Diverse accumulation and distribution of nutrient elements in developing wheat grain studied by laser ablation inductively coupled plasma mass spectrometry imaging.

The present study focused on the elemental distribution in the developing wheat grain by using the laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) imaging technique. Our studies show that the embryo accumulated high concentrations of nutrient elements, such as Fe, K, Cu, and Zn, while Ca was accumulated in the bran of the wheat grain which might be attributed to its function of structural maintenance. In the endosperm the majority of the nutrients were located in the aleurone layer. Within the grain, the embryo could be considered as a nutrient pool for macro- and micro-elements essential for the development of the seedling. Elemental images showed that considerable amounts of nutrients were stored in the scutellum of the embryo, which might be related to the high gene expression of element transporters in the scutellum. Root primordia and leaf primordia were enriched in particular elements, such as Mn and Zn respectively. In total 34 cross sections were analyzed and used for generation of a sequence of elemental distribution images to demonstrate elemental changes along the perpendicular axis of the wheat grain embryo. Further development of three-dimensional modeling will be combined with physiological studies to better understand the mechanisms of elemental distribution and storage in the wheat grain. These studies will provide fundamental knowledge on improving the nutritional value and agronomic practices.

A putative role for amino acid permeases in sink-source communication of barley tissues uncovered by RNA-seq.

BACKGROUND: The majority of nitrogen accumulating in cereal grains originates from proteins remobilised from vegetative organs. However, interactions between grain filling and remobilisation are poorly understood. We used transcriptome large-scale pyrosequencing of flag leaves, glumes and developing grains to identify cysteine peptidase and N transporter genes playing a role in remobilisation and accumulation of nitrogen in barley. RESULTS: Combination of already known and newly derived sequence information reduced redundancy, increased contig length and identified new members of cysteine peptidase and N transporter gene families. The dataset for N transporter genes was aligned with N transporter amino acid sequences of rice and Arabidopsis derived from Aramemnon database. 57 AAT, 45 NRT1/PTR and 22 OPT unigenes identified by this approach cluster to defined subgroups in the respective phylogenetic trees, among them 25 AAT, 8 NRT1/PTR and 5 OPT full-length sequences. Besides, 59 unigenes encoding cysteine peptidases were identified and subdivided into different families of the papain cysteine peptidase clade. Expression profiling of full-length AAT genes highlighted amino acid permeases as the group showing highest transcriptional activity. HvAAP2 and HvAAP6 are highly expressed in vegetative organs whereas HvAAP3 is grain-specific. Sequence similarities cluster HvAAP2 and the putative transporter HvAAP6 together with Arabidopsis transporters, which are involved in long-distance transfer of amino acids. HvAAP3 is closely related to AtAAP1 and AtAAP8 playing a role in supplying N to developing seeds. An important role in amino acid re-translocation can be considered for HvLHT1 and HvLHT2 which are specifically expressed in glumes and flag leaves, respectively. PCA and K-means clustering of AAT transcript data revealed coordinate developmental stages in flag leaves, glumes and grains. Phloem-specific metabolic compounds are proposed that might signal high grain demands for N to distantly located plant organs. CONCLUSIONS: The approach identified cysteine peptidases and specific N transporters of the AAT family as obviously relevant for grain filling and thus, grain yield and quality in barley. Up to now, information is based only on transcript data. To make it relevant for application, the role of identified candidates in sink-source communication has to be analysed in more detail.