Cytological observations in some hybrids between the rice species Oryza sativa L. and O. glaberrima Steud.

Summary Several hybrids between Oryza sativa and O. glaberrima and their backcrosses with O. sativa were studied. Their seed sterility was very different; large differences were also observed in the level of pollen sterility and in the earliness of microspore failure. The proportion of aborted embryo sacs was much lower than the rate of sterile male gametophytes. The backcross populations were much more sterile than the corresponding F₁ hybrids. On the base of our observations and according to the literature, we may conclude that genic unbalance is the main cause of sterility of these hybrids, but that physiological factors may also be involved. Thus a restoration of fertility is generally possible by selection. On the other hand, male-sterile lines could be bred from some of these hybrids.     

Identification and transfer of a new cytoplasmic male sterility source from Oryza perennis into indica rice (O. sativa)

Summary Most of the commercial hybrids of indica rice are based on wild abortive (WA) source of cytoplasmic-genetic male sterility (CMS). Such cytoplasmic uniformity may lead to genetic vulnerability to disease and insect pests. To overcome this problem, diversification of CMS sources is essential. Crosses of 46 accessions of O. perennis and two accessions of O. rufipogon as female parents were made with two restorers (IR54, IR64) of WA cytosterility. Sterile hybrids were backcrossed with the respective recurrent parents. Of all the backcross derivatives, one line having the cytoplasm of O. perennis Acc 104823 and the nuclear background of IR64 was found to be stable for male sterility. The newly developed CMS line has been designated as IR66707A. This line is completely sterile (0% seed set) under selfed conditions. Crosses of IR66707A with 10 restorers of WA cytoplasm showed almost complete (93–100%) pollen sterility, indicating that the male sterility source of IR66707A is different from WA sterility. Southern hybridization of IR66707A, O. perennis (cytoplasmic donor), IR66707B (maintainer) and V20A (WA cytoplasm) using mitochondrial DNA specific probes (5 endonucleases × 8 probes) showed identical banding patterns between IR66707A and O. perennis. However, in more than half of the combinations, different banding patterns were observed between IR66707A and IR66707B and between IR66707A and V20A. The results suggest that IR66707A has the same cytoplasm as the donor (O. perennis), and CMS may not be caused by any major rearrangement or modification of mtDNA. The new CMS source identified will be useful in cytoplasmic diversification in hybrid rice breeding.     

Interspecific cytoplasm substitutions of an Indica strain of Oryza sativa L. and O. Glaberrima Steud

Summary To study differential nucleus-cytoplasm interactions between the two cultivated rice species, Oryza sativa and O. glaberrima, cytoplasmic substitution lines were made by using a glaberrima strain (G) and an Indica strain of sativa (S). The G cytoplasm had no adverse effect on pollen development when combined with the nucleus of S. On the other hand, when the S cytoplasm was combined with the G nucleus, the substitution line showed no seed set because of male sterility although the pollen grains were normally stained with I₂-KI solution. A dominant gene derived from S strain seemed to cause anther indehiscence in the substitution line. Further, a restorer gene (Rf _j)from Akebono of Japonica type was effective on pollen restoration in the male sterile line, suggesting that the S cytoplasm is the same as those of Japonica type in terms of a fertility-restoring system.This paper is Genetic studies of speciation in cultivated rice. 4.     

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.     

Crossability and cytology of hybrid progenies in the cross between Brassica campestris and three wild relatives of B. oleracea,B. bourgeaui,B. cretica and B. montana

Summary Crossability and cytology were examined in F₁, F₂, B₁ and hybridsplants of F₁ hybrids of Brassica campestris and three wild relatives of B. oleracea, B. bourgeaui, B. cretica and B. montana, respectively. The F₂ plants were obtained after self-and open pollination of the F₁ hybrids. The B₁ and hybrid plants were produced after the F₁ hybrids backcrosses with B. campestris and crossed with B. napus, respectively. After crossing the F₁ hybrids, many seeds of the F₂, B₁ and hybrid plants were harvested. Multivalent formation was high in the chromsome configuration for the PMCs of F₂, B₁ and hybrid plants, suggesting that crossing over might occur between them. Many different types of aneuploids were obtained in the progenies of the F₂, B₁ and hybrid plants. It is suggested that different types of normal egg cells may be produced by one-by-one or little-by-little chromosome addition. The possibility is discussed of gene transfer from B. bourgeaui, B. cretica and B. montana, to cultivated plants, B. campestris and B. napus.     

The transfer of characters from Brassica campestris L. to Brassica napus L.: Production of clubroot-resistant oil-seed rape (B. napus ssp oleifera)

Summary Four oil-seed rape lines were crossed with a clubroot resistant Brassica campestris line from the European Clubroot Differential sct. The allotriploid hybrids were backcrossed to the rape lines to introgress clubroot resistance into oil-seed rape. Using a combination of screening for disease resistance and chromosome number, a high proportion of 38-chromosome, clubroot resistant selections were obtained.     

Transfer of resistance to race 2 of Plasmodiophora Brassicae from Brassica napus to cabbage (B. oleracea var. Capitata). I. Interspecific hybridization between B. napus and B. oleracea var. Capitata

Summary Interspecific hybridization between Brassica napus L. (2n=38, a₁a₁c₁c₁) and B. oleracea var. capitata L. (2x- and 4x-cabbage; 2n=2x=18, cc and 2n=4x=36, cccc) was carried out for the purpose of transferring clubroot disease resistance from the amphidiploid species to cabbage. Nineteen hybrids with three different chromosome levels (2n=28, a₁c₁c; 2n=37, a₁c₁cc and 2n=55, a₁c₁cccc) were obtained. The F₁ plants were mostly intermediate between the two parents but as the number of c genomes in the hybrids increased, the more closely the hybrids resembled the cabbage parent. All F₁ hybrids were resistant when tested against race 2 of Plasmodiophora brassicae wor. The complete dominance of resistance over susceptibility suggested that the gene(s) controlling resistance to this particular race of the clubroot pathogen is probably located on a chromosome of the a genome in Brassica.Contribution No. J654.     

Molecular mapping of S-c, an F1 pollen sterility gene in cultivated rice

Hybrids between indica and japonica rice varieties usually show partial sterility, and are a major limiting factor in the utilization of heterosis at subspecific level. When studying male-gamete (pollen) abortion, a possibly important cause for sterility, six loci (S-a, S-b, S-c, S-d, S-e and S-f) for F₁ pollen sterility were identified. Here we report genetic and linkage analysis of S-c locus using molecular markers in a cross between Taichung 65, a japonica variety carrying allele S-c ^j, and its isogenic line TISL5, carrying alleleS-c ^j. Our results show that pollen sterility occurring in the hybrids is controlled by one locus. We used 208 RFLP markers, as well as 500 RAPD primers, to survey the polymorphism between Taichung 65 and TISL5. Six RFLP markers located on a small region of chromosome 3, detected different RFLP patterns. Co-segregation analysis of fertility and RFLP patterns with 123 F₂ plants confirmed that the markers RG227, RG391, R1420 were completely linked with the S-c locus. The genetic distances between the markers C730, RG166 and RG369 and the S-c locus were 0.5 cM, 3.4 cM, and 3.4 cM respectively. Distorted F₂ ratios were also observed for these 4 RFLP markers in the cross. This result suggests that the `one locus sporo-gametophytic' model could explain F₁ hybrid pollen sterility in cultivated rice. RG227, the completely linked marker, has been converted to STS marker for marker-assisted selection. This revised version was published online in August 2006 with corrections to the Cover Date.     

Analysis of genic male sterility in Brassica oleracea

Summary The male sterility system MS-1 of Brassica oleracea was studied in order to elucidate if nucleo-cytoplasmic interactions determine this system. Crosses of male sterile MS-1 genotypes with heterozygous MS-5 genotypes gave fully fertile F₁ progenies. Selfing of seven F₁ plants resulted in five F₂ populations showing a 9:7 segregation ratio and two a 3:1 ratio for fertile and male sterile plants. Two F₂ progenies deviated from the expected 9:7 or 3:1 segregation ratios for fertile and male sterile plants. Thermosensitivity and distortion of the meiosis are suggested as the causal factors underlying the deviation of the segregation ratios. It was concluded that nuclear factors determine the male sterility in the MS-1 system, because the presence of a nucleocytoplasmic interaction in this system should have given only a 3:1 segregation ratio for fertile and male sterile plants in the F₂ generation.     

Screening of Portuguese cole landraces (Brassica oleracea L.) with Leptosphaeria maculans and Xanthomonas campestris pv. campestris

Summary Fifty-six landraces of Portuguese coles (Brassica oleracea l.), representing fourteen morphotypes, were screened for resistance to Leptosphaeria maculans at the cotyledon stage. Three isolates of the pathogen belonging to three pathogenicity groups were used in the screening. The mean disease interaction phenotype of the B. oleracea landraces with the L. maculans isolates confirmed their differences in pathogenicity. When compared to pathogenicity grouping using a B. napus differential set, a different classification was observed on B. oleracea for the three L. maculans isolates, suggesting possible differences in the genetic interaction between the two plant species and the pathogen. Most of the Portuguese cole landraces were found to be susceptible to all three isolates of L. maculans, although a few have shown intermediate interaction phenotype. Some accessions, on the other hand, elicit a differential reaction with the three isolates tested, and can be further selected to be used in host differential sets or in breeding for resistance to black leg.Fifty-four landraces of Portuguese coles, representing eleven morphotypes, were also screened in the field for resistance to Xanthomonas campestris pv. campestris. A good level of resistance to the bacterium was found in several of the morphotypes.Accessions of the Penca type, particularly, were identified as highly resistant. Landraces belonging to the same Portuguese cole group presented a similar range of mean interaction phenotype with the bacterium. Some Portuguese landraces of B. oleracea are potential sources for use in breeding programs for black rot resistance.     

Cytoplasmically controlled cold tolerance in a cytoplasmic-genetic male sterile line of rice

Summary The cytoplasmic-genetic male sterile line, Krishna-A with Kalinga-I cytoplasm was developed in rice through repeated backcrossing the completely pollen sterile F₁ of the cross Kalinga-I/Krishna with the recurrent male parent cv. Krishna. The germination percentage and root-shoot length in Krishna-A at 12°C and 9°C indicated a higher degree of tolerance to low temperature than the cold tolerant female parent Kalinga-I. The high yielding cold tolerant variety Kalinga-I was developed from a cross involving the cold tolerant variety Dunghansali and the high yielding variety IR 8. It is inferred that tolerance to low temperature during germination in Krishna-A was inherited from the cytoplasm of cv. Dunghansali through Kalinga-I besides male sterility. The cytoplasmic control of cold tolerance is reported here for the first time in rice literature. The male sterile line Krishna-A because of its tolerance to low temperature would be suitable for the development of hybrid rice especially for areas where low temperature is a problem during germination.     

Genetic analysis of male sterility in rice mutants with environmentally influenced levels of fertility

Summary Twenty-six male sterile plants grown in the field were recovered in the M7 generation from ethyl methane sulfonate-treated material of the rice cultivar M-201. Fertility increased five-fold when ratooned plants from the field were grown in a growth chamber with a 12 hour daylength. Crosses between mutant and normal fertile cultivars produced fertile F₁ plants. Female fertility was normal as judged by percent seed set from unbagged panicles of parental and recombinant lines. Transgressive segregation for fertility was observed for all crosses in the F₂ and F₃ generations. Five of 37 F₃ male sterile plants showed moderate levels of seed fertility under winter greenhouse conditions and reduced seed set when transplanted to summer field plots. Fertility data from reciprocal crosses suggested cytoplasmic factors had little or no effect on levels of male sterility in the mutant lines. Chi-squared analyses of F₂ and F₃ generation results indicated male sterility of the mutants is conditioned by two nuclear genes with epistatic effects.     

Development of new cytoplasmic genetic male sterile lines through Indica × Japonica hybridization in rice

Summary From 28 Indica-Japonica crosses, two Indica cultivars, V.20B and Sattari were identified to possess male sterile cytoplasm with fertility restoring genes. It was possible to develop a new Japonica cytoplasmic genetic male sterile line (Zhunghua-1) on Indica male sterile cytoplasm (V 20B) by repeated backcrossing the complete pollen sterile plants of V 20B x Zhunghua-1 to the recurring male parent, Zhunghua-1. The study indicated that it would be possible to develop male sterile lines rom indica-japonica crosses only when there is sufficient amount of reciprocal differences with respect to pollen sterility. Further, it was inferred that it would be easier to develop Japonica male sterile lines on Indica cytoplasm than developing Indica male sterile line with japonica cytoplasm.     

Genetic studies on the interspecific cytoplasm substitution lines of japonica varieties of Oryza sativa L. and O. glaberrima Steud.

Summary Interspecific cytoplasm substitution lines of Oryza sativa and O. glaberrima, i.e. (sativa)-glaberrima and (glaberrima)-sativa, have been bred by means of successive backcrosses, using three japonica varieties of sativa and two glaberrima strains.In all the six substitution lines with the cytoplasm of the glaberrima strains, the fertility increased with succeeding backcrosses, and eventually completely fertile plants whith the characteristics of the parental japonica variety appeared. This indicates that the glaberrima cytoplasm exerted no effect on the genome manifestation of these japonica varieties. Of the five substitution lines with the cytoplasm of each of the japonica varieties, four lines produced male sterile (M.S.) plants only in the backcross generations. In the remaining substitution line with the cytoplasm of the japonica variety Akebono, there was simultaneous segregation for male sterile (M.S.) and pollen fertile plants bearing indehiscent anthers (ID.M.F.) in the backcross generations. In the compulsively selfed progeny of ID.M.F. plants, pollen fertile plants with dehiscent anthers (D.M.F.) occurred with M.S- and ID.M.F. plants. Morphologically, these three types were supposed to have the same genetic background as the glaberrima parent. It was established that D.M.F.-and ID.M.F. plants were homozygous and heterozygous for a dominant nuclear gene restoring pollen fertility, respectively, and the M.S. plants and the two glaberrima strains used in this study carried a recessive gene for pollen sterility in homozygous condition. The restorer gene was assumed to derive from the japonica variety Akebono. The expression of the restorer gene was of the sporophytic type. The pollen sterility of the substitution lines that possessed the cytoplasm of the japonica varieties was of cytoplasmon-genic nature.     

Genetics of thermosensitive genic male sterility in rice

Summary Inheritance of thermosensitive genic male sterility (TGMS) in Norin PL12 and IR32364TGMS and their allelic relationship were studied from F₁, F₂ testcross (TC) and F₃ generations of the crosses made with the two mutants and several fertile tester parents. F₂, TC and F₃ segregation behavior for pollen and spikelet fertility indicated that the TGMS trait in the two mutants was controlled by a single recessive gene. Allelic relationship studies indicated that TGMS genes of the two mutants were different. Since TGMS gene in Norin PL12 has been designated as tms ₂ , the TGMS gene present in IR32364TGMS is tentatively designated as tms ₃ (t) until allelic test is done with another TGMS gene (tms ₁ ) reported from China in a line 5460S seeds of which were not available.     

The introgression of S alleles into forage rape,Brassica napus L. from turnip. Brassica campestris L. ssp. Rapifera

Summary Forage rape. B. napus, is self-compatible, the work described illustrates the introgression of functional incompatibility alleles into B. napus from turnips, B. campestris, in two generations. By grading seed, produced by backcrossing turnip/rape hybrids to rape, the frequency of 2n=38 semi articial B. napus recovered, amply justifies the cytological screening involved in such breeding programmes.     

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.     

Transfer of resistance to race 2 of Plasmodiophora brassicae from Brassica napus to cabbage (B. oleracea ssp. capitata). III. First backcross and F2 progenies from interspecific hybrids between B. napus and B. oleracea ssp. capitata

Summary The first backcross and F₂ progenies from triploid F₁ and tetraploid F₁ hybrids between B. napus and 2x and 4x B. oleracea ssp. capitata (cabbage) were studied for their general morphology, resistance to race 2 of the clubroot pathogen, chromosome number and meiotic chromosome behavior. No linkage was apparent between resistance and the major morphological characters. Unreduced gametes played a large part in the successful formation of seed of the B₁ and F₂ progeny. B₁ plants with low chromosome numbers were selected for use in recurrent backcrosses. The potential use of anther culture to extract gametic progenies from resistant B₁ and F₂ plants with higher chromosome numbers was suggested. The presence of homoeologous pairing observed in all the plants is considered advantageous for selecting suitable progeny in later generations.     

Genetics of fertility restoration of ‘Wild Abortive’ cytoplasmic male sterility in rice,Oryza sativa L.

Summary Genetics of fertility restoration in six varieties and breeding lines of rice was studied in Wild Abortive cytoplasmic genetic male sterility system using cytoplasmic male sterile lines V 20 A and IR 54752 A. Fertility evaluation of the plants in F₂ and testcross populations of the crosses of V 20 A with PR 103, PR 106 and PAU 502-94-1, and IR 54752 A with PAU 1124-36-1 and PAU 1126-1-1 revealed that fertility restoration in PR 103, PR 106, PAU 502-94-1, PAU 1124-36-1 and PAU 1126-1-1 was controlled by two independently segregating dominant genes. The two genes appeared to have additive effects; one of them being stronger than the other in imparting fertility restoration. Data on spikelet fertility of the plants in F₂ and testcross populations of V 20 A/UPR 82-1-1 cross showed that fertility restoration in UPR 82-1-1 was controlled by two independently segregating dominant genes which exhibited recessive epistatic interaction.

---

     

Development of new cytoplasmic source of male sterile lines in rice for shallow lowland

Two shallow lowland male sterile lines, MotiA and PadminiAwith Miz.4 cytoplasmic source were developed throughintervarietal hybridization and recurrent backcrossing withtheir respective parents. In all probability, MotiA and PadminiA of Miz.4 source may be genetically different from the Wild abortive CMS source (V.20A). These two CMS lines are semitall in stature, photosensitive, of late duration in flowering with easy fertility restoration ability, and thus could be useful for developing hybrid rice suitable for shallow lowland areas.