Production and identification of doubled haploids in tropical maize.
The production of maize doubled haploid (DH) lines is a technique commonly used by private companies, but not by Brazilian public institutions. Research on this technique is essential to develop and improve the production of DH lines grown under tropical conditions. We assessed the ability of a gynogenetic haploid inducer system to induce haploids in a tropical environment, assessed the induction rate of haploids identified using the R-navajo morphological marker, checked for interference from the generation of hybrid donors on haploid induction, measured the ability of flow cytometry, and simple sequence repeat marker techniques to identify doubled haploids. Seeds from the inducer Krasnodar Embryo Marker Synthetic (KEMS) line were sown in Ponta Grossa, PR, and Cravinhos, SP, and the plants were crossed to produce six hybrids and their F2 generations. The seeds were separated according to the R-navajo morphological marker indicator of haploidy (purple endosperm and white embryo) and germinated in a controlled environment. Chromosomal duplication was performed in seedlings selected as putative haploids. We performed subsequent confirmation of ploidy and the success of duplication using flow cytometry and SSR marker techniques. We concluded that DH lines can be obtained from hybrids crossed with the inducer KEMS line. The generation of inbred hybrids did not affect the induction rate or chromosomal duplication in haploids. The use of flow cytometry and SSR markers was effective in verifying chromosomal duplication in haploids.
Effectiveness of the R1-navajo embryo marker on sorting haploids in tropical maize germplasm
Modern maize breeding is currently based on Doubled Haploids (DH) technology. In following the steps to produce inbred lines by DH, sorting the haploids are the most important of them. Haploid inducers usually carry the R1 gene, responsible of purple pigmentation on seeds. Navajo is a typical phenotyping from this gene and leads to purple coloration on endosperm and embryo of the diploids but any pigmentation on haploid kernels. Aimed to analyze the effectiveness of R1-nj on sorting true haploids over tropical sources, two different experiments were conducted. In the first experiment, kernels analyzed came from the following induction cross (L47 x IL47), (P9 x IP9), (2B707 x IL47), and (2B707 x IP9) in which IL47 and IP9 were the tropical haploid inducers. In the second experiment, KHI (Krasnodar Haploid Inducer) was the haploid inducer and three commercial were adopting as sources. Putative haploids were sorted by R1-nj expression and the haploid level was checked by chromosome counting our phenotyping at the flowering stage. Overall, very low true haploid discovery rates were observed. 
2B707 was the most reliable source in displaying true haploids. The ratio of this effectiveness among the two ex- periments to R1-nj marker ranged from 8.80 to 30.2%. These results can be explained by the presence of inhibitory genes on the tropical sources. Additionally, at the flowering stage, haploids plants had data of spontaneous doubling collected and summarized. Recovery on the female organs seems to be higher than male. Haploids from the IAC125 source exhibited best fully spontaneous doubling among the sources analyzed.
Investigations on the rôle of organic matter in plant nutrition
1. As the result of decomposition of green manure in a swamp soil, the coarse fractions (including coarse and fine sand) of the surface layer (first 3 inches) decreased by 11·2 per cent. while the fine ones (silt+clay) increased by 9·8 per cent. in the course of 43 days. The effect was less pronounced in the manured soil maintained at 60 per cent. saturation, while no alteration was to be noticed in the case of the unmanured specimens. 2. Treatment with dilute hydrochloric acid prior to dispersion had no appreciable effect on the estimates of mechanical composition obtained in the different cases. Loss in solution was also comparatively small. 3. The silica-sesquioxide ratio increased slightly as the result of decomposition of green manure in swamp soil. The composition of the fine fractions was also altered, the total percentages of the oxides of iron, aluminium and silicon increasing from 90·5 to 99·3 in the course of 71 days. 4. The practical significance of the above and related observations has been discussed.