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Home > Virulence Analysis of Xanthomonas Campestris Pv. Sesami and Pseudomonas Syringae Pv. Sesami the Causal Organisms of Sesame Sesamum Indicum L. Bacterial Blight

Virulence Analysis of Xanthomonas Campestris Pv. Sesami and Pseudomonas Syringae Pv. Sesami the Causal Organisms of Sesame Sesamum Indicum L. Bacterial Blight

Thesis Info

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Author

Fardoos, Sadiqa

Program

PhD

Institute

Pir Mehr Ali Shah Arid Agriculture University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2009

Thesis Completion Status

Completed

Subject

Botany

Language

English

Link

http://prr.hec.gov.pk/jspui/handle/123456789/5538

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676727662854

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Sesame (Sesamum indicum L.) locally called as til is an important conventional oilseed crop of Pakistan. Pakistan ranks 14th among major sesame producing countries in the world. Pakistan is facing a chronic shortage in edible oil and the situation is getting serious with alarmingly explosion of population. Its indegenious production is below the utilization level and there exists wide gap between production and utilization. Sesame crop is subjected to various abiotic and biotic stresses in all stages of growth. Two prominent bacterial pathogens associated with sesame are bacterial blight caused by Xanthomonas campestris pv. sesami (Xcs) and bacterial leaf spot caused by Pseudomonas syringae pv. sesami (Psse). These pathogens are responsible for sesame production constraints during monsoon season. Despite the shortage of edible oil, no profound efforts have been made on this important oilseed crop with reference to diseases. To handle the shortage of edible oil, there was an urgent need to explore the basic information on host pathogen interaction. The present work consisted of five experiments. The first study was the standardization of mass culturing of stored Xcs and Psse isolates to enhance their virulence and confirmation of their ability to induce hypersensitive reaction. All isolates were revived on non host plant and confirmation was made on the basis of pigmentation they produced in their respective media and hypersensitive test was performed in tomato and potato plants. The second study was conducted to analyse the virulence of virulent isolates in vitro by comparing symptoms induction and bacterial multiplication in different genotypes. Plants were inoculated by pin prick method and were monitored daily for symptoms development and measurements of lesions were taken until fully symptoms induction. Bacterial populations were determined by counting bacterial colonies. Psse isolates showed necrotic lesions (chl+) surrounded by halos as well as only black necrotic lesions (chl-). Size of the lesions and bacterial population between chl+ and chl- was the same and at maximum at 7 DAI in susceptible genotypes, while tolerant showed delayed in reaction. Similar mode of lesions expansion and rate of bacterial growth between chl+ and chl- isolates of Psse indicated that the virulence factor involved in symptomatology function as pathogenicity factor and only contributed to induction of chlorotic producing symptoms for Psse. Water soaking to blight symptoms along with maximum bacterial growth in all the susceptible and moderately susceptible genotypes by Xcs was recorded at 12 DAI. The third study was conducted to confirm process of infection of these bacterial pathogens in susceptible and tolerant genotypes by light microscopy. Inoculation was done by Injection method (IM) and Bacterial suspension dip method (BSDM). Xcs colonized tracheary elements of xylem vessels through intercellular spaces of the spongy parenchyma at 7 DAI and bacterial masses were identified as dark blue infected structures using toluidine blue O stain. Blight symptoms by Xcs were reported to be due to the blockage of nutrients and water flow. Psse showed thining and disruption of mesophyll tissues on the appearance of chlorotic symptoms 3-4 DAI. There were only empty spaces of tissues were observed 7 DAI. Overall the infection was same but delayed in tolerant genotypes. Disruption of mesophyll tissues might be due to the action of chlorosis producing toxin (coronatine) that degraded chloroplast membrane of host tissues. The forth study was conducted to detect the virulence factors of Xcs and Psse using suitables bioassays such as antibacterial test, induction of potato hypertrophic outgrowth and seedlings assay. Xcs and Psse (chl-) isolates showed zone of inhibition. The zone of inhibition produced by chl- isolates showed that chl- was not the defective mutant of chl+ isolates as reported in third study, but this test confirmed that these isolates produced another class of toxin that showed antibacterial activity. Induction of hypertrophic outgrowth in potato tuber and seedlings inhibition from culture filtrate of chl+ isolates of Psse confirmed that the toxin produced by these isolates was similar to phytotoxin coronatine (a polyketide molecule) and it might mimics the action of one of the phytohormones. The fifth study was conducted to extract the virulence factors as well as their purification and identification was also performed. Identification was made on the basis of reference data. Crude extracts of acetone preparation of Xcs and Psse (chl-) isolates were concentrated on silica TLC plates. Further purification was carried out by HPLC and TLC. The toxic aciticity eluted from the HPLC column after 10 min corresponding with single active peak showed antibacterial activity. Reverse phase HPLC of chl- isolates extracted partially purified produced an elution pattern like reported in mangotoxin from Pss strain UMAF0158. Acetone praperation of cell free culture filtrates of virulent Xcs also showed active peaks having phytotoxic activity obtained from the HPLC column after 10 min.
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مولانا محمد جلیل کیرانوی

مولانا محمد جلیل کیرانوی
افسوس ہے کہ گزشتہ ماہ اگست میں حضرت شیخ الہندؒ کے دومنتسبین،مولانا محمد جلیل کیرانوی استاذ اورمولانا محمد مبارک علی نائب مہتمم دارالعلوم دیوبند واصل بحق ہوکر اس جہانِ فانی کوالوداع کہہ گئے۔اناﷲ وانا الیہ راجعون۔ اوّل الذکر (المتولد۱۳۱۸ھ)نے اگرچہ دورۂ حدیث حضرت الاستاذ مولانا محمد انورشاہ کے عہد میں تمام کیا تھا لیکن درحقیقت پروردہ تھے حضرت شیخ الہند کے گھرانے کے ہی۔ نوبرس کی عمر تھی کہ اُن کے والد حضرت ؒ کے سپردکرگئے تھے۔یہ اس آستانۂ قدس کوایسے چمٹے کہ مرتے دم تک اسے نہ چھوڑا۔اس لیے حضرت شیخ الہند کے خادم خاص اور شریکِ جلوت وخلوت تھے اس بناء پر حضرت شیخ الہند کی مشہور’’ریشمی خطوط‘‘والی تحریک کے جزوکل سے خوب واقف اوراس کے محرمِ اسرار تھے۔اس سلسلہ میں انھوں نے بڑے بڑے مصائب اورشدائد برداشت کئے لیکن تحریک کابھید آشکار نہیں کیا۔حضرت ؒ کی وفات کے بعد ادہر ادہرمدرس رہے۔آخر میں دیوبند آگئے تھے اوردرس کی خدمات انجام دیتے تھے۔
[ستمبر۱۹۶۸ء]

 

نقض امن میں ‘‘ف’’ سے شروع ہونے والے سات عوامل کا کردار

According to the certain teachings of al-Qur’ān mentioned at four different places (4: 1, 6: 98, 7: 189 and 39: 6), all humans have their origin in a single cell or soul. One of the objectives behind these proclamations is perhaps to ensure that the unity of humanity at large and of the Muslims in particular, is never to be compromised and that the differences existing among them are to be resolved through a process of mutual understanding on the basis of the notions derived from the al-Qur’ān (2: 213) and Sunnah. AlQur’ān and Sunnah acknowledge the human diversity, rather, describe it as a functional aspect of existence, but not as structural. Referring to the Quranic verse 5: 48, Allah would have made humanity a single people, but, His plan is to test them in whatever He has given to them, so they should emulate for virtues. The present article is an attempt to shortly describe the role of the seven crucial factors in disruption of peace, all starting with the Arabic alphabet fā, i. E., Fitnah, the false Fatāwā, Fujūr, Fakhr, Furqah, Fisq and Fasād, with the purpose of developing an overall religious harmony for strengthening the inner and the outer peace. These seven factors play significant role in disturbing the stability of society. The Islamic injunctions also stress that these factors should be avoided in order to live a righteous and peaceful life.

Studies on Molecular Determinants of Stem Rust in Wheat Triticum Aestivum

Almost 90% of the wheat is facing the threat of stem rust (Puccinia graminis) worldwide. In Pakistan, most of the farmers tend to grow old wheat varieties, which are susceptible to the disease. Replacement of older varieties with high yielding and modern genetically bred varieties will protect farmers against the inevitable attack of stem rust and other diseases. The inevitability of Puccinia graminis f. sp. tritici (Pgt) migration from Iran to Pakistan, coupled with the presence of dangerous new races of yellow rust and leaf rust, are of such importance that their molecular surveillance and rust resistant varieties are now required to improve genetically. Constantly evolving new variants of plant pathogens pose a threat to wheat production. To overcome this, lot of efforts have been made to better understand molecular aspects of resistance to disease and virulence factors that promote the onset of disease. There are many genes identified and characterized, which have resistance against stem rust disease to various levels. They include Sr gene family. Screening of these Sr gene family and some other genes (RPG genes) was done against wheat germplasm. We screened 108 wheat cultivars for different reported resistant genes. Frequency of Sr45 is highest among all other genes which is 65%. Sr35, RPG1 and Sr22 have gene frequency respectively 58%, 37% and 33%. While Sr33 and RPG5 does not appear in any cultivar. Sr22 was selected for isolation and transformation. Today, many transformation methods of resistant genes to various crop plants including wheat are widely used. Sr22 was triggered by inducing Puccinia graminis on healthy resistant varieties such as Mexi-Pak, Auqab 2000 and AS 2002. After inducing Puccinia graminis on healthy plant total mRNA was isolated which was used to synthesize cDNA and full-length gene. The gene was introduced to a commercial susceptible variety, LASANI 2008. Gene gun method was used for transformation. The pCAMBIA2300 plasmid was used having the Kanamycin resistant gene and Cauliflower Mosaic Virus promoter. After transformation, gene integration and expression studies were carried out.