De novo Transcriptome Assembly and Differential Expression Analysis of Catharanthus roseus in Response to Salicylic acid

Publication history: Accepted - 12 September 2022; Published online - 24 October 2022The anti-cancer vinblastine and vincristine alkaloids can only be naturally found in periwinkle (Catharanthus roseus). Both of these alkaloids' accumulations are known to be influenced by salicylic acid (SA). The transcriptome data to reveal the induction effect (s) of SA, however, seem restricted at this time. In this study, the de novo approach of transcriptome assembly was performed on the RNA-Sequencing (RNA-Seq) data in C. roseus. The outcome demonstrated that SA treatment boosted the expression of all the genes in the Terpenoid Indole Alkaloids (TIAs) pathway that produces the vinblastine and vincristine alkaloids. These outcomes supported the time-course measurements of vincristine alkaloid, the end product of the TIAs pathway, and demonstrated that SA spray had a positive impact on transcription and alkaloid synthesis. Additionally, the abundance of transcription factor families including bHLH, C3H, C2H2, MYB, MYB-related, AP2/ ERF, NAC, bZIP, and WRKY suggests a role for a variety of transcription families in response to the SA stimuli. Di-nucleotide and tri-nucleotide SSRs were the most prevalent SSR markers in microsatellite analyses, making up 39% and 34% of all SSR markers, respectively, out of the 77,192 total SSRs discovered

Potato starch synthases : Functions and relationships

Starch, a very compact form of glucose units, is the most abundant form of storage polyglucan in nature. The starch synthesis pathway is among the central biochemical pathways, however, our understanding of this important pathway regarding genetic elements controlling this pathway, is still insufficient. Starch biosynthesis requires the action of several enzymes. Soluble starch synthases (SSs) are a group of key players in starch biosynthesis which have proven their impact on different aspects of the starch biosynthesis and functionalities. These enzymes have been studied in different plant species and organs in detail, however, there seem to be key differences among species regarding their contributions to the starch synthesis. In this review, we consider an update on various SSs with an emphasis on potato SSs as a model for storage organs. The genetics and regulatory mechanisms of potato starch synthases will be highlighted. Different aspects of various isoforms of SSs are also discussed

Isolation and identification of probiotic lactic acid bacteria from donkey,s milk

Background : The utilization of donkey’s milk is increasing with regard to nutritional values and non-allergenic proteins. Donkey’s milk microbiota, especially lactic acid bacteria (LAB) has not yet been fully identified. This study was conducted to identify and isolate LAB with probiotic properties. Materials and Methods: Specific culture-dependent techniques and standard microbiology methods were used to identify LAB. 250 isolates were randomly picked from different donkey’s milk samples different farms in Alashtar, north of Lorestan province, Iran. Gram-staining, catalase reaction, motility test, growth at different temperatures, pH 9.6 and in presence of 6.5% NaCl as well as the ability to ferment different sugars, sodium hippurate hydrolysis and light microscope analysis were investigated. Results: Out of 250 isolates, 20 isolates were gram-positive, catalase-negative and cocci form. Enterococcus and Streptoccus genera were dominant LAB in donkey’s milk. Entreococcus faecalis (55%) and Streptococcus devriesei (45%) were among the predominant species. The results of this experiment, for the first time, reported the occurrence of Streptococcus devriesei in donkey’s milk in Iran. Conclusion: Despite high lysozyme concentration, donkey’s milk rich in probiotic LAB can be used to feed infants with allergy to cow’s milk proteins. Both Entreococcus and Streptococcus bacteria produce different types of bacteriocins which are effective against a wide range of different type of infections. Occurrence of Streptococcus devriesei in donkey,s milk may provide new research field in Iran

Cloning and in planta expression of an omiganan antimicrobial peptide in tobacco and potato plants to control the growth of human bacterial pathogens

Background: Antimicrobial peptides are one of the vital components of innate immunity in plants and animals. The identification and introduction of novel and effective peptide molecules to plants is a cost-effective way both to improve the resistance of crop plant species to pathogens, and to produce peptides for pharmaceutical applications by using recombinant DNA technology. Material and Methods: An expression construct containing the omiganan (MBI-226) antimicrobial gene sequence was cloned and used for tobacco and potato Agrobacterium tumefaciens-mediated transformation. Following tissue culture, Polymerase chain reaction analysis (PCR) confirmed that some kanamycin resistant plants are transgenic. A number of transgenic plants, along with a non-transgenic control, were selected. Total protein was extracted from the transgenic plants, and the non-transgenic control, and was used for antimicrobial activity assay against some human pathogens, including; Escherchia coli, Staphylococcus epidermis, Pseudomonas aeruginosa, Salmonella typh, Staphylococcus aureus, Bacillus cereus, Candida albicans using the disc diffusion method. Results: Total protein extract from transgenic plants was significantly (P0.05) on human gram-positive pathogenic bacteria. Conclusion: The total protein from the omiganan-expressing peptide had a strong antibacterial activity against some human bacterial pathogens. By expression and purification of the omiganan peptide, the peptide could be used as an antibiotic to destroy pathogenic bacteria. This approach could open an opportunity to produce antibacterial peptides in plants for pharmaceutical applications

The Expression of Terpenoid Indole Alkaloid (TIAs) Pathway Genes in Catharanthus roseus in Response to Salicylic Acid treatment

Publication history: Accepted - 28th August 2020; Published online - 4th September.Vinblastine and vincristine are two important anti-cancer drugs that are synthesized by the Terpenoid Indole Alkaloids (TIAs) pathway in periwinkle (Catharanthus roseus). The major challenge in the pharmaceutical industry is the low production rate of these alkaloids. TIA pathway is affected by elicitors, such as salicylic acid (SA). This study aimed to investigate the expression pattern of some key genes in TIAs pathway under SA treatment. Foliar application of SA (0.01 and 0.1 mM) was used and leaves samples were taken at 0, 12, 18, 24 and 48 hours after the treatment. qRT-PCR was used to investigate the expression pattern of Chorismate mutase (Cm), Tryptophan decarboxylase (Tdc), Geraniol-10-hydroxylase (G10h), Secologanin synthase (Sls), Strictosidine synthase (Str), Desacetoxyvindoline-4-hydroxylase (D4h) and Deacetylvindoline-4-O-acetyltransferase (Dat) genes, following the SA treatment. The results of this experiment showed that transcript levels of Tdc, G10h, Sls, Str, D4h and Dat genes were significantly up-regulated in both SA concentration treatments. Furthermore, the highest transcript levels of Dat was observed after 48 hours of the SA treatments. qRT-PCR results suggests that SA induces transcription of major genes involved in alkaloids biosynthesis in Catharanthus roseus. It can be concluded that up-regulation of Tdc, G10h, Sls, Str, D4h and Dat genes can result in a higher production rate of vinblastine and vincristine alkaloids

Transcriptome response of roots to salt stress in a salinity-tolerant bread wheat cultivar.

Salt stress is one of the major adverse environmental factors limiting crop productivity. Considering Iran as one of the bread wheat origins, we sequenced root transcriptome of an Iranian salt tolerant cultivar, Arg, under salt stress to extend our knowledge of the molecular basis of salinity tolerance in Triticum aestivum. RNA sequencing resulted in more than 113 million reads and about 104013 genes were obtained, among which 26171 novel transcripts were identified. A comparison of abundances showed that 5128 genes were differentially expressed due to salt stress. The differentially expressed genes (DEGs) were annotated with Gene Ontology terms, and the key pathways were identified using Kyoto Encyclopedia of Gene and Genomes (KEGG) pathway mapping. The DEGs could be classified into 227 KEGG pathways among which transporters, phenylpropanoid biosynthesis, transcription factors, glycosyltransferases, glutathione metabolism and plant hormone signal transduction represented the most significant pathways. Furthermore, the expression pattern of nine genes involved in salt stress response was compared between the salt tolerant (Arg) and susceptible (Moghan3) cultivars. A panel of novel genes and transcripts is found in this research to be differentially expressed under salinity in Arg cultivar and a model is proposed for salt stress response in this salt tolerant cultivar of wheat employing the DEGs. The achieved results can be beneficial for better understanding and improvement of salt tolerance in wheat

Fusion proteins comprising the catalytic domain of mutansucrase and a starch-binding domain can alter the morphology of amylose-free potato starch granules during biosynthesis

It has been shown previously that mutan can be co-synthesized with starch when a truncated mutansucrase (GtfICAT) is directed to potato tuber amyloplasts. The mutan seemed to adhere to the isolated starch granules, but it was not incorporated in the starch granules. In this study, GtfICAT was fused to the N- or C-terminus of a starch-binding domain (SBD). These constructs were introduced into two genetically different potato backgrounds (cv. Kardal and amf), in order to bring GtfICAT in more intimate contact with growing starch granules, and to facilitate the incorporation of mutan polymers in starch. Fusion proteins of the appropriate size were evidenced in starch granules, particularly in the amf back- ground. The starches from the various GtfICAT/ SBD transformants seemed to contain less mutan than those from transformants with GtfICAT alone, suggesting that the appended SBD might inhibit the activity of GtfICAT in the engineered fusion proteins. Scanning electron microscopy showed that expression of SBD-GtfICAT resulted in alterations of granule morphology in both genetic backgrounds. Surprisingly, the amf starches con- taining SBD-GtfICAT had a spongeous appearance, i.e., the granule surface contained many small holes and grooves, suggesting that this fusion protein can interfere with the lateral interactions of amylopectin sidechains. No differences in phys- ico-chemical properties of the transgenic starches were observed. Our results show that expression of granule-bound and ‘‘soluble’’ GtfICAT can affect starch biosynthesis differently