Metabolomics & metabolites engineering in plant breeding
Saeed Mollaei; Poopak Farnia; Jalaledin Ghanavi; Mohammad Majidi
Abstract
Herbal plants play significant roles in the treatment of diseases and development of novel drugs. Salsola vermiculata is an annual plant which is broadly distributed in the southwest Asia, and used for the treatment of stomach disorders. This present study aimed at identifying and comparing the metabolic ...
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Herbal plants play significant roles in the treatment of diseases and development of novel drugs. Salsola vermiculata is an annual plant which is broadly distributed in the southwest Asia, and used for the treatment of stomach disorders. This present study aimed at identifying and comparing the metabolic profiles of different parts of S. vermiculata and evaluating the inhibitory potential of their extracts and fractions against acetylcholinesterase and α-glucosidase. LC-ESI-MS, GC, and GC-MS analytical methods were employed for metabolite profiling of the extracts, and their fractions. The inhibitory activities were determined by microplate reader-based colorimetric methods. 44 metabolites were identified in different parts of S. vermiculata. In roots, vanillic acid, rutin, salsoline, salsoline A, palmitic acid, oleic acid, linoleic acid, cumin aldehyde, and carvone; in seeds, vanillic acid, salsoline A, palmitic acid, oleic acid, linoleic acid, carvone, and β-caryophyllene; in leaves, gallic acid, vanillic acid, caffeic acid, rosmaric acid, rutin, quercetin, limonene, and carvone, and in flowers, gallic acid, vanillic acid, cinnamic acid, rosmaric acid, rutin, kaempferol, limonene, linalool, and carvone were reported as major components. According to the inhibitory activities results, the ethyl acetate fractions of leaves and the aqueous-acid fraction of roots displayed the highest inhibitory activity against acetylcholinesterase (IC50: 17.24 µg/mL), and α-glucosidase (IC50: 62.37 µg/mL), respectively. Finally, the leaves and roots of S. vermiculata are rich of phenolic and alkaloid compounds and the findings of this study describe them as a promising acetylcholinesterase and α-glucosidase inhibitors, and therefore, can be utilized for the development of new drugs.
Molecular biology and (trans) gene expression technology in plant breeding
Dariush Taghavi; Mohammad Majidi; Saeed Mollaei; Bahman Panahi
Abstract
Shikonin is a naphthoquinone with some important medicinal properties and is found in gromwell plant. There are so many biotechnological approaches proposed for the productivity enhancement, and elicitation is recognized as one of the most effective strategies for increasing the production of secondary ...
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Shikonin is a naphthoquinone with some important medicinal properties and is found in gromwell plant. There are so many biotechnological approaches proposed for the productivity enhancement, and elicitation is recognized as one of the most effective strategies for increasing the production of secondary metabolites in plant in vitro cultures. Moreover, a deeper understanding of the mechanisms and factors affecting shikonin biosynthesis can led to the design of more intelligent and efficient biological production systems. To this aim, in the present study, the expression of some genes related to the shikonin biosynthetic pathway including PAL, 4CL, HMGR, GPPS and PGT in in vitro cultures of Lithospermum officinale in response to methyl jasmonate (MJ) at different times, were investigated by real-time PCR. The results showed that MJ had a significant effect on increasing gene expression levels in elicited samples compared to control samples. Additionally, we found that the studied genes respond to MJ with different pattern, in which the highest increase in gene expression level was observed for PGT while the lowest increase was observed for GPPS. Maximum and minimum transcript levels were obtained in most genes at 4 and 96 h post-elicitation, respectively. It was also found that phenylpropanoid pathway genes respond better to MJ than terpenoid pathway genes. The results of the present study would increase our knowledge about elicitor signal transduction pathways, and may be particularly useful for enhancement of shikonin production in plant cell cultures of L. officinale.
Genetic engineering & plant breeding
Mohammad Majidi; Yones Bahmani
Abstract
Isolation of high-quality RNA is one of the most crucial methods in molecular biology. RNA extraction from woody plants has been problematic due to the presence of rigid and woody tissues, large amounts of polysaccharides, polyphenols and other secondary metabolites. Here we present a suitable protocol ...
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Isolation of high-quality RNA is one of the most crucial methods in molecular biology. RNA extraction from woody plants has been problematic due to the presence of rigid and woody tissues, large amounts of polysaccharides, polyphenols and other secondary metabolites. Here we present a suitable protocol for RNA isolation from a wide range of woody plants that includes eight gymnosperms and four angiosperms. The method is based on the CTAB protocol which was modified by adding sodium citrate and two helper buffers. Agarose gel electrophoresis showed a good RNA integrity and total RNA profile that includes all expected RNA bands. Also, DNA and protein contaminations were not observed. Spectrophotometric quantification of RNA samples by NanoDrop showed that the average RNA yields ranged from 35.68 to 216.98 µg per gram fresh weight, that is enough to proceed into cDNA synthesis and other RNA-related works. Both the A260/A280 and A260/A230 ratios were in the desired ranges, indicating that RNA was of high purity and without protein, polyphenol, and polysaccharide contamination. The efficiency of isolated RNA for downstream applications was verified by real-time PCR and successful amplification of a long cDNA. Finally, some advantages and possible applications of the method are also mentioned.