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Home > Isloation of Alternaria and Fusarium Spp. , Their Pathogenesis and Pathogenesis Related Pr Protiens Associated With Sesame Sesamum Indicum L.

Isloation of Alternaria and Fusarium Spp. , Their Pathogenesis and Pathogenesis Related Pr Protiens Associated With Sesame Sesamum Indicum L.

Thesis Info

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Author

Nayyar, Brian Gagosh

Program

PhD

Institute

Pir Mehr Ali Shah Arid Agriculture University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2018

Thesis Completion Status

Completed

Subject

Botany

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/10302/1/PhD-Thesis15-with%20ref.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726526926

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Sesame is one of the oldest and an important oil seed crops of the world known to human being, holding special importance due to the high-quality oil produced from the seed. Sesame seeds contain 48-60% oil and 18-23.5% proteins. It is very valued cash crop because of the little expenses of production and great market value. Pakistan ranks fourteenth amongst the major sesame producing countries of the world. In Pakistan, sesame is grown in all four provinces. The Punjab province covers 28.5% of the land area of Pakistan but contributes almost 90% to total sesame production in the country, and is considered a major sesame producing area of South Asia. Despite the multiple uses of sesame, it’s farming in Pakistan is highly disappointing because of the low yields per hectare. The demand to supply ratio of edible oil is raising with the alarming rate of human population increase in Pakistan, but the generation of edible oil is declining each year. There is a wide gap, therefore, between production and consumption of edible oil in the country. The sesame crop is subjected to various biotic and abiotic stresses in all growth stages. Low yields of sesame in Pakistan may be attributed to attacks by various pathogens, amongst which two fungal pathogens predominate: Alternaria and Fusarium, which are responsible for blight and wilt diseases, respectively. Even with low yields and high demand for edible oil, no concerted efforts have been made on this highly valuable cash crop with respect to these two major pathogens. To address this lack of effort, there was an urgent need to explore the plant-pathogen interactions between sesame and Alternaria and Fusarium. The aim of the work presented in this thesis, therefore, was to study the incidence of Alternaria and Fusarium species on sesame seeds and plants, to determine their virulence, and to provide an evaluation of the potential for pathogenesis-related proteins to be used in examining the acquisition of systemic acquired resistance in response to these destructive pathogens. The prevalence of Alternaria and Fusarium species associated with sesame seeds from crops in the Punjab province was determined to illustrate the contributions of these pathogens to yield losses in production in South Asia. For this purpose, 105 seed samples were collected directly from the fields of major sesame producing areas in the Punjab and subjected to the standard blotter paper method for the isolation of fungi. A total of 428 isolates of Alternaria and 520 isolates of Fusarium were recovered and grouped into 36 and 23 distinct taxonomic groups on the basis of growth patterns and morphology in vitro. Three isolates of Alternaria (A13, A47, A215) and three of Fusarium (F01, F98, F153) were chosen as representative of the most common morphological groups recovered from seed, and identified using molecular methods based on DNA barcoding using universal (ITS) and species-specific primers. The identities of these isolates of Alternaria and Fusarium were confirmed as Alternaria alternata and Fusarium proliferatum based on sequencing data obtained major allergen gene (Alt a 1) and translation elongation factor (TEF-1α), respectively, after initial identification using internal transcribed spacer (ITS) region of ribosomal DNA of both genera. This basic work on seed testing demonstrated that seeds can be badly contaminated by pathogenic fungi so the detection of seed-borne pathogens and their accurate identification is an important step in crop disease management. The pathogenicity and virulence of the identified isolates of Alternaria and Fusarium on sesame seeds and plants was investigated using both fungi and culture filtrates. Sesame seeds and intact sesame plants were inoculated or treated to confirm the effects of the pathogens on germination, seedling growth and overall plant health. All isolates tested were pathogenic to sesame, but A13 and F01 were the most virulent, causing the lowest germination and highest disease incidence (DI) and disease severity index (DSI). Culture filtrates of A13 and F01 also reduced seed germination and vigor of sesame seedlings in the absence of fungal cells. Typical symptoms of blight or wilt appeared after application of spore suspensions of these isolates on sesame plants. Alternaria isolates caused foliar blight, leading to browning and defoliation. Fusarium isolates induced wilting symptoms on the plants, with yellowing of leaves, and internal staining of stem and roots, ultimately leading to disintegration. The inoculations confirmed that Alternaria and Fusarium are capable of causing leaf blight and wilt diseases in sesame, and may be amongst the causes of sesame yield losses in the Punjab. The possibility of using exogenous application of chemical and biological elicitors of plant defense responses, for example salicylic acid, methyl jasmonate and Pseudomonas syringae for induction of systemic acquired resistance (SAR) against Alternaria and Fusarium was determined. Induction of SAR was evaluated through assessing the accumulation of Pathogenesis-Related protein family 1 (PR1) by western blotting and determining metabolomic changes in the plants following treatments. Total protein content of extracted plant samples was separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunological detection of PR1 proteins. For investigation of metabolomic changes in primed and control plants, non-targeted metabolites were extracted and metabolite profiling was carried out by Linear Trap Quadrupole (LTQ) mass analyzer attached to Surveyor HPLC system using Flow Infusion Electrospray – Mass Spectrometry. The work demonstrated clearly that SAR occurred in sesame. The mechanisms of SAR included accumulation of PR1 proteins, biomarkers of the phenomenon in primed plants. The metabolomic changes associated with SAR were to metabolites involved in lipid/sterol metabolism, the TCA cycle and the defense based phenylpropanoid pathway. The work presented in this thesis demonstrated unambiguously that species of Alternaria and Fusarium infect sesame in the Punjab, Pakistan, and some isolates of these species are of high virulence. The finding that SAR occurs in sesame provides a potential novel and environmentally-friendly method for the control of fungal pathogens in integrated crop management systems.
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مولانا شاہد فاخری الٰہ آبادی

مولانا شاہد فاخری الہٰ آبادی
افسوس ہے پچھلے دنوں مولانا شاہد فاخری الہٰ آبادی کاانتقال ہوگیا۔مرحوم دائرہ اجملیہ الٰہ آباد کے سجادہ نشین،خلافت تحریک کے عظیم قائد اورشعلہ بیان مقرر مولانا فاخر الٰہ آبادی کے خلف الرشید اوران کی روایات وخصوصیات کے بدرجۂ اتم حامل تھے چنانچہ انھوں نے بھی ساری عمر آزادی سے پہلے اورآزادی کے بعد کی قومی وملی تحریکات میں گزار دی۔
والدؒ کی طرح شعلہ بیان اورنہایت پُرجوش خطیب تھے۔آخر تک کانگریس اورجمعیۃ علمائے ہند سے وابستہ رہے اوراس سلسلہ میں قیدوبند کے محن سے بھی دوچار ہوئے۔ان کی زندگی سراپا ایثار وخلوص تھی۔حق بات کہنے میں نہایت جری اوربیباک تھے۔ ان کاقومی اورملی حلقوں میں بڑا احترام اوروقار تھا۔آزادی کے بعد انھوں نے جس جرأت وجسارت سے مسلم کاز کی حمایت کی وہ ان کا طغرائے امتیاز تھا۔پنڈت جواہر لال نہرو اوران کے خاندان سے ان کے ذاتی اوربے تکلفانہ تعلقات تھے۔مگر پنڈت جی کے سامنے بھی وہ حق بات کہنے میں کبھی نہیں ہچکچائے جس کی وجہ سے پنڈت جی ان کی بڑی قدرکرتے تھے۔اللھم اغفرلہ وارحمہ۔ [اکتوبر۱۹۷۵ء]

مولانا غلام اللہ خان کی تفسیر جواہر القرآن: منہج اور خصوصیات

Maulana Ghulamullah khan is considered one of the best commentators of the Quran from  Punjab. Ghulamullah Khan was born in 1905 in Chaj Darya, Attock district of Punjab. He got his Quranic education from Maulana Rashid Ahmad Gangohi, who was a disciple of Maulana Hussain, a well-known and a leading commentator of the Quran. He studied hadith  from Hussain Ahmad Madni, Maulana Shabir Ahmad Usmani and Anwar Shah Kashmeri. This tafseer consists of fifteen hundred pages. It has a long preface and covred up in three volumes over all. Maulana Hussain Ali named him a true successor of the Quranic studies and acknowledged that he had done this duty well. Among his works, TAFSEER JAWAHIR -UL- QURAN, has its own uniqueness and usefulness. Millions of copies have been published of this Tafsier. This tafseer is a compilation of rabat bain surulayat (connection between sura and ayat) of  Maulana Hussain Ali by Ghulamullah khan and has been revised by Maulana Said Ahmad Hussain Sajad Bukhari.  He is termed as Shaikh ul Quran and was called so by Maulana Hussain Ali.I have hinted at different sources for my article so that readers may expand their knowledge about the Quran and Tafseer. This article is about JAWAHIR -UL- QURAN and it will throw light on its features.

Genomic and Culture Based Microbiological Characterization of Fermented Milk Product Dahi and its Impact on Product Quality, Nutrition and Safety

Traditional fermented milk products are characterized by the presence of complex microbial communities. It has been well established that quality, safety and sensorial attributes of these products are related with type and succession of microbiota. Dahi is widely known for unique organoleptic, nutritional and therapeutic attributes. However, the quality of the product is highly variable due to undefined starter cultures and poor control over safety. Therefore, present study was aimed to understand the complex microbial diversity of Dahi and its impact on product quality. Selected bacterial strains were characterized for technological properties and genetic potential. Moreover, impact of selected Dahi strains was analyzed in milk ecosystem in comparison to commercial starter cultures. In first phase of study microbial diversity of Dahi was determined by using culture dependent and metagenomic approach and its effect on physiochemical, nutritional and organoleptic properties of the product was analyzed. Lactic acid bacteria were found to be dominating with up to 7 logs/gram including species of Lactobacillus, Streptococcus and Enterococcus, while fungal community was represented by Geotrichum candidum and other yeasts species with a relative abundance of nearly 3 logs/gram. Similarly, Gram negative count was also nearly 3 logs/gram. Culture independent metagenomic based diversity analysis had shown the presence of 146 bacterial and 56 fungal species. Dominating microbial species include Lactobacillus delbrueckii (53%), Streptococcus thermophilus (28%), Kluyveromyces marxianus (22%) and Torulaspora delbrueckii (14%) on average in representative Dahi samples. Potential pathogens including Gram negatives were also present relatively at low level; Pseudomonas syringe (1.9%), P. cinnamophila (0.8%), Klebsiella pneumoniae, (0.3%), Escherichia coli (3.6 %) and Staphylococcus aureus (0.15%). Microbial and nutritional profiling of Dahi was found to be comparable to many traditional cheeses and fermented milks. Average pH was 3.8 having strong correlation with lactic acid bacteria, however change in pH during storage was correlated with presence of higher fungal count. The nutritional profiling illustrated the nutraceutical importance of the product. The average fat and protein (crude) content was 3.29 and 4.82 respectively while ash content was up to 0.78% containing significant concentration of essential minerals including iron (Fe), magnesium (Mg) calcium (Ca), potassium (K) zinc xvi (Zn) and sodium (Na). Detailed lipid analysis revealed the higher HDL to LDL ratio. The organoleptic properties had shown the richness of flavoring compounds that was also confirmed by consumer’s acceptability. In second phase of study it was found that isolated lactic acid bacteria by culturing in previous phase; were mostly thermophilic having higher rate of acid production and variable salt tolerance. All lactic acid bacteria shown strong proteolytic and lipolytic activities and few also showed biogenic amine production at ambient temperature. Moreover, de-acidification ability of various G. candidum strains was analysed out of these G. candidum QAUGc01 showed optimum results. G. candidum QAUGc01 also showed remarkable antimicrobial potential against several pathogens. The consistent presence of Enterococcus spp. provoked the genome sequencing of E. mundtii QAUEM2808. Genomic analysis revealed that this bacterium had Mundaticin and Enterocin genes, while no putative genes for pathogenicity, virulence and antibiotic resistance were present which excludes the safety concerns about enterococci in Dahi. In last phase of the study milk fermentation performance of Streptococcus thermophilus QauSt1 and Lactobacillus bulgaricus QauLb01 co-culture was evaluated. Comparatively, physiochemical, nutritional and sensorial properties of fermented milk product were equally good to a commercial starter culture fermentation. In another experiment, use of G. candidum QAUGc01 was found to have a positive influence on the stability and quality of the fermented milk due to less variation in product pH, increase in solid content and decrease in syneresis. In conclusion, the present study highlighted high microbial diversity of Dahi. It also demonstrates a strong correlation of microbial diversity with physiochemical, nutritional and sensorial attributes of the product.