Synthesis of hexaploid (AABBCC) somatic hybrids: a bridging material for transfer of ‘tour’ cytoplasmic male sterility to different Brassica species

Most of the alloplasmic cytoplasmic male sterility (CMS) systems are known to be associated with a number of floral abnormalities that result from nuclear-cytoplasmic incompatibilities. One such system, tour, which is derived from Brassica tournefortii, induces additional floral abnormalities and causes chlorosis in Brassica spp. While the restorer for this CMS has been reported to be present in B. napus, in B. juncea, where the abnormalities are more pronounced, no restorer has yet been identified. Rectification of these floral abnormalities through mitochondrial recombinations and chloroplast replacement might result in the improvement of this CMS system. As organelle recombinations can possibly be achieved only by somatic cell hybridization, fusion experiments were carried out between hygromycin-resistant B. juncea AABB carrying tour cytoplasm and phosphinotricin-resistant, normal B. oleracea CC to generate AABBCC hexaploid somatic hybrids. The presence of selectable marker genes facilitated the selection of hybrids in large numbers. The resulting hybrids showed wide variation in floral morphology and organelle composition. Regenerants with normal, male-sterile flowers having recombinant tour-or oleracea-type mitochondria and oleracea-type chloroplasts were obtained. Hybrids with male-fertile flowers were also obtained that had recombined tour mitochondria. The AABBCC hexaploid hybrids synthesized in the present study were successfully utilized as a bridging material for transferring variability in the organelle genome simultaneously to all the digenomic Brassica species, and all of these hybrids are now being stabilized through repeated backcrosses to the allopolyploid crop brassicas.     

Genetic analysis of brown planthopper resistance in twenty varieties of rice,Oryza saliva L.

Summary The inheritance of resistance to brown planthopper, Nilaparvata lugens (Stol.), of 20 rice cultivars was studied. Single dominant genes that are allelic to Bph 3 condition the resistance in cultivars Ptb 19, Gangala (Acc. 7733), Gangala (Acc. 15207), Horana Mawee, Kuruhondarwala, Mudu Kiriyal and Muthumanikam. Single recessive genes that are allelic to bph 4 govern the resistance in cultivars Gambada Samba, Heenhoranamawee, Hotel Samba, Kahata Samba, Kalukuruwee, Lekam Samba, Senawee, Sulai, Thirissa and Vellai Illankali. The resistance in Ptb 33, Sudu Hondarwala, and Sinna Sivappu is governed by one dominant and one recessive gene which segregate independently of each other. The dominant resistance genes in these cultivars appear allelic to either Bph 1 or Bph 3. Similarly, the recessive genes in these cultivars seem allelic to either bph 2 or bph 4. Further investigations are needed to conclusively determine the allelic relationships of resistance genes in Ptb 33, Sudu Hondarwala and Sinna Sivappu.     

RFLP mapping of three new loci for resistance genes to powdery mildew (Erysiphe graminis f. sp. hordei) in barley

Three new major, race-specific, resistance genes to powdery mildew (Erysiphe graminis f. sp. hordei) were identified in three barley lines, RS42-6*O, RS137-28*E, and HSY-78*A, derived from crosses with wild barley (Hordeum vulgare ssp. spontaneum). The resistance gene origining from wild barley in line RS42-6*O, showed a recessive mode of inheritance, whereas the other wild barley genes were (semi)-dominant. RFLP mapping of these three genes was performed in segregating F₂ populations. The recessive gene in line RS42-6*O, was localized on barley chromosome 1S (7HS), while the (semi)-dominant genes in lines RS137-28*E, and HSY-78*A, were localized on chromosomes 1L (7HL) and 7L (5HL), respectively. Closely linked RFLP clones mapped at distances between 2.6cM and 5.3 cM. Hitherto, specific loci for powdery mildew resistance in barley had not been located on these chromosomes. Furthermore, tests for linkage to the unlocalized resistance gene Mlp revealed free segregation. Therefore, these genes represent new loci and new designations are suggested: mlt (RS42-6*O), Mlf (RS137-28*E), and Mlj (HSY-78*A). Comparisons with mapped QTLs for mildew resistance were made and are discussed in the context of homoeology among the genomes of barley (H-vulgare), wheat (Triticum aestivum), and rye (Secale cereale). Duplications of RFLP bands detected in the neighbourhood of Mlf and mlt might indicate an evolutionary interrelationship to the Mla locus for mildew resistance.     

Monosomic analysis of tissue culture response in wheat (Triticum aestivum L.)

Summary The ability of immature embryos of wheat (Triticum aestivum L.) to respond in cell culture was examined in crosses between the Wichita monosomic series and a highly regenerable line, ND7532. Segregation in disomic controls and 13 monosomic families showed a good fit to a monogenic ratio indicating a qualitative mode of inheritance. Segregation in the cross involving monosomic 2D showed a high frequency of regeneration (93.6%) and high callus growth rate (1.87 g/90 days) indicating that 2D is a critical chromosome. Modifying genes may be located on other chromosomes. Substitution of chromosomes from a low regenerable cultivar Vona further indicated that the group 2 chromosomes, in particular chromosome 2D, possess genetic factors promoting callus growth and regeneration.     

Spontaneous chlorophyll mutants of Pennisetum americanum: Genetics and chlorophyll quantities

Summary Thirteen spontaneously occurring chlorophyll deficient phenotypes have been described and their genetic basis was established. Ten of these — white, white tipped green, patchy white, white virescent, white striping 1, white striping 2, white striping 4, fine striping, chlorina and yellow virescent showed monogenic recessive inheritance and the remaining three — yellow striping, yellow green and light green seedling phenotypes showed digenic recessive inheritance. The genes for (i) white tipped green (wr) and yellow virescent (yv) and (ii) patchy white (pw) and white striping 1 (wst 1) showed independent assortment. Further, the genes for white (w), white tipped green (wr) and yellow virescent (yv) were inherited independently of the gene for hairy leaf margin (Hm).In the mutants — white tipped green, patchy white, white striping 1, white striping 2, fine striping, chlorina, yellow virescent, yellow striping, yellow green and light green phenotypes total quantity of chlorophyll was significantly less than that in the corresponding controls, while in white virescent there was no reduction in the mature stage. For nine of the mutants the quantity of chlorophyll was also estimated in F₁'s (mutant x control green). In F₁'s of six of the mutants — white tip, patchy white, chlorina, yellow virescent, fine striping and yellow striping the quantity of chlorophyll was almost equal to the wild type. In the F₁'s of three of the mutants — white striping 1, white striping 2 and light green an intermediate value between the mutant and wild types was observed. In yellow virescent retarded synthesis of chlorophyll, particularly chlorophyll a was observed in the juvenile stage. Reduced quantity of chlorophyll was associated with defective chloroplasts. In the mutants — white tipped green, white virescent, fine striping, chlorina, yellow striping, yellow green and light green defective plastids were also observed. In patchy white secondary destruction of chlorophylls and the presence of defective plastids were found to be associated with reduced chlorophyll quantity at maturity.Paper chromatographic studies of leaf flavonoids revealed some variation between the inbreds, but there were three common spots, 7, 8 and 9, except for PDP in which the spot 8 was absent. Chlorophyll deficient mutants differed from their respective controls in the absence of one or more of the spots present in the controls and in the presence of new spots in some of the mutants.Most of the chlorophyll mutants showed higher survival rate in the Kharif season than in Rabi season which was attributed to the higher mean day temperature and longer day light period in the Kharif season than in Rabi season.     

Effect of anther orientation on microspore-callus production in barley (Hordeum vulgare L.)

Barley anthers from cold pretreated spikes produced no or few calluses when plated with both loculi in contact with the medium (flat). When anthers were plated with only one loculus in contact with the medium (up), a high proportion of the anthers produced calluses. The top loculus of the up anthers was most productive. Flat anthers, when compared with up anthers, were not only slower to produce multicellular pollen grains (MCPs) and microcalluses, but also produced fewer of them and ceased production earlier. The MCPs and microcalluses in flat anthers grew more slowly and few developed beyond the 30 cell stage. These results establish the importance of anther orientation for barley anther culture.     

Ionic relations of aeroponically-grown olive genotypes,during salt stress

Two olive (Olea europaea L.) genotypes, Frantoio and Leccino, were exposed to increasing concentrations of NaCl (0-30-60-120 mM) in an aeroponic cultivation system for 60 days. Dry weights and sodium and potassium contents of apical and basal leaves, new and old wood, and roots were measured to determine Na uptake rate, Na translocation rate and K-Na selectivity ratio (S_K,Na). Frantoio showed a higher salt resistance than Leccino. Frantoio and Leccino had a similar Na uptake rate, but largely differed for Na translocation to the shoot. Furthermore Frantoio exhibited a higher K-Na selectivity than Leccino at both whole plant level and above all at the level of shoot system. Resistance mechanism of Frantoio is probably related to Na esclusion by roots and to the ability to maintain an appropriate K/Na ratio in actively growing tissues.Research supported by National Research Council of Italy, Special project RAISA.     

Inheritance of resistance to the bean-pod weevil (Apion godmani Wagner) in common beans from Mexico

The bean-pod weevil (BPW), Apion godmani Wagner, often causes heavy losses in crops of common bean (Phaseolus vulgaris L.). Farmers need resistant bean cultivars to minimize losses, cut production costs, stabilize seed yield, and reduce pesticide use and consequent health hazards. To design effective breeding methods, breeders need new and better sources of resistance and increased knowledge of their modes of inheritance. We therefore: (1) compared sources of resistance to BPW, (2) studied the inheritance of resistance, and (3) determined whether the sources possess similar or different genes for BPW resistance. The following sources of resistance, originating from the Mexican highlands, were evaluated for 3 years at INIFAP-Santa Lucía de Prias, Texcoco, Mexico: Amarillo 153, Amarillo 169, Hidalgo 58, J 117, Pinto Texcoco, Pinto 168, and Puebla 36. All except Puebla 36 were crossed with the susceptible cultivar Jamapa. Amarillo 153 and Puebla 36 were crossed with another susceptible cultivar, Bayo Mex. The parents, F₁ hybrids, and F₂ populations were evaluated for BPW damage in 1992. Backcrosses of the F₁ of Jamapa/Pinto 168 to the respective susceptible and resistant parents were also evaluated in 1992. All seven resistant accessions were crossed in all possible combinations, excluding reciprocals. The resulting 21 F₁ hybrids and 21 F₂ populations were evaluated for BPW damage in 1994. J 117 had the highest level of resistance to BPW. Pinto Texcoco and Puebla 36 had the highest mean damage score of all seven sources of resistance. The F₁ hybrids between susceptible parents and resistant sources were generally intermediate. Two genes segregating independently controlled the BPW resistance in each accession. One gene, Agm, has no effect when present alone, whereas the other gene, Agr, alone conferred intermediate resistance. When both genes were present, resistance to BPW was higher. Based on mean BPW damage scores, all 21 F₁ hybrids and their F₂ populations, derived from crosses among seven resistant accessions, were resistant. However, data from individual plant damage scores in F₂ populations of Amarillo 169/Pinto 168 and Pinto Texcoco/Pinto 168 suggested that at least one gene in each of the three accessions was non-allelic. Data also indicated that Amarillo 169 had a dominant gene that conferred high levels of BPW resistance, irrespective of the alleles at the other locus; and that Pinto Texcoco and Pinto 168 possessed two different genes for intermediate resistance.     

Variability and genetics of tolerance for aluminum toxicity in rice (Oryza sativa L.)

A study was undertaken to investigate the variability among lowland rice cultivars and the mode of gene action of aluminum (Al) toxicity tolerance in rice. Pregerminated seeds were grown in a nutrient solution containing 30 ppm Al and in normal nutrient solution, and relative root length (RRL) was determined at the 14-day-old stage to characterize genotypes for tolerance. Sixty-two traditional rice cultivars grown on lowland acid sulfate soil areas of Asia and West Africa were tested. Tolerant varieties Azucena, IRAT104, and Moroberekan, moderately sensitive IR29 and IR43, and sensitive IR45 and IR1552 were used to investigate the genetics of tolerance by diallel analysis. Of the 62 cultivars tested, only 3 were found to be sensitive to A l toxicity. Among the tolerant cultivars identified, 11 (Siyam Kuning, Gudabang Putih, Siyam, Lemo, Khao Daeng, Siyamhalus, Bjm-12, Ketan, Seribu Gantang, Bayer Raden Rati, and Padi Kanji) were found to possess higher levels of tolerance than the improved tolerant upland cultivar IRAT104. Diallel analysis revealed that high RRL is governed by both additive and dominance effects with a preponderance of additive effects. The trait exhibited partial dominance, and one group of genes was detected. Heritability was high, and environmenal effects were low. Findings suggest that when breeding for A1 toxicity tolerance, selection can be made in early generations. The pedigree method of breeding would be suitable. Combining ability analysis revealed the importance of both general combining ability (GCA) and specific combining ability (SCA) in the genetics of A1 toxicity tolerance in rice. GCA was more prevalent than SCA. Tolerant parens Azucena, IRAT104, and Moroberekan were the best general combiners. The presence of reciprocal effects among crosses suggested the proper choice of parents in hybridization programs. Results indicated that Azucena, IRAT 104, and Moroberekan should be used as the female in crosses for A1 toxicity tolerance.     

Characterization of groups of the zygomycete genusRhizopus

Rhizopus is a zygomycetous genus. Several species of this taxon may infect humans and lower animals. Seventeen isolates ofRhizopus species in three distinct morphological groups were studied: the stolonifer group (sporangiophores greater than 1 mm in height, sporangial diameters of 100–275 µm, branched rhizoids); the arrhizus group (sporangiophores greater than 1 mm in height, branched rhizoids, sporangial diameters of 100–240 µm); and the microsporus group (sporangiophores less than 0.8 mm in height, sporangial diameters less than 100 µm, simple rhizoids). Maximal growth temperatures were characteristic: the stolonifer group grew at 30°C, the arrhizus group grew at 36°C, and the microsporus group grew at 45°C. The DNA mol% G + C base composition of all isolates ranged from 34.9 to 40.2% Species within the three groups were grouped by DNA differences. The arrhizus group was most distinctive with a value of 34.9–36.3%; the stolonifer and microsporus groups had G + C values of 37.0–39.3% and 37.8–40.2%, respectively. Our research clarifies and defines the G-C values of the three important groups ofRhizopus species.     

Inheritance of callus formation ability in anther cultures of rice,Oryza sativa L.

Summary Inheritance of ability to form callus in rice anther culture was studied using the diallel technique. Anthers containing uninucleate microspores from two japonica cultivais (Minehikari and Taipei 309), two indica cultivars (Mingolo and Suweon 290), and 12 F₁'s of the diallel crosses involving these four parents were cultured on Chaleffs R₂ medium and evaluated for callus induction. The parents showed significant differences in anther callus formation, from 41.9% (Taipei 309) to 0% (Suweon 290). Callus induction ability was inherited as a recessive character conditioned by a single block of genes. Additive gene effects were predominant. The japonica types seemed to be good combiners for callus induction. The order of dominance among the four parents was Suweon 290, Mingolo, Minehikari and Taipei 309.     

Haploid production in durum wheat by the interaction of Aegilops kotschyi cytoplasm and 1BL/1RS chromosomal interchange

The present study describes the development of an alloplasmic haploid-inducer in durum wheat cv Cando. This cultivar possesses the homozygous wheat-rye translocation 1BL/1RS from the 6x-wheat cv Veery. The nucleus of 4x-Cando-Veery 1BL/1RS was introduced into Aegilops kotschyi cytoplasm by initially using (kotschyi)-Salmon as the maternal parent. In the cross of this alloplasmic durum line with Cando-Veery 1BL/1RS, which was used as the recurrent pollen parent, haploids (n=14) were produced. The frequency of haploids increased from 5.7% in the F₁ generation to 14% in the BC₁ generation. The presence of rye chromosome arm 1RS and the concomitant loss of 1BS in (kotschyi)-Cando-Veery 1BL/1RS are necessary for haploid induction. Proposals are made which may enable the use of haploids produced by nucleo-cytoplasmic interactions in future wheat breeding programs.     

Screening of turfgrass species and cultivars for NaCl tolerance

Summary Information regarding the relative levels of salt tolerance between cultivars of Kentucky bluegrass (Poa pratensis L.) is lacking. The objectives of this study were to 1) develop a simple, quick and sensitive method of screening turfgrass species for NaCl tolerance and 2) to compare the relative salt tolerance of five cultivars of Kentucky bluegrass (Ram I, Adelphi, Baron, Bensun, and Nassau) to other known salt tolerant turfgrass species such as alkalaigrass (Puccinellia distans (L.) Parl. cv. Fults) and two cultivars of red fescue (Festuca rubra L. Dawson, and Checker).Alkalaigrass and both cultivars of red fescue retained a high level of salt tolerance compared to the Kentucky bluegrass cultivars. Significant variability in salt tolerance was apparent among the Kentucky bluegrass cultivars with Adelphi and Ram I exhibiting the best overall tolerance.     

Restriction fragment analysis of ‘null’ forms at the Gli-1 loci of bread and durum wheats

Summary Wheat accessions lacking some of the - and -gliadin components encoded by the Gli-1 loci on the short arm of chromosome 1D in bread wheat and chromosome 1A in durum wheat were studied by two-dimensional polyacrylamide gel electrophoresis and restriction fragment analysis. Digested genomic DNAs of normal and null forms were probed with a cDNA clone related to -/-gliadins and with a genomic clone encoding an LMW subunit of glutenin. The hybridisation patterns with the -/-gliadin probe were similar to those of cvs Chinese Spring and Langdon used as standards for bread and durum wheats, respectively, but several restriction fragments located on the 1D chromosome of bread wheat and the 1A chromosome of durum wheat were absent in the null forms. In addition, specific LMW glutenin fragments encoded by the same chromosomes were also absent in the null forms, suggesting that simultaneous deletions of blocks of genes for both -/-gliadins and LMW glutenins had occurred. Comparisons of the protein and RFLP patterns enabled some proteins to be mapped to specific restriction fragments.     

Minding the Emperor's new mind

This essay equates Penrose's (1989) Emperor with the scientist engaging in mental (Schrödinger's cat) or real experiments.The simultaneous presence of apparently contradictory phase-spatial symmetry conditions on the various hierarchical levels of biological systems are seen as the result of genetic and neurophysiological information that interferes with the physico-chemical vectors between the structural components of the system, the experimenter being an integral part of this informational causality. Equations pertaining to the lowest structural levels of matter, therefore, may not be extendable over higher levels of biological organization, including human science and technology, which are seen as part and parcel of biology. This situation calls for a formal theoretical biophysics which concentrates on macroscopic processes where life and, above all,Homo sapiens, is involved.     

Inheritance of alleles for Cgy1 and Gy4 storage protein genes in soybean

Summary A cultivar lacking the glycinin subunit A₅A₄B₃ (Raiden) was crossed with one lacking the -subunit of -conglycinin (Keburi). Analysis of F₂ and F₃ progeny indicated that the missing bands of the A₅A₄B₃ and the -subunit were each controlled by a recessive allele of two independently segregating genes. Gene symbols Gy ₄/gy ₄ and Cgy ₁/cgy ₁ were proposed for the genes which confer the presence or absence of the glycinin and conglycinin subunits, respectively.Cooperative research of USDA-ARS and the Indiana Agric. Exp. Stn., Purdue Univ., West Lafayette, IN 47907, USA. Indiana Agric. Exp. Stn. Journal Article 9675. Financial support from the American Soybean Association Research Foundation is gratefully acknowledged     

Donor tissue and culture condition effects on mesophyll protoplasts of Medicago sativa

Mesophyll protoplasts were produced from clones of two cultivars of Medicago sativa, Rangelander and Regen S. Protoplasts from the Regen S clone generally gave rise to calli while those from the Rangelander clone would undergo direct embryogenesis. Effects of plant growth conditions, donor tissue pretreatment and protoplast culture conditions on mesophyll protoplast production and subsequent development patterns were investigated. The major factor determining whether or not mesophyll protoplasts would be produced from either of the clones was the pretreatment in water of shoots excised from the donor plants. Pretreatment in water containing growth regulators did not alter protoplast production or development in the Regen S clone. Pretreatment of the Rangelander clone shoots with abscisic acid or naphthaleneacetic acid was slightly beneficial to embryo production while pretreatment with benzylaminopurine was detrimental. Altered leaf morphology induced by growth condition changes did not affect mesophyll protoplast production or subsequent development patterns when shoots were pretreated in water. Culture of protoplasts in liquid droplets or solid agar medium increased low density protoplast survival and subsequent embryo production in the Rangelander clone.     

Hypoxia Tolerance of Two Haplochromine Cichlids: Swamp Leakage and Potential for Interlacustrine Dispersal

The ability to tolerate hypoxia in some haplochromine cichlid fishes contributes to the richness of habitats occupied by the lineage and may be important in interlacustrine dispersal through swampy channels. Lacustrine members of the genus Astatotilapia tend to be ecologically plastic but are rarely encountered in the interior of dense swamps. A notable exception is seen in the swamp corridor that joins Lake Kabaleka with Lake George, Uganda, where one species (Astatotilapia wrought-iron) is abundant, and a second species, A. aeneocolor, is rare. Both species are abundant in the open waters of the main lake. In this paper, we compare physiological (oxygen consumption) and behavioral indicators of hypoxia tolerance between A. wrought-iron from swamp and open-water habitats and between the two species of Astatotilapia. When exposed to progressive hypoxia, all fish used aquatic surface respiration (ASR); however, swamp-dwelling A. wrought-iron showed lower gill ventilation rates prior to the initiation of ASR, higher pre-ASR aggression rates, higher swimming speed during ASR, and a higher rate of bubble exchange than both the open-water group of A. wrought-iron and A. aeneocolor. These differences may reflect interpopulational variation in selection pressure for low-oxygen tolerance between swamp and open-water habitats. Several lines of evidence suggest that A. wrought-iron was in general more hypoxia tolerant than A. aeneocolor. These include a lower ASR₉₀ threshold, a drop in gill ventilation rate with the onset of ASR, and lower rate of equilibrium loss under extreme hypoxia in A. wrought-iron. The routine metabolic rate and critical oxygen tension did not differ between swamp-dwelling and open-water A. wrought-iron, or between A. wrought-iron and A. aeneocolor. Comparative data on the ASR thresholds and critical oxygen tensions of the Astatotilapia species from Lake Kabaleka and other East African cichlids suggest intermediate hypoxia tolerance. Nevertheless, our study suggests that some generalized lacustrine haplochromines may leak through swamp corridors even under relatively extreme conditions.     

A neural coding model for sensory intensity discrimination,to be applied to gustation

Summary This paper presents a model of the neural coding and discrimination of sensory intensity. The model consists of five stages: (1) the coding of stimulus intensity in peripheral receptors or neurons by a rate code. The relevance of comparing different analysis intervals for the response is pointed out; (2) neural processing, according to either labeled-line or across-fiber pattern theory. In addition, two possible non-linearities in the processing are considered: a threshold mechanism, and contrast enhancement by reciprocal inhibition; (3) a neural discriminator, based on signal-detection theory; (4) a memory stage; (5) an effector organ providing a behavioral output. Emphasis is put on stages 2 and 3.The model produces predictions of the differential threshold, which should be directly testable in a behavioral two-alternative forced-choice paradigm. The model will be applied to gustatory intensity discrimination in rat in a subsequent study (Maes and Erickson 1984). The Discussion pays attention to the relative contributions of peripheral and central noise sources. It also compares the present model with Beidler's (1958) approach through just noticeable differences (JND's). The model presented here seems more adequate in providing an understanding of sensory information processing.Abbreviations AFP across fiber pattern - DA discrimination acuity - DT differential threshold - JND just noticeable difference - LL labeled line - NTS nucleus tractus solitarius     

Variation in the karyotype of three cultivars of Narcissus tazetta L. (Amaryllidaceae)

Three cultivated varieties of Narcissus tazetta, Grand Soleil d'Or, Chinese Sacred Lily and Cypri, are triploid (2n = 3x+30) with the basic number 10. Grand Soleil d'Or has three homomorphic sets, each comprising 2 long submetacentrics, 4 long acrocentrics and 4 short acrocentrics. Karyotypes of the other two varieties are heteromorphic. Both possess one telocentric satellited chromosome. In addition, Cypri shows translocation between two chromosomes belonging to the seventh and eighth triplets. The number of secondary constrictions varies between 3 (Chinese Sacred Lily) and 4 (Grand Soleil d'Or and Cypri) which is also the number of nucleoli observed in the respective varieties.