Inheritance of root traits and phosphorus uptake in common bean (Phaseolus vulgaris L.) under limited soil phosphorus supply

Heterosis is an important way to improve yield and quality for many crops. Hybrid rice and hybrid maize contributed to enhanced productivity which is essential to supply enough food for the increasing world population. The success of hybrid rice in China has led to a continuous interest in hybrid wheat, even when most research on hybrid wheat has been discontinued in other countries for various reasons including low heterosis and high seed production costs. The Timopheevii cytoplasmic male sterile system is ideal for producing hybrid wheat seeds when fertility restoration lines with strong fertility restoration ability are available. To develop PCR-based molecular markers for use in marker-assisted selection of fertility restorer lines, two F₂ populations derived from crosses R18/ND36 and R9034/ND36 were used to map fertility restoration genes in the two elite fertility restorer lines (R-lines) R18 and R9034. Over 678 SSR markers were analyzed, and markers closely linked to fertility restoration genes were identified. Using SSR markers, a major fertility restoration gene, Rf3, was located on the 1B chromosome in both populations. This gene was partially dominant in conferring fertility restoration in the two restorer lines. SSR markers Xbarc207, Xgwm131, and Xbarc61 are close to this gene. These markers may be useful in marker-assisted selection of new restorer lines with T. timopheevii cytoplasm. Two minor QTL conferring fertility restoration were also identified on chromosomes 5A (in R18) and 7D (in R9034) in two R-lines.     

Random amplified polymorphic DNA analyses of cytoplasmic male sterile and male fertile Pigeonpea (Cajanus cajan (L.) Millsp.)

Randomly amplified polymorphic DNA (RAPD) markers were used for the identification of two pigeonpea cytoplasmic genic male sterile (CMS) lines derived from crosses between the wild (Cajanus scarabaeoides & C. sericeus) and the cultivated species of Cajanus cajan. The male sterile (A) line and its maintainer (B) line could be easily differentiated with certain random primers. The two male sterile (288 A and 67 A)systems are based on C. scarabaeoidesand the other is based on C. sericeus could also be differentiated. Amplification product of 600bp amplified by primer OPC-11 was observed in both the cytoplasmic male sterile lines (288 A and 67A), which was absent in the maintainer lines (288 B and 67 B) and the putative R-line (TRR 5 and TRR 6). CMS lines, putative R lines, other cultivars and wild species under the study could be easily distinguished with the help of different primers. Dendrogram constructed based on the similarity index showed that considerable genetic variation exist sbetween CMS lines, two putative R line and wild species studied. This revised version was published online in July 2006 with corrections to the Cover Date.     

Transfer of resistance to Verticillium dahliae Kleb. from Solanum torvum S.W. into potato (Solanum tuberosum L.) by protoplast electrofusion

Summary Interspecific somatic hybrid plants were regenerated after electrofusion of mesophyll protoplasts with the objective of transferring resistance to Verticillium dahliae from Solanum torvum into potato. Early selection of the putative hybrids was based on differences in cultural behaviour of the parental and hybrid calli (particularly the ability of the latter to regenerate early) in combination with morphological markers. Four putative hybrids were recovered from hundreds of calli, probably resulting from complementation of the two parental genomes. The regenerates were tetraploids (2n=4×=48 chromosomes) and exhibited intermediate traits including leaf form, plant morphology and the presence of anthocyanin. The hybrid nature of the four selected plants was confirmed by examining isoenzyme patterns for isocitrate dehydrogenase (Idh), malate dehydrogenase (Mdh), phosphoglucoisomerase (Pgi) and 6-phosphogluconate dehydrogenase (6-Pgd). While the hybrid plants rooted readily and grew vigorously under in vitro conditions, in the greenhouse their development and growth were retarded by difficulties in rooting. When grafted on potato or S. torvum rootstocks, the hybrid plants recovered normal development and growth. Again, they exhibited intermediate morphological traits. Tests for resistance realized in vitro with medium containing 50% Verticillium wilt filtrate showed that all the somatic hybrids were resistant to the fungus filtrate.     

C-banded karyotypes and meiotic abnormalities in germplasm derived from interploidy crosses in Avena

C-banded karyotypes of somatic chromosomes and meiotic abnormalities were investigated in four crown rust resistant lines derived from interploidy crosses in Avena.C-banding revealed that line N770-165-2-1 contained a 6C/21translocation while line DCS1789 contained a pair of A. strigosa chromosomes substituted for A. sativachromosome 12. Line JR2-3-3-B contained both the 6C/21 translocation and the pair of substituted A. strigosachromosomes, but line MAM17-4 contained neither. Although meiotic irregularities, including mispositioned bivalents, occurred in all four lines and in their F₁ hybrids with A. sativa, the percentages of normal meiosis ranged from 75.4 to 88.6%. This allowed for stability of line performance and for their use as breeding stocks. This revised version was published online in July 2006 with corrections to the Cover Date.     

Screening for resistance to take-all in wheat,triticale and wheat-triticale hybrid lines

Summary Fifteen triticale and wheat-triticale hybrid lines were evaluated for resistance to the take-all fungus Gaeumannomyces graminis var. tritici and compared with five wheat and two rye lines in inoculated field and pot trials. The triticale and wheat-triticale hybrid lines varied in rye chromosome number and degree of resistance expressed. One line, Venus with seven pairs of rye chromosomes consistently showed levels of resistance intermediate between wheat and rye. A trend was observed where increasing rye chromosome content led to greater resistance but exceptions showed that variation within triticales could not be ascribed to rye chromosome content alone.     

Somatic embryogenesis in pea (Pisum sativum L. and Pisum arvense L.): Diallel analysis and genetic control

Summary Six lines of Pisum were tested in vitro for their ability to produce somatic embryos from apices. Significant quantitative variation was observed. Inheritance of the ability to form somatic embryos was studied using a diallel cross among six different lines. About 80% of the observed genotypic variation was due to additive effects. There is a tendency for the favourable genes to be recessive. It appears that there are two genetic systems involved. Analysis of the distribution of F₃ families means from a cross among two extreme lines seems to indicate the presence of a few major genes in the control of somatic embryogenesis of pea.     

Seed productivity test of CMS lines of Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of a wild species, A. galanthum Kar. et Kir.

In a previous study, we developed male sterile lines of Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of a wild species, A. galanthum Kar. et Kir., by backcrossing. To evaluate seed productivity of the male sterile lines in practise, they were crossed with the male fertile line, cultivar 'Kujyo', using honeybees as pollinators under field conditions. The number of florets and seeds per inflorescence, seed set and seed germination of the material were investigated. Although variation was observed among the male sterile lines, there were several lines having seed productivity equal to cultivar 'Kujyo'. Our data demonstrate that the male sterile lines of A. fistulosum possessing the cytoplasm of A. galanthum are useful as seed parents for the commercial F seed production of A. fistulosum.     

Intraspecific somatic and sexual hybridisation between dihaploid lines of Solanum tuberosum L.: evaluation of morphological traits and resistance to late blight Phytophthora infestans (Mont.) de Bary in the foliage

Intraspecific tetraploid somatic and sexual hybrid plants have been resynthesised following protoplast fusion and by sexual crosses between two dihaploid potato (Solanum tuberosum) lines each possessing complementary agronomic traits. The dihaploid PDH 40 possesses good tuber shape and yield but has foliage susceptibility to late blight (Phytophthora infestans). On the other hand, the dihaploid PDH 727 possesses resistance to blight in the foliage but has a low yield of small and irregular shaped tubers. Since it was only possible to use a partial selection strategy based on culture media to facilitate recovery of somatic hybrid plants-further morphological and esterase isozyme based characterisations were performed to identify somatic hybrid plants from amongst the non-hybrid plant material. When the blight resistance of both the intraspecific somatic and sexual hybrid plants was assessed there was no significant difference in the mean resistance value and it was intermediate between those of their parents. However, the range of resistance was much wider among the sexual hybrids than among the plants derived from somatic fusion. An assessment of tuber yield between tetraploid sexual and somatic hybrids showed no significant difference and it was higher than that of either parent value. The implication of these results in the context of potato genetics and breeding is discussed.     

Amphidiploids between Cyclamen persicum Mill. and C. purpurascens Mill. induced by treating ovules with colchicine in vitro and sesquidiploids between the amphidiploid and the parental species induced by conventional crosses

Summary We cultured colchicine-treated hybrid ovules in vitro to produce fertile amphidiploids of C. persicum (2n=2x=48. referred to as AA) × C. purpurascens (2n=2x=34, referred to as BB). Seedlings and mature plants were obtained from the ovules without colchicine and those exposed to 50 mg/l colchicine for 5, 10 and 15 days, whereas they were not obtained from the ovules exposed to 50 mg/l colchicine for 20 days and 500 mg/l for 5, 10, 15 and 20 days. Although 8 mature hybrids derived from the ovules without colchicine produced a few fertile pollen grains, they failed to produce viable seeds by self-fertilization. The hybrids had 41 somatic chromosomes. Four and 3 mature plants were derived from ovules exposed to 50 mg/l colchicine for 10 and 15 days, respectively. One each among 4 and 3 mature plants showed a high frequency of pollen grain fertility, produced several seeds by self-fertilization, and had 82 somatic chromosomes which is twice the number of hybrid chromosomes (2n=41, AB). These findings indicated that these plants are amphidiploids (2n=82, AABB) between C. persicum and C. purpurascens. Three and 2 viable seeds were derived by the conventional crosses of diploid C. persicum × the amphidiploid and the amphidiploid × C. purpurascens, respectively. Flowering plants that developed from the seeds of diploid C. persicum × the amphidiploid were barely fertile and had 65 somatic chromosomes (2n=65, AAB), whereas those that developed from the seeds of the amphidiploid × C. purpurascens were barely fertile and had 58 somatic chromosomes (2n=58, ABB). The somatic chromosomes indicated that these plants are probably sesquidiploids between the amphidiploid and either C. persicum or C. purpurascens. The interspecific cross-breeding of cyclamen using the amphidiploids and the sesquidiploids is discussed.     

Genes Lr48 and Lr49 for hypersensitive adult plant leaf rust resistance in wheat (Triticum aestivum L.)

The genetic bases of leaf rust resistance in wheat (Triticum aestivum L.) line CSP44, selected from the Australian cultivar Condor, and Indian cultivar VL404, were studied. The reaction patterns of CSP44 and VL404 against Indian races 12, 77, 77-1, 77-2, 77-3, 77-4, 77-5 and 108 were different from reaction patterns shown by near-isogenic lines with known adult plant resistance (APR) genes, viz. Lr12, Lr13, Lr22b and Lr34. Although the reaction patterns of CSP44 and VL404 were similar to the near-isogenic line Tc+Lr22a, tests of allelism indicated absence of Lr22a in both CSP44 and VL404. On the basis of genetic studies, their resistances in field tests against race 77-5, the most virulent race from the Indian sub-continent, were each ascribed to two genes. One of the two genes in each wheat was identified to be the non-hypersensitive APR gene Lr34. The second APR genes in CSP44 and VL404 gave hypersensitive reaction types and were recessive and dominant, respectively. The gene in CSP44 was designated Lr48and the gene in VL404, Lr49. This revised version was published online in August 2006 with corrections to the Cover Date.     

Cytoplasmic male sterility with self-incompatibility,a novel approach to utilizing heterosis in rapeseed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

The Polima cytoplasmic male sterility (CMS) system has been successfully used in three/two-line hybrid production in rapeseed (Brassica napus L.). However, the sterility of the Polima (pol) CMS lines is sensitive to temperature fluctuations. Also, traces of pollen can cause self-pollination within the CMS lines, which results in reduced levels of F₁ hybrid seed purity and leads to a significant yield loss. Self-incompatibility (SI) is another important approach for hybrid seed production in rapeseed. Despite having a wide range of restorers and being easily selected in a breeding program, SI system has some drawbacks. In this study, SI genes from a self-incompatible line of Brassica napus were transferred to a pol CMS line and S372A, a novel line of combined cytoplasmic male sterility with self-incompatibility was bred. Due to the SI genes, this line produced very few seeds when it was selfed at low temperature and no seeds at high temperature. This suggested that the line with CMS + SI had combined the advantages and overcome the disadvantages of both the pol CMS and SI systems. Furthermore, our results showed that most of the maintainers and all the restorers of the pol CMS system were also maintainers and restorers of the CMS + SI line, respectively. This indicates that the CMS + SI system can be easily used to establish three-line hybrids of rapeseed, and we believe this novel system could be extended to other species of Brassica.     

Intergeneric symmetric and asymmetric somatic hybridization in Festuca and Lolium

Summary Intergeneric symmetric and asymmetric somatic hybrids have been obtained by fusion of metabolically inactivated protoplasts from embryogenic suspension cultures of tall fescue (Festuca arundinacea Schreb.) and unirradiated or 10–500 Gy-irradiated protoplasts from non-morphogenic cell suspensions of Italian ryegrass (Lolium multiflorum Lam.). Genotypically and phenotypically different somatic hybrid Festulolium mature flowering plants were regenerated.Species-specific sequences from F. arundinacea and L. multiflorum being dispersed and evenly-represented in the corresponding genomes were isolated and used for the molecular characterization of the nuclear make-up of the intergeneric, somatic Festulolium plants recovered. The irradiation of Italian ryegrass protoplasts with 250 Gy X-rays prior to fusogenic treatment favoured the unidirectional elimination of most or part of the donor chromosomes. Irradiation of L. multiflorum protoplasts with 500 Gy produced highly asymmetric (over 80% donor genome elimination) nuclear hybrids and clones showing a complete loss of donor chromosomes.The RFLP analysis of the organellar composition in symmetric and asymmetric tall fescue (+) Italian ryegrass regenerants confirmed their somatic hybrid character and revealed a bias towards recipient-type organelles when extensive donor nuclear genome elimination had occurred.Approaches aimed at improving persistence of ryegrasses based on asymmetric somatic hybridization with largely sexually-incompatible grass species (F. rubra and Alopecurus pratensis), and at transferring the cytoplasmic male sterility trait by intra- and inter-specific hybridization in L. multiflorum and L. perenne, have been undertaken.Abbreviations cpDNA chloroplast DNA - CMS cytoplasmic male sterility - 2,4-D 2,4-dichlorophenoxy-acetic acid - IOA iodoacetamide - mtDNA mitochondrial DNA - RFLP restriction fragment length polymorphism     

Segregation of 12 isozyme genes among doubled haploid lines derived from a japonica x indica cross of rice (Oryza sativa L.)

Summary The segregation of 12 heterozygous isozyme markers was analyzed among F₂ plants and 51 anther culture (AC)-derived lines obtained from the japonica × indica cross of rice, IRAT 177 × Apura. All the lines except two were homozygous products of recombination of the two parental phenotypes. Doubled haploid (DH) lines derived from plants regenerated from the same callus were identical, confirming previously obtained results in rice. Surprisingly, some lines derived from different calli were also identical, suggesting a phenomenon of early callus fragmentation. All these observations at the isozyme level were confirmed by field evaluation. Deviations of segregations from the expected 1 : 1 ratio were observed at 4 loci among the DH lines. Among these, two were also noted among the F₂ plants. The two other distortions, both in favor of the japonica allele, were observed specifically in the AC-derived materials.Although this concerns a small proportion of the genes under study, it suggests that the embryogenic microsporal population does not represent a random gametic array. On the other hand, evaluation of recombination between isozyme genes located on chromosome 6 appears consistent with F₂ data and data previously recorded on the other japonica × indica crosses. The potential use of isozymes in breeding doubled haploids derived from remote crosses in rice is discussed.Abbreviations MCPA = 2-methyl-4-chlorophenoxyacetic acid - IAA = indolacetic acid - AC plant or line = anther culture-derived plant or line - DH line = doubled haploid line     

Somatic hybrids between Gossypium hirsutum L. (4×) and G. davidsonii Kellog (2×) produced by protoplast fusion

Somatic hybrids were obtained from electrofused protoplasts derived from embryogenic suspension cultures of tetraploid cotton (G. hirsutum L. cv. Coker 201) and embryogenic callus of diploid wild cotton G. davidsonii. The regenerants were initially identified as hybrids by RAPD (random amplified polymorphic DNA) analysis. Subsequently, observation on chromosome counting, morphology and SSR (simple sequence repeat) confirmed the hybrid status. Cytological investigation of the metaphase root-tip cells of the regenerated plants revealed there were 74 to 84 chromosomes in the plants, close to the expected 78 chromosomes. SSR analysis revealed the regenerated plants contained specific genomic fragments from both fusion partners, further confirmed their hybridity. The morphology of the plants was intermediate between the two fusion partners. The regenerants were difficult to develop into mature plants because their roots browned and they wilted from the stem apex before forming 3 to 5 true leaves. The hybrid plants were transferred to soil by grafting in vitro onto rootstocks.     

A gene for hybrid necrosis in an inbred line of rye (Secale cereale L.)

Summary An inbred line of rye (Secale cereale L.) has been found to carry a gene for hybrid necrosis. This gene was detected in crosses with a highly crossable wheat (Triticum aestivum L.) genotype which carries the gene Ne₂. This appears to be the first report of a gene for hybrid necrosis being present in the rye genome.     

Breeding for hard red winter wheat cultivars adapted to conventional-till and no-till systems in northern latitudes

Summary Monosomic alien addition lines combining individual F. drymeja chromosomes and the L. multiflorum complement were isolated from the cross between the triploid hybrid L. multiflorum (4x) × F. drymeja (2x) and diploid L. multiflorum (2x). Chromosome pairing in the addition lines was studied at metaphase 1 of meiosis and the relationship between single F. drymeja chromosomes and the corresponding homologous pair in L. multiflorum is discussed. Trivalent frequency in the addition lines was higher than expected from observations of chromosome pairing in the triploid hybrid and there were differences between lines in the number of trivalent associations formed. There is some evidence to suggest that trivalent frequency is not entirely dependent on chromosome length and that transmission of the alien chromosome in the female is dependent on the size of the added chromosome. Morphological studies were made to assess the phenotypic effects of the addition of single F. drymeja chromosomes to the L. multiflorum complement. Two plants (2n=14) with recombination between a L. multiflorum and a F. drymeja chromosome were identified.     

Heterosis and inbreeding depression in forage rape (Brassica napus L.)

Summary Heterosis and inbreeding depression for total fresh and dry weights were studied over two seasons in 15 crosses of six cultivars of forage rape (Brassica napus L.). Some of the hybrids exhibited considerable heterosis over both mid-parent and better parent; followed by depression in the F₂. The cross Windal × Canard had 62% and 25% greater dry matter yield over the best cultivar used in this experiment in 1978 and 1979, respectively. Possibilities of the utilization of this hybrid superiority are discussed.     

Response to selection for plant height in X-ray treated population of jute (Corchorus capsularis L.) cv. JRC 212

Summary X irradiation of dry seeds of Corchorus capsularis L. cv. JRC 212 induced variations in plant height as evidenced by the high values of genotypic variance and heritability estimate in M₃ generation. Selection on the basis of line mean of plant height in M₃ generation was effective in increasing plant height as well as fibre yield in M₃ generation. A higher response for plant height rather than for fibre yield itself was observed in lower irradiation dose while responses for these two traits were almost similar in higher dose. An average of 12% increase in fibre yield was achieved in M₆ generation when all top five lines were selected. The top two lines, had 17% and 16% higher fibre yield than that of control.     

Hybrid plant of Phaseolus vulgaris L. and P. lunatus L. obtained by means of embryo rescue and confirmed by restriction endonuclease analysis of rDNA

Summary Hybridizations between three lines of the common bean (Phaseolus vulgaris L.) and two lines of the lima bean (P. lunatus L.) were attempted in order to transfer characters from the lima bean to the common bean. A total of 115 interspecific hybrid embryos were rescued and cultured, and 7 plantlets were eventually transferred to soil. The most compatible cross was Edogawa XPI164891, which had a high proportion of expanded ovules, and from which we obtained one mature interspecific hybrid. In general, morphological characters of the hybrid were intermediate between the parents, and the chromosome number of the hybrid was 2n=22 the same as that of both parents. The hybrid nature of the progeny plant was confirmed by restriction endonuclease analysis of rDNA. Species specific fragments were detected when total DNA was digested by BamHI, and BamHI-digested total DNA of the hybrid contained all fragments from both parents. Selves and backcrosses were attempted, but no progeny were obtained. The only hybrid obtained was completely sterile and meiosis highly irregular.     

Effect of common bunt on the frost resistance and winter hardiness of wheat (Triticum aestivum L.) lines containing Bt genes

In order to determine the effects of bunt inoculation on frost resistance and winter hardiness in lines containing resistance genes, the bunt [Tilletia foetida (Wallroth) Liro, T. caries (DC.) Tulasne] susceptibility of wheat lines containing bunt resistance genesBt1 to Bt10 and the effect of the year on the degree of infection were studied over six years from 1991 to 1997 in an artificial inoculation nursery. Uninoculated and artificially inoculated wheat plants were tested for frost resistance in the phytotron in 1995 and in the field in boxes in three years from 1994/95 to 1996/97. The line withBt10 was very resistant, lines with Bt5, Bt6, Bt8 and Bt9 were resistant, the line with Bt4 was moderately resistant, those with Bt2 and Bt3 were moderately susceptible, the line with Bt1 was susceptible and the line with Bt7 was very susceptible to the local bunt population in Hungary. Bunt incidence also varied over years. The frost resistance of the Bt lines was generally lower after bunt inoculation than that of uninoculated plants. The increased frost kill in inoculated plants was not correlated with the extent of varietal susceptibility to bunt. Some lines with resistance, namely those with Bt5 (1.6% infection), Bt8 (0.6%) and Bt10 (0.0%), suffered significantly greater frost kill in the young plant stage as the result of bunt inoculation. By contrast, the Bt7line had excellent frost resistance and winter hardiness but suffered the greatest extent of bunt infection, whereas the Bt6 line had good frost resistance and good bunt resistance. This revised version was published online in July 2006 with corrections to the Cover Date.