Search or add a thesis

Advanced Search (Beta)
Home > Agrobacterium Mediated Transformation and Secondary Metabolite Analysis of Ajuga Bracteosa Wall. Ex. Benth

Agrobacterium Mediated Transformation and Secondary Metabolite Analysis of Ajuga Bracteosa Wall. Ex. Benth

Thesis Info

Access Option

External Link

Author

Rani, Rehana

Program

PhD

Institute

Quaid-I-Azam University

City

Islamabad

Province

Islamabad.

Country

Pakistan

Thesis Completing Year

2015

Thesis Completion Status

Completed

Subject

Natural Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/7753/1/Dr%20Rehana%20thesis.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725444161

Asian Research Index Whatsapp Chanel
Asian Research Index Whatsapp Chanel

Join our Whatsapp Channel to get regular updates.

Similar


Ajuga bracteosa is a valuable yet endangered medicinal plant containing many important compounds, including withanolides, phytoecdysteroids, neo-clerodane and iridoid glycosides. These compounds possess a broad spectrum of biological, pharmacological and medicinal properties, but their yield is very low in wild A. bracteosa and chemical synthesis is not viable. The increased demand of natural products for medicinal purposes coupled with the low product yields and supply concerns of plant harvest has renewed interest in large-scale in vitro plant culture technology. A.bracteosa plant was subjected to in vitro culturing with the aim to study the effect of plant growth regulators on somatic embryogenesis for mass production and uniform plantlets for further genetic transformation. Leaf explants cultured on B5 medium containing 1mg/L 1-naphthaleneaceatic acid (NAA) + 0.5 mg/L 6-Benzyl amine purines (BAP) proved to be the most effective combination for somatic embryo induction. Agrobacterium tumefaciens strain GV3101 harboring pPCV002-A and pPCV002-CaMVC was used to obtain transgenic A. bracteosa plants. Transformation efficiency of 38% for rolA gene and 31% for rolC was obtained. Molecular analysis such as PCR and Southern blotting confirmed the integration of the rolA and rolC genes in the putative transgenic lines. The transcripts of these two genes were monitored by semi-quantitative RT-PCR. Furthermore, a significant difference in phytochemistry and the pharmacological activities (antioxidant, antibacterial, antiproliferative and antidiabetic) was observed not only between the transgenic and control/untransformed plants but also among the transgenic lines themselves. The rolC transgenic lines in general showed better results compared to rolA. Quantitative analysis of Electro-spray ionization mass spectrometer (ESI-TOF) revealed relatively higher contents of 20 hydroxyecdysone (20-HE) in transgenic plant lines of rolA and rolC compared to control/untransformed plant. Effect of low temperature stress on 20-HE biosynthesis revealed relatively high 20-HE content at 15°C compared to control (25°C). Principal component analysis (PCA) of the metabolic profiles obtained with non-targeted (ESI-MS) approach successfully distinguished the control A. bracteosa from the transgenic rolA and rolC population. Defense related secondary metabolites (tryptophan, mannitol, Cinnamic acid, and Shikimic acid) were identified in transgenic A. bracteosa lines. Taken together, our study suggests that low temperature stress and transformation with rol genes can be used for increased, constant and permanent production of valuable secondary metabolites from A. bracteosa plant.
Loading...
Loading...

Similar Books

Loading...

Similar Chapters

Loading...

Similar News

Loading...

Similar Articles

Loading...

Similar Article Headings

Loading...

66. Al-Tahreem/The Prohibition

66. Al-Tahreem/The Prohibition

I/We begin by the Blessed Name of Allah

The Immensely Merciful to all, The Infinitely Compassionate to everyone.

66:01
a. O The Prophet!
b. Why should you prohibit for yourself that Allah has allowed to you just to please your wives?
c. Allah is Ever-Forgiving, Ever-Merciful.

66:02
a. Allah has already prescribed a way regarding the absolution of your oaths.
b. And Allah is your Protector, and
c. HE is the All-Knowing, the All-Wise.

66:03
a. And when The Prophet confided a matter to one of his wives, then she disclosed it to another and Allah made it known to him – The Prophet.
b. He made some of it known to her and passed over some of it.
c. So when he told her of it, she asked:
d. ‘Who told you of this?’
e. He responded:
f. ‘The All-Knowing, the All-Wise told me.’

66:04
a. If both of you would repent to Allah it will be good for you as your hearts are inclined towards it.
b. But if you both would support each other against him – The Prophet - then it is Allah WHO is his Protector, and so are Gabriel and the righteous among the believers, and also the angels, are his supporters.

66:05
a. O Wives of The Prophet!
b. It may well be that if he were to divorce you all, then his Rabb - The Lord would give him wives in your place - better than you: submitting, believing, dutiful, repentant, worshiping, devout - previously married as well as virgins.

66:06
a. O The Faithful!
b. Guard yourselves and your families from...

قرآنی اصولوں کی روشنی میں معاشرتی استحکام

Islam is the religion of nature. It not only approves the social interaction among the masses, but also helps in its development towards positive ends. Islam has given natural and universal principles which help its followers to develop a harmonious society, discouraging all the attempts to divide the society into different sections. Islamic society is based upon the following fundamental principles i. E equality, harmony of thoughts, justice, amar-bilmaaruf-wa-nahi-anilmunkar (ask for good and forbid from evil), unity, sense of responsibility, virtue and evil, abolition of sectarianism & fulfillment of promises, reflecting the universality of the religion, Islam. Pakistan today, is facing various social problems like terrorism, corruption, poverty, unemployment, broken families, sectarianism, onslaught of western culture and demand of unrestricted liberty by womenfolk. The moral degradation of the society is due to the fact that the true Islamic spirit and moral teachings and trainings of Islam have not been applied with true mind and honest intentions. The moral values are ignored by the media which is the cause of great concern.

Ecology, Diversity and Metabolites Analysis of Extremophiles

Our earth is naturally bestowed with extreme ecological niches which rarely support any life form to exist there. Microbes harboring such habitats are unusual and exhibit specific characters to cope with stress conditions. Present research is conducted to study the diversity, ecology and metabolites of such microbes i.e. extremophiles. Water, soil and sediment samples were collected from selected extreme sites i.e. salt mine, acid seep, and hot water spring. Total DNA was extracted from soil and sediment samples for metagenome sequencing. Microbial diversity was investigated by barcoded amplicon sequencing of metagenome. Sequencing output files were submitted in the National Centre for Biotechnology Information (NCBI) database under the accession numbers SAMN08026743, SAMN08026744, SAMN08398365, and SAMN08398366 for saline, hot spring, and acid seep samples respectively. Metagenomic analysis revealed that microbial communities were dominated by Proteobacteria in all samples. Other than Proteobacteria, Firmicutes constituted the most abundant population in hot spring, Euryarchaeota were abundant in saline soil and Cyanobacteria dominated the acid seep sample. Owing to the complexities of extreme sites, some unidentified sequences were also found in samples. For isolation and characterization of extremophiles, cultivation based method was used. Eleven halophilic bacteria i.e. MB588, MB589, MB590, MB591, MB592, MB593, MB594, MB595, MB596, MB597, and MB598 were isolated from saline soil samples on LB medium with 4M NaCl concentration. Isolates were characterized morphologically, biochemically and physiologically. On the basis of 16S rDNA gene analysis, MB590, MB591, MB593, MB594, MB595, and MB596 were identified as Halomonas elongata. Whereas, MB588, MB589, MB592, MB597 and MB598 were identified as Halobacillus karajiensis, Alkalibacillus almallahensis, Aquisalibacillus elongatus, Salinicoccus sesuvii and Halomonas aquamarina respectively. Most of the strains exhibited multiple heavy metal and antibiotic resistance. Multiple antibiotics and heavy metal resistance indices i.e. MAR and MHR, revealed Halomonas elongata MB593 as a most resistant strain. Strong association among heavy metal resistance and antibiotic tolerance was observed in halophilic bacteria. No significant extremozyme production was observed in halophilic bacteria. However, biosurfactant production was indicated in nine halophilic strains.Most of the strains significantly reduced surface tension up to 35 dynes/cm and explicated emulsification activities of different hydrocarbons i.e. hexane, canola, xylene and petrol. Moreover, all the strains exhibited xviii oil displacement. Structural characterization of biosurfactants revealed mixture of glycolipids. Three bacterial strains i.e. MB592, MB597 and MB598 exhibited pigment production. Characterization of pigments using different analytical techniques i.e. UV spectroscopy, IR spectroscopy, and HPLC analysis revealed bacterioruberin carotenoids. Methanolic extracts of pigments showed strong antibacterial and antioxidant activities. From hot spring sample, seven hyper-thermophilic bacteria were isolated at 80˚C. Isolates were characterized morphologically, biochemically and physiologically. On the basis of 16S rDNA sequence analysis, isolates were identified as Geobacillus jurassicus MB599, Geobacillus jurassicus MB605, Geobacillus sp. MB601, Geobacillus sp. MB604, Geobacillus stearothermophilus MB600, Geobacillus thermocatenulatus MB602 and Geobacillus thermoparaffinivorans MB603. All the strains exhibited significant protease and amylase production. From acid seep sample, a gram negative, iron(II) oxidizing, acidophilic bacterium IO2C was isolated at pH 2 and 25˚C. On the basis of 16S rDNA analysis, it was identified as Acidithiobacillus ferrooxidans. Strain produced biominerals by oxidation of iron substrates like iron sulfate and iron ammonium sulfate. SEM, XRD and FTIR analysis of bio-minerals indicated schwertmannite and jarosite. Whole genome sequence analysis of Acidithiobacillus ferrooxidans IO-2C revealed 58.73% of G+C content, 2,716,894 base pairs in 23 contigs and 2,927 protein coding sequences. In addition to this, it contained functional genes for iron and sulfur metabolism, multidrug resistance, and heavy metal resistance. Present study provided a comprehensive examination of microbial diversity of extremophiles by consolidating culture dependent and independent techniques to get more realistic portrayal of microbial communities. This study also identified bacterial metabolites and enzymes which will provide industry with more cost-effective biomolecules for specific purposes. Future in depth studies on genomics and metabolomics of extremophilic bacterial strains identified in this study will provide insight to their adaptive mechanisms and their potential roles in bioremediation of toxic pollutants, food processing, and biomedicine.