Cell-fate specification is typically thought to precede and determine cell-cycle regulation during differentiation. Here we show that endoreplication, also known as endoreduplication, a specialized cell-cycle variant often associated with cell differentiation but also frequently occurring in malignant cells, plays a role in maintaining cell fate. For our study we have used Arabidopsis trichomes as a model system and have manipulated endoreplication levels via mutants of cell-cycle regulators and overexpression of cell-cycle inhibitors under a trichome-specific promoter. Strikingly, a reduction of endoreplication resulted in reduced trichome numbers and caused trichomes to lose their identity. Live observations of young Arabidopsis leaves revealed that dedifferentiating trichomes re-entered mitosis and were re-integrated into the epidermal pavement-cell layer, acquiring the typical characteristics of the surrounding epidermal cells. Conversely, when we promoted endoreplication in glabrous patterning mutants, trichome fate could be restored, demonstrating that endoreplication is an important determinant of cell identity. Our data lead to a new model of cell-fate control and tissue integrity during development by revealing a cell-fate quality control system at the tissue level.

BackgroundLake Magadi and little Magadi are hypersaline, alkaline lakes situated in the southern part of Kenyan Rift Valley. Solutes are supplied mainly by a series of alkaline hot springs with temperatures as high as 86 °C. Previous culture-dependent and culture-independent studies have revealed diverse groups of microorganisms thriving under these conditions. Previous culture independent studies were based on the analysis of 16S rDNA but were done on less saline lakes. For the first time, this study combined illumina sequencing and analysis of amplicons of both total community rDNA and 16S rRNA cDNA to determine the diversity and community structure of bacteria and archaea within 3 hot springs of L. Magadi and little Magadi.MethodsWater, wet sediments and microbial mats were collected from springs in the main lake at a temperature of 45.1 °C and from Little Magadi “Nasikie eng’ida” (temperature of 81 °C and 83.6 °C). Total community DNA and RNA were extracted from samples using phenol-chloroform and Trizol RNA extraction protocols respectively. The 16S rRNA gene variable region (V4 – V7) of the extracted DNA and RNA were amplified and library construction performed following Illumina sequencing protocol. Sequences were analyzed done using QIIME while calculation of Bray-Curtis dissimilarities between datasets, hierarchical clustering, Non Metric Dimensional Scaling (NMDS) redundancy analysis (RDA) and diversity indices were carried out using the R programming language and the Vegan package.ResultsThree thousand four hundred twenty-six and one thousand nine hundred thirteen OTUs were recovered from 16S rDNA and 16S rRNA cDNA respectively. Uncultured diversity accounted for 89.35 % 16S rDNA and 87.61 % 16S rRNA cDNA reads. The most abundant phyla in both the 16S rDNA and 16S rRNA cDNA datasets included: Proteobacteria (8.33–50 %), Firmicutes 3.52–28.92 %, Bacteroidetes (3.45–26.44 %), Actinobacteria (0.98–28.57 %) and Euryarchaeota (3.55–34.48 %) in all samples. NMDS analyses of taxonomic composition clustered the taxa into three groups according to sample types (i.e. wet sediments, mats and water samples) with evident overlap of clusters between wet sediments and microbial mats from the three sample types in both DNA and cDNA datasets. The hot spring (45.1 °C) contained less diverse populations compared to those in Little Magadi (81–83 °C).ConclusionThere were significant differences in microbial community structure at 95 % level of confidence for both total diversity (P value, 0.009) based on 16S rDNA analysis and active microbial diversity (P value, 0.01) based on 16S rRNA cDNA analysis, within the three hot springs. Differences in microbial composition and structure were observed as a function of sample type and temperature, with wet sediments harboring the highest diversity.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-016-0748-x) contains supplementary material, which is available to authorized users.

Endoreplication, also known as endoreduplication, is a phyogenetically widespread modified version of the cell cycle in which DNA replication is not followed by cell division. The SIAMESE (SIM) gene of Arabidopsis thaliana encodes the founding member of a novel class of plant-specific cyclin-dependent kinase (CDK) inhibitors and is a key regulator of endoreplication during the development of trichomes (shoot epidermal hairs). Here, we have identified mutations in the CCS52A1 gene as genetic modifiers of the multicellular trichome phenotype of sim mutants. Loss-of-function ccs52A1 mutations dramatically enhance the multicellularity of sim mutants trichomes in double mutants, whereas overexpression of CCS52A1 completely suppresses the sim mutant phenotype. CCS52A1 encodes a CDH1/FZR-like protein, a class of proteins that function as activators of the anaphase-promoting complex. Unicellular ccs52A1 trichomes become multicellular upon overexpression of B-type cyclin, consistent with repression of the accumulation of mitotic cyclins in the developing trichome by CCS52A1. As these M-phase-specific cyclins are known to accumulate in sim mutant trichomes, our data suggest that CCS52A1 and SIM cooperate in repressing accumulation of mitotic cyclins to establish the trichome endocycle. Comparison with endoreplication pathways in Drosophila and mammals indicates that while these organisms all use similar components to initiate endoreplication, the components are deployed differently in each organism.

BackgroundThe Arabidopsis thaliana CONSTITUTIVE EXPRESSOR OF PATHOGENESIS-RELATED GENES5 (CPR5) gene has been previously implicated in disease resistance, cell proliferation, cell death, and sugar sensing, and encodes a putative membrane protein of unknown biochemical function. Trichome development is also affected in cpr5 plants, which have leaf trichomes that are reduced in size and branch number.ResultsIn the work presented here, the role of CPR5 in trichome development was examined. Trichomes on cpr5 mutants had reduced birefringence, suggesting a difference in cell wall structure between cpr5 and wild-type trichomes. Consistent with this, leaf cell walls of cpr5 plants contained significantly less paracrystalline cellulose and had an altered wall carbohydrate composition. We also found that the effects of cpr5 on trichome size and endoreplication of trichome nuclear DNA were epistatic to the effects of mutations in triptychon (try) or overexpression of GLABRA3, indicating that these trichome developmental regulators are dependant on CPR5 function for their effects on trichome expansion and endoreplication.ConclusionOur results suggest that CPR5 is unlikely to be a specific regulator of pathogen response pathways or senescence, but rather functions either in cell wall biogenesis or in multiple cell signaling or transcription response pathways.

SUMMARYEndoreplication, also called endoreduplication, is a modified cell cycle in which DNA is repeatedly replicated without subsequent cell division. Endoreplication is often associated with increased cell size and specialized cell shapes, but the mechanism coordinating DNA content with shape and size remains obscure. Here we identify the product of the BRANCHLESS TRICHOMES (BLT) gene, a protein of hitherto unknown function that has been conserved throughout angiosperm evolution, as a link in coordinating cell shape and nuclear DNA content in endoreplicated Arabidopsis trichomes. Loss-of-function mutations in BLT were found to enhance the multicellular trichome phenotype of mutants in the SIAMESE (SIM) gene, which encodes a repressor of endoreplication. Epistasis and overexpression experiments revealed that BLT encodes a key regulator of trichome branching. Additional experiments showed that BLT interacts both genetically and physically with STICHEL, another key regulator of trichome branching. Although blt mutants have normal trichome DNA content, overexpression of BLT results in an additional round of endoreplication, and blt mutants uncouple DNA content from morphogenesis in mutants with increased trichome branching, further emphasizing its role in linking cell shape and endoreplication.

Sorghum (Sorghum bicolour (L.) Moench) grown under rain-fed conditions is usually affected by drought stress at different stages, resulting in reduced yield. The assessment of variation in morpho-physiological traits contributing towards drought tolerance at these stages is of vital importance. This study was conducted using a split plot design with three replications to evaluate 25 sorghum accessions at post flowering stage under well watered and drought stress conditions at Hamelmalo Agricultural College. The data of 14 different morpho-physiological traits were subjected to analysis of variance, estimation of genetic variability and heritability and principal component analysis. We analyzed variance for seedling vigor, number of leaves, leaf area, stay-green, peduncle exsertion, panicle length and width, plant height, days to flowering and maturity, grain yield, biomass and harvest index under drought stress and irrigated conditions. The results showed that genotypic differences were significant at P < 0.05 -< 0.001. High magnitude of phenotypic and genotypic coefficient of variations for plant height, harvest index and biomass as well as high heritability for days to flowering, panicle length, days to maturity and over all agronomic score were recorded. Principal component (PC) analysis showed that the first 4 PCs had Eigen value >1 explaining 74.6% of the total variation with grain yield, biomass, stay-green, leaf area, peduncle exsertion and days to flowering and maturity being the most important characters in PC1 and PC2. This research demonstrated high diversity for the characters studied. Moreover, the result showed that drought stress reduced the yield of some genotypes, though others were tolerant to drought. Accessions EG 885, EG 469, EG 481, EG 849, Hamelmalo, EG 836 and EG 711 were * Corresponding author.Tesfamichael et al. 1411identified as superior for post-flowering drought tolerance and could be used by breeders in improvement programs.

Cowpea (Vigna unguiculata) plays an important role in sustainable food security and livelihood improvement in Sub-Saharan Africa (SSA). The crop is rich in phytonutrients and minerals, which are key in solving malnutrition and hunger crisis, a major challenge in SSA. However, physiological status, storage temperature and duration affect phytonutrient levels and postharvest life of the leafy vegetable. Despite the significant importance of cowpeas, the maturity and postharvest storage effects on quality of the leafy vegetable remains unrevealed. The aim of this study was to analyze the dynamics of phytonutrients in cowpea leaves during development under field conditions in Kenya and in storage. The total carbohydrates (glucose, fructose, sucrose and starch) were highest at 90 d after planting (105.9 ± 2.5 g kg-1) compared to 30, 60 and 120 d. The total Phenolics (Gallic acid equivalents) increased gradually with age up to 12.0 ± 0.2 g kg-1 by 120 d. Catechin equivalent flavonoids, trolox equivalent antioxidants (TEA) and chlorophyll were highest in concentrations at 60 d after planting with 8.0 ± 0.5 g kg-1, 26.19 ± 0.5 g kg-1 and 5.7 ± 0.4 g kg-1, respectively. Quercetin equivalent flavonoids and total carotenoids did not show significant changes with age, while mineral concentration dynamics were specific for each element. Storage of cowpea leaves at room temperature (50–55 % relative humidity) led to a stronger decline of phytonutrients after 4 d, but mostly they remained stable at cold storage (5 °C). Results of this study highlight the importance of developmental stage at harvest, storage conditions and duration for the optimal availability of phytonutrients in freshly consumed leaves and for postharvest management strategies.

Mango fruits are highly nutritious and economically important to Kenyan farmers, who cultivate three categories of cultivars/landraces; local small-fruited, local big-fruited and improved, introduced cultivars. The small-fruited landraces are said to be well adapted to the local environment but are being replaced by introduced cultivars before their diversity has been documented. This study aimed at assessing morphological and genetic diversity of 36 local mango landraces from 35 randomly selected farms in Eastern Kenya. Fruits were collected from three locations for morphological characterization using the 'Descriptors for Mango' of the International Plant Genetic Resources Institute. Leaves of the same accessions were sampled for genetic diversity assessment using microsatellites. Morphological characterization showed that mean fruit length was 5.6-12.5 cm, while mean fruit weight was 93-578 g. Fruit shape was mostly 'roundish' , while fruit ground colour 'green'. Hierarchical cluster analysis with seven discriminant morphological variables resulted in four clusters. Analysis of molecular variance (AMOVA) indicated that variation was high (97%) among, but low (3%) within groups. Phylogenetic analysis using Neighbor Joining method resulted in three clusters that lacked consistency with the morphological clusters. Findings from this study may assist to select superior local mango accessions for future breeding programmes and to develop 'conservation through use' strategies for Kenyan local mangoes to retain their valuable genetic resources.