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Potential of Phytohormones Producing Endophytic Fungi As Biofertilizers

Thesis Info

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Author

Lubna

Program

PhD

Institute

Abdul Wali Khan University

City

Mardan

Province

KPK

Country

Pakistan

Thesis Completing Year

2018

Thesis Completion Status

Completed

Subject

Botany

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/10836/1/Lubna_Botany_2018_AWKU_PRR.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726979241

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Being a good source of phytohormones fungal endophytes have been considered as potent plant growth promoters. The present report elucidates the isolation of endophytic fungi from medicinal as well as crop plants. In current study 46 endophytic fungal isolates were isolated from medicinal (Cannabis sativa, Chenopodium album, Oxalis corniculata, Euphorbia helioscopia, Amaranthus viridis, And Parthenium hysterophorus) and crop (Triticum asativum, Brassica compestris) plants. Culture filtrated (CF) and biomass was initially screened on maize in order to find the plant growth promoting endophytic fungi and for indole acetic acid. Among these isolate culture filtrate of 13 strains showed promotary effect on maize seedling. The isolates produced IAA via L-tryptophan-independent pathway in rage of 1.254ug/mL to 26.73ug/mL and tryptophan dependent pathway in the range of 0.301ug/mL to 35.434ug/mL. Other important secondary metabolites including phenolic, flavonoid and sugars were also detected in the culture media of these endophytes. The isolated endophytes also supported growth of maize seedlings grown in fungal biomass supplemented soil. Based on their plant growth promoting potential, the endophytes CSR1, CSR3, TS2, CSL1, CHS1, EUR1, TS1 and BRL1 were selected for further study. Culture filtrates of the selected endophytes were screened for gibberellins (GAs) by using GAs deficient mutant rice Waito-C. The results revealed that different plant growth characteristic such as chlorophyll content, root-shoot length, and biomass production of Waito-C rice was significantly promoted during endophyts treatment showing the presence of GAs in their culture filtrate. Majority of these endophytes produced iron chelating siderophores and solubilized phosphate which is among the known phytostimulant tools of the endophytes. These morphologically representative strains were identified further by means of molecular phylogenetic analysis based on ITS and D1/D2 regions. Molecular identification using ITS and partial 28S ribosomal DNA (D1/D2) sequences from the isolates revealed 4 genera including Aspergillus, Fusarium, Bioplaris and Curvularia. These fungi are curvularia lunata TS2, Aspergillus fumigatus TS1, Aspergillus terreus EUR1, Aspergillus flavus CHS1, Aspergillus niger CSR3, Fusarium proliferatum x BRL1, Fusarium oxysporum CSR1 and Bipolaris sp CSL1. The isolated strains belonged to division Ascomycetes and among these 2 species belong to order Pleosporales, 2 belong to hypocreals and 4 belong to eurotiales. The selected endophytes were further characterized by determining phytohormones including IAA and gibberellins (GAs) using GC/MS SIM. This study is the first report related to curvularia lunata, Aspergillus terreus and Fusarium proliferatum to produce IAA. This growth promotion was due to various types of GAs and presence of IAA in endophyte culture filtrate. The gas chromatography/mass spectrometry (GC/MS) analysis showed the presence of different gibberellins in various quantities (ng/ml). Nine kinds of GAs tested were biologically active: GA1, GA3, GA4, GA7, GA8, GA9, GA12, GA20 and GA24. Biologically active gibberellins include GA1, GA3, GA4, and GA7 were identified in almost all fungi. Moreover, GA1 was detected in high quantity in CSR1 (0.638±0.019 ng/mL), followed by BRL1 (0.392±0.007ng/mL), CSL1 (0.753±0.005 ng/mL) and CHS1 (0.133±0.012ng/mL) respectively. Similarly, GA3 was found in high concentration in two fungi EUR1 (0.352±0.012ng/mL) and TS1 (0.324±0.077ng/mL). Notably, GA4, important bioactive GA, was detected in almost all fungal cultures in significant amount, However, the highest concentrations were found in CSL1 (0.943±0.081ng/mL), CSR3 (0.479±0.01ng/mL) and CHS1 (0.435±0.016ng/mL). Similarly, the fungal strains CSL1, CHS1, EUR1, and BRL1 produced 0.638±0.012, 0.476±0.082, 0.423±0.003 and 0.492±0.005 ng/mL of GA7. Furthermore, Inactive types of GA present in the FCF were GA8, GA9, GA12, GA20 and GA24. The ABA contents were also checked in these isolates and among these fungal strains, CSR1 had the highest concentration of ABA (0.0903ng/ml) in its CF. Upon inoculation, these endophytes contributed significant amount of GAs to the endogenous pool of Waito-C rice. Among these endophytes, CSL1 increased GA1, GA3, GA4 and GA12 concentration up to several fold peaking at 18.37ng/g DW, 11.37ng/g DW, 33.23ng/g DW and 25.534ng/g DW respectively. The isolate CSR3 enhanced endogenous level of GA4, GA7 and GA12 by up to 20.34ng/g DW, 28.264ng/g DW and 34.23ng/g DW as compared to the non-endophytes seedlings. Similarly, TS2 significantly increased the concentration of endogenous GA4 and GA12 of the mutant rice. Along with GAs, endogenous ABA level was also significantly enhanced in Wiato-C rice inoculated with the isolated endophytes than the control. The percent increase by different strains TS2, TS1, BRL1, CSR1, EUR1, CHS1, CSL1 and CSR3 is, 14%, 46.6%, 6.5%, 14%, 67%, 39%, 60.7%, 77.9% respectively. Contrary to this, endogenous concentration of JA dropped significantly in the endophytes associated seedlings indicating involvement of fungal GA and IAA. The percent decrease of JA in treated seedlings as compared to non-treated Waito-C rice seedlings is TS2 (27%), TS1 (53.8%), BRL1 (23.9%), CSR1 (33.5%), EUR1 (61%), CHS1 (49.9%), CSL1 (56%), CSR3 (68%). Furthermore, RT-PCR confirmed the presence of GA and IAA pathways genes (P50-1, P450-4, ggs2, des and iaaH) in the selected endophytic strains by positive expression. Moreover, the application of these fungal spore suspensions with uniconazole and yucasin on maize seedling revealed that like exogenous GA3 and IAA, the endophytic fungal strains CF application mitigated the inhibitory effect of both yucasin and uniconazole and promote growth attributes of maize seedling. Also, secondary metabolites of maize seedlings associated with the endophytic fungi were significantly enhanced recovering the seedlings from the suppressing effects of both uniconazole and yucasin. Besides, the selected strains were checked on cucumber cotyledon to check its cytokine like activity. In these strains some increase chlorophyll contents as compare to control which indicate the presence of cytokinin like compound in them. These finding suggest that these gibberellins and IAA producing endophytic strains plays important roles in plant growth promotion, which could be used for the improvement of crop growth under diverse environmental condition to mitigate the stresses.
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بھٹو مرتا کیوں نہیں

بھٹو مرتا کیوں نہیں ؟

بے نظیر اپنی کتاب میں لکھتی ہیں کہ جب لاہور ہائی کورٹ نے انہیں سزائے موت سنائی تو وہ کوٹ لکھپت جیل گئیں بھٹو صاحب کو لوہے کی تاروں سے بنی چارپائی پر لٹا کر ان کے بازو اور پائوں کو زنجیروں سے باندھا ہو ا تھا ۔چارپائی پر چٹائی بھی نہیں تھی ۔مچھروں کے کاٹنے کی وجہ سے ان کے ہاتھ پائوں اور چہرہ سرخ ہو رہا تھا ۔بے نظیر بھٹو پر نظر پڑتے ہی کڑکدار ا آواز میں بولے "Hi pinci how are you"اور پھر کہنے لگے آپ کو اندر سے توڑنے کے لیے مجھے اس طرح باندھا گیا ہے لیکن آپ نے ٹوٹنا نہیںہے ۔

پھر جب سپریم کورٹ کے چاروں ججوں نے سزائے موت سنائی (تین ججوں نے انیں بری کیا کل سات جج تھے )تو انہیں جیل کی کال کوٹھری میں رکھا گیا ۔انہوں نے بھوک ہڑتال کی جو گیارہ روز جاری رہی اس عرصہ میں کال کوٹھری کی چھت پر لوگ بڑے بوٹ پہن کر ناچتے رہتے تھے تا کہ بھٹو سو نہ سکے اس کے باوجود بھٹو بیمار نہیں ہوئے اور بارہویں رات کو پھانسی چڑھ گئے ۔

جو دوست اب سوال پوچھتے ہیں کہ بھٹو مرتا کیوں نہیں ہے تو گزارش ہے کہ ایسے بہادر انسان کا جسم تو مر جا تا ہے لیکن نام رہتی دنیا تک زندہ رہتا ہے ۔جیے بھٹو ۔

 

Laws of Inheritance in Islam and Hinduism: A Comparative Study

Laws of inheritance and succession exist since time immemorial in every human society. The estate of the deceased person is distributed amongst his/her legal heirs according to the laws of inheritance. Based on religions i.e. Islam and Hinduism-two different societies exist in the Indo-Pak sub-continent, having different frames of rules regarding socio-religious life. Coupled with a long history of closeness and co-existence, have deep religio-moral and socio-cultural diffusions and infusions with one another. The same impact permeates the laws of inheritance as well. This paper is an analysis of this permeation by focusing on the points of comparison and contrast in the light of the Holy Qur’ān, Sunn’ah and the sacred books of Hindūism.  

Tunable Metamaterials

Metamaterials (MTM) are composite materials that provide some unique characteristics which are not available by nature. But there is one major drawback or limitation of MTM which is its fixed and narrow working frequency region. To overcome this limitation, there are few techniques, presented in this thesis, to vary the working frequency band of MTM. As MTM are artificial materials, so there are various resonators available in literature, presented for MTM. In this thesis, three resonators, split ring resonator (SRR), S-shaped resonator (SSR) and single side paired S-ring resonator (SSPSRR) have been chosen from literature and different techniques to shift their resonant frequencies are presented. For SRR, a tunable case i.e., inner ring shorted split ring resonator (IRS-SRR) is pre- sented in this thesis. The analytical expression of the effective permeability of IRS-SRR is calculated and for the verification of the analytical work, the experimental results are also included in this thesis. The analytical expression for the effective permeability of SSR is also presented in this thesis. Two tunable or modified cases of SSR, i.e., bottom metallic strips shorted S-shaped resonator (BSSR) and top-bottom metallic strips shorted S-shaped resonator (TBSSR) are presented in this thesis. The analytical expressions of the effective permeabilities of BSSR and TBSSR are also calculated. For verification of the analytical work of SSR, BSSR and TBSSR, experimental results are included in this thesis. Similar to SSR, the analytical expression of the effective permeability of SSPSRR is presented in this thesis and the experimental results are also included to verify the analytical work. Two tunable cases of SSPSRR, i.e., bottom metallic strips shorted single side paired S- ring resonator (B-SSPSRR) and top-bottom metallic strips shorted single side paired S-ring resonator (TB-SSPSRR), are also presented in this thesis along with experimental results. Other than the tunable cases of various resonators, rotation of resonators is presented in this thesis as a technique to vary the working frequency of MTM. This technique has been applied on SSR, SSPSRR and their respective tunable cases in this thesis and the vexperimental results are presented for justification of this technique. There are number of devices (made using MTM), are available in literature. All these devices can be upgraded to work as tunable devices using the techniques presented in this thesis.