Comparative mapping of the Oregon Wolfe Barley using doubled haploid lines derived from female and male gametes.

The Oregon Wolfe Barley mapping population is a resource for genetics research and instruction. Prior reports are based on a population of doubled haploid (DH) lines developed by the Hordeum bulbosum (H.b.) method, which samples female gametes. We developed new DH lines from the same cross using anther culture (A.C.), which samples male gametes. Linkage maps were generated in each of the two subpopulations using the same 1,328 single nucleotide polymorphism markers. The linkage maps based on DH lines derived from the products of megasporogeneis and microsporogenesis revealed minor differences in terms of estimated recombination rates. There were no differences in locus ordering. There was greater segregation distortion in the A.C.-derived subpopulation than in the H.b.-derived subpopulation, but in the region showing the greatest distortion, the cause was more likely allelic variation at the ZEO1 plant height locus rather than to DH production method. The effects of segregation distortion and pleiotropy had greater impacts on estimates of quantitative trait locus effect than population size for reproductive fitness traits assayed under greenhouse conditions. The Oregon Wolfe Barley (OWB) population and data are community resources. Seed is available from three distribution centers located in North America, Europe, and Asia. Details on ordering seed sets, as well as complete genotype and phenotype data files, are available at http://wheat.pw.usda.gov/ggpages/maps/OWB/ .

Improvements in the production of doubled haploids in durum wheat (Triticum turgidum L.) through isolated microspore culture.

The objective of this study was to produce durum wheat doubled haploid (DH) plants through the induction of microspore embryogenesis. The microspore culture technique was improved to maximize production of green plants per spike using three commercial cultivars. Studies on factors such as induction media composition, induction media support and the stage and growth of donor plants were carried out in order to develop an efficient protocol to regenerate green and fertile DH plants. Microspores were plated on a C(17) induction culture medium with ovary co-culture and a supplement of glutathione plus glutamine; 300 g/l Ficoll Type-400 was incorporated to the induction medium support. Donor plants were fertilized with a combination of macro and microelements. With the cultivars 'Ciccio' and 'Claudio' an average of 36.5 and 148.5 fertile plants were produced, respectively, from 1,000 anthers inoculated. This technique was then used to produce fertile DH plants of potential agronomic interest from a collection of ten F(1) crosses involving cultivars of high breeding value. From these crosses 849 green plants were obtained and seed was harvested from 702 plants indicating that 83% of green plants were fertile and therefore were spontaneously DHs. No aneuploid plant was obtained. The 702 plants yielded enough seeds to be field tested. One of the DH lines obtained by microspore embryogenesis, named 'Lanuza', has been sent to the Spanish Plant Variety Office for Registration by the Batlle Seed Company. This protocol can be used instead of the labor-intensive inter-generic crossing with maize as an economically feasible method to obtain DHs for most crosses involving the durum wheat cultivars grown in Spain.

Production of doubled haploids in durum wheat (Triticum turgidum L.) through isolated microspore culture.

The objective of this work was to produce doubled haploid plants from durum wheat through the induction of androgenesis. A microspore culture technique was developed and used to produce fertile doubled haploid plants of agronomic interest. Five cultivars, one selected line, plus a collection of 20 F(1) crosses between different genotypes of high breeding value were used. Studies on several factors such as pre-treatments and media components were carried out in order to develop a protocol to regenerate green haploid plantlets. Anthers were pre-treated in 0.7 M mannitol. Microspores, from anther maceration, were plated on a C(17) induction culture medium with ovary co-culture. The optimum regeneration medium J25-8 was used. From 35 microspore isolations, 407 green plantlets were obtained. With this technique mature embryos were obtained. Green plants were regenerated from all genotypes used and approximately 67% of them were spontaneously doubled haploids. Some haploids and a very few polyploids plants were obtained. From the 407 plants, 275 were completely fertile and gave enough seeds to be assayed in the field. This protocol could be used complementary to or instead of the intergeneric crossing with maize as an economically feasible method to obtain doubled haploids from most durum wheat genotypes.

[Pneumonia diagnosed with bronchoscopy in HIV-positive patients]. / Neumonías diagnosticadas por broncoscopia en pacientes HIV-positivos.

BACKGROUND: The object of our research is to analyse the microbiological results of the samples which have been obtained by means of fibronchoscopy (FB) from HIV positive patients from 1991 until 1993. METHODS: Sixty fibrobronchoscopies were carried out on fifty-seven HIV positive patients. In every case, samples of bronchoaspirate (BAS), bronchoalveolar lavage (BAL) and telescoping plugged catheter (TPC) were cultured; the last two in a quantitative way. Pneumocystis carinii was investigated in BAL by means of immunofluorescence with monoclonal antibodies. RESULTS: Some microorganisms were isolated in forty-seven bronchoscopies. Thirteen episodes resulted negative. The most frequent etiologic agent was Pneumocystis carinii (seventeen cases). The etiology of fifteen episodes was polymicrobial. The intersticial radiological pattern was the predominant one. It was observed in twenty-seven cases. With regard to immunity, 91% of the patients showed CD4 < 200. CONCLUSIONS: In our research work, the samples that have been obtained by means of FB showed a high percentage of diagnoses; that is the reason why we regard this technique as very useful for the diagnosis of pneumonia in patients with AIDS. Due to the large number of bacterian pneumonia, we consider necessary not only the use of BAL, but also that of TPC in these processes.