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1.
Tissue electroporation was applied to a member of the Triticeae family, namely tritordeum (Hordeum chilense Roem.×Triticum turgidum L. Conv. durum), for the generation of fertile transgenic plants. Two transgenic plants were recovered following the treatment of 361 explants
of immature inflorescences (although they were subsequently found to result from the same transformation event). The expression
of both inserted marker genes (uidA and bar) was confirmed using standard assays, while transgene integration was confirmed using PCR and Southern hybridization analyses.
Integration pattern, segregation ratio and the inheritance of transgene expression in T1 progeny were consistent for the presence of a single transgene locus containing five to ten plasmid insertions. Although
this procedure has been applied to other cereal species, stable transformation of the Triticeae using tissue electroporation
has not previously been reported.
Received: 28 October 1999 / Revision received: 25 August 2000 / Accepted: 29 August 2000 相似文献
2.
Selection and molecular characterization of imidazolinone resistant mutation-derived lines of Tritordeum HT621 总被引:1,自引:0,他引:1
C. Rodríguez-Suárez M. C. Ramírez C. Martínez S. Nadal A. Martín S. G. Atienza 《Molecular breeding : new strategies in plant improvement》2009,23(4):565-572
Imidazolinone herbicides resistant varieties, induced by mutations at the AHAS gene (acetohydroxyacid synthase), have been
developed in many crops. Hexaploid tritordeum (Tritordeum Asch. & Graebn.) is the amphiploid derived from the cross between Hordeum chilense (HchHch) and durum wheat Triticum turgidum L. (Thell) (AABB). Tritordeums have the potential to become a new crop with high added-value for food or feed. Mutagenesis
with EMS was conducted to obtain imidazolinone resistant lines derived of the tritordeum HT621. Eleven M3 plants were selected after imidazolinone treatment and five descendants of two of these lines (HT621-M3R1-3 and HT621-M3R10-1)
were analyzed at the molecular level. Partial sequences of the three homologous AHAS loci in genomes A, B, and Hch were obtained as well as those of HT621. A partial sequence of the AHAS gene in Hordeum chilense is first described in this work, and the designation ahasL-H
ch
1 is proposed. A single Ser-Asn627 substitution at the AHAS locus in the B genome is responsible of resistance in both lines. We propose the name AhasL-B2 for this resistance allele. This is the first report of the selection of imidazolinone resistant lines of tritordeum and
the molecular characterization of the mutation conferring this resistance. 相似文献
3.
J. A. Fernandez J. M. Gonzalez N. Jouve 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1985,70(1):85-91
Summary The meiotic behaviour of the amphiploid Hordeum chilense X Triticum turgidum conv. durum using a C-banding staining method is studied. Nine pairs of chromosomes at metaphase-1 (4A, 7A and the seven of the B genome) were identified and the remaining wheat chromosomes (1A, 2A, 3A, 5A and 6A) and seven of the chilense (1 to 7 H
ch
chromosomes) were assigned to its particular genome. A similar mean number of univalents from parental genomes (wheat and wild barley) were found. No meiotic pairing between chilense and turgidum chromosomes was detected. Differences in the meiotic behaviour per chromosome and amongst genomes are explained on the basis of cytomorphological and heterochromatin characteristics. 相似文献
4.
J. A. Fernández N. Jouve 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1987,73(5):690-698
Summary Polyacrylamide and starch gel electrophoresis of esterase (EST), glutamate oxaloacetate transaminase (GOT) and phosphoglucomutase (PGM) isozymes in Hordeum chilense, Triticum turgidum conv. durum, the amphiploid H. chilense X T. turgidum (Tritordeum), and the durum wheat/H. chilense monosomic addition lines revealed the chromosomal location of one EST locus, two GOT loci and one PGM locus. Loci Est-H
ch1 and Got-H
ch2 were found on chromosome 6Hch,Got-H
ch3 on chromosome 3Hch, and Pgm-H
ch1 on chromosome 4Hch. These results lend evidence for the assumed homoeology relationships between chromosomes of Triticeae species. 相似文献
5.
P. Hernández M. Hemmat N. F. Weeden G. Dorado A. Martín 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,98(5):721-727
RAPD markers were developed for octoploid×Tritordeum (amphiploid Hordeum chilense×Triticum aestivum) and its parents. Addition lines were used to identify specific RAPD markers for the Hordeum chilense chromosomes detectable in a wheat background. Twelve RAPD fragments have been cloned, sequenced and converted into STS markers.
Eleven of these STSs have maintained both the chromosome specificity and the possibility of detection in a wheat background.
The use of these markers in multiplexed PCRs facilitates both the efficient and reliable screening of new addition lines as
well as the monitoring of introgression of H. chilense in bread and durum wheat.
Received: 5 June 1998 / Accepted: 17 September 1998 相似文献
6.
7.
J. A. Fernández N. Jouve 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1987,73(3):433-439
Summary A study of 6-phosphogluconate dehydrogenase and malate dehydrogenase isozyme expression in Triticum turgidum conv. durum /Hordeum chilense monosomic addition lines has revealed the location of two structural genes, 6-pgd-H
ch
2 and Mdh-H
ch
1, on chromosome 1Hch of H. chilense. The homoeology between 1Hch and other chromosome of Triticeae related species is discussed on the basis of isozyme gene analysis. 相似文献
8.
Hernández P Laurie DA Martín A Snape JW 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2002,104(4):735-739
A selection of 36 wheat and 35 barley simple sequence repeat markers (SSRs) were studied for their utility in Hordeum chilense. Nineteen wheat and nineteen barley primer pairs amplified consistent H. chilense products. Nine wheat and two barley SSRs were polymorphic in a H. chilense mapping population, producing codominant markers that mapped to the expected homoeologous linkage groups in all but one case.
Thirteen wheat and 10 barley primer pairs were suitable for studying the introgression of H. chilense into wheat because they amplified H. chilense products of distinct size. Analysis of wheat/H. chilense addition lines showed that the H. chilense products derived from the expected homoeologous linkage groups. The results showed that wheat and barley SSRs provide a valuable
resource for the genetic characterization of H. chilense, tritordeums and derived introgression lines.
Received: 20 November 2000 / Accepted: 12 April 2001 相似文献
9.
Analysis and optimisation of DNA delivery into wheat scutellum and Tritordeum inflorescence explants by tissue electroporation 总被引:4,自引:0,他引:4
Wheat scutella and tritordeum inflorescences were transformed by tissue electroporation with plasmid DNA containing a β-glucuronidase (GUS) gene (gus A) under the control of the rice actin1 promoter. Factors affecting electroporation efficiency
were analysed. Important factors were electroporation voltage and pulse length, the volume of electroporation buffer, the
osmoticum of electroporation buffer and medium, the osmoticum of pre-electroporation culture medium, and pre-electroporation
incubation time and temperature. Maximum transient gene expression was obtained with a single pulse of 550 V/cm from a 960-μF
capacitor, using 200 μl of electroporation buffer, after 2–3 h culture on media with 357 mOsm for wheat scutella or 1 day
on media with 222 mOsm for tritordeum inflorescences, and 0.5–1 h pre-electroporation incubation with DNA at 24 °C. Under
these conditions, up to 90% of the explants showed GUS expression, and up to 149 expression signals were recorded per replicate.
Electroporated explants showed high rates of survival and retained the ability to regenerate plants via somatic embryogenesis.
Received: 9 August 1996 / Revision received: 11 September 1997 / Accepted: 2 July 1998 相似文献
10.
P. Barceló A. Cabrera C. Hagel H. Lörz 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1994,87(6):741-745
The effects of different media and cold pretreatment of spikes on the androgenic response and regeneration capacity from anther culture of tritordeum was studied. L5 medium gave the highest frequency of anther response. The frequency of cultures regenerating green or albino plantlets was not affected by the composition of the medium tested. Cold pretreatment of the spikes significantly increased the frequency of anther response and also the percentage of cultures giving albino plantlets. A mean of four green plants was obtained per 100 subcultured calli/embryos. The percentage of spontaneous chromosome doubling was only 1%. The addition of colchicine at 0.02% to the induction medium significantly increased the frequency of doubled haploids regenerated without any effect on regeneration capacity. This technique proved more efficient than a conventional chromosome-doubling method. 相似文献