Optimization of embryogenic-callus induction and embryogenesis of Glycyrrhiza glabra

Glabridin is a major biologically active flavonoid isolated specifically from the root of Glycyrrhiza glabra, which has many pharmacological activities. The production of the wild G. glabra was sharply decreased due to immoderate and ruinous utilization. In vitro regeneration via somatic embryogenesis is important for clonal propagation and genetic transformation. In this paper, factors affecting the embryogenic calli and embryo induction, maintenance and multiplication of G. glabra are assessed. The results showed that the explants of hypocotyl give the highest calli formation frequency of 93.3% on Murashige and Skoog (MS) medium containing 2.0 mg/L 6-benzylaminopurine (6-BA) and 0.5 mg/L 2,4- dichlorophenoxyacetic acid (2,4-D). The maximum efficiency of embryo were obtained on MS medium with 0.5 mg/L 6-BA + 0.5 mg/L kinetin zeatin (KT) + 0.1 mg/L indole-3-butyric acid (IBA); the embryos could develop further on medium with 1000 mg/L malt extract (ME). The occurrence of the embryogenic calli and proglobular embryo were studied by histological section, indicating the single cell origin of the embryogensis of G. glabra. With the protocol reported herein, some green embryo-like cultures were obtained, from which shoots were successfully regenerated in the germinated medium after 10 months of subculture.Keywords: Glycyrrhiza glabra L., callus induction, embryogenesis, cell culture, histological sectionAfrican Journal of Biotechnology Vol. 9(36), pp. 5823-5829, 6 September, 201

Transcriptomic and Proteomic Analysis of Shaan2A Cytoplasmic Male Sterility and Its Maintainer Line in Brassica napus

Cytoplasmic male sterility (CMS) lines are widely used for hybrid production in Brassica napus. The Shaan2A CMS system is one of the most important in China and has been used for decades; however, the male sterility mechanism underlying Shaan2A CMS remains unknown. Here, we performed transcriptomic and proteomic analysis, combined with additional morphological observation, in the Shaan2A CMS. Sporogenous cells, endothecium, middle layer, and tapetum could not be clearly distinguished in Shaan2A anthers. Furthermore, Shaan2A anther chloroplasts contained fewer starch grains than those in Shaan2B (a near-isogenic line of Shaan2A), and the lamella structure of chloroplasts in Shaan2A anther wall cells was obviously aberrant. Transcriptomic analysis revealed differentially expressed genes (DEGs) mainly related to carbon metabolism, lipid and flavonoid metabolism, and the mitochondrial electron transport/ATP synthesis pathway. Proteomic results showed that differentially expressed proteins were mainly associated with carbohydrate metabolism, energy metabolism, and genetic information processing pathways. Importantly, nine gene ontology categories associated with anther and pollen development were enriched among down-regulated DEGs at the young bud (YB) stage, including microsporogenesis, sporopollenin biosynthetic process, and tapetal layer development. Additionally, 464 down-regulated transcription factor (TF) genes were identified at the YB stage, including some related to early anther differentiation such as SPOROCYTELESS (SPL, also named NOZZLE, NZZ), DYSFUNCTIONAL TAPETUM 1 (DYT1), MYB80 (formerly named MYB103), and ABORTED MICROSPORES (AMS). These results suggested that the sterility gene in the Shaan2A mitochondrion might suppress expression of these TF genes in the nucleus, affecting early anther development. Finally, we constructed an interaction network of candidate proteins based on integrative analysis. The present study provides new insights into the molecular mechanism of Shaan2A CMS in B. napus

Stable, Environmental Specific and Novel QTL Identification as Well as Genetic Dissection of Fatty Acid Metabolism in Brassica napus

Fatty acid (FA) composition is the typical quantitative trait in oil seed crops, of which study is not only closely related to oil content, but is also more critical for the quality improvement of seed oil. The double haploid (DH) population named KN with a high density SNP linkage map was applied for quantitative trait loci (QTL) analysis of FA composition in this study. A total of 406 identified QTL were detected for eight FA components with an average confidence interval (CI) of 2.92 cM, the explained phenotypic variation (PV) value ranged from 1.49 to 45.05%. Totally, 204 consensus and 91 unique QTL were further obtained via meta-analysis method for the purpose of detecting multiple environment expressed and pleiotropic QTL, respectively. Of which, 74 stable expressed and 22 environmental specific QTL were also revealed, respectively. In order to make clear the genetic mechanism of FA metabolism at individual QTL level, conditional QTL analysis was also conducted and more than two thousand conditional QTL which could not be detected under the unconditional mapping were detected, which indicated the complex interrelationship of the QTL controlling FA content in rapeseed. Through comparative genomic analysis and homologous gene annotation, 61 candidates related to acyl lipid metabolism were identified underlying the CI of FA QTL. To further visualize the genetic mechanism of FA metabolism, an intuitive and meticulous network about acyl lipid metabolism was constructed and some closely related candidates were positioned. This study provided a more accurate localization for stable and pleiotropic QTL, and a deeper dissection of the molecular regulatory mechanism of FA metabolism in rapeseed

An Effective Screening Method and a Reliable Screening Trait for Salt Tolerance of Brassica napus at the Germination Stage

Salinity is a major and complex abiotic stress that inhibits plant growth and reduces crop yield. Given the global increase in soil salinity, there is a need to develop salt-tolerant species. Brassica napus L. is an important oilseed crop with some level of salt tolerance. However, few studies have evaluated its salt tolerance thoroughly or screened for traits that can be reliably evaluated for salt tolerance. Here, we evaluated salt tolerance in 549 B. napus inbred lines with different genetic backgrounds using the membership function value (MFV) of certain traits, including the germination rate, root and shoot length, root and shoot fresh weight, and total fresh weight. According to the evaluation criteria-mean MFV, 50 highly salt-tolerant, 115 salt-tolerant, 71 moderately salt-tolerant, 202 salt-sensitive, and 111 highly salt-sensitive inbred lines were screened at the germination stage. We also developed a mathematical evaluation model and identified that the salt tolerance index of shoot fresh weight is a single trait that reliably represents the salt tolerance of B. napus germplasm at the germination stage. These results are useful for evaluating and breeding salt-tolerant B. napus germplasm

Development of iFOX-hunting as a functional genomic tool and demonstration of its use to identify early senescence-related genes in the polyploid \u3ci\u3eBrassica napus\u3c/i\u3e

Functional genomic studies of many polyploid crops, including rapeseed (Brassica napus), are constrained by limited tool sets. Here we report development of a gain-of-function platform, termed ‘iFOX (inducible Full-length cDNA OvereXpressor gene)-Hunting’, for inducible expression of B. napus seed cDNAs in Arabidopsis. A Gateway-compatible plant gene expression vector containing a methoxyfenozide-inducible constitutive promoter for transgene expression was developed. This vector was used for cloning of random cDNAs from developing B. napus seeds and subsequent Agrobacterium-mediated transformation of Arabidopsis. The inducible promoter of this vector enabled identification of genes upon induction that are otherwise lethal when constitutively overexpressed and to control developmental timing of transgene expression. Evaluation of a subset of the resulting ~6000 Arabidopsis transformants revealed a high percentage of lines with full-length B. napus transgene insertions. Upon induction, numerous iFOX lines with visible phenotypes were identified, including one that displayed early leaf senescence. Phenotypic analysis of this line (rsl-1327) after methoxyfenozide induction indicated high degree of leaf chlorosis. The integrated B. napus cDNA was identified as a homolog of an Arabidopsis acyl-CoA binding protein (ACBP) gene designated BnACBP1-like. The early senescence phenotype conferred by BnACBP1-like was confirmed by constitutive expression of this gene in Arabidopsis and B. napus. Use of the inducible promoter in the iFOX line coupled with RNA-Seq analyses allowed mechanistic clues and a working model for the phenotype associated with BnACBP1-like expression. Our results demonstrate the utility of iFOX-Hunting as a tool for gene discovery and functional characterization of Brassica napus genome

Large expert-curated database for benchmarking document similarity detection in biomedical literature search

Document recommendation systems for locating relevant literature have mostly relied on methods developed a decade ago. This is largely due to the lack of a large offline gold-standard benchmark of relevant documents that cover a variety of research fields such that newly developed literature search techniques can be compared, improved and translated into practice. To overcome this bottleneck, we have established the RElevant LIterature SearcH consortium consisting of more than 1500 scientists from 84 countries, who have collectively annotated the relevance of over 180 000 PubMed-listed articles with regard to their respective seed (input) article/s. The majority of annotations were contributed by highly experienced, original authors of the seed articles. The collected data cover 76% of all unique PubMed Medical Subject Headings descriptors. No systematic biases were observed across different experience levels, research fields or time spent on annotations. More importantly, annotations of the same document pairs contributed by different scientists were highly concordant. We further show that the three representative baseline methods used to generate recommended articles for evaluation (Okapi Best Matching 25, Term Frequency-Inverse Document Frequency and PubMed Related Articles) had similar overall performances. Additionally, we found that these methods each tend to produce distinct collections of recommended articles, suggesting that a hybrid method may be required to completely capture all relevant articles. The established database server located at https://relishdb.ict.griffith.edu.au is freely available for the downloading of annotation data and the blind testing of new methods. We expect that this benchmark will be useful for stimulating the development of new powerful techniques for title and title/abstract-based search engines for relevant articles in biomedical research.Peer reviewe

Functional and Structural Diversity of Acyl-coA Binding Proteins in Oil Crops

Diversities in structure and function of ACBP were discussed in this review. ACBP are important proteins that could transport newly synthesized fatty acid, activated into -coA, from plastid to endoplasmic reticulum, where oil in the form of triacylglycerol occurs. ACBP were detected in various animal and plants species, which indicated their importance in biological function. In fact, involvement of ACBP in important process such as lipid metabolism, regulation of enzyme and gene expression, and in response to plant stresses has been proven in several studies. In this review, findings on ACBP of 11 well-known oil crops were reviewed to comprehend diversity, comparative analyses on ACBP structure were made, and link between structure and function, tissue expression and subcellular location of ACBP were also observed. Incomplete reports in some species were mentioned, which might be encouraging to start or to perform deeper studies. Similar characteristics were found in paralogs ACBP, and orthologs ACBP had different functions, despite the high identity in amino acid sequence. At the end, it is confirmed that ortholog proteins could not necessarily display the same function, even from closely related species

Sensory Integration Training Tool Design for Children with Autism Spectrum Disorder

This study aims to design a training tool for therapy of children with autism spectrum disorder (ASDs). Typically, ASDs pass through obstacle track several times with sandbags, which should be picked up from starting point and threw into a box at the end during sensory integration therapy. Counting the sandbags can help ASDs to have concept about the progress of mission. We redesign the counting box named “Skybox” which can help counting by playing sound after detect something throw in it. Aims to probe into the sound preference of two main subjects, an experiment with four kinds of sounds is conducted in this paper by using the method of paired comparisons. The result shows they like animals most, followed by human voice and nature sounds, and music instrument is the last. The material preference experiment shows two subjects like acrylic most, wood and paper are secondary while furry is the last. Skybox shortens their training time for 23.53%, 29.87% and 37.37% in three different projects. We consider that Skybox attracts ASDs therefore reduces their distraction and improves their performance in the usability test