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Incidence and Characterization of Major Fungal Pathogens of Strawberry Diseases

Thesis Info

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Author

Mehmood, Nasir

Program

PhD

Institute

Pir Mehr Ali Shah Arid Agriculture University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Plant Pathology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/10218/1/Nasir%20Mehmood_Plant%20Pathology_2019_PMAS_PRR.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726446995

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Strawberry (Fragaria x ananassa Duch.) is a member of family Rosaceae and is among the most widely consumed fruit throughout the world. Its fruit ripens in early spring which gives good economic returns to the farmers. Strawberry crop suffers from numerous diseases and among them, fungal diseases are highly destructive. As no systematic research, so far has been conducted in Pakistan regarding strawberry diseases, the present study was conducted to determine disease incidence and prevalence of important fungal diseases of the strawberry crop and the morpho-molecular characterization of associated fungal pathogens. For field-based disease assessment, a two year (2014-15 and 2015-16) disease survey of farmer fields was conducted in 12 important strawberry producing districts of Punjab (Rawalpindi, Sargodha, Gujranwala, Sialkot, Narowal, Sheikhupura, Lahore and Multan), Khyber Pakhtunkhwa (KPK) (Mardan, Charsadda and Swat) and important areas of Islamabad (ICT). On the basis of these surveys four major fungal diseases viz. Alternaira leaf spot (ALS), Fusarium fruit rot (FFR), Anthracnose (Colletotrichum) fruit rot (AFR) and Bortyis fruit rot (BFR) or gray mold were found to be prevalent, with no prevalence in district Swat during both years in case of FFR while maximum of 100 % were observed in case of all fungal diseases. Disease incidence of ALS was recorded from 17.25 % to 55 %, followed by no disease to 59 % in case of FFR while 14.13% to 44.71 % of AFR and 17.13 to 48.88 % as of BFR. Pathogens were identified on the basis of morpho-molecular characters. The morphological characterization was done on pathogenic isolates of 4 fungal pathogens viz. 82 isolates of Alternaria alternata, 77 isolates of Fusarium solani, 90 isolates of Colletotrichum spp. (68 isolates of C. acutatum and 21 isolates of C. xvi 3 gloeosporioides) and 92 isolates of Botrytis cinerea. These isolates were identified based on standard identification keys and results in identification of these pathogens. A total of fifty four (54) highly virulent and representative isolates from each district were subjected to molecular characterization. Of these, 12 isolates were each of A. alternata and B. cinerea while 19 isolates were Colletotrichum spp. (12 of C. acutatum and 7 of C. gloeosporioides) and 11 isolates were F. solani. These isolates were amplified with ITS gene primers (ITS1/ITS4), endopolygalacturonase (endoPG) gene primers (PG3/PG2b) for A. alternata, Beta (β)-tubulin (TUB2) primers (BT2a/BT2b) for Colletotrichum spp., translation elongation factor 1-alpha (TEF-1α) primer (ef1/ef2) for F. solani and Glyceraldehyde-3-phosphate dehydrogenase (G3PDH) primers (G3PDH_for/G3PDH_rev) was for B. cinerea. The nucleotide sequences further analyzed by phylogenetic software and resulted in genetic homology of current study isolates with previously reported isolates and hence confirmed the morphological identification. This research work provided the first comprehensive factual picture of fungal diseases of strawberry from Pakistan and proper morpho-molecular characterization of associated destructive pathogens and is expected to play a central function in future studies.
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پروفیسر سید نواب علی

پروفیسر سید نواب علی
ہماری پرانی علمی شخصیتیں ایک ایک کرکے اٹھتی جاتی ہیں، اور ان کا بدل نظر نہیں آتا، جون کی آخری تاریخوں میں مشہور اہل قلم اور نامور فاضل پروفیسر سید نواب علی صاحب ایم اے نے انتقال کیا، ان کا اصل وطن نیوتنی ضلع اوناؤ تھا، لیکن ملازمت کے سلسلے میں ان کا قیام زیادہ تر گجرات میں رہا، وہاں وہ مختلف بڑے بڑے تعلیمی عہدوں پر ممتاز رہے، سرکاری ملازمت سے ریٹائر ہونے کے بعد ریاست جوناگڑھ کے وزیر تعلیم ہوگئے تھے، اس سے سبکدوش ہونے کے بعد وطن لوٹ آئے تھے، پھر قیام پاکستان کے بعد کراچی چلے گئے اور وہیں گذشتہ ۳۰؍ جون کو وفات پائی۔
مرحوم، حضرت سید صاحب رحمۃ اﷲ علیہ کے ہمعصر تھے، ان کی طالب علمی کا زمانہ لکھنؤ میں گزرا تھا، اس زمانہ میں وہ مولانا شبلی کی صحبت سے مستفید ہوئے، اس لیے دارالمصنفین اور اس کے کارکنوں سے ان کے تعلقات بہت قدیم تھے، اور وہ اس کے ابتدائی ارکان میں تھے، جدید علوم کے ساتھ عربی سے بھی واقف تھے اور اسلامی علوم پر بھی ان کی نظر تھی، ان کا علمی ذوق بہت بلند تھا۔ وہ متعدد بلند پایہ کتابوں کے مصنف تھے، اور اپنی تصانیف کے ذریعہ انھوں نے دین کی بڑی خدمت انجام دی، ان کی تصانیف کی تعداد ایک درجن کے قریب ہوگی ان میں ’’سیرۃ الرسول‘‘ اور ’’تاریخ صحف سماوی‘‘ اور ’’معارج الدین‘‘ زیادہ اہم ہیں، اب ایسے محقق فاضل مسلمانوں میں مشکل سے پیدا ہوں گے۔ (شاہ معین الدین ندوی،اگست ۱۹۶۱ء)

 

Second Wave of COVID-19 in Pakistan: Our Responsibilities and Challenges as A Nation

The Corona virus (SARS-CoV2) pandemic initiated in late December 2019 in Wuhan city of Hubei, China, which has rapidly progressedinvolving more than 215 countries of the world. It was caused by novel SARS-COV2 coronavirus with Huanan seafood wholesale market as the possible point of origin. In past two decades, coronaviruses epidemic of Middle East Respiratory Syndrome (MERS-COV) had 37% mortality rate and Severe Acute Respiratory Syndrome (SARS-COV) had 10% affecting more than 10,000 population together. World Health Organization (WHO) declared it as the sixth Public Health Emergency of International Concern (PHEIC) on January 30, 2020 and later on March 11, 2020, the WHO labeledit as a pandemic. The first case of COVID-19 from Pakistan was reported on 26th February, 2020 and has affected over 354,000 people with a mortality of over 7000 patients. Many countries of the world have seen second wave of this pandemic. Government ofPakistan has also declared a second waveon October 28, 2020, after the rise in cases from 500 to 750 per day. Now it has crossed 2000 cases. The data released by the National Command and Operation Centre (NCOC) shows that the current percent positivity rate is close to 3 compared to the previous figure of lesser than 2 making it a bigger challenge than first wave in Pakistan. The patients now presenting in hospitals are all in critical condition. Lack of a specific vaccine or antiviral drug and non-compliance to the standard preventive measures is the major reason of initiation of a second wave of this viral infection in Pakistan. Being a nation we need to be responsible. Our country may go into economic crisis & our health facilities may choke. We have to understand how to live with this virus till the availability of vaccine or Curative antiviral drug. TheGovernment of Pakistan is creating awareness in the public for the second wave as the situation of pandemic is getting worse. Smart lock downs are being implementedbut people are not following preventive measures that are leading to infection spread at a very alarming speed. At the moment preventive measures are the only way to stop the spread of disease. Preventive measures should be adopted to contain this deadly disease. Wearing masks, using hand sanitizers, washing hands with soap for 20 sec, keeping social distance of 6 feet are mandatory preventive strategies. Social, political, business, recreationaland religious gatherings, should be avoided. Educational institutesshould follow strict standard operating procedures. Most of the people in Pakistan are not considering this disease as a matter of serious concern due to unawareness, poverty, beliefs and lack of resources. People should ignore such disbeliefs and should start considering it as a great health concern. They should follow the preventive measures in true sense.

Environmental Concerns of Poultry Manure Applied into Agriculture Fields of Khyber Pakhtunkhwa: Special Emphasis on Antibiotics and Heavy Metals

Heavy metals and antibiotics are growing environmental contaminants leading to public health concern.Poultry manure is enriched with antibiotics and heavy metals that are commonly used as organic fertilizer for agricultural crops. Antibiotics (Suphonamide, lincomycine, erythromycin, oxytetracycline and tylosine) and heavy metals (Pb, Cr, Cd, As Ni, Co and Cu) are used as growth promoters and therapeutic agents in poultry feed. Some of these antibiotics and all heavy metals are not biodegradable in the body tissues of chicken, get deposited in meat as parent compounds and ultimately excreted via poultry droppings into the environment.In the first study an attempt have been done to present a better understanding of emerging contaminants in environment and their associated health effects. In this part we investigated the use of antibiotics in poultry sector and their release into the environment which develop antibiotics resistance bacteria (ARB) along with antibiotics resistance genes (ARGs). The development of ARGs and ARB might lead to huge physical and economic losses, as these bacteria cannot be treated with commonly used antibiotics, proving fatal for thousands of people every year in the world. These antibiotics after entering into food chains seriously affect human immune system, growth and metabolism of the body. Therefore, remediating the ARB, ARGs and antibiotics from the agriculture soil by environmental friendly technique is important to protect the future human health risks. Manure composting and biochar application might be the possible ways to reduce the risk and spread of ARGs in environment. In the second study, heavy metals were quantified in poultry manure, soil and food plants of agriculture fields from eleven districts of Khyber Pakhtunkhwa province. The concentrations of selected metals (Cd, Cr, Ni, Co, Cu, Ba, Pb) in most of the soils (ranged 0.036-0.3±0.086, 1.05-54.1±17.3, 1.8-35.2±9.1, 0.47-16.9±4.1, 0.056-32.5±9.8, 48.5-200.8±45, 0.26-15.7±4.57 mg kg-1, respectively) and manure (ranged 0.1-0.3±0.06, 6.75-52.4±11.9, 9.18 23.6±4.5, 2.86-16.7±4.4, 15.0-82.6±19.7, 24.7-86.9±20.9, 2.67-11.1±2.4 mg kg-1, respectively) samples were observed below the maximum allowable viii limits set by United States, Environmental Protection Agency (USEPA). The concentrations of Cu (0.81 to 247 mg kg-1) and Cr (0.18 to 9.81 mg kg-1) in most of the food plants were significantly (Cu-P<0.05) (Cr-P<0.01) higher and crossed their respective permissible limits (Cu-20 mg kg-1 and Cr-0.5 mg kg-1) set by SEPA, China. The mean concentrations of Pb (ranged 0.41-11.4 mg kg1) in all the food plants were significantly (P<0.01) higher and crossed the limit (0.3 mg kg-1) of FAO and WHO (2001). The values of Cd in radish (0.45 mg kg-1), clover (0.31 mg kg-1), lettuce (0.34 mg kg-1) and argula (0.28 mg kg1) were also above the allowable limits (0.2 mg kg-1) of SEPA. Metals like Pb, Cr, Cd, Cr and Cu were shown higher (>1) bioaccumulation values in the selected plants like radish, garlic, barley and wheat.Among selected heavy metals, Co, Ni and Ba were shown lower BCF values and assumed to have no serious health risk.The health risk index (HRI) values were found >1 in barley (for Cu and Cd), wheat and radish (for Cu, Cd and Pb) and pea and onion (for Cd), which were considered, that may cause potential health risks to the local community. Therefore, heavy metal concentrations in poultry manure must be reduced before its application into cultivated fields through safe and environmentally friendly technology such as biochar and composting. The third study determined the health risk of total As (TAs) and its species in selected food plants grown on poultry manure-based agriculture soils of Khyber Pakhtunkhwa, Pakistan. The concentrations of total arsenic (TAs), arsenobetaine (AsB), monomethyarsenate (MMA), dimethylarsenate (DMA), arsenite (As3+) and arsenate (As5+) were investigated in soil, poultry manure, food plants and poultry feed. The TAs concentration in soil of the study area ranged from 4.56 to 9.98 mg kg−1, showing the high variability in the selected sites. Inorganic arsenic (iAs) was found highest in Charsadda (9.9 mg kg−1) and lowest in district Swat (4.02 mg kg−1) The mean TAs concentrations in the poultry manure and feed ranged from 0.64 to 1.60 mg kg−1 and 0.06 mg kg−1 to 0.13 mg kg−1, respectively. The mean TAs concentrations in the edible parts of the food plants was in range from0.096 to 1.25 mg kg−1, which were found above the safe permissible limits (0.1 mg kg−1) of FAO/WHO (2011) in 83% offood plants, except in pea (0.096 mg kg−1) and mint (0.093 mg kg−1). The ix maximum concentration was observed for wheat (1.25 mg kg−1), cucumber (0.67 mg kg−1) and mallow (0.63 mg kg−1). Furthermore, human health risk was assessed through health risk index (HRI), hazard quotient (HQ), dialy intake rate (ADI) and life time cancer (LTCR). The ADI of As in the different food plants was ranged from 1.42×10−6 mg kg−1 d−1 to 6.62 ×10−5 mg kg−1 d−1 in the adults and 4.07×10−6 mg kg−1 d−1 to 1.89 ×10−4 mg kg−1 d−1 in the children with highest value for Malva neglecta (leafy plants) and lowest for Allium sativum. The total ADI for children (6.33×10-4 mg kg−1day−1) was higher than the USEPA oral reference dose (3 × 10-4 mg kg−1day−1) posing non cancer risk. The HQ values for TAs were less than 1 for the ingestion of the tested food plants, revealing less non-carcinogenic risk in the area. The results revealed that HI value for children (2.1×10+0) were three times higher than the value for adults (7.87×10−1), posing non cancer health risks (HI>1) in the children. The long term potential cancer risk (LTCR) was found slightly higher for children (1.53 × 10-4) than the permissible limits of USEPA, while minimal cancer risk was observed for adults via consumption of selected food plants.This research highlights a need to reduce the level of As in the poultry feed and further suggests to treat poultry manure before their application to the crop field. A fourth study was carried out on the development of ARG and ARB due to manure application in agriculture field. Three different samples of poultry manure (M), soil amended manure (S+M) and soil without manure (S) were collected for assessment of ARGs in the agriculture field area of Khyber Pakhtunkhwa, Pakistan. Total number of 285 ARGs was observed by means of HT-qPCR (High Throghput quantitative PCR), which targeted most common classes of resistance genes. Poultry manure addition significantly (P<0.05) raised the abundance copies of ARGs in manure based soil and have changed composition of bacterial community. The detected resistance genes in manure amended soil gave resistance to antibiotics such as multidrug, aminoglycoside, MLSB, beta-lactmase, tetracycline, Vancomycine, tansposase and sulphoniamide. Total 130 unique ARGs were developed, 76 ARGs were shared by all three groups, 47 x ARGs were transferred from manure to soil and 49 ARGs were shared by manure and manure-based soil. Gene sequencing of 16S rRNA was carried out for the samples, which concluded that Proteobacteria, Bacteroidetes Actinobacteria, Fermicutes, Acidobacteria and Chloroflexi were the common phyla of microbial community, having about 87% of total microbes of 16S rRNA gene sequences. Manure has increased Bacteroidetes, Actinobacteria and Fermicutes in the soil. Hence manure has elevated the ARGs and antibiotic resistance bacteria (ARB) by horizontal gene transfer from manure based soil to indigenous soil microbiota. This study will help us to know the dissemination and fate of ARGs in this ecosystem to efficiently use poultry manure in crop land and highlight antibiotics and ARGs risks managements in the agriculture area of Pakistan.