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Coconut (Cocos nucifera) is an economically important palm tree with diverse applications. However, limited scientific research exists on coconut diseases in the Kenyan coastal region. This cross‐sectional study aimed to investigate the genetic diversity of latent bacteria associated with yellowing symptoms in coconut plants along the Kenyan coast. Sixty‐two samples with symptoms were collected and their bacterial diversity assessed using culture‐independent methods. DNA was extracted from controls and symptomatic samples using the cetyl trimethyl ammonium bromide (CTAB) method. Next‐generation sequencing with Illumina MiSeq was used to profile the bacterial communities using amplicons of 16S ribosomal RNA sequences (V4 region). Sequence data were analysed using the Quantitative Insights Into Microbial Ecology 2 (QIIME 2) pipeline. A total of 113,330 reads were obtained, which clustered into 285 Operational Taxonomic Units. Bacterial diversity was highest in Kilifi, followed by Kwale and Lamu, while control samples exhibited low bacterial diversity. Actinobacteria was the predominant phylum across all counties, and Streptomyces was the most abundant genus. Kilifi and Kwale counties were more affected than Lamu. This is a pioneer study that provides insight into the bacterial diversity associated with yellowing disease symptoms in coconut plants in Kenya and will help with future elucidation of the agents causing or exacerbating coconut disease symptoms. [ABSTRACT FROM AUTHOR]

Background: Doum palms (Hyphaene compressa) perform a crucial starring role in the lives of Kenya's arid and semi-arid people for empowerment and sustenance. Despite the crop's potential for economic gain, there is a lack of genetic resources and detailed information about its domestication at the molecular level. Given the doum palm's vast potential as a widely distributed plant in semi-arid and arid climates and a source of many applications, coupled with the current changing climate scenario, it is essential to understand the molecular processes that provide drought resistance to this plant. Results: Assembly of the first transcriptome of doum palms subjected to water stress generated about 39.97 Gb of RNA-Seq data. The assembled transcriptome revealed 193,167 unigenes with an average length of 1655 bp, with 128,708 (66.63%) successfully annotated in seven public databases. Unigenes exhibited significant differentially expressed genes (DEGs) in well-watered and stressed-treated plants, with 45071 and 42457 accounting for up-regulated and down-regulated DEGs, respectively. GO term, KEGG, and KOG analysis showed that DEGs were functionally enriched cellular processes, metabolic processes, cellular and catalytic activity, metabolism, genetic information processing, signal transduction mechanisms, and posttranslational modification pathways. Transcription factors (TF), such as the MYB, WRKY, NAC family, FAR1, B3, bHLH, and bZIP, were the prominent TF families identified as doum palm DEGs encoding drought stress tolerance. Conclusions: This study provides a complete understanding of DEGs involved in drought stress at the transcriptome level in doum palms. This research is, therefore, the foundation for the characterization of potential genes, leading to a clear understanding of its drought stress responses and providing resources for improved genetic modification. [ABSTRACT FROM AUTHOR]

A high yield of isolated protoplast and reliable regeneration system are prerequisite for successful somatic hybridization and genome editing research. However, reproducible plant regeneration from protoplasts remains a bottleneck for many crops, including cassava. We evaluated several factors that influence isolation of viable protoplasts form leaf mesophyll, induction of embryogenic calli, and regeneration of plants in three cassava cultivars; Muchericheri, TMS60444 and Karibuni. A relatively higher protoplast yield was obtained with enzyme mixture containing 5 g/L Macerozyme and 10 g/L cellulase. Muchericheri recorded relatively higher protoplast yield of 20.50±0.50×106 whereas TMS60444 (10.25±0.25×106) had the least protoplast yield in 10 g/L cellulase and 4 g/L cellulase. Freshly isolated protoplast cells were plated on callus induction medium (CIM) solid medium containing MS basal salt, 60 g/L D-glucose, 30 g/L sucrose, B5 vitamins, 100 mg/L myo-inositol, 0.5 mg/L copper sulphate, 100 mg/L casein hydrolysate, 4.55 g/L mannitol, 0.1 g/L MES, 10 mg/L picloram and 3 g/L gelrite to induce protoplast growth and development. The three cultivars reached colony formation but no further development was observed in this culture method. Protoplast growth and development was further evaluated in suspension culture using varying cell densities (1, 2 and 3× 105 p/mL). Development with highest number of minicalli was observed in cell density of 3× 105 p/mL. Minicalli obtained were cultured on CIM supplemented with 10mg/L picloram. Callus induction was observed in all cell densities with the cultivars. Highest somatic embryogenesis was observed in 2× 105 p/ml while no somatic embryogenesis was observed in cell density of 1×105 p/mL. Somatic embryos were matured in EMM medium supplemented with 1 mg/L BAP, 0.02 mg/L NAA and 1.5 mg/L GA3 then germinated in hormone free medium for plant regeneration. This protocol which used simple mixture of commercial enzymes is highly reproducible and can be applied in biotechnology research on cassava. [ABSTRACT FROM AUTHOR]

Cassava is the world's most essential food root crop, generating calories to millions of Sub-Saharan African subsistence farmers. Cassava leaves and roots contain toxic quantities of the cyanogenic glycoside linamarin. Consumption of residual cyanogens results in cyanide poisoning due to conversion of the cyanogens to cyanide in the body. There is a need for acyanogenic cassava cultivars in order for it to become a consistently safe and acceptable food, and commercial crop. In recent years, the CRISPR/Cas system, has proven to be the most effective and successful genome editing tool for gene function studies and crop improvement. In this study, we performed targeted mutagenesis of the MeCYP79D1 gene in exon 3, using CRISPR/Cas9, via Agrobacterium-mediated transformation. The vector design resulted in knockout in cotyledon-stage somatic embryos regenerated under hygromycin selection. Eight plants were recovered and genotyped. DNA sequencing analysis revealed that the tested putative transgenic plants carried mutations within the MeCYP79D1 locus, with deletions and substitutions being reported upstream and downstream of the PAM sequence, respectively. The levels of linamarin and evolved cyanide present in the leaves of mecyp79d1 lines were reduced up to seven-fold. Nevertheless, the cassava linamarin and cyanide were not completely eliminated by the MeCYP79D1 knockout. Our results indicate that CRISPR/Cas9-mediated mutagenesis is as an alternative approach for development of cassava plants with lowered cyanide content. [ABSTRACT FROM AUTHOR]

Doum palm (Hyphaene compressa) is a perennial economic plant primarily growing in Kenya's Arid and Semi-Arid Lands (ASALs). It is heavily relied upon for food, animal feed, construction materials and medicine, making it an ideal plant for resource sustainability. However, the limited information on its genetic resources has hindered its breeding and conservation studies. This study used the genotyping by sequencing approach to identify Single Nucleotide Polymorphisms. These SNPs were further used to assess the genetic diversity and population structure of 96 H. compressa accessions from Coastal, Northern and Eastern ASAL regions of Kenya using two approaches; reference-based and de novo -based assemblies. STRUCTURE analysis grouped the sampled accessions into two genetic clusters (Cluster 1 and Cluster 2). Cluster 1 included accessions from the Northern region, whereas Cluster 2 included all accessions from Eastern and Coastal regions. Accessions from Kwale (Coastal) had mixed ancestry from both Cluster 1 and Cluster 2. These STRUCTURE findings were further supported by principal components analysis, discriminant analysis of principal components and phylogenetic analysis. Analysis of molecular variance indicated greater genetic variation within populations (92.7%) than among populations (7.3%). An overall FST of 0.074 was observed, signifying moderate genetic differentiation among populations. The results of this study will provide information useful in breeding, marker-assisted selection and conservation management of H. compressa. [ABSTRACT FROM AUTHOR]

Hyphaene compressa is an economically important palm in Africa. Despite its significant role in the livelihoods of rural communities, the diversity of doum palm is poorly documented and studied. In addition, it has no model descriptor that can aid such studies. Ninety H. compressa accessions collected from Northern, Eastern, and Coastal regions of Kenya were examined to determine the morphological variability of the vegetative and fruit traits of H. compressa and to identify its morphotypes for improvement. A total of 19 morphological characters including seven quantitative and 12 qualitative traits of fruit and vegetative traits were selected. Linear mixed-effects models, principal component analysis, and linear discriminant analyses were used to assess the variation in the morphological traits of doum palm based on the regions. Hierarchical clustering was performed to identify the morphotypes of H. compressa. There was variability in H. compressa morphological traits, particularly at the Kenyan Coast. All seven quantitative traits were able to effectively discriminate doum palm phenotypically p ≤ 0.001 . The 90 accessions clustered into five morphotypes designated as 1, 2, 3, 4, and 5. Morphotype 4 was specific only to the Coastal region. Morphotype 5 had the tallest trees with the biggest fruits and included palms from Eastern and Coastal regions making it the best morphotype for fruit traits. This study will inform the domestication, improvement, and conservation of H. compressa by selecting elite accessions. [ABSTRACT FROM AUTHOR]