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Identification and Functional Characterization of P2- Type Calcium Atpases in Selected Species of Gramineae

Thesis Info

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Author

Roohi. Aslam

Program

PhD

Institute

National University of Sciences & Technology

City

Islamabad

Province

Islamabad

Country

Pakistan

Thesis Completing Year

2018

Thesis Completion Status

Completed

Subject

Natural Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/9486/1/Roohi_Aslam_Viorlogy_%26_Immunology_HSR_2018_NUST_01.08.2018.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726389324

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Calcium and manganese perform various important biological functions within the plant body, including signal transduction, proper cell wall formation, photosynthesis, respiration and nitrogen assimilation etc. Calcium and manganese exist in the form of ions within cells. P2- type calcium ATPases are responsible for maintaining homeostasis of these ions in both domains of life. P2- type ATPases are further divided into P2A (ECAs) and P2B- types (ACAs). In present research work, BLAST searches in different databases were conducted for the retrieval of calcium ATPase sequences of selected monocots and a phylogenetic analysis was performed. Furthermore, Triticum aestivum plants were grown under calcium and manganese stress using hydroponic culture. The tissue generated was used to find out the expression of selected P2A and P2B- type calcium ATPases under manganese and calcium stress. Moreover, an ECA3 construct was generated and it was transformed into yeast model (Pmr1) to find out the possible role of this gene in manganese nutrition. Overall, phylogenetic analysis revealed a high degree of evolutionary relatedness among P2- type calcium ATPases. High expression of selected ACAs was observed in T. aestivum plants grown under calcium stress, which gave clues to the role of ACAs in signal transduction in T. aestivum. High expression of selected T. aestivum ECAs under calcium and manganese stress was also observed which may indicate the potential roles of these ATPases in calcium and manganese nutrition in T. aestivum. Additionally, ECA3 gene (which is a P2A- type ATPase) was amplified using T. aestivum cDNA and sequenced subsequently. The data indicated that it retained introns in some of the transcripts. The qRT-PCR analysis revealed that events of intron retention in ECA3 transcripts were much enhanced when plants were exposed to calcium stress. ECA3 construct was further transformed into yeast mutant Pmr1, which is a null mutant for xi its own manganese pump and cannot grow on high manganese concentration in the medium. Interestingly, ECA3 rescued the phenotype and helped the mutant to grow successfully on manganese toxic media. This further indicated, the importance of ECA3 in manganese translocation in T. aestivum. Hence, ECA3 gene is reported here as first P2A- type pump identified in T. aestivum and is required for manganese management in this species. its own manganese pump and cannot grow on high manganese concentration in the medium. Interestingly, ECA3 rescued the phenotype and helped the mutant to grow successfully on manganese toxic media. This further indicated, the importance of ECA3 in manganese translocation in T. aestivum. Hence, ECA3 gene is reported here as first P2A- type pump identified in T. aestivum and is required for manganese management in this species.
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مفتی محمد شفیع ؒ

مفتی محمد شفیع ؒ

 مفتی شفیعؒ کا آبائی وطن دیوبند ہے، جو ضلع سہارنپور یوپی میں برصغیر کا مشہور ترین قصبہ ہے یہیں آپ کی ولادت۱۳۱۴ھ میں ہوئی شمسی حساب سے یہ جنوری ۱۸۹۷ء تھا آپ کے والد محمد یسین بہت بڑے عالم دین تھے آپ کے دادا نے آپکا نام محمد مبین رکھا لیکن آپ کے والد بزرگوار محمد یسین نے ولادت کی اطلاع کا خط اپنے شیخ گنگوہیؒ کو لکھا تو انہوں نے جواب میں نام محمد شفیع تجویز کیا۔

تعلیم و تربیت

”قرآن کی تعلیم سے فراغت کے بعد دارالعلوم ہی میں خط و املاء کی مشق اور فارسی کی تمام مروجہ کتابوں کی تعلیم اپنے والد سے حاصل کی حساب اور فنون ریاضی اقلیدس وغیرہ اپنے چچا منشی منظور احمد ؒ سے پڑھے عربی نحو و صرف اور فقہ کی ابتدائی کتابیں بھی فصول اکبری،ہدایۃ النحو اورمنیۃ المصلی اپنے والد سے ہی پڑھیں اس طرح تعلیم قرآن کے بعد تقریباً پانچ سال فارسی، ریاضی وغیرہ کی پوری تعلیم اور عربی کی ابتدائی کتب میں صرف ہوئے۔“ [[1]]

۱۳۳۰ھ میں سولہ سال کی عمر میں اصول فقہ اور ادب کی کتابیں دارالعلوم کے درجہ عربی میں باقاعدہ داخلہ لے کر شروع کیں۔

شیخ الہند سے استفادہ اور اصلاحی تعلق

 مفتی شفیع اپنے والد کے ساتھ بچپن سے ہی ان کو خدمت میں حاضر ہوا کرتے تھے۔

 

انور شاہ کشمیری سے استفادہ اور دورہ حدیث

 ۱۳۳۵ھ میں مفتی شفیع نے دورہ حدیث انور شاہ کشمیری ؒ کے سامنے کیا فلسفہ کی بعض کتابیں بھی انہی سے پڑھیں رد قادیانیت کا جو کام شاہ صاحب نے شروع کیا اس میں بھی بڑھ چڑھ کر حصہ لیا اور اسی موضوع پر کئی کتابیں ختم نبوت مسیح و موعود کی پہچان بھی تالیف...

Completeness Analysis of Completeness Filling and Time of Returning The Medical Record for Inpatient Patients at Regional General Hospital of Makassar City

At Makassar City Hospital, one of the service indicators that has not been achieved is in incomplete medical record files and medical record files that are returned more than 2x24 hours after service. This study aims to analyze the implementation of the completeness of filling in and the timeliness of returning inpatient medical record files at the Makassar City Hospital. This type of research is mixed methods research. The study design used a cross-sectional approach. The study was conducted in September - October 2020. The results showed that the implementation of completeness of filling in and the timeliness of returning medical record files was still low, this has led to the accumulation of medical record files in the treatment room and delays in returning the files of inpatients to the medical record installation of the City Hospital Makassar. Training on the implementation of medical records has not been comprehensive for all officers at the Makassar City Hospital. The result of the delay in returning the documents is the delay in payment of insurance claims to the hospital. Accumulation of medical records in the treatment room from incomplete medical records and returned to the treatment room. Health workers who forget to fill in complete medical records are only given a sanction in the form of a warning during a meeting with the medical committee. Availability of SOP on filling and returning medical record files at the hospital. The facilities and infrastructure in the implementation of medical records are still insufficient for medical record employees at Makassar City Hospital. It is recommended that the hospital improve the implementation of the completeness and timeliness of returning medical record files, provide incentives or rewards for completing filling in, increase the number of computers and expand the room in the medical record installation, and review the medical record format at Makassar City Hospital

Understanding the Molecular Basis of Cotton Leaf Curl Disease Resistance in Cotton Germplasm.

The production and processing of cotton is a major source of foreign exchange for the economy of Pakistan. The majority of cotton fiber produced in the region comes from the tetraploid Gossypium hirsutum, although some is still produced from the cotton species native to the region, the diploid G. arboreum. Since the early 1990s, cotton production in Pakistan and northwestern India has been adversely affected by cotton leaf curl disease (CLCuD). The disease is caused by single-stranded DNA viruses of the genus Begomovirus (family Geminiviridae) in association with a specific satellite, Cotton leaf curl Multan betasatellite (CLCuMuB). At this time only a single virus, Cotton leaf curl Burewala virus (CLCuBuV), is associated with CLCuD across most of Pakistan. This virus is resistance breaking, overcoming resistance to the previous begomoviruses/satellite complex that was introduced into cotton by conventional breeding. At this time there are no commercially available G. hirsutum lines that are resistant to CLCuBuV/CLCuMuB. However, all lines of G. arboreum are “immune” to CLCuD and plant breeders have long been trying to introduce the “resistance” from this species into the more desirable G. hirsutum lines. In addition, recently two lines of G. hirsutum originating from France (cvs. Dominique and Haiti) have shown promise in field screening for resistance against CLCuD. The study described here was designed to investigate the nature of the resistance of G. arboreum cv. Ravi and the French G. hirsutum cultivars, Dominique and Haiti, using whitefly-mediated and graft inoculation of the CLCuD virus complex. Additionally the possibility of using biolistic inoculation of viral DNA was investigated as a possible means of experimentally introducing the virus complex causing CLCuD into cotton. In large scale field screening of G. arboreum cv. Ravi over a period of two years, no symptoms of virus infection were detected under inoculation pressure conditions where 79-89% of the susceptible control (G. hirsutum cv. CIM 496) plants were symptomatic. Rolling circle amplification/polymerase chain reaction xiii (RCA/PCR) diagnostics, the most sensitive diagnostic method available to detect geminiviruses in plants, did not detect either CLCuBuV or CLCuMuB in field grown G. arboreum cv. Ravi plants; consistent with the idea that G. arboreum is immune to the virus complex. However, graft inoculation with scions from CLCuD affected G. hirsutum showed firstly that the virus complex can move systemically in the plant and that G. arboreum can respond to virus infections by the production of symptoms. Surprisingly, in a few cases, the disappearance of established symptoms was seen following removal of the graft. In all graft inoculated Ravi plants, after removal of the graft, newly emerging tissues were non-symptomatic and no virus could be detected. These results show that, rather than being immune, G. arboreum is highly resistant to the CLCuD complex and has a high virus/satellite threshold for the induction of symptoms, which whitefly inoculation likely is not able to achieve. The low virus levels detected in G. arboreum suggest that possibly the resistance targets virus/satellite replication and, without a continual source (such as from a graft), the virus/satellite complex is rapidly lost. In small-scale, glasshouse-based insect transmission studies, plants of G. hirsutum cvs. Dominique and Haiti remained symptomless under conditions where all G. hirsutum cv. CIM 496 plants became infected. Graft inoculation showed the Dominique and Haiti plants to be susceptible but showing only mild symptoms, slightly higher than in grafted G. arboreum cv. Ravi plants. The virus/satellite levels in such plants were lower than in the susceptible control but higher than detected in G. arboreum cv. Ravi. Upon removal of the graft, newly developing leaves did not show symptoms and no virus/satellite could be detected. The response to infection seen in G. hirsutum cvs. Dominique and Haiti very much mirrors what was seen for G. arboreum cv. Ravi. Recovery from infection has, for other viruses, been shown to be an RNA interference phenomenon and the results are discussed in light of this possibility. G. hirsutum cvs. Dominique, Haiti, Coker and S-12, as well as G. arboreum cv. Ravi plants were biolistically inoculated with cloned CLCuBuV/CLCuMuB, Cotton leaf curl Kokhran virus (CLCuKoV; a begomovirus prevalent in cotton in Pakistan in the 1990s)/CLCuMuB and with RCA products from field-infected G. hirsutum cv. CIM 496 plants shown to be infected with CLCuBuV/CLCuMuB. Only a small number of Coker and S-12 plants, inoculated with cloned xiv CLCuKoV/CLCuMuB, became infected and showed the typical symptoms of CLCuD. Overall the findings indicate that G. hirsutum cvs. Dominique and Haiti harbor a useful resistance to the virus(es) causing CLCuD which should be used for introgression into elite cotton varieties. The results obtained with G. arboreum cv. Ravi indicate that, rather than being a non-host, this harbors extreme resistance to the viruses causing CLCuD and further efforts should be made to characterize the molecular basis for the resistance. Finally the biolistic studies indicate that this can potentially be a useful method for experimentally introducing begomoviruses/satellites into which should be investigated further.