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Home > Mocrobial Diversity and Metagenomic Analysis of Rhizosphere of Plants Growing in Extremely Halophytic Environment Khewra Salt Mines

Mocrobial Diversity and Metagenomic Analysis of Rhizosphere of Plants Growing in Extremely Halophytic Environment Khewra Salt Mines

Thesis Info

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Author

Mukhtar, Salma

Program

PhD

Institute

Forman Christian College

City

Lahore

Province

Punjab

Country

Pakistan

Thesis Completing Year

2018

Thesis Completion Status

Completed

Subject

Biotechnology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/10130/1/PhD%20thesis%20%28Dr.%20Salma%20Mukhtar%2c%20FC%20College%2c%20Lahore%29.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726650504

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Natural hypersaline environments such as Khewra Salt Mines seem to be a promising source of bacteria able to alleviate salt stress in important crops such as wheat, rice and maize. Bacteria belonging to various genera have been able to impart salt tolerance to plants growing in such places. Saline soils get more attention these days as a result of shortage of arable lands and demands for environmental restoration. Therefore it is important to get better understanding of the microbial diversity and their ecological role in saline environments. The goal of the present study was to evaluate the diversity of bacterial communities in the rhizosphere of halophytes (Salsola stocksii and Atriplex amnicola), non-rhizospheric soil and hypersaline lake-bank soil samples by culture dependent and metagenomic approaches. This study included assessment of microbial diversity by isolation of bacteria from the different fractions of the rhizosphere of halophytes, to unlock the massive uncultured microbial diversity present in the rhizosphere of halophytes from moderately and highly saline environments by pyrosequencing analysis of 16S rRNA gene, comparison of culturable and non-culturable microbial diversity from the rhizosphere of halophytes. Additionally, megaplasmids were isolated from different halophilic bacterial strains and plasmids conferring salt tolerant genes were characterized. To study culturable microbial diversity from the rhizosphere of halophytes (S. stocksii and A. amnicola), non-rhizospheric soil and hypersaline lake-bank soil samples, two media were used; (1) HaP (halophilic medium) with 2M NaCl concentration and (2) hypersaline MGM (minimal growth medium) specific for haloarchaea with 4M NaCl concentrations. On the basis of salt tolerance ability, 47 halophilic isolates from the rhizosphere of Salsola, 42 isolates from Atriplex, 27 isolates from non-rhizospheric and 27 isolates from hypersaline lake-bank soils were identified by using 16S rRNA gene analysis. Phylogenetic analysis showed that bacterial strains belonging to Bacillus, Pseudomonas, Halobacillus and Kocuria were dominant in the rhizosphere of halophytes (Salsola and Atriplex) and non-rhizospheric and hypersaline lake-bank soils. Halobacterium and Halococcus were dominant archaeal genera identified from all soils. To characterize plasmid conferring salt tolerant genes, selected halophilic strains with salt tolerance greater than 2.5M NaCl were plasmid cured by using heat shock method, 3% SDS and sodium benzoate. These plasmids were isolated and transformed into E. coli strains (Top10, DH10α and BL21). The growth response of wild type, plasmid cured and transformed E. coli strains was compared at 1.5-4M NaCl concentration. Plasmids from halophilic Bacillus strain HL2HP6 (Bacillus endophyticus) were sequenced by using whole genome sequencing technique at Hirsch’s lab, UCLA in collaboration with Joint Genome Institute lab (JGI), USA. Results of plasmid curing showed that some halophilic bacterial strains such as Bacillus strain HL2HP6, Oceanobacillus strain AT3HP15 and Oceanobacillus strain LK3HaP7 lost their ability to grow in halophilic medium but they grew well on LB medium. Functional analysis of plasmid sequences showed different proteins and enzymes which are known to be involved in genetic information processing, carbohydrate metabolism, amino acid metabolism, xenobiotics biodegradation and metabolism, glycan biosynthesis and metabolism and human diseases. To study the plant growth promoting effect of phosphate solubilizing bacteria, five bacterial isolates; three Bacillus (HL1HP11, HL3RS14 and LK1HaP9), one Enterobacter aerogenes (AT1HP4) and one Aeromonas veronii (AT1RP10) strains were used as inoculants; in the form of seed coat and enriched soil based phosphate biofertilizers. All bacterial strains positively affected the plant growth as compared to non-inoculated control plants. Plants inoculated with Bacillus strain HL3RS14 based soil biofertilizers showed maximum increase in dry weights of root (51-104%) and shoot (35-114%) as compared to control (soil + rock phosphate, no inoculum). Rhizosphere microbiomes of halophytes Urochloa, Kochia, Salsola, and Atriplex living in moderately and highly saline environments and non-halophyte Triticum were analysed by using pyrosequencing technique. Metagenomic analysis of soil microbiomes indicated that Actinobacteria were dominant in the saline soils whereas Proteobacteria predominated in non-saline soils. Firmicutes, Acidobacteria, Bacteriodetes, Planctomycetes and Thaumarchaeota were predominant phyla in saline and non-saline soils, whereas Cyanobacteria, Verrucomicrobia, Choroflexi, Gemmatimonadetes and the unclassified WPS-2 were less abundant. Plant microbiome of halophyte (Salsola) and non-halophyte (wheat) was also studied through metagenomics approach. Proteobacteria and Actinobacteria were the most abundant phyla in the rhizosphere, root endosphere and phyllosphere of Salsola and wheat. However, Firmicutes, Acidobacteria, Bacteriodetes, Planctomycetes, Cyanobacteria, Verrucomicrobia, Choroflexi and Euryarchaeota were predominant groups from halophyte whereas Actinobacteria, Proteobacteria, Firmicutes, Cyanobacteria, Acidobacteria, Bacteriodetes, Planctomycetes and Verrucomicrobia were predominant phyla of wheat samples. Diversity and differences of microbial flora of Salsola and wheat suggested that functional interactions between plants and microorganisms contribute to salt stress tolerance.
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یاقوت کے ورق

یاقوت کے ورق

                ناطق کی نظموں کا دوسرا مجموعہ ہے۔اس مجموعہ میں بھی انہوں نے نظموں کے ذریعے دل سے دل تک کا سفر طے کیا ہے۔ان کا کہناہے کہ وہ تب تک شعر نہیں کہتے جب تک ان کے پاس لکھنے کیلئے مسئلہ نہ ہو ۔وہ شاعری  میں صرف احساسات کا اظہار کرکے لفظی ڈرامہ بازی نہیں چاہتے ہیں بلکہ وہ حقیقت کا رنگ بھرتے ہیں۔وہ لفظوں کے ساتھ انصاف کرتے ہیں اور صاف گوئی سے کام لیتے ہیں۔اس کتاب کا انتساب آپ نے اپنی بیوی رفیعہ پروین اور بیٹی وجیہہ بتول کے نام لکھا ہے۔کل 36 نظموں پر مشتمل یہ مجموعہ ہے اور اس کتاب میں ناطق نے نظم’’سفیر لیلی‘‘  لکھی جو پندرہ صفحات پر مشتمل ہیاور کل 4حصوں میں اس کو تقسیم کیا گیاہے۔

                ان کا کہنا ہے کہ وہ داستان کے آدمی ہیں،ماضی، حال اور مستقبل کے آدمی ہیں۔ وہ کھنڈرات کو دیکھتے ہیں تو اندازہ کرتے ہیں کہ کیا وجہ ہے کہ یہ تباہ وبرباد ہوئے۔ انہیں ماضی سے لگاؤہے کیونکہ یہ ماضی ہمیں ہمارے حال مستقبل کی آگاہی دیتاہے لیکن انسان کا مسلسل ماضی میں رہناہی انسان کو پیچھے دھکیل لے جاتا ہے۔وہ بتاتے ہیں کہ جب ’’سفیر لیلی‘‘ نظم ان کے ذہن میں آئی تب ہی انھوں نے سوچا کہ انھوں نے ایک عظیم داستان سنانے کی ذمہ داری لے لی ہے۔

’’سفیرلیلیٰ یہی کھنڈر ہیں جہاں سے آغاز داستان ہے

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فصیل قریہ کے سرخ پتھر اوران پہ اڑدرنشان برجیں گواہ‘‘(4)

                نظم کے شروع میں ہی پڑھتے ہوئے قاری کو محسوس ہوتا ہے کہ اس سے اچھا آغاز اور کوئی ہو نہیں سکتا۔یہی وہ کھنڈر ہیں جس سے...

Determinant Factors Affecting The Value Of Telecommunication Sub-Sector Companies Listed On The Indonesia Stock Exchange For The Period 2018-2022

The purpose of this study is to determine whether earnings per share, profitability, leverage, sales growth and research and development intensity have a significant effect on firm value. This research method is quantitative research by taking samples using a purposive sampling technique based on predetermined characteristics of 7 telecommunications sub-sector companies listed on the IDX for the 2018- 2022 period. The type of data used is secondary data and the method of analysis used is panel data regression using Eviews. The results showed that the calculation of the hypothesis that is earnings per share has no significant effect on firm value with a significant level of 0.1905 > 0.05. Profitability has a significant positive effect on firm value with a significant level of 0.0015 <0.05. Leverage has no significant effect on firm value with a significant level of 0.1873 <0.05. Sales growth has a significant positive effect on firm value with a significant level of 0.0276 <0.05 and the intensity of research and development has no effect on firm value with a significant level of 0.2800 > 0.05. For simultaneous testing, it is obtained F count of 2.202910 with a probability of 0.000159 <0.05 meaning that earnings per share, profitability, leverage, sales growth and research and development intensity influence simultaneously on firm value.

Effect of Chemically Treated Wood Coal on the Physiochemical Properties of Tomato Riogrande

In a pot experiment, wood coal was applied to tomato plants of Rio Grande cultivar in the form of 32 different treatments, which were made by treating the same number of powdered wood coal portions (250g each ) with one liter each of 32 different acid base solutions i.e. 4% HCl, 4% H2SO4, 15% HNO3, 5% NaOH, 5% KOH abbreviated as ''C'', ''F'',''N'', ''S'', ''P'' and their combinations CF, CN, CS, CP, FN, FS, FP, NS, NP, SP, CFN, CFS, CFP, FNS, FNP, FSP, CNS, CNP, CSP, FSP, NSP, CFNS, CFNP, CFSP, CNSP, FNSP, CFNSP in one ratio one respectively. Quality of the treated wood coal and its role as fertilizer was studied in term of physicochemical parameters of pots soil and tomato plants in comparison to controls i.e. NPK, humic acid, F.Y.M, wood coal and untreated soil. pH and EC of the treated wood coal showed significant difference (P<0.05) with ranges of 1.48 to 11.9 and 3.45 to 28.4µs/dl respectively, where most of pH values were in acidic region, except S, P, CS, FP, SP, CSP and FSP treatments while EC was highest in acidic or acid containing treatments. Soil parameters showed significant variation (P<0.05) where the least EC 96.60 µS/dl was found in untreated soil while highest 190.10 µS/dl in NSP treated soil.Acidity of soil ranged from 0.97x10-2g/100mL in S to 1.41x10-2g/100mL in FP treated soils. The highest value 6.58x10-3% of inorganic nitrogen was observed in CFNSP treated soil. An increase in soil pH from 7.85 to 8.61 was observed after treatments where treatment wise comparison showed a range from 8.10 to 8.32 in CFP and CNS respectively. The effect of treatments on soil minerals in mg/kg indicated greatest variation in Fe from 2.759 to 5.358, Mn from 0.324 to 0.73, Na from 3.9 to 11.9 and K from 8.60 to 15.30 while other minerals like Cu, Zn, P, Pb and Ni showed moderate variability. Higher trend of soil minerals were found in sulfuric or nitric acids containing treatments. Data of tomato plants physical parameters,taken in intervals each of 15 days, varied significantly (P<0.05) where higher values of plant height 51.53cm was found in CN, leaf area 26.77 cm2 in FSP, No. of leaves plant-113.13 in CFN, No. of leaflets plant-1 53.93 in NP and No. of flower plant-1week-1 13.80 in SP treated plants with lower values of these parameters25.20 cm, 15.94cm2, 8.07, 33.87 and 1.42 in untreated soil respectively, while No. of branches 4.56 of CNP were higher than humic acid 1.56 and No. of fruits plant-1week-1 4.75 of FNS than 1.00 of CSP. Proximate composition of plant parts taken in percent showed the higher values of moisture 96.28, crude protein 1.30 and NFE 88.36 in roots, ash 4.75 and crude fiber 7.32 in stem and crude fat 2.64 in young leaves. Percent proximate composition of treated plants varied from 0.5 to 3, where maximum values ofmoisture 95.67, ash 4.03, crude fat 2.05, crude protein 1.21, crude fiber 6.05 and NFE 87.94 were detected in C, CFP CNS, CNSP, CFNP and FN treated plants respectively. Except Na and K with highest values of 16.10 and 37.63, other minerals of plant parts, like Fe, Zn, Mn, P etc., varied from 0.01 to 3.00mgKg-1 where most of them were higher in roots and leaves of tomato. Treatment variation of these mineral were also significant at P=0.05 where highest values of Fe was found in CNSP, Zn, Cu and in C, Mn in FNSP, Na and P in NSP, K in N and Ni was highest in FSP treated plants. Most of the fruits proximate parameters except moisture 94.82% and NFE 96.88% ranged from 0.02 to 3% with higher values in KOH containing treatments. The lowest values of these minerals were present in untreated soils. Among the mineral content of fruits Fe content of FNSP, Zn of CFSP, Cu and Na of CFNS, Mn and P of CS, Pb of CNP, Ni of SP were highest compared to other treatments. Highest values of Carbon 84.16% and lycophene 4.50mgKg-1 were found in CFP and FSP treatments, while chlorophyll was high at day 45of transplantation and in CFNSP treated plant. From the results it was conjectured that treated wood coal may be developed as fertilizer for tomato and other crops where further research particularly under field condition are needed.