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Home > Expression Studies of Viral Vector System Against Sugarcane Mosaic Virus in Sugarcane

Expression Studies of Viral Vector System Against Sugarcane Mosaic Virus in Sugarcane

Thesis Info

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Author

Sajed Ali

Program

PhD

Institute

University of the Punjab

City

Lahore

Province

Punjab

Country

Pakistan

Thesis Completing Year

2015

Thesis Completion Status

Completed

Subject

Molecular Biology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/13089/1/Sajed_Ali_Molecular_biology_HSR_2015_UoP_Lahore_25.10.2016.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726174298

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Sugarcane is one of the major cash crops in Pakistan which has a prime contribution in the GDP. Average yield is very low due to pathogenic epidemics particularly sugarcane mosaic virus. Molecular biology tools have been adopted to increase the defense mechanism of sugarcane against viral attacks. Reverse transcription polymerase chain reaction (RT-PCR) was developed for the detection of sugarcane mosaic virus (SCMV) from the sugarcane samples collected from all major sugarcane cultivation districts in the Punjab, Pakistan. The segments of SCMV-CP gene were amplified from SCMV infected plants and sequenced. The sequencing results did not show 100% homology with any already reported isolate in the GeneBank data base. The sequences were submitted to the NCBI GeneBank and got the accession number KC200152 and KC249907 to KC249917. VIGS is a powerful tool used in functional genomics. Mechanism of VIGS is referred as post transcriptional gene silencing (PTGS) which involves the introduction of exogenous gene into the host plant as a result small interfering RNAs (siRNA) are produced which make a RISC assembly and the DICER like proteins chop the invading RNA as a defense mechanism. All plants naturally produce siRNAs as a defense against viruses and exogenous genes. If they are triggered as exogenous genes, the endogenous genes are silenced. In the present study, VIGS vector pBINTRA6 derived from TRV was modified by inserting SCMV-CP gene (amplified from infected sugarcane plants), GFP gene (amplified from Nicotiana benthamiana 16c GFP transgenic plant) and ChlI gene (amplified from Nicotiana benthamiana plant) for expression in N. benthamiana and sugarcane. Two SCMV-CP gene segments, 313 and 454 bp were cloned in pBINTRA6 to produce two VIGS constructs SCMV-CP/pBINTRA6-313 and SCMV-CP/pBINTRA6-454. Transient expression of GFP and ChlI genes silencing was studies in N. benthamiana GFP transgenic line 16c. SCMV-CP/pBINTRA6-313 has resulted relatively better silencing of GFP and ChlI genes in N. benthamiana than SCMV-CP/pBINTRA6-454. Two sugarcane cultivars HSF-242 and HSF-245 were selected for VIGS vector transformation of SCMV-CP/pBINTRA6-313 construct based on the results and observations of relatively more silencing of GFP and ChlI genes in N. benthamiana. The transformed (transgenic) plants were confirmed by PCR amplification of GUS reporter gene. Moreover, GUS assay was performed to confirm the transformation of plasmids as integral part of plant genome. Knockdown expression was studied by amplification of SCMV-CP gene by real time PCR. No amplification of SCMV-CP gene was seen in the transgenic sugarcane plants which confirmed the virus induced gene silencing of SCMV in sugarcane. Among the two sugarcane cultivars, HSF-242 demonstrated a strong expression for GUS assay. Bioassay i.e. transfection with virulent aphids (Rhopalosiphum maidis) was done on 4-6 fully developed leaves of transgenic sugarcane plants after shifting and acclimatization in soil pots under greenhouse conditions followed by molecular analyses. Results of bioassay determined that transgenic plants exhibited slightly slow growth and weak chlorotic phenotype when compared with the non-transgenic control plants. Results clearly demonstrated that transgenic plants showed resistance developed against sugarcane mosaic virus as a result of initiation of VIGS mechanism. The developed transgenic plants may be subjected to the biosafety studies and variety development in future studies. Moreover, VIGS optimized in the current study can be helpful for identification of new traits and control of other pathogenic diseases through PTGS which will play a key role in value addition and economy of the country.
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111. Al-Masad/Twisted strands

111. Al-Masad/Twisted strands

I/We begin by the Blessed Name of Allah

The Immensely Merciful to all, The Infinitely Compassionate to everyone.

111:01
a. Doomed be the hands of Abu Lahab,
b. and doomed he be!

111:02
a. Neither his wealth be of any benefit to him,
b. nor his earnings be against Allah’s Wrath.

111:03
a. In the Hereafter, he will definitely be flung into Fire of Blazing Flames.

111:04
a. And with him, his wife will enter the Fire as well,
b. as she was a carrier of firewood and thorn branches and of evil tales and slander The Prophet.

111:05
a. She will have twisted strands of palm-fiber around her neck.

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

Outcomes of Extracorporeal Induced Destruction of Urinary Tract Stones by Shock Waves Lithotripsy and its Associated Factors Among Patients at French Medical Institute for Children Fmic , Afghanistan

Background: Extracorporeal Shock Wave Lithotripsy (ESWL) has now been in clinical use for years and it has replaced other treatment techniques for the treatment of Urinary Tract Calculi. ESWL is a non-invasive method for the treatment of renal and ureteral stones. The current range of its indications includes majority of non-selected urinary stone treatment. It is used among the patients with more complex stones in the urinary tract. It has revolutionized the treatment of urinary calculi and it should be considered the treatment of choice for renal stones smaller than 2 cms. The success rate of this procedure is ranging from 60 to 90% in the available literature. To the best of researcher’s knowledge, no published data related to the outcomes and factors associated with Lithotripsy among patients with Urinary Tract stones is available in Afghanistan context. Objective: The current study is aimed to determine the outcomes of Extracorporeal Shock Wave Lithotripsy in the treatment of urinary tract stones and its associated factors at French Medical Institute for Children, Kabul Afghanistan.Methodology: An analytical hospital based retrospective study design was employed. Records of 293 participants who had undergone Lithotripsy procedure at FMIC hospital with the help of a structured check list was extracted by using purposive sampling. The data was collected from the available records during the period of 2008 to 2015. Chi-square test XII of indepedence was run to identify the factors associated with outcomes of Lithotripsy procedure. Results: Our study findings have identified the overall success rate of Lithotripsy procedure as 68.4%. The findings of the study have also revealed that majority of the participants 51.36% had successful, 5.12% of participants had partially and 43.20% of participants’ stones had incomplete broken stones as an outcome of first time Lithotripsy. Among patients, who had undergone second time Lithotripsy procedure, 60.97% had successful and 39.02 % had unsuccessful broken stones. Factors like gender and size of stone were significantly associated with outcomes of first time lithotripsy in this population. On the other hand, we could not determine any association between successful outcomes and second time lithotripsy. Conclusion: Extracorporeal Shock Wave Lithotripsy is one of the cheap and non-invasive, procedures which induce fewer complications which can be managed spontaneously in short period of time. The findings of this study and the available literature shows that ESWL is an effective procedure for the management of Urinary Calculi. Our study has reported a success