Genome-wide identification and characterization of functionally relevant microsatellite markers from transcription factor genes of Tea (Camellia sinensis (L.) O. Kuntze)

Tea, being one of the most popular beverages requires large set of molecular markers for genetic improvement of quality, yield and stress tolerance. Identification of functionally relevant microsatellite or simple sequence repeat (SSR) marker resources from regulatory “Transcription factor (TF) genes” can be potential targets to expedite molecular breeding efforts. In current study, 2776 transcripts encoding TFs harbouring 3687 SSR loci yielding 1843 flanking markers were identified from traits specific transcriptome resource of 20 popular tea cultivars. Of these, 689 functionally relevant SSR markers were successfully validated and assigned to 15 chromosomes (Chr) of CSS genome. Interestingly, 589 polymorphic markers including 403 core-set of TF-SSR markers amplified 2864 alleles in key TF families (bHLH, WRKY, MYB-related, C2H2, ERF, C3H, NAC, FAR1, MYB and G2-like). Their significant network interactions with key genes corresponding to aroma, quality and stress tolerance suggests their potential implications in traits dissection. Furthermore, single amino acid repeat reiteration in CDS revealed presence of favoured and hydrophobic amino acids. Successful deployment of markers for genetic diversity characterization of 135 popular tea cultivars and segregation in bi-parental population suggests their wider utility in high-throughput genotyping studies in tea.

Characterization of transcription factor genes related to cold tolerance in Brassica napus

Brassica napus is the third most important oilseed crop in the world; however, in Korea, it is greatly affected by cold stress, limiting seed growth and production. Plants have developed specific stress responses that are generally divided into three categories: cold-stress signaling, transcriptional/post-transcriptional regulation, and stress-response mechanisms. Large numbers of functional and regulatory proteins are involved in these processes when triggered by cold stress. Here, our objective was to investigate the different genetic factors involved in the cold-stress responses of B. napus. Consequently, we treated the Korean B. napus cultivar Naehan at the 4-week stage in cold chambers under different conditions, and RNA and cDNA were obtained. An in silico analysis included 80 cold-responsive genes downloaded from the National Center for Biotechnology Information (NCBI) database. Expression levels were assessed by reverse transcription polymerase chain reaction, and 14 cold-triggered genes were identified under cold-stress conditions. The most significant genes encoded zinc-finger proteins (33.7%), followed by MYB transcription factors (7.5%). In the future, we will select genes appropriate for improving the cold tolerance of B. napus.

Analisis Single Nucleotide Polymorphism Gen Faktor Transkripsi MYB dalam Biosintesis Antosianin Kulit Buah Mangga (Analysis of Single Nucleotide Polymorphism Gene Transcription Factor MYB in Mango Skin Anthocyanin Biosynthesis)

<p>Perkembangan pasar bebas berdampak terhadap selera produk-produk pertanian, seperti warna buah mangga. Antosianin merupakan senyawa yang bertanggung jawab dalam menginduksi warna pada buah. Senyawa ini diregulasi oleh gen faktor transkripsi MYB. Mutasi Single Nucleotide Polymorphism (SNP) daerah ekson gen MYB dapat mengubah asam amino yang memengaruhi aktivitas enzim yang mengakibatkan munculnya variasi fenotipe warna buah di antara individu-individu dalam spesies yang sama. Penelitian ini bertujuan menemukan SNP pada gen MYB dari kulit buah mangga varietas Arum Manis, Gedong Gincu, Manalagi, Golek, dan Cengkir. Penelitian dilakukan dengan tiga tahap utama, yaitu isolasi DNA kulit buah mangga, Polymerase Chain Reaction (PCR), dan proses sekuensing oleh Macrogen Inc. (Singapore). Hasil multiple sequence alignment asam amino gen faktor transkripsi MYB menunjukkan adanya perbedaan basa yang mengakibatkan munculnya stop codon dari SNP 337 A→T dan SNP 338 A→G yang memengaruhi fenotipe warna kulit buah. SNP yang memunculkan stop codon dapat direkomendasikan untuk membedakan fenotipe pigmentasi antosianin pada kulit buah mangga Gedong Gincu yang bewarna merah dengan warna kulit buah mangga yang lainnya. Adanya SNP menyebabkan prematur stop codon yang terjadi pada gen faktor transkripsi MYB dan diduga berpengaruh terhadap pigmentasi antosianin.</p><p><strong>Keywords</strong></p><p>Mangga; SNP; Faktor transkripsi; Antosianin; MYB</p><p><strong>Abstract </strong></p><p>The development of free markets gives an impact on appetite for agricultural products, such as the color of mangoes fruit skin. Anthocyanins are compounds that are responsible for giving color to the fruit skin, these compounds are regulated by the MYB transcription factor genes. Single Nucleotide Polymorphism (SNP) mutations in the exon region of the MYB gene can change amino acids that affect enzyme activity, resulting in phenotypic variations in fruit color among individuals in the same species. This study aims to find SNP in MYB genes from mango peel varieties Arum Manis, Gedong Gincu, Manalagi, Golek, and Cengkir. The research was conducted in three main stages, namely isolation of mango peel DNA, Polymerase Chain Reaction (PCR), and sequencing process by Macrogen Inc (Singapore). The results of multiple sequence alignment of the amino acid MYB transcription factor genes showed a base difference which resulted in the appearance of a stop codon from SNP 337 A→T and SNP 338 A→G which affected the phenotype of fruit skin color. The SNP that raises the stop codon can be recommended to differentiate the anthocyanin pigmentation phenotype on the red skin of the mango Gedong Gincu from the skin color of other mangoes. The presence of SNP causes premature stop codon that occurs in the MYB transcription factor gene and is thought to have an effect on anthocyanin pigmentation.</p><p> </p>

GhAIL6, a Member of the AP2 Family Gene, Promotes Somatic Embryogenesis in Cotton.

Background Somatic embryogenesis (SE) is the process by which plant somatic cells are cultured in vitro without fertilization to regenerate embryos and develop into intact plants, the difficulty of cotton regeneration has severely limited functional gene research and transgenic breeding. The AP2 family is a relatively large family of transcription factor genes that regulate the process of growth and development, but the role of Aintegumenta-Like6 ( AIL6) in cotton SE has not been reported. Methods The 35S::AIL6:GR vector was constructed and transformed into cotton JH713 by Agrobacterium-mediated method, after 3 years of self-breeding, stable genetic T3 generation positive plants were obtained, identified by Southern, and three lines were selected for the following regeneration experiments.Results The results showed that overexpression of GhAIL6 significantly inhibited the proliferation of callus during the first 30 days, and promoted the embryogenic callus production at about 45 days.Couclusion Our results indicated that GhAIL6 was a key regulator of cotton SE, overexpression of GhAIL6 helped to improve the regeneration efficiency of cotton SE

Identification of the Yield Traits Related Haplotype Combinations of Transcription Factor Genes TaHDZ34 in Common Wheat

Improvement of yield-traits is one of the predominating objectives in wheat breeding. Homeodomain-leucine zipper (HD-ZIP) transcription factor plays significant roles in plant growth and development. The TaHDZ34 (A, B and D sub-genomics) genes consisting of three members of the HD-ZIP IV transcription factor gene subfamily in wheat (Triticum aestivum L.) were cloned. Two haplotypes of TaHDZ34-7A, TaHDZ34-7B or TaHDZ34-7D were respectively identified after the sequence polymorphism analysis, and three functional molecular markers were developed. The TaHDZ34 genes were divided into eight haplotype combinations. Association analysis and distinct population validation jointly indicated that TaHDZ34 had the function of modulating grain number per spike, effective spikelet number per spike, 1,000 kernel weight, and flag leaf area per plant in wheat. Among all haplotype combinations of TaHDZ34, Hap-ABD was the most excellent one. Subcelluar localization showed that TaHDZ34-7A was localized in the nucleus. Interaction proteins of TaHDZ34-7A protein proved to be involved in protein synthesis/degradation, energy production and transportation, and photosynthesis processes. Geographic distribution and frequencies of TaHDZ34 haplotype combinations suggested that the Hap-Abd and Hap-AbD were preferential selection in Chinese wheat breeding programs. The high-yield related haplotype combinations Hap-ABD of TaHDZ34 provided beneficial genetic resources for marker-assisted selection of new wheat cultivars.

A novel approach to co-expression network analysis identifies modules and genes relevant for moulting and development in the Atlantic salmon louse (Lepeophtheirus salmonis)

Background The salmon louse (Lepeophtheirus salmonis) is an obligate ectoparasitic copepod living on Atlantic salmon and other salmonids in the marine environment. Salmon lice cause a number of environmental problems and lead to large economical losses in aquaculture every year. In order to develop novel parasite control strategies, a better understanding of the mechanisms of moulting and development of the salmon louse at the transcriptional level is required. Methods Three weighted gene co-expression networks were constructed based on the pairwise correlations of salmon louse gene expression profiles at different life stages. Network-based approaches and gene annotation information were applied to identify genes that might be important for the moulting and development of the salmon louse. RNA interference was performed for validation. Regulatory impact factors were calculated for all the transcription factor genes by examining the changes in co-expression patterns between transcription factor genes and deferentially expressed genes in middle stages and moulting stages. Results Eight gene modules were predicted as important, and 10 genes from six of the eight modules have been found to show observable phenotypes in RNA interference experiments. We knocked down five hub genes from three modules and observed phenotypic consequences in all experiments. In the infection trial, no copepodids with a RAB1A-like gene knocked down were found on fish, while control samples developed to chalimus-1 larvae. Also, a FOXO-like transcription factor obtained highest scores in the regulatory impact factor calculation. Conclusions We propose a gene co-expression network-based approach to identify genes playing an important role in the moulting and development of salmon louse. The RNA interference experiments confirm the effectiveness of our approach and demonstrated the indispensable role of a RAB1A-like gene in the development of the salmon louse. We propose that our approach could be generalized to identify important genes associated with a phenotype of interest in other organisms.

Analyses of Key Gene Networks Controlling Carotenoid Metabolism in Xiangfen 1 Banana

Background: Banana fruits are rich in various high-value metabolites and play a key role in the human diet. Of these components, carotenoids have attracted considerable attention due to their physiological role and human health care functions. However, the accumulation patterns of carotenoids and genome-wide analysis of gene expression during banana fruit development have not been comprehensively evaluated. Results: In the present study, an integrative analysis of metabolome and transcriptome profiles in banana fruit with three different development stages was performed. A total of 11 carotenoid compounds were identified, and most of these compounds showed markedly higher abundances in mature green and/or mature fruit than in young fruit. Results were linked to the high expression of carotenoid synthesis and regulatory genes in the middle and late stages of fruit development. Co-expression network analysis revealed that 79 differentially expressed transcription factor genes may be responsible for the regulation of LCYB, a key enzyme catalyzing the biosynthesis of α- and β-carotene. Conclusions: Collectively, the study provided new insights into the understanding of dynamic changes in carotenoid content and gene expression level during banana fruit development.

The legume-specific transcription factor E1 controls leaf morphology in soybean

Background The leaf is a determinate organ essential for photosynthesis, whose size and shape determine plant architecture and strongly affect agronomic traits. In soybean, the molecular mechanism of leaf development is not well understood. The flowering repressor gene E1, which encodes a legume-specific B3-like protein, is known to be the gene with the largest influence on soybean flowering and maturity. However, knowledge of its potential other functions remains poor. Results Here, we identified a novel function of E1 protein in leaf development. Unifoliolate leaves of E1-overexpression (E1-OE) lines were smaller and curlier than those of wild type DongNong 50 (DN50) and Williams 82 (W82). Transverse histological sections showed disorganized cells and significantly elevated palisade tissue number, spongy tissue number, and bulliform cell number in E1-OE lines. Our results indicate that E1 binds to the promoters of the leaf- development-related CINCINNATA (CIN)-like TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR (TCP) transcription factor genes to negatively regulate their expression. Conclusions Our findings identify E1 as an important new factor in soybean leaf development.