Cytology and leafspot resistance in Arachis hypogaea x wild species hybrids

Summary Introgression of germplasm from diploid wild Arachis species to A. hypogaea has great potential for improving pest resistance in cultivated peanuts. This investigation evaluated methods for incorporating exotic germplasm into cultivated peanuts, especially for Cercospora arachidicola Hori resistance. Interspecific hybrids between A. hypogaea (cvs. NC 2 and NC 5) and the wild species A. cardenasii Krap. et Greg. nom. nud. and A. chacoense Krap. et Greg. nom. nud. were analyzed cytologically and for leafspot resistance. All F₁ hybrids were sterile, had irregular meiosis, and very few multivalents. They were highly resistant to C. arachidicola in field tests and had a 10-fold reduction of conidia per lesion in the greenhouse as compared to A. hypogaea cultivars. After colchicine treatments of F₁ hybrids, hexaploids (2n=60) and aneuploids (2n=54, 56, 63) were observed. The hexaploids had up to 18 univalents per pollen mother cell and very few multivalents, indicating a low frequency of intergenomic chromosome pairing. For C. arachidicola resistance, significant differences were not found among wild species parents, F₁ hybrids and two generations of hexaploids. Most hexaploids were stable at 2n=60 and embryos aborted when backcrosses with the respective wild species were attempted. However, when hexaploids were backcrossed to A. hypogaea, several fertile pentaploid (2n=50) offspring were obtained. Use of self-pollinating pentaploids is believed to be the quickest method to recover 40-chromosome hybrid derivatives in these hybrids.     

Morphological, cytological and disease resistance studies of the intersectional hybrid between Arachis hypogaea L. and A. glabrata Benth.

Arachis glabrata Benth, variety glabrata collector GK 10596 (PI276233; ICG 8176) belonging to section Rhizomatosae has resistance to rust, late leaf spot and viral diseases. A. hypogaea L. cv MK 374 (section Arachis) is susceptible to rust, late leaf spot and to the viral diseases peanut stripe, peanut mottle and peanut bud necrosis. Hybrids between A. hypogaea cv MK 374 and A. glabrata were produced after inter specific pollinations and embryo culture. The hybrids produced had morphological characters of both parents plus floral abnormalities not seen in either parent. It was possible to identify the hybrids by esterase isozyme analysis when still in culture. Cytological research showed variable chromosome association and also homeology between the genomes of A. hypogaea and A. glabrata. The hybrids inherited resistance to rust, late leaf spot, peanut bud necrosis and peanut stripe diseases from the pollen parent A. glabrata. This revised version was published online in August 2006 with corrections to the Cover Date.     

Production of interspecific hybrids between Alstroemeria pelegrina L. var. rosea and A. magenta Bayer by ovule culture

Interspecific hybrids were efficiently produced in the cross-incompatible combination between Alstroemeria pelegrina L. var. rosea and A. magenta Bayer by culturing immature ovules with placenta 7–14 days after pollination on 2 g/l Gelrite-solidified MS medium containing 3% (w/v)sucrose. The plants showed intermediate characteristics between the parents and their hybridity was confirmed by karyotype and DNA analyses. The mean number of chromosome association per PMC at metaphase I was 2.60I+6.70II, pollen stainability was20.8%, and they produced viable seeds after self-pollination. Furthermore, mature plants were obtained when the hybrids were backcrossed as male parents with both the parents. The backcross-progeny from A. pelegrina var. rosea × hybrids exhibited 3.8 to 79.7% pollen stainability and that from A. magenta × hybrids 78.8 to 98.3%. Almost all of these plants produced viable seeds after self-pollination, which implies that they can beutilized for breeding of novel cultivars of Alstroemeria. This revised version was published online in July 2006 with corrections to the Cover Date.     

Interspecific hybridization in the genus Arachis through embryo culture

Summary The embryos of a cultivated tetraploid peanut (Arachis hypogaea), a wild diploid species (A. villosa), and their hybrid embryos, which generally abort in nature, were cultured in vitro and the plants have been successfully transferred to the soil. The hybrids showed triploid chromosome number (3x=30). The significance of wide hybridization in peanut-improvement programs is discussed.     

Ribosomal DNA repeat unit polymorphism and heritability in peanut (Arachis hypogaea L.) accessions and related wild species

Repeat unit length and restriction site variation in ribosomal RNA geneclusters (rDNA) was surveyed in 77 Arachis accessions, includingsamples from 39 accessions of cultivated Arachis hypogaea(2n=4x=40), 36 accessions representing 15 related tetraploid and diploidwild species, and two synthetic amphidiploids. Total genomic DNA wasdigested with five restriction enzymes, and probed with three heterologousribosomal clones of wheat and broad bean. Four rDNA repeat unit lengthclasses were recognized in the Arachis species. Restriction site analysisshowed that some SacI, BamHI and TaqI cleavage sites in rDNA unit werehighly conserved. With few exceptions, the variable BamHI and EcoRV siteswere able to differentiate the taxonomic sections and species, respectively.Arachis hypogaea and A. duranensis accessions produced fourrDNA length classes. Among these, three were identical with those of otherArachis species. A SacI restriction site (s) from probe (Ver6-5) cangenerally distinguish the two subspecies A. hypogaea ssp. hypogaea and A. hypogaea ssp. fastigiata. Forty nine per centof bands were polymorphic across the A. hypogaea accessionsanalysed. This study does not support A. batizocoi to be a progenitorof A. hypogaea. For the gene array, the contribution from eachparental genome can be detected in the two synthetic amphidiploids.     

Variation in progenies of an Arachis hypogaea x diploid wild species hybrid

Summary Derivatives of a cross between cultivated peanuts, Arachis hypogaea L. (2n=40), and the wild species collection GKP 10017 (2n=20) were compared morphologically, for leafspot resistance and for yield. The objective of the study was to determine the effects of wild species germplasm on the A. hypogaea genome. The sterile F₁ hybrid which resulted from crossing the two species was treated with colchicine to restore fertility at the 6x ploidy level. The resulting hexaploid was cytologically unstable and progeny lost chromosomes until stability was regained at the 2n=40 chromosome level. Forty-seven characters were used to analyze the variation among plants in the tetraploid interspecific hybrid population. The plants were compared to four cultivated lines plus GKP 10017. Many hybrids were intermediate to the two parents in morphology. Individual traits such as growth habit, pod and seed size, elongation of the constricted area between pods, nodulation and leaflet size were altered by the presence of GKP 10017 germplasm in many of the hybrid plants. Cercospora arachidicola Hori and Cercosporidium personatum (Berk. & Curt.) Deighton resistances were evaluated for all plants. Several hybrids had few lesions due to either leafspot pathogen. In addition, 24 largeseeded interspecific hybrid selections were compared to the cultivated variety NC 5 for yield. Five selections were superior to both parents at p=0.01. Morphology, disease resistance and yields appeared to be greatly influenced by the wild species GKP 10017 germplasm in plants of the interspecific hybrid population. The potentials of using wild species for improvement of the cultivated peanut are discussed.Paper number 5948 of the journal series of the North Carolina Agricultural Research Service, Raleigh, NC 27650. The investigation was supported in part by ICRISAT and SEA-CR grant no. 701-15-51.     

Rapid estimation of ploidy levels in in vitro-regenerated interspecific Arachis hybrids and fertile triploids

Summary A significant correlation among chromosome number, chloroplast number and pollen grain size was observed using interspecific hybrids (2n=30,60), Arachis hypogaea (2n=40), A. stenosperma (PI 338280) (2n=20), A. batizocoi (PI 468329) (2n=20) and A. villosulicarpa Hoehne (PI 336984) (2n=20), representing four ploidy groups. Both chloroplast number in guard cells and pollen grain size were found to be positively correlated with ploidy, and provided a reliable method to distinguish diploid, triploid, tetraploid and hexaploid plants from each other regardless of their taxonomic backgrounds. These methods in combination with root-tip chromosome counts enabled us to confirm ploidy determinations on all three dissue layers, L₁, L₂ and L₃ (guard cells, microsporocytes and root meristematic cells, respectively), and to detect the chimeric nature of some colchicine-treated tissue culture-derived plants. Screening pollen grains by size and the detection of highly stainable and viable, 30-chromosome pollen grains have enhanced the use of triploid hybrids in peanut breeding programs.     

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.     

Strain improvement in the cultivated mushroom Agaricus bisporus

Summary Early attempts at genetic improvement in the cultivated mushroom Agaricus bisporus (Lange) Imbach were empirical, for little was understood of its natural breeding system. The mushroom is now known to be a secondarily homothallic species with a single multiallelic mating-type factor. This better understanding makes it possible to evaluate those breeding methods previously used and to suggest alternatives.Strain selection alone based on single spores, multispores or tissue culture may give improvement in the short term but it is unlikely to be as effective as methods involving controlled crossing. Mixing fertile strains may produce hybrids but it is difficult to identify them. It is better to use non-fertile isolates because only hybrids fruit. The earlier recognition of hybrids can be achieved using markers which are expressed in culture and genetic resistances may be especially useful in this respect.There is also a possible role for other Agaricus species which may be grown commercially and are more amenable to genetic manipulation than is A. bisporus.     

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.     

The transfer of triazine resistance from Brassica napus L. to B. oleracea L. III. First backcross to parental species

Summary Atrazine resistant Brassica napus × B. oleracea F₁ hybrids were backcrossed to both parental species. The backcrosses to B. napus produced seeds in both directions but results were much better when the F₁ hybrid was the pollen parent. Backcrosses to B. oleracea failed completely but BC₁s were rescued by embryo culture both from a tetraploid hybrid (2n = 4x = 37; A₁C₁CC) and sesquidiploid hybrids (2n = 3x = 8; A₁C₁C). Progeny of crosses between the tetraploid hybrid and B. oleracea had between 25 and 28 chromosomes. That of crosses between the sesquidiploid hybrid and B. oleracea had between 21 and 27. A few plants that had chromosome counts outside the expected range may have originated from either diploid parthenogenesis, unreduced gametes or spontaneous chromosome doubling during in vitro culture. Pollen stainability of the BC₁s ranged from 0% to 91.5%. All the BC₁s to B. oleracea were resistant to atrazine.     

Relationships between African species of the genus Cucumis L. Estimated by the production,vigour and fertility of F1 hybrids

Summary The production, vigour, and fertility of F₁ hybrids between nine African species of the genus Cucumis L. were studied as a measure of the relationships between the species. Hybrid plants were obtained from 29 out of the 72 possible cross combinations. Two F₁ hybrids died as seedlings, and 27 hybrids were raised to maturity. Pollen production and stainability varied greatly amongst these hybrids, as did fruit and seed set following self pollination and backcrossing with either parental species. The fruit shape of the hybrids was always intermediate between that of both parental species.Two species appeared to be closely related: C. prophetarum L. and C. anguria var. longipes A. Meeuse. Most other species produced highly to moderately fertile F₁ hybrids with at least one other species. C. metuliferus Naud. produced only sterile hybrids with C. zeyheri Sond. 2x. The results sustain the recent taxonomic classification of the genus (Jeffrey, 1980).     

Chromosome association and pollen fertility in some interspecific hybrids of Lilium

Summary Meiotic observations in PMCs were made in eight kinds of interspecific hybrids in Lilium. Three hybrids, 6134-S × L. cernuum. L. regale × L. leichtlinii maximowiczii and L. auratum platyphyllum × L. henryi showed respectively the mean chromosome association frequencies of 1.4_II, 0.8_II and 1.5_II per cell at MI which were the lowest values hitherto reported in Lilium hybrids. In L. longiflorum × L. henryi, 33.3° of the cells had 1 or 2 multivalents of three to five chromosomes in addition to uni- and bivalents. 12° bivalents and 12 univalents were invariably observed in the triploid hybrid L. longiflorum × L. cernuum. The hybrids between the species belonging to the different sections of the genus generally showed high pollen sterility, with some exceptions. A more or less remote genomic homology was found between the different sections in Lilium.     

The probable genome donors to Arachis hypogaea L. based on arachin seed storage protein

Summary Interrelationships among six diploid species (A. chacoense, A. villosulicarpa, A. batizocoi, A. correntina, A. duranensis and A. cardenasii), tetraploid wild groundnut A. monticola and four accessions of A. hypogaea were studied by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of total seed proteins and arachin immunoprecipitates. Arachin in A. monticola and A. hypogaea cv. Trombay Groundnut 9 (TG-9) was composed of four acidic subunits (47.5 kd, 45.1 kd, 42.6 kd and 41.2 kd) and one major basic subunit (21.4 kd). The arachin subunit pattern in cv. Spanish Improved (SP) and TG-1 was almost similar to the pattern observed in A. monticola with the exception that the SP lacked the 41.2 kd and TG-1 lacked the 42.6 kd acidic subunits of A. monticola. Seed extracts of all diploid species studied reacted with the anti-arachin antibodies raised against SP arachin. The electrophoretic analysis of immunoprecipitates of diploid species showed a range of acidic subunits from 46.2 kd to 41.2 kd and one or two basic subunits of 21.4 kd and 20.2 kd. None of the diploid species showed the 47.5 kd subunit found in A. monticola or A. hypogaea. Of the diploid species, A. duranensis and A. cardenasii demonstrated two acidic subunits of 45.1 kd and 41.2 kd and a basic subunit of 21.4 kd as found in A. monticola. Likewise A. batizocoi showed two acidic subunits of 45.1 kd and 42.6 kd and the basic subunit of 21.4 kd as was observed in A. monticola. Based on electrophoretic data, our research supports the earlier conclusion that the probable genome donors to A. monticola are A. batizocoi and A. duranensis or A. cardenasii.     

Aseptic culture of gynophores to obtain peanut intersectional hybrids

Summary Cross-incompatibility between cultivated peanuts and their wild relatives outside the section Arachis has impeded the utilization of many species possessing high resistances or good qualities. Despite the great efforts made to culture immature ovules or embryos, few hybrid offspring have been obtained. In this study, gynophores from Arachis hypogaea L. pollinated with A. glabrata Benth. were cultured and F₁ hybrids seeds were harvested, and F₂ and F₃ generations produced. The characters of F₂ generation exhibited a wide range of segregation. Leaf peroxidase isozyme PAGE analysis revealed that the hybrids were quite different from their parents in relation to band number, width and isozyme activity. The zymograms of the hybrids and their parents were partially alike. This verified the authenticity of the hybrids.     

Chromosome association and fertility in hybrids of Iris laevigata Fisch. x I. ensata Thunb.

Summary Chromosome association at MI of PMCs in hybrids of I. laevigata x I. ensata was examined and compared with that of parental species. The mean chromosome association per cell in the parental species was 0.02_I+11.99_II for I. ensata (Kachô, 2n=24), 0.11_I+15.94_II and 0.15_I+15.92_II for I. laevigata (Yukidôrô and wild type, 2n=32), respectively. On the other hand, the mean chromosome association per cell in the hybrids (2n=28) was 20.22_I+3.88_II+0.0046_IV for Yukidôrô × Kachô and 20.92_I+3.54_II for wild type × Kachô. Thus, the hybrids indicated a low homology between I. ensata and I. laevigata, and did not exhibit pollen and seed fertility. The utility of the hybrids was discussed.     

A cluster analysis of wild and domesticated soybean phenotypes

Summary Cluster analysis techniques were used to examine phenotypic variation within the USDA Soybean Germplasm Collection. This study included ten Plant Introductions each of the domesticated soybean, Glycine max (L). Merr., the wild soybean, G. soja Sieb. & Zucc., and a disputed species, G. gracilis Skvortz. G. max and G. soja were found to be morphologically distinct entities and G. gracilis was found to be conspecific with G. max. Because G. gracilis probably represents weedy races associated with the cultivated soybean and because gracilis phenotypes can be distinguished from G. max, the designation, G. max forma gracilis, is recommended.Joint contribution: North Central Region, Agricultural Research, Science and Education Administration, U.S. Department of Agriculture, and Journal Paper No. J-9389 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa 50011; Project 2107.     

Unilateral ‘eclipse sterility’ in reciprocal crosses between Solanum verrucosum Schlechtd. and diploid S. Tuberosum L.

Summary F₁ hybrids from Solanum verrucosum CPC 2247 x diploid S. tuberosum exhibit a type of male sterility which after staining with lactophenol acid fuchsin gives the pollen grains an eclipse-like appearance, but size and shape are fully normal. The reciprocal cross, which could be made using some exceptional diploid S. tuberosum clones that did not show unilateral incompatibility with S. verrucosum, gives rise to F₁ hybrids without this eclipse sterility (Fig. 1 and 2).Pollen quality of the reciprocal hybrids was compared applying several direct and indirect methods. A critical cytological study revealed that meiosis in pollen mother cells is completely normal. When eclipse sterility is involved, the nucleus in the young microspore does not divide any more after meiosis and some kind of plasmolytic process occurs in the microspore leading to the characteristic eclipselike appearance.     

The origin,evolution, cultivation,dissemination, and diversification of Asian and African rices

Summary Available evidences drawn from biosystematics, evolutionary biology, biogeography, archaeology, history, anthropology, paleo-geology and paleo-meteorology are pooled to reconstruct the series of events that led to the cosmopolitan cultivation of the Asian cultivated rice (O. sativa) and the regionalized planting of the African cultigen (O. glaberrima) in West Africa. The genus Oryza originated in the Gondwanaland continents and, following the fracture of the supercontinent, became widely distributed in the humid tropics of Africa, South America, South and Southeast Asia, and Oceania. The two cultivated species have had a common progenitor in the distant past. Parallel and independent evolutionary processes occurred in Africa and in Asia, following the sequence of: wild perennialwild annualcultivated annual. The weed races also contributed to the differentiation of the cultivated annuals. The corresponding members of the above series are O. longistaminata Chev. et Roehr., O. barthii A. Chev., O. glaberrima Steud., and the stapfii forms of O. glaberrima in Africa; O. rufipogon Griff., O. nivara Sharma et Shastry, O. sativa L., and the spontanea forms of O. sativa in Asia.The differentiation and diversification of the annuals in South Asia were accelerated by marked climatic changes following the last glacial age, dispersal of plants over latitude or altitude, human selection, and manipulation of the cultural environment.Cultivation of rice began in many parts of South and Southeast Asia, probably first in Ancient India. Cultural techniques such as puddling and transplanting were first developed in north and central China and later transmitted to Southeast Asia. Wetland culture preceded dryland culture in China, but in hilly areas of Southeast Asia, dryland cultivation is older than lowland culture. The planting method progressed from shifting cultivation to direct sowing in permanent fields, then to transplanting in bunded fields.Widespread dispersal of the Asian cultigen led to the formation of three eco-geographic races (Indica. Sinica or Japonica, and Javanica) and distinct cultural types in monsoon Asia (upland, lowland, and deep water). Varietal types changed readily within the span of a millenium, largely due to cultivators' preferences, socio-religious traditions, and population pressure. Genetic differentiation developed parallel to the ecologic diversification process.The African cultigen developed later than the Asian cultigen and has undergone less diversification. The wild races in South America and Oceania retain their primitive features mainly due to lack of cultivation pressure or dispersal.Both the African and Asian rices are still undergoing evolutionary changes at habitats where the wild, weed, and cultivated races co-exist.     

Intergeneric hybrids and an amphiploid between Elymus canadensis and Psathyrostachys juncea

Summary Fifty four hybrid plants between Elymus canadensis and Psathyrostachys juncea were obtained by handpollination and embryo culture. The average cross compatibility between both species was 31.2 percent. One amphiploid plant was induced by colchicine treatment. The hybrid and amphiploid plants resembled P. juncea in appearance but showed a higher plant height and dry matter yield than the parents. The hybrids showed extremely low pollen stainability and were completely sterile. With the exception of one plant (2n=3x+1=22), all hybrid plants were allotriploids (SHN, 2n=3x=21). The amphiploid plant (SSHHNN, 2n=6x=42) showed 58.9% pollen stainability and 11.6% seed fertility.Mean chromosome associations of the hybrids and amphiploid at metaphase I were 0.02IV+0.06III+2.03II+16.91I and 0.07III+18.00II+5.85I, respectively. Lagging chromosomes, chromosome bridges, abnormal cytokinesis, and micronuclei were occasionally observed at the anaphase, telophase, or tetrad stage.