Tomato (Solanum lycopersicum L.) is most nutritionally and economically important crop in Pakistan and around the world. Early blight (EB) in one of tomato dreadful disease caused by fungal pathogen Alternaria solani (Ellis and Martin, 1882) causes major yield losses. Prolonged humidity due to extensive rain and warm conditions during growing season of tomato make this disease unmanageable through sanitation, crop rotation, fungicide application etc. leaving cultivation of resistant varieties as the most appropriate control measure. This study focused mainly to screen phenotypic and molecular marker assisted R gene analogues against A. solani for EB in tomato • Initially twenty nine tomato germlines/varieties were evaluated for their resistance against A. solani by artificially inoculating 15-days old tomato seedlings grown in potted soil in green house. Among them, 8 germlines/varieties were grouped as resistant (RR), seven as moderately resistant (MR); six as moderately susceptible (MS) and eight as susceptible (SS) on the basis of percent disease index (PDI) and growth inhibition index (GII) at 30 DAI (days after inoculation) and 60 DAI. Physiological parameters i.e. total chlorophyll content and carotenoids decreased in all inoculated germlines/varieties with most significant reduction in moderately susceptible and susceptible germlines/varieties at both 30 DAI and 60 DAI. However, biochemical attributes i.e. total phenolics, total protein content and activities of antioxidant enzymes [(peroxidase (POX), polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL) and catalase (CAT)] increased in all inoculated germlines/varieties with respect to corresponding control. Total phenolics, total protein content, POX, PPO and PAL activities were higher in susceptible than in resistant germlines/varieties. Whereas, activity of CAT was the highest in resistant and least in susceptible groups. • RAPD assay using 50 RAPD decamers reveled polymorphism with 32 decamers. The polymorphic pattern acquired by 32 RAPD decamers produced 181 loci (5.7 loci per primer), 157 were polymorphic (86%) and 24 were monomorphic (14%) in all 29 germlines/varieties. Marker A-04, A-10, B-05, L-15, L-17, M-04 and M-07 produced bands ranges from 500-1031 bp specifically in RR group and M-08 and M-10 generated loci (500-800 bp) exclusively in MR. Similarly, A-18, L-06, L-09, L-11, M-09 and OPJ- 10 produced bands from 200-1500 bp in MS and L-09 amplified the loci of 1200 bp only in completely SS. The cluster analysis revealed 60% homology among all germlines/varieties and segregated them into two major groups. Group I was further divided into 3 sub-groups including RR (575, Zeba, KHT-105, KHT-106-G, Advanta- 1206, Mishal, Namadar and Savana); MR (Advanta-1225, Baby red, Yaqoot, Maharaja, Commander, KHT-101 B and TO-1057) and MS (FMC-339, Shangrilla, Thorgal, Rio Grande, Roma and Surkhab). Group 2 comprised of SS germlines (AS-2565, Cluster- 809, GHT-1, Naqeeb, GHT-2, Mongal, Cluster-805 and Roshan). • Resistance gene analogues (RGAs) screening was conducted with 12 tomato germlines/varieties i.e. 4 from RR, 3 from MR, 3 from MS and 2 from SS categorized through PDI, GII and RAPD analysis. DNA of selected 12 germlines/varieties was amplified with 10 RGA primers pairs from conserved region of leucine-rich repeat (LRR), nucleotide binding site (NBS) and protein kinase (Ptokin) domains. Only 3 RGAs xv primer pairs i.e. PtoFen (S+AS), Ptokin (3+4) and Ptokin (1+2) successfully generated discrete PCR products ranges from 200-1100 bp. Primer pair PtoFen (S+AS) produced bands in all germlines/varieties, Band of 533 bp was amplified only in SS and in one variety (Roma) in MS, while band of 511 bp was amplified in the remaining germlines/varieties. The sequence of PtoFen RGAs’ from RR, MR and MS showed the maximum homology of 97-100% with serine/threonine protein kinase protein and had Pkc domain encoding region at 121 to 510 nucleotide. Primers Ptokin 3 and Ptokin 4 generated PCR product of ≃130 bp in SS, while band of ≃208 bp was produced in other germlines/varieties. Moreover, only the band sequenced from RR, MR and MS showed homology of 97-99% with Lycopersicum hirsutum clone RGA sequences. Primers Ptokin (1+2) produced two discrete bands of ≃ 1 kb and 1.3 kb only in RR, MR and MS germlines/varieties. No band was generated in SS germlines. However, sequence and cluster analysis dichotomize the bands of ≃ 1 kb in 2 MR + 2 MS (TO-1057, Yaqoot, Surkhab, FMC-339) and≃ 1.3 kb in 3 RR (Advanta-1209, Zeba and 575) into two divisions with 46% homology and 0.26 genetic distance. It was concluded that combination of phenotypic and genotypic (RGAs) approaches with bioinformatics tools could be used to identify EB resistance in tomato. This study would be a guideline towards solution to devastating fungal pathogen through developing resistant cultivars that could be later used in breeding program for sustainable crop production.
آہ ! پاسبان حرم ملک فہد یکم اگست کو دنیائے اسلام پر یہ خبر بجلی بن کر گری کہ ملک فہد بن عبدالعزیز سعودی حکومت کا تاج و تخت چھوڑ کر اس ملک الملوک کی بارگاہ کبریا میں پہنچ گئے جس کے ملک و سلطنت کو کبھی زوال نہ ہوگا اور وہ ہمیشہ قائم و باقی رہے گا، کل من علیھا فان ویبقیٰ وجہ ربک ذوالجلال والاکرام۔ [الرحمن:۲۶۔۲۷] وہ ۱۹۹۵ء ہی سے بیمار چل رہے تھے، ان کی معذوری کی وجہ سے حکومت کا کاروبار بڑی حد تک ان کے بھائی اور ولی عہد عبداﷲ بن عبدالعزیز انجام دینے لگے تھے، اس سال ملک فہد کی بیماری نے شدت اختیار کرلی تو ۲۷؍ مئی ۲۰۰۵ء کو ریاض کے خاص شاہ فیصل اسپتال میں علاج کے لیے داخل ہوئے، مرض میں تخفیف و اضافہ ہوتا رہتا تھا، آخر یکم اگست بروز دوشنبہ داعی اجل کا پیغام آگیا، اناﷲ وانا الیہ راجعون۔ عالم اسلام اور پوری دنیا کے مسلمان ان کے انتقال سے غم زدہ اور سوگوار ہیں، ان کی ذات بڑی فیض بخش تھی، اور ان کے دریائے کرم اور جودوسخا کی بارش عام تھی، اس لیے ان کے غم میں سب کی آنکھیں اشک بار ہیں، ع عمت فواضلہ فعم مصابہ اب ان کے بھائی عبدالعزیز نے حکومت کی باگ ڈور سنبھال لی ہے، اور لوگوں نے ان سے بیعت کرلی ہے، انہوں نے اپنے بھائی سلطان بن عبدالعزیز کو ولی عہد مقرر کیا ہے جو اس وقت تک دفاع اور سیاحت کے وزیر تھے، اﷲ تعالیٰ ان کو اپنی بھاری ذمہ داری اٹھانے کی قوت دے اور ان کی مدد کرے، ملک فہد کی تدفین منگل کے روز ۲؍ اگست کو ہوئی جس میں دنیا کے اکثر ملکوں کے سربراہوں اور نمائندوں نے شرکت کی، ہندوستان سے بھی ایک وفد جنازے میں شریک ہوا تھا۔ ملک فہد...
This article is an attempt to elaborate the phenomenon of equilibrium prevailing everywhere in the microcosmic and macrocosmic systems associated with universal stability and rhythm. It has been accentuated what is beautifully illuminated in the beginning verses of Sūrah Al-Rahmān successively regarding balance that invites one’s thought towards the well controlled cosmic system as well as with the concept of balance by means of different manners as identified by exegetes and scholars concerning daily life i.e. From simple and common to that of complex issues and intricacies. Qualitative method has been employed for this research while some aspects have also been handled in accordance with quantitative approach. Article also emphasized the series of cause and effect nexus may be declared as self explanatory episode an indication towards a Supreme Force whose widespread control and interference can’t be denied rationally. Sūrah beautifully pictured the physical and metaphysical sketch of balance having many dimensions from man to universe and vice versa as well as from both towards Ultimate Reality whose dominion is limitless where one has to follow peaceful living and to put oneself aside indulging any disruption from ordinary clash to that of nuclear war to be waged.
Background: Oral cancer is a major problem globally and more so in Pakistan as it ranks as the second most common malignancy with an aggressive nature and a high mortality despite modern, advanced treatment options. The strong causal association with smoked and chewed tobacco and its substitutes, prevalent in this region makes it imperative to consider the mutation analysis of molecular genetic profile and the role of HPV in oral oncogenesis. It would help in early and accurate detection of targets for therapeutic implementations. Objectives: To identify the genetic mutations in a subset of Pakistani patients of oral cancer and precancer with dominant exposure to a different set of environmental carcinogens as betel quid, arecanut and its substitutes. Moreover to identify the role of HPV in oral carcinogenesis with interactive relationship of chemical and viral carcinogens. Methodology: Hundred clinically diagnosed and histologically confirmed cases of OSCC and 50 cases of oral premalignant lesions were included in the study. Details of demographic data alongwith personal habits concerning tobacco related carcinogen exposure were noted. A meticulous local and general examination was conducted. Controls were included for histological and molecular comparisons. The tissues obtained at biopsy or surgical resections were subjected for routine histopathological reporting followed by Immunohistochemical analysis of commonly reported mutated oral cancer genes in oral cancer viz p53, p16, H ras, CyclinD1, C Myc, and EGFR. HPV 16 and18 status was detected by Q- PCR. Statistical evaluation was done by SPSS version 16. Results: The ages of 100 patients of OSCC ranged from 25-80 years and 50 preneoplasia from 26 – 65 years with the mean age being 47.84+/- 12.18 and 40.22+/- 9.66 respectively. In OSCC group 74 were males and 26 were females, in preneoplasia 35 were males and 15 females; the male to female ratio being 2.84:1 and 2.33:1, respectively. Ninety one patients of OSCC and 46 (92%) of oral preneoplasia were exposed to tobacco, BQ and BQS, 29 and 32 were exposed to a combination of these. Cheek was the most common site for OSCC (50%) and OPL (42%). Nearly half (48%) of OSCC were well differentiated and majority (74%) presented in advanced stages III and IV. In oral preneoplasia cases 80% presented as leukoplakia and 50% showed mild dysplasia. HR- HPV 16 and 18 were found to be positive in 15 (15%) cases of OSCC and 3 (6%) cases of oral preneoplasia presented with greater prevalence of HPV 16. p53 nuclear protein positivity was seen in 70 (70 %) cases of OSCC and 27 (54 %) of OPL significant correlation (p <0.05) with chemical risk factors was found. Lack of immunoexpression of p16 was observed in 82 (82 %) cases of OSCC and 43(86%) of OPL. EGFR revealed membranous staining in 68 ( 68 %) of cancer cases and in 23 (46 %) dysplastic oral lesions. A strong association was seen with the most common BQ and tobacco chewing habit and significant correlation was observed with stage of oral cancers. H ras cytoplasmic immunoexpression was detectable in a significantly high proportion (50%) of oral cancers and premalignant lesions 17 (34 %). C myc nuclear positivity was observed in 36 /100 (36%) cases of Invasive oral squamous carcinomas and 11 / 50 (22%) cases of premalignant dysplasias. Cyclin D1 gene overexpression with evidence of nuclear positivity was detected in biopsy materials of 40 (40%) oral cancers and 14 (28%) dysplastic oral precancerous lesions. Highly significant coexpression (p<0.05) of HPV was observed with p16 and negatively significant with p53. A significant negative association (p < 0.05) was seen among p16 positive oral cancer cases and p53 and EGFR. Conclusion: A strong etiologic role of chemical carcinogens as smoked and chewed tobacco, betel quid and its substitutes is confirmed in Pakistani population. A frequency of molecular alterations was detected in p53, p16, EGFR and H ras; in early stages of oncogenesis. HR- HPV was detected in a substantial number of oral cancers and preneoplasias but chemical carcinogens have a dominant role. Overexpression of p16 was found in HR HPV positive cases. Key words: Oral Squamous cell carcinoma, Oral premalignant lesions, Betel quid, tobacco, areca nut, immunohistochemistry, molecular markers, HPV, PCR.