In this paper, a micropropagation protocol of sugar palm (Arenga pinnata Wurmb Merr) through callogenesis and somatic embryogenesis was examined. Callus induction frequency and somatic embryogenesis response were dependent on plant growth regulators (PGRs) and genotype. Semi-compact and compact embryogenic calluses were induced from excised immature zygotic embryo (IZE) cultured on semi-solid MS (Murashige & Skoog, 1962) medium supplemented with various concentration and combination of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyl aminopurine acid (BAP). MS medium supplemented with 0.4 mg/L 2,4-D and 0.5 mg/L BAP was found optimum to induce 100% rate of embryogenic calluses and maximum degree of callus formation after 8 and 12 weeks of culture. The incorporation of increased sucrose concentration (60.0 g/L) and 2.0 g/L casein hydrolysate (CH) to the culture medium with similar PGRs composition enhanced the induction of globular somatic embryos (SEs), while addition of silver nitrate (AgNO3) produced SEs of different stages. SEs maturated in MS medium containing 1.0 mg/L BAP and 1.0 mg/L naphthalene-acetic acid (NAA) formed cotyledon-stage embryos. Clonal roots regeneration was obtained on half-strength MS devoid of PGRs after 4 months of culture. Frequent subcultures increased embryogenesis rate favourably.
Rafflesia possesses unique biological features and known primarily for producing the world's largest and existing as a single flower. However, to date, little is known about key regulators participating in Rafflesia flower development. In order to further understand the molecular mechanism that regulates Rafflesia cantleyi flower development, RNA-seq data from three developmental stages of floral bud, representing the floral organ primordia initiation, floral organ differentiation, and floral bud outgrowth, were analysed. A total of 89,890 transcripts were assembled of which up to 35% could be annotated based on homology search. Advanced transcriptome analysis using K-mean clustering on the differentially expressed genes (DEGs) was able to identify 12 expression clusters that reflect major trends and key transitional states, which correlate to specific developmental stages. Through this, comparative gene expression analysis of different floral bud stages identified various transcription factors related to flower development. The members of WRKY, NAC, bHLH, and MYB families are the most represented among the DEGs, suggesting their important function in flower development. Furthermore, pathway enrichment analysis also revealed DEGs that are involved in various phytohormone signal transduction events such as auxin and auxin transport, cytokinin and gibberellin biosynthesis. Results of this study imply that transcription factors and phytohormone signalling pathways play major role in Rafflesia floral bud development. This study provides an invaluable resource for molecular studies of the flower development process in Rafflesia and other plant species.
Gymnema sylvestre (R.Br.) is an important diabetic medicinal plant which yields pharmaceutically active compounds called gymnemic acid (GA). The present study describes callus induction and the subsequent batch culture optimization and GA quantification determined by linearity, precision, accuracy, and recovery. Best callus induction of GA was noticed in MS medium combined with 2,4-D (1.5 mg/L) and KN (0.5 mg/L). Evaluation and isolation of GA from the calluses derived from different plant parts, namely, leaf, stem and petioles have been done in the present case for the first time. Factors such as light, temperature, sucrose, and photoperiod were studied to observe their effect on GA production. Temperature conditions completely inhibited GA production. Out of the different sucrose concentrations tested, the highest yield (35.4 mg/g d.w) was found at 5% sucrose followed by 12 h photoperiod (26.86 mg/g d.w). Maximum GA production (58.28 mg/g d.w) was observed in blue light. The results showed that physical and chemical factors greatly influence the production of GA in callus cultures of G. sylvestre. The factors optimized for in vitro production of GA during the present study can successfully be employed for their large-scale production in bioreactors.
The SCF complex is a widely studied multi-subunit ring E3 ubiquitin ligase that tags targeted proteins with ubiquitin for protein degradation by the ubiquitin 26S-proteasome system (UPS). The UPS is an important system that generally keeps cellular events tightly regulated by purging misfolded or damaged proteins and selectively degrading important regulatory proteins. The specificity of this post-translational regulation is controlled by F-box proteins (FBPs) via selective recognition of a protein-protein interaction motif at the C-terminal domain. Hence, FBPs are pivotal proteins in determining the plant response in multiple scenarios. It is not surprising that the FBP family is one of the largest protein families in the plant kingdom. In this review, the roles of FBPs, specifically in plants, are compiled to provide insights into their involvement in secondary metabolites, plant stresses, phytohormone signalling, plant developmental processes and miRNA biogenesis.