Galactone-γ-lactone-dependent ascorbate biosynthesis alters wheat kernel maturation

Kernel development and maturation involve several well-characterised events, such as changes in ascorbate (ASC) metabolism, protein synthesis and storage, programmed cell death (PCD) of starchy endosperm and tissue dehydration. Despite many studies focusing on these events, whether and how they are metabolically related to each other, remains to be elucidated. In the present investigation, the changes in ASC-related metabolism, PCD occurrence, kernel filling and dehydration have been analysed during kernel maturation, over a 3-year period in plants grown under normal conditions and in plants displaying modified ASC synthesis. The obtained results suggest that ASC plays a pivotal role in the network of events characterising kernel maturation. During this process, a decrease in ASC content occurs. When ASC biosynthesis is improved in the kernel, by feeding the plants with its immediate precursor, L-galactone-γ-lactone (GL), the decrease in ASC, observed during kernel maturation, is delayed. As a consequence, ascorbate peroxidase (APX) activity is also enhanced. Moreover, a delay in the ASC decrease permits a delay in PCD occurring in kernel storage tissues and in kernel dehydration. Interestingly, the data emerging from the present investigation suggest that the delay in the decrease in ASC content and APX activity also improves kernel filling. The relevance of the ascorbate-dependent redox regulation for kernel productivity is discussed.     

Does natal habitat preference modulate cereal kernel preferences in the rice weevil?

The influence of larval diet source in Sitophilus oryzae (L.), a primary grain feeder, on the choice of a new cereal host by young adult was investigated through a multiple choice comparative bioassay. For this purpose, virgin male and female adults, which had been reared as immatures in different grain commodities, were monitored according to their food preference in cylindrical arenas containing whole kernels of maize, rice, barley and wheat. The adults showed a clear individual preference for maize kernels, regardless of the previous food experience, sex and also the interval on which the adults are exposed to food. Males were much more mobile than females and visited more vigorously different commodities before the final choice. Female response was found to be related with male response to specific food sources. Hence, our results indicate that there was no natal habitat preference induction, since maize was, in most cases, the final commodity choice.     

When will plant morphology affect the shape of a seed dispersal "kernel"?

Most models of dispersal assume that plants are point sources. In reality, the scale in height over which seed sources are distributed is often of the same order as the scale in distance over which most individual seeds are dispersed. But is this sufficient to affect the fundamental shapes of dispersal frequency distributions? Most published conclusions about the effects of canopy structure on dispersal are subjective. A model is developed to explore the consequences of plant canopies for the shapes of whole-plant seed dispersal "kernels". The canopies were described by simple geometric shapes, while an empirical probability density function (PDF) was used for dispersal from a point source. It was found that the resulting whole-plant PDF for dispersal distance was almost invariably peaked, whereas the PDF for the density of seed rain (as would be measured by pitfall traps) could either be peaked or monotonic according to the canopy shape, position of seeds in the canopy, and mean dispersal distance. The shapes of kernels from whole plants (distributed seed sources) can be very different from those derived from a point source under certain circumstances.     

Do all inter-patch movements represent dispersal? A mixed kernel study of butterfly mobility in fragmented landscapes

1. In times of ongoing habitat fragmentation, the persistence of many species is determined by their dispersal abilities. Consequently, understanding the rules underlying movement between habitat patches is a key issue in conservation ecology. 2. We have analysed mark-release-recapture (MRR) data on inter-patches movements of the Dusky Large Blue butterfly Maculinea nausithous in a fragmented landscape in northern Bavaria, Germany. The aim of the analysis was to quantify distance dependence of dispersal as well as to evaluate the effect of target patch area on immigration probability. For statistical evaluation, we apply a 'reduced version' of the virtual migration model (VM), only fitting parameters for dispersal distance and immigration. In contrast to other analyses, we fit a mixed dispersal kernel to the MRR data. 3. A large fraction of recaptures happened in other habitat patches than those where individuals were initially caught. Further, we found significant evidence for the presence of a mixed dispersal kernel. The results indicate that individuals follow different strategies in their movements. Most movements are performed over small distances, nonetheless involving travelling between nearby habitat patches (median distance c. 480 m). A small fraction (c. 0·025) of the population has a tendency to move over larger distances (median distance c. 3800 m). Further, immigration was positively affected by patch area (I～A(ζ) ), with the scaling parameter ζ = 0·5. 4. Our findings should help to resolve the long-lasting dispute over the suitability of the negative exponential function vs. inverse-power one for modelling dispersal. Previous studies on various organisms found that the former typically gives better overall fit to empirical distance distributions, but that the latter better represents long-distance movement probabilities. As long-distance movements are more important for landscape-level effects and thus, e.g. for conservation-oriented analyses like PVAs, fitting inverse-power kernels has often been preferred. 5. We conclude that the above discrepancy may simply stem from the fact that recorded inter-patch movements are an outcome of two different processes: daily routine movements and genuine dispersal. Consequently, applying mixed dispersal kernels to disentangle the two processes is recommended.     

Marker–trait association analysis of kernel hardness and related agronomic traits in a core collection of wheat lines

Identification of marker–trait associations is the first step towards marker-assisted selection in plant breeding. Here we assess genetic diversity and population structure of 94 diverse wheat elite lines and use genome-wide association mapping to identify marker–trait associations for five important traits: kernel hardness (KHA), thousand-kernel weight, grain protein content, test weight (TWT), and plant height (PHT). The 94 accessions employed in this study were grouped into three subpopulations based on the first three principal components, which accounted for 51.5 % of the variations. A mixed linear model was used to detect marker–trait associations incorporating covariance of population structure and relative kinship. A total of six marker loci was significantly associated with KHA, TWT, and PHT after the correction of false discovery rate (α _c  = 0.05). The gene pinB was found to be highly associated with KHA, and is reported to be a major determinant of KHA together with the gene pinA at the Ha locus on chromosome 5D. Marker XwPt-7187 on chromosome 2A was also significantly associated with KHA, two Diversity Arrays Technology markers XwPt-1250 and XwPt-4628 with TWT, and marker Xgwm512 with PHT, making the first report of marker–trait associations in these genomic regions.     

Sequence variation in 3′UTR region of crtRB1 gene and its effect on β-carotene accumulation in maize kernel

β-carotene fortification of maize has emerged as a potential, long-term and sustainable approach to alleviate vitamin A deficiency in humans. Among the several genes involved in the carotenoid biosynthetic pathway, the 543 bp allele at crtRB1 3′TE (Transposable Element) gene (allele 1, without insertion) is associated with higher β-carotene accumulation. Estimation of β-carotene through high performance liquid chromatography showed that the CIMMYT genotypes with allele 1 had high kernel β-carotene content whereas the Indian inbreds with the same allele had low β-carotene content. To know the reason for this variation, allele 1 of crtRB1 3′TE gene was sequenced from a set of 11 diverse maize inbreds collected from CIMMYT and Indian germplasm. The sequence data of the allele 1 revealed the presence of 13 single nucleotide polymorphisms (SNPs) and 7 insertions and deletions (InDels). Exonic region had two SNPs, intronic region had one SNP and one InDel, whereas 3′-untranslated region (UTR) region of the gene showed 10 SNPs and 6 InDels. Among the several SNPs and InDels, SNP4, SNP13, InDel6 and InDel7 identified in the 3′-UTR region clearly differentiated the high and the low β-carotene genotypes. These 3′-UTR polymorphisms in allele 1 of the crtRB1 3′TE gene could be associated with the variation in kernel β-carotene accumulation by regulating the translation and stability of the mRNA. The SNPs and the InDels associated with higher level of β-carotene will be used as a gene-based marker(s) in selection of genotypes and to develop biofortified maize hybrids to alleviate vitamin A deficiency in humans.     

Molecular diversity and genetic variability of kernel tocopherols among maize inbreds possessing favourable haplotypes of γ-tocopherol methyl transferase (ZmVTE4)

Journal of Plant Biochemistry and Biotechnology - Vitamin E deficiency is a serious health concern in humans. Biofortification of maize kernel with high vitamin E (α-tocopherol) provides...     

Overexpression of Puroindoline a gene in transgenic durum wheat (Triticum turgidum ssp. durum) leads to a medium–hard kernel texture

Durum wheat is the second-most widely grown wheat species, and is primarily used in the production of pasta and couscous. The grain utilization of durum wheat is partly related to its very hard kernel texture because of the lack of the D genome and consequentially the Puroindoline genes. Our previous study reported the transformation of durum wheat with the Puroindoline a (Pina) gene. Here, we characterized the transgenic durum wheat lines expressing the Pina gene, and studied the effects of PINA on grain texture and other kernel characteristics. SDS-PAGE and Western blotting results demonstrated that starch-bound PINA levels of Pina-overexpressing lines were lower than that of Pina-positive control, common wheat cv. Chinese Spring, suggesting a weak association of PINA protein with starch granules in the absence of Pinb. Grain hardness analysis and flour milling tests indicated that the overexpression of PINA resulted in decreased grain hardness and increased flour yield in transgenic durum wheat lines. The agronomic performance of the transgenic and control lines was also examined and it was found that no significant differences in measured traits were observed between Pina-overexpressing and control lines in the 2-year field trials. Since grain hardness strongly affects milling and end-use qualities, the development of medium–hard-textured durum wheat lines is not only of significance for our knowledge of grain hardness and Puroindolines, but also has practical implications for plant breeders and food technologists for the expansion of utilization of durum wheat.     

Utilization of palm kernel cake for production of β-mannanase by Aspergillus niger FTCC 5003 in solid substrate fermentation using an aerated column bioreactor

The production of β-mannanase from palm kernel cake (PKC) as a substrate in solid substrate fermentation (SSF) was studied using a laboratory column bioreactor. The simultaneous effects of three independent variables, namely incubation temperature, initial moisture content of substrate and airflow rate, on β-mannanase production were evaluated by response surface methodology (RSM) on the basis of a central composite face-centered (CCF) design. Eighteen trials were conducted in which Aspergillus niger FTCC 5003 was cultivated on PKC in an aerated column bioreactor for seven days under SSF process. The highest level of β-mannanase (2117.89 U/g) was obtained when SSF process was performed at incubation temperature, initial moisture level and aeration rate of 32.5°C, 60% and 0.5 l/min, respectively. Statistical analysis revealed that the quadratic terms of incubation temperature and initial moisture content had significant effects on the production of β-mannanase (P < 0.01). A similar analysis also demonstrated that the linear effect of initial moisture level and an interaction effect between the initial moisture content and aeration rate significantly influenced the production of β-mannanase (P < 0.01). The statistical model suggested that the optimal conditions for attaining the highest level of β-mannanase were incubation temperature of 32°C, initial moisture level of 59% and aeration rate of 0.5 l/min. A β-mannanase yield of 2231.26 U/g was obtained when SSF process was carried out under the optimal conditions described above.     

A novel thermophilic β-mannanase with broad-range pH stability from Lichtheimia ramosa and its synergistic effect with α-galactosidase on hydrolyzing palm kernel meal

β-Mannanase is the key enzyme in the hydrolysis of mannan which has been widely applied in diverse industrial fields such as biobleaching pulps, food and feed industry, bioethanol and pharmaceutical applications. In this study, a novel GH5 family β-mannanase gene (LrMan5B) with 381 amino acid residues was identified from Lichtheimia ramosa, and highly expressed in Pichia pastoris X33. The amino acid sequence shares the highest identity (64%) with the β-mannanase from Rhizomucor miehei. Purified recombinant LrMan5B showed the optimal activity at pH 5.0 and 65 °C. It had broad-range pH stability (retaining >65% activity after incubation at pH 3.0–8.0 at 37 °C for 24 h) and was highly thermostable (retaining >80% activity after incubation at 60 °C for 30 min). LrMan5B displayed the highest catalytic efficiency for locust bean gum and the k_cat/K_m value was 1357.47 mL·mg⁻¹·s⁻¹, followed by guar gum (512.82 mL·mg⁻¹·s⁻¹), konjac glucomannan (454.21 mL·mg⁻¹·s⁻¹), and palm kernel meal (137.00 mL·mg⁻¹·s⁻¹). In order to evaluate the synergistic effect of LrMan5B and α-galactosidase LrAgal36A from L. ramosa, LrAgal36A was supplemented to hydrolyze palm kernel meal with LrMan5B together, showing that the reducing sugar release significantly increased by 21% (compared with the sum of that by hydrolysis of single Lrman5B or LrAgal36A). Due to its favorable enzymatic properties, LrMan5B might own potential applications in the area of food and feed processing.     

Identification of genetic factors controlling kernel hardness and related traits in a recombinant inbred population derived from a soft × ‘extra-soft’ wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) cross

Kernel hardness or texture, used to classify wheat (Triticum aestivum L.) into soft and hard classes, is a major determinant of milling and baking quality. Wheat genotypes in the soft class that are termed ‘extra-soft’ (with kernel hardness in the lower end of the spectrum) have been associated with superior end-use quality. In order to better understand the relationship between kernel hardness, milling yield, and various agronomic traits, we performed quantitative trait mapping using a recombinant inbred line population derived from a cross between a common soft wheat line and a genotype classified as an ‘extra-soft’ line. A total of 47 significant quantitative trait loci (QTL) (LOD ≥ 3.0) were identified for nine traits with the number of QTL affecting each trait ranging from three to nine. The percentage of phenotypic variance explained by these QTL ranged from 3.7 to 50.3%. Six QTL associated with kernel hardness and break flour yield were detected on chromosomes 1BS, 4BS, 5BS, 2DS, 4DS, and 5DL. The two most important QTL were mapped onto orthologous regions on chromosomes 4DS (Xbarc1118–Rht-D1) and 4BS (Xwmc617–Rht-B1). These results indicated that the ‘extra-soft’ characteristic was not controlled by the Hardness (Ha) locus on chromosome 5DS. QTL for eight agronomic traits occupied two genomic regions near semi-dwarf genes Rht-D1 on chromosome 4DS and Rht-B1 on chromosome 4BS. The clustering of these QTL is either due to the pleiotropic effects of single genes or tight linkage of genes controlling these various traits.     

Starch-bound 2S proteins and kernel texture in einkorn, <Emphasis Type="Italic">Triticum</Emphasis> <Emphasis Type="Italic">monococcum</Emphasis> ssp <Emphasis Type="Italic">monococcum</Emphasis>

The starch granule proteins from 113 einkorn wheat (Triticum monococcum ssp monococcum) accessions were analyzed by acidic, polyacrylamide gel electrophoresis (A-PAGE), and two-dimensional A-PAGE x SDS-PAGE. All accessions were confirmed to contain equal amounts of two polypeptide chains corresponding to puroindoline B (Pin-B), as well as a prominent component plus a faint band corresponding to puroindoline A (Pin-A). When compared with soft-textured common wheat, “monococcum” accessions showed an increase of 3.2- and 2.7-fold in Pin-A and Pin-B levels on the starch granules, respectively. In addition, all accessions contained a novel component of the 2S super-family of seed proteins named Einkorn Trypsin Inhibitor (ETI), which was found to be encoded as a pre-protein 148 residues long. Wild-type ETI encoded by allele Eti-A ^m 1a and “valine-type” ETI encoded by allele Eti-A ^m 1b, which occurred in 107 and six einkorn accessions, respectively, were found to accumulate on starch granules as a mature protein of 121 amino acids with a hydrophobic central domain. The einkorn accessions exhibited an average SKCS index as low as −2.05 ± 11.4, which is typical of extra-soft kernels. The total surface area of starch granules in “monococcum” wheat, as determined by visual assessments in counting chambers, was estimated at 764 mm²/mg of starch, and was about 1.5 times higher than that for common wheat. The results are discussed in relation to the identification of factors that cause the extra-soft texture of einkorn kernels.