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Isolation, Screening and Characterization of Plant Growth Promoting Rhizobacteria from Legumes Rhizosphere to Improve Crop Growth and N2 Fixation

Thesis Info

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Author

Amjad Ali

Program

PhD

Institute

Pir Mehr Ali Shah Arid Agriculture University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2015

Thesis Completion Status

Completed

Subject

Applied Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/6821/1/Amjad_Ali_Soil_Science_2015_PMAS_Rwp.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726558846

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Plant-microbe interaction in the rhizosphere is the determinant of soil fertility and plant health. The presence of beneficial bacteria in the vicinity of roots stimulates plant growth. In this way, soil bacteria play very important role in improving plant nutrition and have been utilized for agriculture for long times. However, little has been done at molecular level in Pakistan to explore their potential. The present study was designed with the objectives to isolate bacterial strains from legume rhizospheric soil and nodules, to characterize and identify potential bacterial strains by using molecular tagging of 16S rRNA gene sequencing and to asses rhizobacterial impact on yield, nodulation and N2-fixation of legume crops under controlled and field conditions. Extensive survey was carried out in Pothwar (District Rawalpindi, Attock and Chakwal) for collection of legumes (mash bean and chickpea) nodules and rhizospheric soil. Five samples of each legume crop were collected from each district. Rhizospheric soil bacteria were isolated through dilution plate technique using Phosphate Buffer Saline solution (PBS; 1X) and nutrient media i.e. Tryptic Soya Agar (TSA; Difco). Root nodules for Rhizobium isolation were washed, crushed and directly streaked on yeast extract mannitol (YEM) plates supplemented with Congo red. About 100 bacterial strains of different genera included Rhizobium were isolated and designated as AM-1, AM-2 to AM-100. These isolated bacterial strains were characterized for plant growth promoting (PGP) properties like auxin i.e. indole acetic acid (IAA), P solubilization and production of NH3. On the basis of PGP traits, 10 most potential strains were selected and identified using molecular techniques i.e. 16S rRNA sequencing. The DNA of each strain was amplified using universal primers 9F: (´GAGTTTGATCCTGGCTCAG´) and 1510R: (´GGCTACCTTGTTACGA´).
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کتنا تنہا تنہا سا ہے

کتنا تَنہا تَنہا سا ہے
دل کی بات نہیں کہتا ہے

یہ کیسی اُس کی عادت ہے
وہ تَنہا سب کچھ سہتا ہے

یاد مُجھے جب تیری آئے
دل ٹھنڈی آہیں بھَرتا ہے

رُوٹھ نہ جائے یار ہَمارا
ہوا چلے تو ، دل ڈرتا ہے

یہ خالی باتیں ہوتی ہیں
کون کسی بن اب مرتا ہے

اِک دن مجھ سے کہا تھا اُس نے
تُو مجھ کو اچھّا لگتا ہے

اِک دن میں نے خواب میں دیکھا
اُس نے مجھ کو مار دیا ہے

اب میں گھر لے آیا ہوں جو
میرؔ اُداسی چھوڑ گیا ہے

لوگ ندیم ندیم کہے ہیں
کون ندیم یہاں بنتا ہے

Discrimination against Religious Minorities in Nigeria: An Analysis with Reference to Human Development in the 21st Century

  This study examines religious discrimination against religious minorities like Muslims living in Christian populated areas in the south east, Christians are as well living in Muslim dominated areas. Minority Traditional worshippers in either Muslim or Christian majority areas, private institution, companies owned by Christians or Muslims etc. The discrimination against religious minorities has mitigated the peaceful co-existence among religious identities and other major life events which has culminated national development in all spheres of human engagement such as economic, social, political, security, etc. The researchers have tried to provide an analytical study of the empirical data as well as of the existing literature. The result of our findings shows that many religious identities have been denied of securing job opportunities, professing religion of their choice, finding it difficult to receive health care services, managing religious institutions, denied of equal rights of citizens, get political appointments, among others. The study recommends that people of different religions should embrace and tolerate one another, avoid the use of fanaticism, allow religious minorities to practice religion of their choice in order to dislodge prejudices from the society.

Genetic Diversity of Begomoviruses Affecting Diverse Host Plants in Periurban Areas of Lahore

Plant foliage exhibiting symptoms indicative of begomovirus infection, veinthickening, leaf curling, yellowing and chlorosis, dwarfing and mosaic were collected from around two km away of Peri-urban areas of Lahore from Multan road, University of the Punjab, Quaid-e-Azam campus, Shikhupura, Ferozwala, Wahga border, Sharaqpur and Ferozpur roads in Pakisatan during 2013-2016. Total plant genomic DNA was isolated from leaf tissues by Cetyl Trimethyl Ammonium Bromide (CTAB) method described by Doyle and Doyle, (1990). Universal primers were used to identify the presence of begomovirus and associated DNA-satellite complex (betasatellite, geminivirus associated alphasatellite) and were subjected to rolling circular amplification (RCA). The expected size of PCR products were cloned and sequenced. Specific abutting primers were designed from the available sequences to amplify the full-length begomoviruses. These full-length PCR entities were cloned and sequenced in their entirety. The isolates of Chickpea chlorotic dwarf virus (CpCDV) and Mesta yellow vein virus (MeYVV) with non-cognate Cotton leaf curl Multan betasatellite (CLCuMuB) were used to produce partial repeat constructs for agro-inoculation. All reported samples were Old World (OW) monopartite begomoviruses, showing recombination and were associated with DNA-satellite complex. In this study the OW monopartite begomovirus Cherry tomato leaf curl virus (CToLCV) was associated with Papaya leaf curl betasatellite (PaLCuB) and Tobacco curly shoot alphasatellite, first time isolated from Parthenium hysterophorus in Pakistan (Qurashi et al., 2017). Similarly ornamental infecting begomovirus associated with DNA- satellite complex also reported from Malva parviflora, a new strain of Hollyhock leaf curl virus (HoLCV-Mal) associated with Kenaf leaf curl betasatellite (KLCuB) and two geminivirus associated alphasatellites species; Ageratum enation alphasatellite (AEA) and Ageratum yellow vein India alphasatellite (AYVIA) in the sub family Geminialphasatellitinae and the genus Colecusatellite (Briddion et al., 2018) identified from Malva parviflora. According to old alphasatellite classification (Mubin et al., 2009) Ageratum enation alphasatellite specie was known as Ageratum conyzoides alphasatellite (Sattar et al., 2017). Another weed infecting begomovirus was associated with DNA-satellite complex also reported from woody plant mulbery isolated as Ageratum enation virus (AEV) associated with Papaya leaf curl betasatellite (PLCuB) and associated geminivirus alphasatellite; Guar leaf curl alphasatellite, but according to new alphasatellite classification this geminivirus associated alphasatellite specie is known as Ageratum enation alphasatellite (AEA) in the genus Colecusatellite described in detailed in this study. All these begomoviruses associated with DNA-satellite complex are reported for the first time in Pakistan. In this study there was prevalence of Pedilanthus leaf curl virus (PeLCV) infecting diverse host plants; ornamental Cestrum nocturnum and reported for first time from vegetables Trigonella foenum and Piper nigrum woody plant Albizia lebbeck in Pakistan. Pedilanthus leaf curl virus (PeLCV) with geminivirus associated alphasatellites; Ageratum enation alphasatellite also reported from Trigonella foenum Albizia lebbeck, respectively. Infectivity assay of the partial repeat constructs of mastrevirus; Chickpea chlorotic dwarf virus (CpCDV) and begomovirus; Mesta yellow vein virus (MeYVV) with non-cognate Cotton leaf curl Multan betasatellite (CLCuMuB) were checked. These were infectious to experimental host plant Nicotiana benthamiana, the Koch’s postulates for CpCDV alone, MeYVV alone and with non-cognate CLCuMuB showed severe symptoms. All the experimental results were satisfied and confirmed with PCR, real-time PCR and Southern blot hybridization. Furthermore, infectivity assay of recombinant PeLCV alone was also checked through gen gun method but PeLCV did not produce begomoviral symptoms in Nicotiana benthamiana, PeLCV need associated betasatellite for begomoviral infection.