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Transformation, Heterologous Expresssion and Functional Evaluation of Germin-Like Protein Gene in Model Legume Medicago Truncatula

Thesis Info

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Author

Sultana, Tasawar

Program

PhD

Institute

Pir Mehr Ali Shah Arid Agriculture University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2016

Thesis Completion Status

Completed

Subject

Natural Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/7433/1/Tasawar%20Sultana%20PhD%20final%20Thesis%2015-11-2016.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676727628387

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Germin-like proteins are widely distributed in various plant tissues like leaves, roots, seeds, fruits and nectar glands, etc. They are expressed during different developmental stages of plants as well as during different biotic and abiotic stress interactions. The current study was planned to explore the function of an Oryza sativa rice root expressed Germin-like protein, OsRGLP1 gene by heterologous expression in Medicago truncatula. For genetic transformation, recombinant vectors pCOsRGLP1 and pGOsRGLP1 both harboring OsRGLP1 gene under transcriptional control of CaMVS35 promoter and Agrobacterium tumefaciens strain AGL-1 were used. Different transgenic lines were produced through Agrobacterium mediated transformation, and confirmed through PCR amplification of OsRGLP1 gene. Reverse transcriptase PCR analysis revealed differential expression of transgene in different transformed lines. Inheritance pattern analysis of seed collected from T0 progeny revealed that transgenic lines followed a Mendelian inheritance pattern. Morphologically under normal growth regime, difference was observed between T1 transgenic and wild type plants for various characters that included the number of leaves, length of stem and number of flowers developed, while non-significant difference observed for seed germination percentage, number of days taken for seed germination and pod formation. Enzymatic characterization of T1 transgenic progeny exhibited significantly increased superoxide dismutase activity (SOD) whereas no oxalate oxidase activity was observed. It was also noted that additional SOD activity attributed to OsRGLP1 transgenic lines was heat resistant. Further, this additional added SOD activity in transgenic lines was insensitive to KCN and sensitive to H2O2 indicating its resemblance to FeSOD rather than Mn or CuSOD. The functionally active status of transgene was also monitored by high accumulation of H2O2 in xviii transformed progeny. Some selected transgenic lines were also tested for their reaction against a highly pathogenic strain of Fusarium oxysporum, a serious soil borne root affecting pathogen active against a wide range of agricultural crops including legumes. Wild type plants were highly affected by pathogen infection showing a percent disease index value of 50 and bore only 2-3 pods per plant, whereas transgenic lines demonstrated high level of tolerance and provided protection against damaging effect of infection with PDA value ranging from 10 to 18 with 14 to 16 pods formed per plant. Significant loss of fresh weight was also observed in infected wild type as compared to transgenic plants. Microscopic observation of roots of infected transgenic plants demonstrated reduction in the attachment and penetrance of fungal hyphae to plant root cells. Transcriptional analysis of subset of defense-related proteins revealed that the expression of OsRGLP1 gene was responsible for activation in the expression of PR-1, PR-1580, PR-1A670 and Def-1 and down regulation for PR-10 in transgenic plants. The findings of the study may suggest that OsRGLP1 is a heat resistant FeSOD-like protein responsible for providing protection against fungal pathogen Fusarium oxysporum, as one of its functions.
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ڈاکٹر عبدالرب عرفان

آہ ڈاکٹر عبدالرّب عرفان!
قارئین معارف کو یہ سن کر بڑا افسوس ہوگا کہ ڈاکٹر عبدالرب عرفان ۳۰؍ اپریل کو انتقال کرگئے، وہ معارف کے بڑے قدر دانوں اور خاص مضمون نگار اور دارالمصنفین شبلی اکیڈمی سے گہرا تعلق رکھتے تھے، اردو، فارسی کے عالم اور ان زبانوں کی ادبیات کی تاریخ سے اچھی واقفیت رکھتے تھے، ہندوستان کے عہد اسلامی کی تاریخ پر ان کی نظر تھی، اس عہد کے سلاطین و مشائخ اور علماء و فضلاء کے حالات و تراجم سے ان کو بڑی دلچسپی تھی اور اس پر ان کے مضامین ملک کے بلند پایہ رسالوں میں شائع ہوتے تھے مگر معارف پر ان کی نظر عنایت زیادہ تھی۔ اس شمارے میں بھی ان کا ایک مضمون شامل ہے اور ابھی دو ایک اور مضامین میری فائل میں ہوں گے۔
میری ان کی ملاقات کبھی نہیں ہوئی مگر خط و کتابت رہتی تھی، ان کے خطوط سے اندازہ ہوتا تھا کہ ان کے دل میں میری کتنی قدر و محبت اور معارف سے ان کو کیسا والہانہ لگاؤ تھا۔ مجھے ان کے حالات زندگی سے کوئی واقفیت نہیں تھی ان کے ایک ہم وطن جناب فیروز حیدری کے خط سے معلوم ہوا کہ انہوں نے کامٹی کے ایم۔ایم ربانی ہائی اسکول سے میٹرک کا امتحان امتیازی نمبر سے پاس کیا تھا۔ اور مارس کالج ناگپور سے بی۔اے کیا تھا۔ پھر فارسی زبان و ادب میں ایم۔اے میں امتیازی نمبر ہی نہیں بلکہ گولڈ میڈل کے بھی حق دار قرار پائے تھے، ۱۹۶۱؁ء میں ودبھ مہاودیالیہ امراؤتی میں فارسی کے استاد کی حیثیت سے ان کا تقرر ہوا اور بہت جلد شعبہ فارسی کے صدر بنائے گئے، ۱۹۸۲؁ء میں ان کا تبادلہ ناگپور میں وسنت راؤناٹک گورنمنٹ انسٹی ٹیوٹ آف آرٹس اینڈ سوشل سائنسز (سابق مارس کالج) ہوگیا۔ ڈاکٹر عبدالرب عرفان طبعاً شریف، سادہ مزاج،...

Analysis of Factors Affecting Nursing Services with Inpatient Satisfaction at Harapan / Pematangsiantar Hospital

Improving quality of nursing services became a major issue in health development both in national and global, because of growing demands on health services organization to give satisfaction with nursing services maximally by providing the best service to facilitate the ease of fulfilling the needs and realize the satisfaction. This study was a quantitative research design survey analytic used cross-sectional approach. It was conducted at Harapan Hospital Pematangsiantar. The population were 280 respondents  and  a sample obtained by 74 respondents. Data analysis of univariate, bivariate using chi - square and multivariate using logistic regression at 95% confidence level (α =.05). The result showed that more respondents who gave a rating of good in the aspect of reliability, followed by the aspect of responsiveness, tangible, assurance, and empathy the influence aspects in this study were reliability        (p=.002), assurance (p=.014), tangible (p=.011), empathy (p=.030), responsiveness (p=.024). The most influential aspects of this study was  the reliability with  Exp(B)/OR=20.667 that aspect of reliability that respondents perceived to have the opportunity 20.667 times to produce patient satisfaction. It is concluded that there was an influence of reliability, assurance, tangible, empathy, responsiveness to patient satisfaction in Harapan Hospital. It is recommended to hospital to further improve the quality of nursing service, to improve the welfare of nurse through provision of appropriate  incentives hope the nurse, provide training, and installing  CCTV in every  room and monitoring of the level of patient satisfaction on a regular basis  through a survey so that can improve the quality of  hospital services as a whole.

Molecular Phylogeny of Artemisia Subgenus Seriphidium Asteracea

Members of Artemisia (Asteraceae) are of great economic, ethnobotanical and medicinal importance. There are about 500 species distributed throughout the world except for Antarctica. Morphological, anatomical, cytological, palynological, genomic and physiological techniques are being used from ancient times for identification and natural classification of these plants but still have restrictions. The current study will help out in resolving the taxonomic complexities of the genus Artemisia sensu lato with an emphasis on subgenus Seriphidium sensu stricto and point out the potential molecular taxonomic markers. Artemisia subg. Seriphidium is one of the largest groups within Artemisia, encompassing more than one hundred species, some of them having considerable ecological and economical importance. However, the evolution of subg. Seriphidium has received less attention in comparison to other subgenera of Artemisia, probably, apart from the difficulty of sampling throughout its very large distribution area, because of the low molecular and morphological variability observed in previous studies. Here, thorough taxonomic sampling within both Artemisia and subg. Seriphidium is used to reconstruct the evolutionary history of the subgenus, employing nuclear and plastid DNA sequences as well as various phylogenetic, biogeographic and diversification dynamics tools to analyze the data. The results show that subg. Seriphidium is not monophyletic but segregated into two main clades: one large monophyletic group corresponding to the formerly recognized sect. Seriphidium and a second, small clade, phylogenetically distant from the first. Biogeographic and diversification analyses indicate that rapid radiation of species within the sect. Seriphidium occurred in Central Asia during the Miocene-Pliocene transition. The results of our biogeographic analysis suggest that this diversification process started around the Tian-Shan, Pamir and Hindu Kush mountain ranges, subsequently expanding into the Eurasian continent. Finally, the study uncovered numerous incongruences between taxonomic and genetic information in several sect. Seriphidium species, which could be explained by morphological uniformity, hybridization and/or incomplete lineage sorting processes. Moreover, the study researches the systematic position of members of Artemisia from Pakistan using molecular data. Furthermore, the research strongly supported the inclusion of Seriphidium within Artemisia. More investigation encompassing extensive sampling from numerous biogeographic regions and markers that are more variable would be necessary to disentangle systematic affinities within the genus.