between the yellow-seeded types indicated that the four var. yellow sarson accessions of Indian origin had the same genotype for seed colour but were different from the Swedish yellow-seeded breeding line. Black seed colour was dominant over yellow. The segregation patterns for seed colour in F₂ (Including reciprocals) and BC₁ (backcross of F₁ to the yellow-seeded parent) indicated that the black seed colour was conditioned by a single dominant gene. Seed colour was mainly controlled by the maternal genotype but influenced by the interplay between the maternal and endosperm and/or embryonic genotypes. For developing yellow-seeded B. napus genotypes, resynthesized B. napus lines containing genes for yellow seed (Chen et al., 1988) were crossed with B. napus of yellow/brown seeds, or with yellow-seeded B. carinata. Yellow-seeded F₂ plants were found in the crosses that involved the B. napus breeding line. However, this yellow-seeded character did not breed true up to F₄. Crosses between a yellow-seeded F₃ plant and a monogenomically controlled black-seeded B. napus line of resynthesized origin revealed that the black-seeded trait in the B. alboglabra genome was possibly governed by two independently dominant genes with duplicated effect. Crossability between the resynthesized B. napus lines as female and B. carinata as male was fairly high. The sterility of the F₁ plants prevented further breeding progress for developing yellow-seeded B. napus by this strategy.     

Variation in pollen and ovule fertility in varieties of cassava,and the effect of interspecific crossing on fertility

Records of the set of seeds obtained from intraspecific cassava crosses, from interspecific crosses between other Manihot species and cassava, and from backcrosses to cassava of three generations of interspecific hybrids are used to assess the variation in pollen and ovule fertility. It is concluded that the capacity of cassava varieties to set seeds has been reduced since they have evolved from non-tuberous wild forms and have been propagated vegetatively. Fertility was found to be very variable, and the capacity of the pollen of a variety to promote seed-set was not related to the fertility of the variety's female flowers. This situation would be expected if reduction in the fertilities of the two sexes were caused either by loss of balance in the genes controlling the functioning of the male gametophyte, or by unbalance of those concerned with the functioning of the female gametophyte or the tissues which nurture the embryo. Additional causes of sterility, probably including meiotic irregularities, appeared to operate in the F₁ interspecific hybrids, but such factors were probably not important after the first backcross generation. Manihot melanobasis, a wild form which is normally propagated by seed and whose relationship to cassava should be regarded as subspecific, contributed factors which enhanced the fertility of its hybrids with cassava. This form could be used in cassava breeding as a donor of seed fertility, but use could also be made of some existing varieties which still possess a moderately high capacity to set seed. It is desirable to select these as one parent when making difficult crosses.This paper is based almost entirely on the records of the late R. F. W. Nichols, who preceded the writer as cassava breeder with the East African Agriculture & Forestry Research Organisation, at Amani, Tanganyika.     

Development of a complete set of disomic rape-radish chromosome-addition lines

After crosses of multiple rape-radish chromosome additions with rape, nine different monosomic additions (2n = 4x = 38 + 1) of individual radish chromosomes, a–i, were isolated, having the genomic background of winter oil seed rape and radish cytoplasm. Extra chromosomes were identified with RAPD markers, specific for individual radish chromosomes. All radish-plasmic monosomic additions, except that of chromosome f, had low seed production due to pistilloid stamens. Rape cytoplasm was substituted by crossing the monosomic additions as pollinators to rape followed by reselection of the radish chromosome additions. The monosomic additions in rape cytoplasm showed normal male fertility. Average transmission rates of radish chromosomes via egg cells and pollen cells were 37% and 27%, respectively. Lowest male transmission was found for chromosomes e and f with 0.04 and 0.01. Monosomic additions were self-pollinated to produce disomic additions. Monosomic and disomic addition progenies were discriminated by a prescreening with quantitative double primer (dp) RAPDs and cytological confirmation of preselected candidates by FISH analysis using a Raphanus-specific probe. Usability of this complete set of nine disomic rape-radish additions (2n = 4x = 38 + 2) was discussed.     

Methods of overcoming interspecific barriers in Trifolium

Summary Barriers to interspecific hybridization in Trifolium were investigated by manipulation of mentor pollen treatments, ploidy levels, and compatibility and male sterility systems. Crosses involving the addition of mentor pollen produced fewer seeds and hybrids than crosses involving normal pollination. Lower seed set with mentor pollen was deduced to result from the use of less viable pollen, approximately half the pollen having been killed by alcohol. Pollinations at the diploid level resulted in more hybrids than at the tetraploid level, perhaps because genes for male sterility produced higher female sterility in the tetraploids. The self-compatible stock produced more seeds, mostly selfs, than the self-incompatible stock, but produced more hybrids only in one cross, T. pratense L. × T. diffusum Ehrh. The use of male-sterile female parents reduced selfing but produced fewer hybrids than male-fertile female parents. Techniques of this study were designed to affect prefertilization barriers, but the lack of effect may indicate that postfertilization barriers in Trifolium are of greater importance.Journal Article No. 98-3-208 of Kentucky Agricultural Experiment Station. Published with approval of the Director.