Maize miRNAs and their putative target genes involved in chilling stress response in 5-day old seedlings.

BACKGROUND: In the context of early sowing of maize as a promising adaptation strategy that could significantly reduce the negative effects of climate change, an in-depth understanding of mechanisms underlying plant response to low-temperature stress is demanded. Although microRNAs (miRNAs) have been recognized as key regulators of plant stress response, research on their role in chilling tolerance of maize during early seedling stages is scarce. Therefore, it is of great significance to explore chilling-responsive miRNAs, reveal their expression patterns and associated target genes, as well as to examine the possible functions of the conserved and novel miRNAs. In this study, the role of miRNAs was examined in 5d-old maize seedlings of one tolerant and one sensitive inbred line exposed to chilling (10/8 °C) stress for 6 h and 24 h, by applying high throughput sequencing. RESULTS: A total of 145 annotated known miRNAs belonging to 30 families and 876 potentially novel miRNAs were identified. Differential expression (DE) analysis between control and stress conditions identified 98 common miRNAs for both genotypes at one time point and eight miRNAs at both time points. Target prediction and enrichment analysis showed that the DE zma-miR396, zma-miR156, zma-miR319, and zma-miR159 miRNAs modulate growth and development. Furthermore, it was found that several other DE miRNAs were involved in abiotic stress response: antioxidative mechanisms (zma-miR398), signal transduction (zma-miR156, zma-miR167, zma-miR169) and regulation of water content (zma-miR164, zma-miR394, zma-miR396). The results underline the zma-miRNAs involvement in the modulation of their target genes expression as an important aspect of the plant's survival strategy and acclimation to chilling stress conditions. CONCLUSIONS: To our understanding, this is the first study on miRNAs in 5-d old seedlings' response to chilling stress, providing data on the role of known and novel miRNAs post-transcriptional regulation of expressed genes and contributing a possible platform for further network and functional analysis.

Characterization of proteins from grain of different bread and durum wheat genotypes.

The classical Osborne wheat protein fractions (albumins, globulins, gliadins, and glutenins), as well as several proteins from each of the four subunits of gliadin using SDS-PAGE analyses, were determined in the grain of five bread (T. aestivum L.) and five durum wheat (T. durum Desf.) genotypes. In addition, content of tryptophan and wet gluten were analyzed. Gliadins and glutenins comprise from 58.17% to 65.27% and 56.25% to 64.48% of total proteins and as such account for both quantity and quality of the bread and durum wheat grain proteins, respectively. The ratio of gliadin/total glutenin varied from 0.49 to 1.01 and 0.57 to 1.06 among the bread and durum genotypes, respectively. According to SDS-PAGE analysis, bread wheat genotypes had a higher concentration of α + ß + γ-subunits of gliadin (on average 61.54% of extractable proteins) than durum wheat (on average 55.32% of extractable proteins). However, low concentration of ω-subunit was found in both bread (0.50% to 2.53% of extractable proteins) and durum (3.65% to 6.99% of extractable proteins) wheat genotypes. On average, durum wheat contained significantly higher amounts of tryptophan and wet gluten (0.163% dry weight (d.w.) and 26.96% d.w., respectively) than bread wheat (0.147% d.w. and 24.18% d.w., respectively).

Characterization of proteins from kernel of different soybean varieties.

BACKGROUND: Total soybean proteins, storage proteins, glycinin (11S) and ß-conglycinin (7S) fractions and their respective subunits in seven soybean varieties were analyzed. In this work we also present the correlation between concentration and activity of bioactive proteins, lipoxygenase and proteinase inhibitors. RESULTS: Glycinin and ß-conglycinin comprise about 750 g kg(-1) of the bean storage protein and as such account for both quantity and quality of the kernel protein. The 11S concentration of the varieties studied ranged from 503.4 to 602.9 g kg(-1) and those of 7S varied from 178.2 to 230.6 g kg(-1) of total extractable proteins. The ratio of 11S/7S proteins varied from 2.43 to 3.29 among the varieties. A very strong positive correlation was found between the concentration of Kunitz trypsin inhibitor and activity of total trypsin inhibitor (r = 0.96). However, lipoxygenase concentration did not show a strong correlation with lipoxygenase activity. CONCLUSION: It appears that among the seven ZP soybean genotypes there are genotypes with different amounts of subunits that should be bred in the future for a desired level of protein components.

Dynamics of Maize Vegetative Growth and Drought Adaptability Using Image-Based Phenotyping Under Controlled Conditions.

Changes in climate are likely to have a negative impact on water availability and soil fertility in many maize-growing agricultural areas. The development of high-throughput phenotyping platforms provides a new prospect for dissecting the dynamic complex plant traits such as abiotic stress tolerance into simple components. The growth phenotypes of 20 maize (Zea mays L.) inbred lines were monitored in a non-invasive way under control, nitrogen, and water limitation as well as under combined nitrogen and water stress using an automated phenotyping system in greenhouse conditions. Thirteen biomass-related and morphophysiological traits were extracted from RGB images acquired at 33 time points covering developmental stages from leaf count 5 at the first imaging date to leaf count 10-13 at the final harvest. For these traits, genetic differences were identified and dynamic developmental trends during different maize growth stages were analyzed. The difference between control and water stress was detectable 3-10 days after the beginning of stress depending on the genotype, while the effect of limited nitrogen supply only induced subtle phenotypic effects. Phenotypic traits showed different response dynamics as well as multiple and changing interaction patterns with stress progression. The estimated biovolume, leaf area index, and color ratios were found to be stress-responsive at different stages of drought stress progression and thereby represent valuable reference indicators in the selection of drought-adaptive genotypes. Furthermore, genotypes could be grouped according to two typical growth dynamic patterns in water stress treatments by c-means clustering analysis. Inbred lines with high drought adaptability across time and development were identified and could serve as a basis for designing novel genotypes with desired, stage-specific growth phenotypes under water stress through pyramiding. Drought recovery potential may play an equal role as drought tolerance in plant drought adaptation.

Image-Derived Traits Related to Mid-Season Growth Performance of Maize Under Nitrogen and Water Stress.

Phenotypic measurements under controlled cultivation conditions are essential to gain a mechanistic understanding of plant responses to environmental impacts and thus for knowledge-based improvement of their performance under natural field conditions. Twenty maize inbred lines (ILs) were phenotyped in response to two levels of water and nitrogen supply (control and stress) and combined nitrogen and water deficit. Over a course of 5 weeks (from about 4-leaf stage to the beginning of the reproductive stage), maize phenology and growth were monitored by using a high-throughput phenotyping platform for daily acquisition of images in different spectral ranges. The focus of the present study is on the measurements taken at the time of maximum water stress (for traits that reflect plant physiological properties) and at the end of the experiment (for traits that reflect plant architectural and biomass-related traits). Twenty-five phenotypic traits extracted from the digital image data that support biological interpretation of plant growth were selected for their predictive value for mid-season shoot biomass accumulation. Measured fresh and dry weights after harvest were used to calculate various indices (water-use efficiency, physiological nitrogen-use efficiency, specific plant weight) and to establish correlations with image-derived phenotypic features. Also, score indices based on dry weight were used to identify contrasting ILs in terms of productivity and tolerance to stress, and their means for image-derived and manually measured traits were compared. Color-related traits appear to be indicative of plant performance and photosystem II operating efficiency might be an importance physiological parameter of biomass accumulation, particularly under severe stress conditions. Also, genotypes showing greater leaf area may be better adapted to abiotic stress conditions.

Development of High Tryptophan Maize Near Isogenic Lines Adapted to Temperate Regions through Marker Assisted Selection - Impediments and Benefits.

Breeding program aimed at converting standard maize inbred lines to their quality protein maize (QPM) counterparts for growing in temperate climate is being conducted at Maize Research Institute (MRI). The objective of the research presented herein was to develop QPM versions of two commercial ZP inbreds through marker assisted selection (MAS) with opaque2 specific molecular markers, while maintaining their good agronomic performances and combining abilities. Donor line was a tropical QPM line CML 144. After two backcross and three selfing generations, six near isogenic lines (NILs) with 93% recovery of the recurrent parent genome were created from one cross. Average increments of 30% in tryptophan content and 36% in quality index were obtained, as well as kernels with less than 25% opaque endosperm. Grain yield was increased by 11-31% and combining abilities of the improved lines were on a par with the original line. Correlations between biochemical and agronomic parameters revealed that selection for plant height, ear length and kernel row number together with tryptophan content could be recommended for development of QPM with this material. However, several impediments emerged during selection. Major drawbacks in NIL development were small number of opaque2 recessive homozygotes (4.5% and 7.6% in BC2F2 of two crosses) and poor seed set throughout selection, which led to the loss of one cross. Moreover, in the other cross many plants in different generations had to be omitted from further selection due to the insufficient number of kernels. This phenomenon could be explained by incompatibility between pollen and style, possibly due to the exotic donor germplasm. Overall, it could be expected that the use of NILs, which are adapted to temperate climate and have high percentage of domestic germplasm, would outbalance the noted impediments and increase MAS efficiency in different breeding programs.

Grain Nutrient Composition of Maize (Zea mays L.) Drought-Tolerant Populations.

A total of 13 maize populations from the drought-tolerant mini core collection from Maize Research Institute gene bank were evaluated for oil, protein, and tryptophan contents, fatty acid (FA) composition, and kernel characteristics. All accessions are high oil (5.8-7.9%) and protein (10.58-12.45%) genotypes. Most of the accessions showed high contents of tryptophan (0.070-0.081%) and saturated (12.65-17.91%) and monounsaturated (24.19-45.52%) FAs. Significant positive correlations were found between oil and protein and between oil and tryptophan contents (p < 0.01). Correlations between oil and principal FA were non-significant. Several accessions showed multiple nutritional advantages. For example, IP6428 had high oil (7.3%), tryptophan (0.081%), and saturated FA (17.9%) contents. Moreover, a positive correlation (p < 0.01) between palmitic (13.68%) and oleic (34.74%) acids enables the use of IP6428 for developing lines high in these FAs. Because drought-tolerant accessions were selected in both subtropical and temperate zones, they could be used for breeding value-added maize adapted to both environments.