Among abiotic stresses, heat stress has the most devastating impact on plant growth. In the present study, heat shock transcription factor HsfA1d was isolated from Arabidopsis thaliana and cloned into an in-house constructed gateway compatible cloning vector pUC57GW-CmRccdB through infusion cloning. Entry clone was confirmed through restriction digestion and sequencing analysis. For subcellular localization, HsfA1d was cloned in-frame with yellow florescent protein (YFP) in pGWB442 through LR clonase reaction resulted in YFP-HsfA1d chimeric gene construct. Homodimerization of HsfA1d was studied using Bimolecular Florescence complementation (BIFC) assay. For BIFC, HsfA1d was cloned in-frame with sequences that codes for N and C termini of yellow florescent protein, into 2 in-house constructed vectors pGSA002-nYFPn and pGSA002-nYFPc respectively. After confirmation through restriction enzyme digestion, the newly constructed vectors were transformed into Agrobacterium tumefaciens GV3101 for transient expression in tobacco (Nicotiana benthamiana) through syringe-infiltration. Subcellular localization of YFP-HsfA1d in DAPI-stained cells was evaluated using confocal microscopy. For plant transformation, HsfA1d was cloned into 2 plant expression vectors pGWB402Ω and pGWB442 through LR clonase reaction. After confirmation through RFLP analysis, the vectors were transformed into Agrobacterium GV3101 for tobacco leaf disc infection. Putative transgenic plants were selected and regenerated on appropriate selection media using different growth regulators. After PCR and confocal-based confirmation, transgenic plants were evaluated for their response to heat stress. HsfA1d, HSP70 and HSP90 exhibited 4.8, 2.8 and 2 folds increase respectively in their expression under heat stress compare to room temperature. HsfA1d was found to significantly enhance the expression of downstream gene HSP70 while no such effect on the expression of HSP90 was recorded. The plants transformed with HsfA1d were found to retain more water and accumulate more proline under heat stress. The transgenic plants exhibited efficient protective system, causing less 8 electrolyte leakage and less chlorophyll damage under heat stress. It is concluded that HsfA1d plays a vital role in thermotolerance enhancement and hence recommended for increasing plant thermotolerance.
موضوع2:تدوینی اصطلاحات متن: • تدوین کے لیے وہ تحریر جسے کوئی ترتیب دینا چاہے۔ • متن اس عبارت کو کہتے ہیں جسے آسانی سے پڑھا جا سکے اور قرات کے دوران اس کی تفہیم ہو سکے۔ متن سے ہم تاریخ یا ماضی سے تعلق رکھنے والی وہ عبارت • مراد لیتے ہیں جس کی ترتیب مقصود ہو۔جسے دریافت کیا گیا ہو اور جو تحقیقی اہمیت کی حامل ہو۔ تدوین : • کسی تصنیف کے مختلف نسخوں کا مقابلہ کرکے درست متن تیار کرنا • کسی مصنف کی منتشر تخلیقات یا کسی تخلیق کے منتشر اجزائ کو صحیح ترتیب سے جمع کرنا مدون: تدوین کا کام کرنے والا مرتب: کتابوں کی ترتیب دینے والا دیوان: وہ مجموعہ کلام جس میں شعراء کی نظمیں اور غزلیں جمع کی جاتی ہیں۔ مخطوطہ: کسی مصنف کے ہاتھ سے لکھی ہوئی تحریر کو کہتے ہیں۔ منسوخ/قلم زد: وہ تخلیقات یا تخلیق کا حصہ جسے مصنف نے خارج کردیا ہو۔ موازنہ: ایک متن کے مختلف نسخوں کے اندراجات کا تقابلی مطالعہ کرکے مناسب ترین کا تعین اختلاف نسخ: تدوین کرتے وقت مختلف نسخوں میں جو اختلافات ملتے ہیں انھیں اختلاف نسخ کہا جاتا ہے، ان کو یک جا کردیا جاتا ہے تاکہ قاری کے سامنے دونوں صورتیں آجائیں۔ نسخہ: کسی قلمی یا مطبوعہ کتاب کی ایک جلد کو نسخہ کہتے ہیں۔
اساسی نسخہ/بنیادی نسخہ: کسی پرانی قلمی کتاب یا مخطوطے کے جب کئی نسخے موجود ہوں تو ان میں سے وہ نسخہ جسے بنیاد مان کر تدوین متن کی جائے اساسی نسخہ کہلاتا ہے۔ خطی نسخہ/قلمی نسخہ/دستخطی نسخہ: مصنف کے ہاتھ کا لکھا یا ٹائپ کیا ہوا نسخہ دستخطی نسخہ کہلاتا ہے۔ آمیختہ نسخہ: وہ نسخہ جس کا متن پہلے کے دو نسخوں سے ملا کر تیار کیا گیا ہو۔
The present study was conducted to probe the cleanliness in the tertiary healthcare hospitals in Peshawar-Pakistan. The data was collected through a structured questionnaire from 600 sampled respondents admitted in three tertiary healthcare hospitals in Peshawar i.e. Khyber Teaching hospital, Hayatabad Medical Complex and Lady Reading Hospital through proportional allocation method. The patients were very critical about the healthcare and cleanliness in the above three major hospitals of Khyber Pakhtunkhwa. The Bi-variate analysis was made and the results reveal that hospital has clean and hygienic environment (P=0.006), Medical OPD, s were clean and tidy (P=0.000), wards were clean and well maintained (P=0.000), Toilets were clean and well maintained (P=0.000). The study recommended that hospital administration should keep focus on cleanliness in washroom, clean water and clean food to the patients.
Lady’s finger or Okra is a member of family malvacae. It is multipurpose crop; its fibrous tender fruits called pods are used as vegetable; seeds, stem and roots used for industrials purposes. In past it was considered as minor crop and no care was taken for its improvement at national and international level in research programs. It grows well at temperature ranges of 18 to 35oC and gives highest yield. Heat and drought events will be intensified due change in climate, activating modifications in the ecosystem and failure or low productivity of crops which are prone to abiotic stresses and same case is with okra. Major constraint which affects vegetative and reproductive phase of okra is heat stress. Heat stress damage quality as well as yield of okra. The research was conducted with the aim to screen out genetically diverse and improved germplasm while eradicating physiological and genetic basis of better adaptation under thermal stress and enhancement of heat tolerance by foliar application of proline. In first experiment comparative performance of one hundred okra genotypes was investigated under heat stress condition. For this purpose one hundred (100) okra genotypes were grown under control environmental conditions of high temperature 45/35°C (day/night). Data for morphological attributes (root length, shoot length, shoot and root fresh weight, number of leaves and root and shoot dry weight) and physiological attributes (transpiration rate, stomatal conductance to water, photosynthetic rate, leaf surface temperature, chlorophyll contents, sub-stomatal CO2 and water use efficiency) was recorded. According to recorded data genotypes were classified on the basis of their performance against heat stress conditions. Under heat stress conditions genotypes showed significant different response and genotypes were divided into heat sensitive and heat tolerant ones. VI051062 and VI060131 were categorized as most heat tolerant and VI046554, while VI048594 were categorized as most heat sensitive ones. In second experiment twenty five okra genotypes screened out from one hundred okra genotypes in experiment # 1 in growth room, (twenty heat tolerant and five heat sensitive genotypes) were sown in summer 2014, in the field conditions at vegetable research area of Institute of Horticultural Sciences, University of Agriculture, Faisalabad. Genotypes were sown at three different sowing dates (02, April, 12, April and 22, April) to check the effect of heat stress on different morpho-physiological and yield attributes. All the cultural practices were kept same for all sowing dates and for all genotypes. There were four replications and each replication contained five plants. In this experiment genotypes VI051062 and VI060131 proved to be most heat tolerant while VI046554 and VI048594 proved to be most heat sensitive under field conditions on the basis of morpho-physiological and yield parameters In third experiment four okra genotypes, two tolerant (VI051062 and VI060131) and two sensitive (VI046554 and VI048594) selected from experiment # 2, were exposed to heat stress (45/35°C day/night temperature) under controlled environmental conditions, two weeks after exposure to heat stress plants were sprayed with proline (control, 1.0, 1.5, 2.0, 2.5 and 3.0mM) to optimize best dose of proline for enhancement the heat tolerance in okra VII genotypes. Morphological and physiological were studied to optimize the best dose of proline for enhancement of heat stress tolerance in okra genotypes. Results revealed that proline application @ 2.5 mM is best for enhancing the heat tolerance potential of okra. Fourth experiment was carried out to check the effect of optimized dose of proline (in experiment # 3) on the morphological, physiological and biochemical attributes of four okra genotypes two heat tolerant and two heat sensitive, under controlled environmental conditions at high temperature 45/35°C (day/night). It was noted that exogenous application of proline @ 2.5 mM significantly affected morphological, physiological, biochemical, water related and enzymatic attributeswhich in turn enhanced the heat tolerance potential of okra genotypes. It can be concluded from the study that by sowing the heat tolerant genotypes, identified in research, the growth period of okra can be extended. Exofenous application of proline @ 2.5 mM can further alleviate the drastic effects of high temperature and growing period can be extended.