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.
Genomic-based-breeding tools for tropical maize improvement
Maize has traditionally been the main staple diet in the Southern Asia and Sub-Saharan Africa and widely grown by millions of resource poor small scale farmers. Approximately, 35.4 million hectares are sown to tropical maize, constituting around 59% of the developing worlds. Tropical maize encounters tremendous challenges besides poor agro-climatic situations with average yields recorded <3 tones/hectare that is far less than the average of developed countries. On the contrary to poor yields, the demand for maize as food, feed, and fuel is continuously increasing in these regions. Heterosis breeding introduced in early 90 s improved maize yields significantly, but genetic gains is still a mirage, particularly for crop growing under marginal environments. Application of molecular markers has accelerated the pace of maize breeding to some extent. The availability of array of sequencing and genotyping technologies offers unrivalled service to improve precision in maize-breeding programs through modern approaches such as genomic selection, genome-wide association studies, bulk segregant analysis-based sequencing approaches, etc. Superior alleles underlying complex traits can easily be identified and introgressed efficiently using these sequence-based approaches. Integration of genomic tools and techniques with advanced genetic resources such as nested association mapping and backcross nested association mapping could certainly address the genetic issues in maize improvement programs in developing countries. Huge diversity in tropical maize and its inherent capacity for doubled haploid technology offers advantage to apply the next generation genomic tools for accelerating production in marginal environments of tropical and subtropical world. Precision in phenotyping is the key for success of any molecular-breeding approach. This article reviews genomic technologies and their application to improve agronomic traits in tropical maize breeding has been reviewed in detail.
Development of new analytical methods for the determination of caffeine content in aqueous solution of green coffee beans
BackgroundThis study was conducted to develop fast and cost effective methods for the determination of caffeine in green coffee beans. In the present work direct determination of caffeine in aqueous solution of green coffee bean was performed using FT-IR-ATR and fluorescence spectrophotometry. Caffeine was also directly determined in dimethylformamide solution using NIR spectroscopy with univariate calibration technique.ResultsThe percentage of caffeine for the same sample of green coffee beans was determined using the three newly developed methods. The caffeine content of the green coffee beans was found to be 1.52 ± 0.09 (% w/w) using FT-IR-ATR, 1.50 ± 0.14 (% w/w) using NIR and 1.50 ± 0.05 (% w/w) using fluorescence spectroscopy. The means of the three methods were compared by applying one way analysis of variance and at p = 0.05 significance level the means were not significantly different. The percentage of caffeine in the same sample of green coffee bean was also determined by using the literature reported UV/Vis spectrophotometric method for comparison and found to be 1.40 ± 0.02 (% w/w).ConclusionNew simple, rapid and inexpensive methods were developed for direct determination of caffeine content in aqueous solution of green coffee beans using FT-IR-ATR and fluorescence spectrophotometries. NIR spectrophotometry can also be used as alternative choice of caffeine determination using reduced amount of organic solvent (dimethylformamide) and univariate calibration technique. These analytical methods may therefore, be recommended for the rapid, simple, safe and cost effective determination of caffeine in green coffee beans.