Production and genetic evaluation of interspecific hybrids within the genus Sambucus

Attempts were made to produce hybrids among various Sambucus species in order to change the plant structure of S. nigra, a medicinal plant of increasing agricultural importance. The first successful genetic recombination, involving four species, was conducted after the introduction of S. javanica from a South Pacific island. A series of genetic analyses of the involved species and some relatives was performed in order to obtain reliable data on genetic similarity among parental species and to confirm the hybrid nature of putative hybrids. The nuclear 2C DNA content revealed by flow cytometry of nine species showed limited variation, the greatest being in S. nigra (28.6 pg) and the smallest in S. caerulea (23.9 pg). Maximum parsimony analysis of seven Sambucus species was used to form two dendrograms, one based on the sequenced nuclear ribosomal DNA (nrDNA) internal transcribed spacer (ITS) region and the other based on the cpDNA trnT‐trnF region. The results showed similar clustering into distinct groups; on both dendrograms S. nigra and S. javanica ES were closely related. Analysis of hybrid origin was based on the sequenced cpDNA region, genome size differences and PCR–RFLP analysis of the nrDNA ITS region. The seed parent and maternal origin of cpDNA was confirmed for all hybrids, while the pollen parent was confirmed in 21 of 22 putative hybrid plants tested.     

Meiosis and pollen grain viability in Helianthus mollis,Helianthus salicifolius,Helianthus maximiliani and their F1 hybrids with cultivated sunflower

Three diploid perennial sunflower species are useful for variety improvement: Helianthus mollis, because of sessile leaves, H. salicifolius, because of a high oil concentration, and H. maximiliani, a potential source of resistance to Sclerotinia sclerotiorum. The crossability of these species to cultivated sunflower was examined.Hybrids were obtained from eight combinations, with 3–15 F₁ plants per combination. The F₁'s exhibited the dominant phenotype of the wild species. Pollen viability varied between 32.1 and 69.9%. Meiosis was irregular in the F₁ hybrids. At diakinesis, bivalents (62.7–97.9% of meiocytes), univalents (0–31.23%), and multivalents (3.84–7.68%) were detected. At anaphase I, chromosome bridges were detected in 6.77 to 11.44% of meiocytes. Fast chromosomes in metaphase I, and lagging chromosomes in anaphase I and telophase II were evidenced in a high percentage of meiocytes.     

Moricandia arvensis cytoplasm based system of cytoplasmic male sterility in Brassica juncea: Reappraisal of fertility restoration and agronomic potential

Cytoplasmic male sterility (CMS) system based on the cytoplasm from Moricandia arvensis (mori) was investigated for fertility restoration and agronomic potential. Fertility restorer gene for mori CMS was introgressed from cytoplasm donor species as all the evaluated Brassica juncea genotypes (155) acted as sterility maintainers. The allosyndetic pairing between M^a and the A/B genome chromosomes in the monosomic addition plants (2n= 18II + 1M^a) facilitated the gene introgression. Partial fertility restoration (43–52% pollen grain stainability) in F₁ hybrids and absence of segregation for male sterility in F₂ progenies suggested gametophytic control of fertility restoration. The pollen fertility in the F₁ hybrids was, however, sufficient to ensure complete seed set upon bag selfing. Introgression from M. arvensis also helped in correction of chlorosis associated with mori cytoplasm in CMS and fertile alloplasmic B. juncea plants. Yield evaluation of thirty F₁ hybrids having the same nuclear genotype but varied male sterilizing cytoplasms (mori, oxy, lyr, refined ogu), in comparison to respective euplasmic hand bred control hybrids, allowed an estimate of yield penalty associated with different CMS systems. It ranged from 1.8% to 61.6%. Hybrids based on cytoplasmically refined ogu were most productive followed by those based on cytoplasmically refined mori CMS. The male sterility systems emanating from somatic hybridization were found superior than those developed from sexual hybridization.     

Production and characterization of hybrids between Cajanus cajan × C. reticulatus var. grandifolius

Cajanus reticulatus var. grandifolius, endemic to Australia and a wild relative of the cultivated species, C. cajan, was successfully crossed with the latter as the female parent. The major wild species characters such as persistent stipules, long pod hairs, pod shattering, brown seeds with grey speckles, and presence of seed strophiole were dominant in the hybrid. For growth and branching habit, and leaflet, flower, pod, and seed size, the hybrid was intermediate between the parents. The meiotic cells of the hybrid were found to have quadrivalents, trivalents, univalents,and showed reduced chromosome pairing as revealed by the increased number of rod bivalents per cell at metaphase-I, and stickiness and precocious movement of chromosomes to poles in the second division. In comparison to the parents, the hybrid had fewer pods and seeds. However,these anomalies in the interspecific hybrid are not significant enough to preclude the gene transfer from C. reticulatus to the cultivated species through a sexual route. This revised version was published online in August 2006 with corrections to the Cover Date.     

Identifying the alien chromosomes in wheat – Leymus multicaulis derivatives using GISH and RFLP techniques

Genomic in situ hybridization (GISH) and restriction fragment length polymorphism (RFLP) were used to identify the Leymus multicaulis (XXNN, 2n = 28) chromosomes in wheat-L. muliticaulis derivatives. Fifteen lines containing L. multicaulis alien chromosomes or chromosomal fragments were identified. All alien chromosomes or fragments in these 15 lines were from the X genome and none were from the N genome. Eleven L. multicaulis disomic addition lines and four translocation-addition lines were identified with chromosome rearrangements among homoeologous groups 2, 3, 6 and 7. Only homoeologous group 1 lacked rearrangements in addition or translocation chromosomes. The results revealed that translocation in non-homoeologous chromosomes widely exists in the Triticeae and therefore it is necessary to identify the alien chromosomes (segments) in a wheat background using these combined techniques. During the course of the work, probe PSR112, was found to detect X genome addition lines involving L. multicaulischromosomes. This may prove to be a valuable probe for the identification of alien chromosomes in a wheat background. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

Assignment of Endosperm Balance Numbers to the tuber-bearing Solanums and their close non-tuber-bearing relatives

The Endosperm Balance Number (EBN) of over 80 species and subspecies of the tuber-bearing Solanums and their close non-tuber-bearing relatives representing 13 taxonomic series has been determined, with no species assigned to more than one EBN level. Among North American species, most diploids are 1EBN, most tetraploids are 2EBN and all hexaploids are 4EBN; however, among South American species most diploids are 2EBN, most tetraploids are 4EBN and again all hexaploids are 4EBN. Thus species may be isolated from others of the same ploidy level by EBN differences, e.g., 4×(2EBN) from 4×(4EBN), while other species differing in ploidy but having the same EBN may be intercrossed, e.g., 4×(2EBN) and 2×(2EBN). Chromosome doubling or 2n gametes can be used to make a lower EBN species compatible with a higher EBN species. These findings also explain the major crossing difficulties previously inherent in the use of North American species in potato improvement. They also have direct implications for potato improvement, barring the occurrence of other incompatibility barriers. Any 4×(4EBN) cultivar is endosperm compatible and thus will cross with 4×(4EBN) and 6×(4EBN) species. The 2×(2EBN) haploids of 4×(4EBN) cultivars likewise will hybridize with 2×(2EBN) and 4×(4EBN) species. All 2×(1EBN) species are crossable with 2×(2EBN) haploids through 2n gametes or chromosome doubling.Cooperative investigation of the U.S. Department of Agriculture, Agriculture Research Service, and the Wisconsin Experiment Station.     

Breaking the crossability barriers between disomic tetraploid Solanum acaule and tetrasomic tetraploid S. tuberosum

Summary A combination of compatible second pollinations and embryo rescue was applied for systematic production of true tetraploid hybrids from crosses between disomic tetraploid Solanum acaule and tetrasomic tetraploid potato, S. tuberosum. Several genotypes of tetraploid potatoes were pollinated with S. acaule, and the compatible second pollinations were made on the following day, with a genotype of S. phureja, IvP 35 to promote fruit development. Embryo rescue was carried out in 21 families, 14 to 27 days after the first pollination. A total of eight plants were obtained from the embryo rescue and their chromosome numbers were counted in the root tips. Three of the eight plants were identified as tetraploid, and five others as diploid. Morphology, isozyme banding patterns, and pollen stainability, as well as potato spindle tuber viroid (PSTVd) resistance, indicated the hybrid nature of the three plants. This is the first report of successful tetraploid hybrid production between disomic tetraploid S. acaule (4x) and tetrasomic tetraploid potatoes. Seed set from the crosses between one of hybrids and diploid potatoes indicated workable levels of both male and female fertility for introgression of valuable genes from S. acaule into the cultivated potato gene pool. The methodology used may be applied to other disomic tetraploid tuber-bearing Solanum species and with some modifications also to distantly related solanaceous species and genera.     

Protoplast Fusion in the Genus Medicago and Isoenzyme Analysis of Parental and Somatic Hybrid Cell Lines

Leaf mesophyll protoplasts of Medicago arborea (2n = 32) were electrofused with cell suspension and/or callus protoplasts of Medicago sativa. Heterokaryons were identified in agarose beds by their dual fluorescein isothiocianate—chlorophyll fluorescence and their coordinates were recorded. Hybrid minicalli were manually picked up and grown first in nurse culture and then in callus induction medium. Hybrid nature of the selected calli was confirmed by isoenzyme analyses. In order to verify whether morphogenesis of somatic hybrid calli was affected by cell incompatibility, mesophyll and cell suspension protoplasts, derived from the same plant of M. sativa with high embryogenic capacity, were fused. Only callus tissues derived from mesophyll protoplasts retained the highly embryogenic character of the M. sativa genotype, while hybrid cell lines were non-morphogenic and showed isoenzyme patterns similar to tissues derived from cell suspension protoplasts. The achievement of somatic hybrid plants in the genus Medicago is discussed.     

Effect of mentor pollen on pistil-pollen incongruities among species of Cucumis L.

Summary The merits of mentor pollen in certain interspecific hybridizations in the genus Cucumis L. were evaluated. African wild cucumber species Cucumis metuliferus Naud. and C. africanus L. and Asian species C. sativus var. hardwickii Alef and var. sikkimensis Hook, were reciprocally crossed. Pollen tube growth was arrested halfway down the style in all combinations except in C. sativus × C. africanus. Mentor pollen irradiated with 100 and 200 krad grew through the style and into the ovules in all species. Fruitset was obtained in all cases when mentor pollen was used either alone or in mixtures. Fruits set after self pollination with mentor pollen yielded only seeds without embryos.In several crosses, aided by mentor pollen, a number of relatively large seeds (with enlarged embryosac) were obtained. A sample of these embryosacs contained a globular structure like an embryo. Embryosacs with embryo-like structures were explanted on several different media but no development was obtained.