Cytological and molecular characterization of a chromosome interchange and addition lines in Cadet involving chromosome 5B of wheat and 6Ag of Lophopyrum ponticum

Efforts to transfer wheat curl mite (Eriophyes tulipae Keifer) resistance from Lophopyrum ponticum 10X (Podb.) Love to bread wheat (Triticum aestivum L.) have resulted in the production of a number of cytogenetic stocks, including an addition line of 6Ag, a ditelo addition line, and a wheat-Lophopyrum translocation line. Characterization of these lines with C-banding, in situ hybridization with a Lophopyrum species-specific repetitive DNA probe (pLeUCD2), and Southern blotting with pLeUCD2 and a 5S ribosomal DNA probe (pScT7) confirmed that the distal portion of the short arm of 6Ag was translocated onto the distal portion of 5BS (5BL. 5BS-6AgS). It was also determined that the ditelo addition was an acrocentric chromosome of 6AgS.     

Introgression and characterization of barley yellow dwarf virus resistance from Thinopyrum intermedium into wheat.

Wheatgrasses (species of Agropyron complex) have previously been reported to be resistant to barley yellow dwarf virus (BYDV). To introgress this resistance into wheat, Triticum aestivum x Thinopyrum (Agropyron) intermedium hybrids were advanced through a backcrossing program and reaction to BYDV, as determined by enzyme-linked immunosorbent assay (ELISA), is reported for the first time in backcross populations of wide hybrids between wheat and wheatgrasses. ELISA values revealed highly resistant to highly susceptible segregants in backcrosses. BYDV resistance was expressed in some backcross derivatives. Continued selection, based on cytology and ELISA in each generation, eliminated most of the unwanted wheatgrass chromosomes and produced self-fertile BYDV resistant wheat lines. The BYDV resistant lines with 2n = 42 had normal chromosome pairing similar to wheat, and their F1 hybrids with wheat had two univalents. DNA analyses showed that the source of alien chromatin in these BYDV resistant wheat lines is distinguishable from that in other Th. intermedium derived BYDV resistant wheat lines. Chromosome pairing and restriction fragment length polymorphism analyses indicated that the 42 chromosome resistant Purdue wheat lines are substitution lines in which chromosome 7D was replaced by a chromosome from Th. intermedium that was carrying gene(s) for BYDV resistance.     

Cytological and molecular characterization of oat x maize partial hybrids

In cereals, interspecific and intergeneric hybridizations (wide crosses) which yield karyotypically stable hybrid plants have been used as starting points to widen the genetic base of a crop and to construct stocks for genetic analysis. Also, uniparental genome elimination in karyotypically unstable hybrids has been utilized for cereal haploid production. We have crossed hexaploid oat (2n=6x=42, Avena sativa L.) and maize (2n=2x=20, Zea mays L.) and recovered 90 progenies through embryo rescue. Fifty-two plants (58%) produced from oatxmaize hybridization were oat haploids (2n=3x=21) following maize chromosome elimination. Twenty-eight plants (31%) were found to be stable partial hybrids with 1–4 maize chromosomes in addition to a haploid set of 21 oat chromosomes (2n=21+1 to 2n=21+4). Ten of the ninety plants produced were found to be apparent chromosomal chimeras, where some tissues in a given plant contained maize chromosomes while other tissues did not, or else different tissues contained a different number of maize chromosomes. DNA restriction fragment length polymorphisms (RFLPs) were used to identify the maize chromosome(s) present in the various oat-maize progenies. Maize chromosomes 2, 3, 4, 5, 6, 7, 8, and 9 were detected in partial hybrids and chromosomal chimeras. Maize chromosomes 1 and 10 were not detected in the plants analyzed to-date. Furthermore, partial self-fertility, which is common in oat haploids, was also observed in some oat-maize hybrids. Upon selfing, partial hybrids with one or two maize chromosomes showed nearly complete transmission of the maize chromosome to give self-fertile maize-chromosome-addition oat plants. Fertile lines were recovered that contained an added maize chromosome or chromosome pair representing six of the ten maize chromosomes. Four independently derived disomic maize chromosome addition lines contained chromosome 4, one line carried chromosome 7, two lines had chromosome 9, one had chromosome 2, and one had chromosome 3. One maize chromosome-8 monosomic addition line was also identified. We also identified a double disomic addition line containing both maize chromosomes 4 and 7. This constitutes the first report of the production of karyotypically stable partial hybrids involving highly unrelated species from two subfamilies of the Gramineae (Pooideae — oat, and Panicoideae — maize) and the subsequent recovery of fertile oat-maize chromosome addition lines. These represent novel material for gene/ marker mapping, maize chromosome manipulation, the study of maize gene expression in oat, and the transfer of maize DNA, genes, or active transposons to oat.Joint contribution of the Minnesota Agricultural Experiment Station and USDA-ARS. Scientific journal series paper No. 21 859 of the Minnesota Agricultural Experiment Station. Mention of a trademark or proprietary product does not constitute a guarantee or warranty by the USDA-ARS or the University of Minnesota and does not imply approval over other products that also may be suitable     

Identification and characterization of wheat-wheatgrass translocation lines and localization of barley yellow dwarf virus resistance.

Stable introgression of agronomically important traits into crop plants through wide crossing often requires the generation and identification of translocation lines. However, the low efficiency of identifying lines containing translocations is a significant limitation in utilizing valuable alien chromatin-derived traits. Selection of putative wheatgrass-wheat translocation lines based on segregation ratios of progeny from gamma-irradiated seed using a standard phenotypic analysis resulted in a low 4% success rate of identifying barley yellow dwarf virus (BYDV) resistant and susceptible translocation lines. However, 58% of the susceptible progeny of this irradiated seed contained a Thinopyrum intermedium chromosome-specific repetitive sequence, which indicated that gamma-irradiation-induced translocations occurred at high rate. Restriction fragment length polymorphism (RFLP) analysis of susceptible lines containing alien chromatin, their resistant sister lines and other resistant lines showed that more than one third of the progeny of gamma-irradiated double monosomic seeds contained wheatgrass-wheat translocations. Genomic in situ hybridization (GISH) analysis of selected lines confirmed that these were wheatgrass-wheat translocation lines. This approach of initially identifying BYDV susceptible deletion lines using an alien chromosome-specific repetitive sequence followed by RFLP analysis of their resistant sister lines efficiently identified resistant translocation lines and localized the BYDV resistance to the distal end of the introgressed Th. intermedium chromosome.     

Properties and isolates of barley yellow dwarf virus

Barley yellow dwarf virus is persistently transmitted by a number of aphid species of which three, Rhopalosiphum padi, Sitobion avenae and Metopolophium dirhodum, are common in most years. Other aphids may be locally important. Isolates of the virus differ in their virulence and geographical distribution and are not transmitted equally well by all aphid vectors. Isolates with similar properties are grouped into strains according to their transmission by vectors and their severity. Changes in strain and aphid occurrence from year to year alter the incidence of virus and its effect on yield. These changes emphasize the need for detailed knowledge of cereal aphid biology and epidemiology of BYDV before effective control can be used.     

Development and characterization of a new barley yellow dwarf virus (BYDV)-resistant wheat germplasm

A barley yellow dwarf virus (BYDV)-resistant line HG295 was selected from a cross between cv. 77-5433 and Zhong 5 after extensive investigation in field, greenhouse and ELISA. Cytological analysis revealed that it was an euploid line and genetically stable. The existence of alien DNA in HG295 was identified by RAPD and Southern hybridization analyses showed that the alien DNAs came from Zhong 5 or Th. intermedium. The differences of BYDV resistance between L1 and HG295 are discussed.     

Sequence and organization of barley yellow dwarf virus genomic RNA.

The nucleotide sequence of the genomic RNA of barley yellow dwarf virus, PAV serotype was determined, except for the 5'-terminal base, and its genome organization deduced. The 5,677 nucleotide genome contains five large open reading frames (ORFs). The genes for the coat protein (1) and the putative viral RNA-dependent RNA polymerase were identified. The latter shows a striking degree of similarity to that of carnation mottle virus (CarMV). By comparison with corona- and retrovirus RNAs, it is proposed that a translational frameshift is involved in expression of the polymerase. An ORF encoding an Mr 49,797 protein (50K ORF) may be translated by in-frame readthrough of the coat protein stop codon. The coat protein, an overlapping 17K ORF, and a 3'6.7K ORF are likely to be expressed via subgenomic mRNAs.     

The effect of increasing dosage of barley yellow dwarf virus on some resistant and susceptible barleys

Five spring barleys, grown either in pots out of doors or in the field, were inoculated with barley yellow dwarf virus (BYDV) using 5, 10, 20 or 50 infective aphids (Rhopalosiphum padi) per plant. Control plants of each variety received no aphids. Infection with all aphid numbers had highly significant adverse effects on all varieties except Cb 1029, an early maturing BYDV-resistant barley of Ethiopian origin. 12583 Co, a locally bred, late maturing barley possessing the same resistance gene as Cb 1029 suffered more in a pot experiment, but less than three susceptible varieties all of which were severely damaged even when few infective aphids were used. Progressive effects with increasing aphid numbers, indicative of dosage response, occurred in some varieties. These effects included delay in heading and increased stunting, but not less yield. In Cb 1029, BYDV infection caused a reduction in the number of heads per plant, but this was partly compensated for by an increase in the number of grains per head. Conversely, BYDV infection in 12583 Co caused an increase in the number of heads, partly offset by a decrease in the number of Brains tier head.     

Inheritance and effectiveness of genes in barley that condition tolerance to barley yellow dwarf virus

Tolerance to barley yellow dwarf virus (BYDV) in five Ethiopian barley genotypes was conditioned by major genes which apparently occurred at, or near, the same chromosome locus in each genotype. The genes or alleles differed in effectiveness, one providing a high, three an intermediate and one a low level of tolerance. In plants having a slow rate of growth, the effectiveness of the tolerance genes tended to be masked, but the effectiveness of the gene or allele which provided the highest level of tolerance was masked to a lesser extent than that of genes or alleles providing low or intermediate levels of tolerance.     

The cytogenetics of a Triticum turgidum x Psathyrostachys juncea hybrid and its backcross derivatives

Psathyrostachys juncea (2n = 2x = 14, NN), a source of barley yellow dwarf (BYDV) virus resistance with tolerance to drought and salinity, has been successfully hybridized in its autotetraploid form (2n = 4x = 28, NNNN) as the pollen parent to durum wheat (Triticum turgidum L.). The 2n = 4x = 28 (ABNN) F₁ hybrid has a mean meiotic metaphase-I configuration of 20.29 univalents + 0.29 ring bivalents + 3.36 rod bivalents + 0.14 trivalents. Spike length, internode length, glume awn length and lemma awn length, as well as the general spike morphology of the F₁ hybrid, are intermediate with those of the two parents. Pollinating the ABNN F₁ hybrid has given backcross (BC)-I derivatives of an amphiploid (AABBNN) that expresses limited self-fertility. BC-2 derivatives have been obtained from these plants. Direct transfers of useful genes from Ps. juncea to wheat would require substantial genetic manipulation strategies. Both conventional and novel methodologies, which may complement each other, and so facilitate reaching an agricultural objective end point, are addressed.     

Visualization of A- and B-genome chromosomes in wheat (Triticum aestivum L.) x jointed goatgrass (Aegilops cylindrica Host) backcross progenies.

Wheat (Triticum aestivum) and jointed goatgrass (Aegilops cylindrica) can cross with each other, and their self-fertile backcross progenies frequently have extra chromosomes and chromosome segments, presumably retained from wheat, raising the possibility that a herbicide resistance gene might transfer from wheat to jointed goatgrass. Genomic in situ hybridization (GISH) was used to clarify the origin of these extra chromosomes. By using T. durum DNA (AABB genome) as a probe and jointed goatgrass DNA (CCDD genome) as blocking DNA, one, two, and three A- or B-genome chromosomes were identified in three BC2S2 individuals where 2n = 29, 30, and 31 chromosomes, respectively. A translocation between wheat and jointed goatgrass chromosomes was also detected in an individual with 30 chromosomes. In pollen mother cells with meiotic configuration of 14 II + 2 I, the two univalents were identified as being retained from the A or B genome of wheat. By using Ae. markgrafii DNA (CC genome) as a probe and wheat DNA (AABBDD genome) as blocking DNA. 14 C-genome chromosomes were visualized in all BC2S2 individuals. The GISH procedure provides a powerful tool to detect the A or B-genome chromatin in a jointed goatgrass background, making it possible to assess the risk of transfer of herbicide resistance genes located on the A or B genome of wheat to jointed goatgrass.     

Molecular characterization of a Thinopyrum intermedium group 2 chromosome (2Ai-2) conferring resistance to barley yellow dwarf virus.

The wheat--Thinopyrum intermedium addition lines Z1 and Z2 carry 21 pairs of wheat chromosomes and one pair of Th. intermedium chromosomes (2Ai-2) conferring resistance to barley yellow dwarf virus (BYDV). GISH results using the genomic DNA of Pseudoroegneria strigosa (S genome) as the probe indicated that the 2Ai-2 chromosome in Z1 and Z2 is an S-J intercalary translocation. Most of the 2Ai-2 chromosome belongs to the S genome, except for about one third in the middle region of the long arm that belongs to the J genome. The results of detailed RFLP analyses confirmed that the 2Ai-2 chromosome is extensively homoeologous to wheat group 2 chromosomes. Some new RFLP markers specific to the 2Ai-2 chromosome were identified. A RAPD marker, OP-R16(340), specific to the 2Ai-2 chromosome, was screened. We converted the RAPD marker into a sequence-characterized amplified region (SCAR) marker (designated SC-R16). The study establishes the basis for selecting translocation lines with small segments of the 2Ai-2 chromosome and localizing the BYDV resistance gene when introgressed into a wheat background.     

Feeding responses of two grain aphids to barley yellow dwarf virus-infected oats

The probing behavior of greenbugs, Schizaphis graminum (Rondani), and oat-bird cherry aphids, Rhopalosiphum padi (L.), was electronically monitored on healthy oats, Avena sativa (L.), and on oats infected with the RPV-NY isolate of barley yellow dwarf virus (BYDV). S. graminum fed better on infected oats than on healthy oats. This was manifested by a shorter time before initiation of committed phloem ingestion, and by increased duration of ingestion from phloem of infected compared to healthy plants. In addition, on infected oats S. graminum made fewer interruptions in their probing once their stylets were inserted into tissues, including phloem. R. padi fed similarly on infected and healthy oats, except that these aphids made fewer short probes to the phloem (lasting <15 min) on infected compared to healthy oats. BYDV infection of oats increased the rate of population growth of S. graminum relative to that on healthy oats but had no effect on the population growth of R. padi. The proportion of aphids of R. padi which developed into alatae on BYDV-infected oats was significantly greater than on healthy oats, but S. graminum showed no such response.

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Résumé Le comportement de sondage de S. graminum (Rond.) et de R. padi (L.) a été suivi électroniquement sur avoine (Avena sativa L.) saine ou contaminée par l'extrait RPV-NY du virus jaune du nanisme de l'orge (BYDV). S. graminum s'est mieux alimenté sur avoine contaminée que saine; ceci se traduisait par un temps de latence inférieur avant l'ingestion de phloème et par une prolongation de la période d'ingestion. De plus, sur avoine contaminée, S. graminum a moins souvent interrompu le sondage, une fois que les stylets avaient étè insérés dans les tissus, y compris le phloème. Les résultats avec R. padi présentaient les mêmes tendances, mais les améliorations des performances alimentaires de cette espèce sur avoine contaminée ont été moins nettes que pour S. graminum. La contamination de l'avoine par BYDV a accru le taux de croissance de la population de S. graminum par rapport à ce qui a été observé sur avoine saine; l'effet était moins prononcé pour R. padi. La proportion de pucerons devenant aliés était plus élevée sur avoine contaminée que saine, rien de tel n'a été observé avec S. graminum.

     

The relationship between growth rate and the expression of tolerance to barley yellow dwarf virus in barley

Barley varieties were most tolerant to infection with barley yellow dwarf virus (BYDV) when they grew rapidly, whether the rate of growth was determined by manipulation of the environment or by the innate genetic constitution of the host. A specific, incompletely dominant gene conditioning a high level of tolerance to the virus in certain rapidly growing genotypes in which it occurred naturally, failed to do so when transferred to slower growing genotypes. However, reintroduction into genotypes capable of rapid growth led to full restoration of the gene's effectiveness. Virus-free aphids recovered BYDV less readily from quick- than from slow-growing genotypes, all homozygous for the tolerance gene.     

Production and cytogenetics of hybrids of Triticum aestivum x Leymus innovatus

Summary Hybrid plants were obtained between Triticum aestivum (2n=6x=42, AABBDD) and Leymus innovatus (2n=4x=28, JJNN) at a frequency varying from 0.4% to 1.2% of the pollinated florets. Improvement of the embryo culture medium resulted in a higher frequency of embryo rescue. Eight of ten hybrids had the expected chromosome number of 35 (ABDJN). Meiotic analysis indicated that there was no homology between the genomes of the two species. Two hybrids had only 28 chromosomes. Comparison of chromosome pairing between the two types of hybrids suggested that Leymus innovatus carries genes that affect chromosome pairing and behavior. The relatively high occurrence of spontaneous doubling in the meiocytes of these hybrids may indicate that backcrossing of the hybrids to wheat should be possible, although frequent chromosome irregularities observed in the meiocytes of the hybrids may decrease the probability of success of this step, which is essential to the process of gene transfer from L. innovatus to wheat.Contrib. no. 366     

Production of cloned cDNA from a Swedish barley yellow dwarf virus isolate

A cDNA library was produced from the RNA of a Swedish MAV-like isolate of barley yellow dwarf virus (BYDV). The procedure involved random priming and the ds cDNA was cloned into the EcoRl site of the plasmid pUC19. Among the clones obtained some hybridised specifically with MAV-like isolates whereas others also hybridised with PAV-like isolates. Only very weak hybridisation was observed with an RPV-like isolate. An Australian cDNA clone, reported to be PAV-specific (pBY82, Waterhouse, Gerlach & Miller, 1986), hybridised with Swedish MAV-like but not with PAV-like isolates. Probes prepared from the clones detected virus in plant extracts by dot-blot hybridisation with sensitivity greater than that of ELISA. Virus was also readily detected in extracts of viruliferous aphids.