Fertility,plant morphology,and transmission rates of the extra chromosome in single and double trisomics of Solanum tuberosum L. Group Tuberosum

Summary Initial single and double trisomics of S. tuberosum Group Tuberosum cv. Gineke were successfully crossed with and backcrossed to clones obtained through inbreeding of self-compatible dihaploids of the same variety. Some trisomics showed reduced flowering or flower abnormalities and contributed therefore only to a small extent. Berry-set and seed-set varied considerably. There was no relation between these phenomena, nor was there any relation between either berry-or seed-set and the identity of the extra chromosome in the female parent. Seed-set seemed to be influenced by the male parent. When a more inbred male parent was used in the crosses with trisomics this often resulted in less seed per cross. There was no clear relation between seed-set and stainability of the pollen grains, but both the strong selection for good pollen and the high level of self-compatibility, have apparently contributed to the success of the crosses. The double trisomic plants yielded less seed than most single trisomics. Only three of the single trisomics produced stainable pollen and two of them yielded progeny when used as male parent in crosses or selfings.In the progenies several of the trisomic types could be distinguished on the basis of plant morphology in the young seedling stage. For ten different trisomics the morphotype is being described. The average rate of female transmission of the extra chromosome was about twenty per cent, but there was a wide variation, both between and within the various trisomic types. The results indicated that for chromosomes 3 and 8 the rate of transmission was higher than the average value. In one out of two trisomic types male transmission of the extra chromosome occurred, with a rate of 4.7%. The possible causes for the observed differences in female transmission of the extra chromosome are being discussed.     

Application of C-banding and fluorescence in situ hybridization for the identification of the trisomics of Hordeum chilense

Hordeum chilense is a wild barley species that has a high degree of genetic variability and significant potential for use in plant breeding. To establish a series of trisomics in H. chilense (2n = 14), plants with 2n + 1 chromosome numbers were isolated from the progenies of selfed triploid plants. Based on both fluorescent in situ hybridization with pAs1 and pTa71 repetitive DNA probes and C-banding patterns, seven different trisomics were tentatively identified. Primary trisomic plants were for chromosomes 1H^ch, 4H^ch, 5H^ch, 6H^ch and 7H^ch. A secondary trisomic carrying a 5H^chS-5H^chS isochromosome as the extra chromosome and a trisomic for chromosome 3H^ch heterozygous for the 3H^chS-4H^chL and 4H^chS–3H^chL interchange were identified. The trisomic for chromosome 1H^ch cannot be phenotypically distinguished from the diploid. The rest of the trisomic types were distinguishable from the diploid by their morphological characteristics (relatively poor vigour, decreased size and shorter spikes) but they were morphologically indistinguishable from each other. The frequencies of trisomics among the progenies derived from self-fertilization of these aneuploids ranged from 10.7% to 37.5%, with an average frequency of 26.1%. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evolving gene pools in crop plants

Summary The three gene pool system can be applied usefully to the whole range of crop plants. The primary gene pool (GP1) show varying degrees of fragmentation. No morphological discontinuity is apparent between wild and cultivated Lathyrus sativus but clear discontinuities have developed between conspecific wild and cultivated Phaseolus and Vigna species. Further discontinuities can develop in cultigens where disruptive selection has been practiced, as in Beta vulgaris, Linum usitatissimum and Brassica oleracea where more than one distinct crop has evolved within a single biological species. Each such crop has developed its own distinctive gene pool.     

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.     

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     

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.     

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.     

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.     

Interspecific hybrids between helianthus PetiolarisNutt. and H. Annuus L.: Effect of backcrossing on meiosis

Summary Interspecific substitutions of the nucleus of Helianthus annuus (2n=34) into the cytoplasm of H. petiolaris (2n=34) were obtained by successive backcrossing using cultivated sunflower, H. annuus, as the recurrent pollen parent.Meiosis in the F₁ was characterized by multivalents, suggesting that 10 of the 34 chromosomes were heterozygous for chromosomal interchanges. An additional pair of chromosomes also contained a paracentric inversion. Continued backcrossing resulted in rapid elimination of the meiotic aberrations evident in the F₁. In the BC₁, 1 of 11 plants had normal meiosis and by the BC₂, only 13 of 54 plants had meiotic aberrations similar to those of the F₁. However, trisomic progeny (2n=35) were found in 3 of the 11 BC₁ plants and 20 of the 54 BC₂ plants. No meiotic aberrations were observed in BC₃ or BC₄ plants. Plants with indehiscent anthers, and considered to be male sterile (M.S.), first occurred in the BC₁ and, by the BC₂, 51 of 54 plants were M.S. All 19 BC₃ and 16 BC₄ plants were M.S. Preliminary investigations suggest that the pollen from such plants is sterile and that the sterility is cytoplasmic rather than genetic.Disc-flower measurements were a useful technique for selecting samples at the correct stage of microsporogenesis, but could not be used to distinguish between successive backcrosses.     

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.     

Trisomic analysis of new gene for late heading in rice, Oryza sativa L

A near isogenic line, T65-LH7 bred from a rice variety, Ketan Nangka by five times of successive backcrossing with Taichung 65 (T65) as recurrent parent was found to carry a recessive lateness gene tentatively designated as ef6(t). The present study was performed to investigate the allelic relationships between ef6(t) and other heading time genes, Ef1, Efx, ef2(t), ef3(t), ef4(t) and ef5 by allelism test and to locate the chromosomal location of ef6(t) by trisomic analysis. In allelism test, six testers carrying each of heading time genes, Ef1, Efx, ef2(t), ef3(t), ef4(t) and ef5 were used. Those testers were the near isogenic lines of T65. T65-LH7 was crossed with respective testers. Heading times in F₂ and/or B₁F₁ plants were examined. All F₂ and/or B₁F₁ populations derived from those crosses exhibited digenic segregations, respectively. These results suggested that ef6(t) was independent of Ef1, Efx, ef2(t), ef3(t), ef4(t) and ef5. Sub sequently, trisomic analysis of ef6(t) was performed using seven Triplo lines having extra chromosomes, 2, 4, 5, 7, 9, 10 and 12. These Triplo lines were the near isogenic lines of T65. They were used as maternal parent to cross with T65-LH7. Heading times in F₂ plants obtained from self-pollination of F₁ plants were observed. Among F₂ plants examined only those derived from a cross between Triplo-7 and T65-LH7 showed a typical trisomic segregation manner, suggesting that ef6(t) was located on chromosome 7. Consequently, the nomenclature of the present gene should be designated as ef6.     

Resistance to the South American tomato pinworm <Emphasis Type="Italic">Tuta absoluta</Emphasis> in high acylsugar and/or high zingiberene tomato genotypes

Zingiberene (ZGB) and acylsugars (AS) are allelochemicals responsible for high levels of arthropod resistance found in Solanum habrochaites (= Lycopersicon hirsutum) var. hirsutum ‘PI 127826’ and S. (= L.) pennelli ‘LA 716’, respectively. These accessions were used to develop commercial lines with good levels of pest resistance. The objective of the present work was to assess the ZGB and AS contents and the levels of resistance to Tuta absoluta in tomato hybrids between high ZGB × high AS lines, as compared with their parental lines and with commercial checks. High AS homozygous lines [TOM-688 and TOM-689, both originated from the interspecific cross S. lycopersicum (= L. esculentum) × S. pennelli], high ZGB homozygous lines (ZGB-703 and ZGB-704, both derived from the interspecific cross S. lycopersicum × S. habrochaites var. hirsutum), double heterozygotes for both ZGB and AS, single heterozygotes for ZGB, and single heterozygotes for AS were assessed for AS and ZGB contents. Low-ZGB low-AS checks ‘Débora Max’ and ‘TOM-684’ were used, as well as checks with high ZGB (PI 127826) and high AS (LA 716). The genotypes were submitted to infestation with South American tomato pinworm adults in a screenhouse, and oviposition counts were taken 10 days after the initial infestation date. Plants were scored for overall plant damage and percent leaflets attacked up to the 38th day after infestation. Genotypes heterozygous for ZGB or AS showed allelochemical contents intermediate to those of their high and low content parents, indicating incompletely dominant gene action for contents of each of the allelochemicals. There were no significant differences in T. absoluta oviposition between high-AS homozygous genotypes, high-ZGB homozygotes, single heterozygotes for AS, single heterozygotes for ZGB and double heterozygotes for ZGB and AS, but all these genotypes showed egg counts significantly lower than the low-ZGB low-AS checks. Feeding damage of T. absoluta was higher in the low-ZGB low-AS checks than in any other ZGB-rich or AS-rich tomato genotype. Relative to ZGB or AS single heterozygotes, the heterozygotes for both ZGB and AS showed higher levels of resistance to the insect, as measured by overall plant damage, indicating a synergic effect of the allelochemicals on resistance.     

Isoenzyme aided selection in the transfer of mildew (Podosphaera leucotricha) resistance fromMalus hupehensis to the cultivated apple

Summary The isoenzymic geneGOT-1 was used as an indicator of ploidy levels and hybridity in the progeny of a 4x × 2x (A878/5,GOT-1 abdn × ‘Gloster 69’,GOT-lcc) cross, derived from the apomictic species (Malus hupehensis), which segregated for mildew resistance. The distinction between resistant and susceptible plants was sharp. After 3 months in a greenhouse, resistant plants showed no signs of sporulation. Of the hybrid seedlings (diploid and triploid) 78% were mildew resistant. This analysis was facilitated through the use of theGOT-lc allele, as a marker of hybridity. It appears that the prospects of transferring high mildew resistance fromM. hupehensis to the cultivated apple are promising; of the 65 hybrid resistant seedlings 18 were diploid. A similar approach in earlier generations might have facilitated the earlier transfer of this resistance to the cultivated apple.     

Ploidy Manipulation and Introgression of Resistance to Alternaria Helianthi from Wild Hexaploid Helianthus Species to Cultivated Sunflower (H. annuus L.) Aided by Anther Culture

The present investigation discusses the scope for transferring of resistance to leaf spot disease incited by Alternaria helianthi from two hexaploid wild species (H. tuberosus and H. resinosus) to diploid cultivated sunflower. Interspecific hybrids produced between sunflower and these two hexaploid species were partially fertile with tetraploid chromosome status. Backcrosses of these interspecific hybrids with cultivated sunflower resulted in the formation of sterile triploid plants. To overcome the problem of sterility and facilitate backcrosses with cultivated sunflower, anther culture of the tetraploid interspecific hybrids was carried out to bring down their chromosome number to diploid status. Anthers from both interspecific hybrids were cultured on basal Murashige and Skoog media supplemented with varying concentrations of organics and the growth regulators benzyladenine and naphthaleneacetic acid. Anthers of interspecific hybrids involving H. resinosus responded well and regenerated through an embryogenic route at a frequency of 98.7%. But in interspecific hybrids with H. tuberosus, anthers formed callus and subsequently regenerated shoots through an organogenic pathway. DNA ploidy analysis of anther culture plants of interspecific hybrids derived from H. tuberosus crosses was carried out to identify plants with desired diploid status. In vitro screening of parents, interspecific hybrids and anther culture plantlets against A. helianthi showed resistance in 68.5% of the anther culture plants of interspecific hybrids from H. tuberosus and in 24.3% of the plants derived from interspecific hybrids involving H. resinosus.     

Genetic segregation of the deformed flower gene in trisomics of Solanum chacoense

Summary The gene df (deformed flower), which is expressed as short anther in sensitive cytoplasm, has been incorporated into the trisomic series of S. chacoense Bitt. The cytoplasmic sensitivity and genotypes for the gene df were tested in the original trisomics and their parents. Two diploid parents were found to be homozygous dominant for Df, while one was homozygous recessive. All the parents and trisomics tested had resistant cytoplasm. Six trisomics were heterozygous for df, segregating with a 1:1 ratio when they were crossed to the homozygous testers ([df ^s]dfdf). When the F₁ trisomics (trisomics x [Df ^r]dfdf) were crossed to the homozygous testers, nine of the 13 trisomics segregated with a 1:1 ratio, three with a 0:1 ratio (all deformed) and one trisomic (V1682.3) with a 2:1 ratio. All the parents of the trisomic V1682.3 were homozygous dominant for Df. This clone is believed to be trisomic for the gene df.Cooperative investigation of the Agricultural Research Service, U.S. Department of Agriculture and the Wisconsin Experiment Station, Madison.     

Introduction of aromatic fragrance into cultivated tomato from the peruvianum complex'

This study was performed to introduce the distinct aromatic fragrance of Lycopersicon peruvianum LA 1554 into the cultivated tomato, Lycopersicon esculentum. The strong breeding barriers existing between these two distantly related species were circumvented by the ovule selection and culture method. A large BC₁F₁ population was developed and among 127 plants, 36 were self‐compatible and yielded fruits. Fruits of some of these selected plants were found to be enriched with a sweet aromatic flavour. Sensory evaluation of the fruit aroma of these selected plants was performed by a panel of 12 members against one of the best consumer‐rated Japanese commercial tomato cultivars, ‘Momotaro’. Although extensive variation was observed in fruit‐aroma in the BC₁F₁ population, panel opinion on ‘flavour‐desirability’ significantly favoured the BC₁F₁ fruits of some selected plants over the cv. ‘Momotaro’. Therefore, it can be concluded that the aromatic fragrance of a ‘L. peruvianum’ accession has successfully been introduced into the cultivated tomato gene pool.     

Inheritance and selection of some fruit characters of apple. I. Inheritance of low and high acidity

Summary Fruit acidity as such appears to be gouverned by a single major gene, low acidity being a recessive character. The additional genetic variation is explainable by the additive gene action model. This mode of inheritance causes the pH distribution of seedlings in progenies to shift with a higher mid-parent pH towards more low-acid cum sweet (pH 3.8) and fewer high-acid cum sour (pH 2.9) seedlings or vice versa with a lower midparent pH. As a result, in crosses between heterozygotes (Mama) the total proportion of undesirable seedlings -low plus high-acid ones-, remains similar, irrespective of the mid-parent pH. This proportion constituted about one third of the seedlings. A same proportion, but of mainly sour plants, was found in crosses between relatively acid homozygotes (Mama) or between these and similar heterozygotes. It was less than one quarter if the heterozygous partner was medium-acid and a few percentages only when both the homozygous and heterozygous partner were medium-acid. Choosing parents in the absence of information, chances are that the very large majority of the medium-acid types are heterozygotes, that among the relatively acid ones one in two is homozygous and that the odds to find homozygotes among the sour types are probably best. Attention was drawn to the importance of a high sugar content — which is quantitatively inherited — in parents with a relatively low pH to increase the chances of selecting seedlings with a good sugar/acid ratio.     

Breeding for resistance to cereal cyst nematode in wheat II. Use of test tube cultures in selection

Summary The use of soil. naturally infested with Heterodera avenae, to select resistant heterozygotes in backcross progenies of wheat, was tested for reliability. Selfed progenies from plants selected as resistant were cultured monoxenically in test tubes with nematodes hatched from single cysts, while backcross progenies from the same parent plants were grown in pots of naturally infested soil. Cyst counts were made after two months' growth. The results showed that over 50% of the backcross lines, screened in previous generations with naturally infested soil, had been erroneously selected as resistant. The test tube cultures clearly differentiated lines carrying resistance from those which were susceptible and corroborated results from pot tests.     

Utilization of Asymmetric Somatic Hybridization for the Transfer of Disease Resistance from Brassica nigra to Brassica napus

Asymmetric somatic hybrid calli were produced between Brassica napus and a transgenic (Hyg ^R) line of B. nigra using a donor recipient fusion method for the production of cybrids. The transgenic line of B. nigra used as a donor also possessed genetic resistance to the pathogenic fungi Phoma lingam and Plasmodiophora brassicae. Using hygromycin for selection, 332 hybrid calli were obtained from which 30 produced shoots (1—-20 per callus) which were rooted on a hormone-free culture medium. The rooted shoots were transferred into soil and cultivated in a growth chamber where the plants were tested for resistance against the two pathogens. Out of 129 hybrid plants tested for resistance against P. brassicae, 30 (23.3 %) plants proved to be resistant and from 78 plants tested for resistance against P. lingam, 41 (52.6 %) plants remained disease-free after infection.     

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.