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Home > In Vitro Regeneration and Agrobacterium Mediated Transformation of Soybean Glycine Max L.

In Vitro Regeneration and Agrobacterium Mediated Transformation of Soybean Glycine Max L.

Thesis Info

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Author

Zia, Muhammad

Program

PhD

Institute

Quaid-I-Azam University

City

Islamabad

Province

Islamabad.

Country

Pakistan

Thesis Completing Year

2010

Thesis Completion Status

Completed

Subject

Natural Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/handle/123456789/663

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726445179

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Agrobacterium mediated transformation of soybean has successfully been achieved. However, the efficiency is usually low indicating that Agrobacterium mediated transformation of soybean demands optimization of more suitable conditions for transfer of T-DNA from plasmid to plant cell; beside these constraints; transformation is also dependent upon plant genotype, Agrobacterium strain and type of plasmid. Present investigation was aimed to find suitable regeneration protocol for two soybean cultivars, NARC-4 and NARC-7 and to standardize some transformation conditions. Based on optimized regeneration and transformation scheme, LFY gene, for early flowering and rol genes (A, B & C) for enhanced rooting were introduced in soybean genome. For transfer of LFY gene EHA105 harboring pROKIILFYGUSintnptII and for rol gene, LBA4404 harboring pLBR were used. Cotyledonary node method was found suitable for direct organogenesis of soybean NARC-4 and NARC-7 cultivars. However, presence of different plant growth regulators in media resulted in variation in number of shoot produced, shoot length and in percentage response. Statistical analysis reveals that BAP resulted in higher frequency of shoot regeneration and number of shoots per explant while mean shoot length was found higher when ZTR was used in the medium. Out of both soybean cultivars, NARC-4 showed better response than NARC-7. For standardization of transformation conditions, different parameters were studied. It was observed that explant cutting in Agro-suspension culture and infection for 1hr resulted in the highest GUS expression (48.3% and 55.9%, respectively). While 5 days co-cultivation resulted in 55.17% GUS response and washing for 2hr in washing medium containing 1g/L cefotaxime was better. It was found that 30mg/L kanamycin was sufficient in selection medium. At this concentration number of GUS positive shoots were maximum (63.6% response). The overall percentage transformation efficiency of both soybean cultivars NARC- 4 and NARC-7 was 24.16% and 15.71% respectively, when tested at best conditions. 11Soybean cultivar NARC-4 was used for transformation of LFY and rol genes. EHA 105 containing LFY gene along with GUS as reporter and nptII as selectable marker showed transformation efficiency 2.2%. Only 4 plants were recovered and confirmed for presence of LFY gene by PCR. These plants were highly dwarfed with small leaves and short nodal distance. Early flowering was observed by these transforments. The flowers turned brown and died after few days. Few of them turned into pods but no seed formation was observed. All the rol transforments produced enhanced rooting as compared to control plants. However, plants morphology varied depending upon rol gene. RolA transforments were small in size and mildly shrubby with ovate to elliptical leaf shape while rolC transforments were also dwarf with divided stem at the base with ovate to slightly globular leaf shape. Soybean rolB transformed plants showed variation in morphology. These plants were dwarf to shrubby with variation in leaf shape. The shrubby plants had reduced nodal distance with a little more zigzag pattern as compared to non-transformed plants. Rol transforments produced flowers in less time period as compared to control plants. These flowers converted into pods and set seeds. PCR analysis confirmed presence of respective gene in these plants. Southern blot analysis confirmed insertion of T-DNA in soybean genome as single copy number to multiple copies in rol gene transforments. In this study rol and LFY genes were efficiently introduced in soybean cultivar NARC-4 after optimization of regeneration and some transformation conditions.
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النتائج

النتائج

1۔ أولاً:

 تعرفنا علی بدایۃ الشعر الحر وما ھي حقیقتہُ وکیف کان التجدید في الشعر ولا یقصد بذلک التنکر لقوانینہِ إنما یکون الابتکار في المعاني۔

2۔ ثانیاً:

 من ھم أھم شعراء وشاعرات العصر الجديد ؟ وما ھي مکانۃ نازک الملائکۃ بین شاعرات عصرھا ؟

3۔ ثالثاً:

 إتفاق الشعراء حول نازک الملائکۃ بأنھا شاعرة ممتازۃ لھا ممیزات أدبیۃ وشعریۃ رائعۃ وأنھا تستجیب لشعورھا وإحساسھا قبل کل شيء وتعتبر ھي رائدۃ الشعر العربي الحر و تعرفنا على بعض شعراء و شاعرات عصرها .

4۔ رابعاً:

 تحدثنا عن الشعر الجدید في شبہ القارۃ الھندیۃ وأن حالي أعطی لغزل ( اللغة الأردية) الأسلوب الجدید واستوعب النقاد من التجارب علی أن ذلک شعر علی شکل نثر کما نقولُہ في العربیۃ الشعر الحُر۔

5۔ خامساً:

 الأدب النسائي في الباكستان وما ھي دور المرأۃ بین الرجال من البدایۃ وحتی العھد الحاضر ، و التعرف على بعض الشعراء والشاعرات العصر الجديد .

6۔ سادساً:

 مکانۃ بروین شاکر في الساحۃ الأدبیۃ وخاصۃً في الغزل وھي من بعض ألمع نجوم السماء علی الأرض في الغزل الأردو والشعر الحر في اللغة الأردية۔

 

 

Impact of Exposure of Chemical Fumes on Blood Pressure and Peak Expiratory Flow Rate in Industrial Workers of Faisalabad

Objectives: 1. To determine the impact of duration of exposure to industrial chemical fumes on Peak Expiratory Flow Rate (PEFR) and blood pressure of the industrial workers. 2. To find out the association between changes in blood pressure and PEFR due to exposure to industrial chemical fumes in these subjects. Methods: This cross-sectional study was performed at Aziz Fatimah Medical and Dental College, Faisalabad. The study participants were 151 males working in the chemical industries. The study was approved by institutional ethical committee and informed consent was taken from the participants. Free camp was arranged for three days in September 2020 in the industrial area of Faisalabad. Thorough history of exposure to chemicals was taken using structured proforma. PEFR values were recorded using Wrights handheld peak flow meter. Blood pressure was taken by auscultatory method using mercury sphygmomanometer. Data was analyzed using SPSS version 22. Results: Systolic and diastolic blood pressures were significantly raised with increase in duration of exposure.  PEFR levels were significantly declined with increase in duration of exposure to chemicals. Significant negative association was noted between diastolic blood pressure and PEFR (p value = 0.003). Negative correlation was observed between PEFR and systolic Blood pressure, however it was not statistically significant (p value = 0.92). Conclusions: PEFR decreased while Systolic and diastolic blood pressures increased significantly with increase in the duration of exposure to chemicals. There was a significant negative correlation between PEFR and diastolic blood pressure while there was no association between PEFR and systolic blood pressure.

Ontology Based Personalized and Adaptive E-Learning Systems

The role of e-learning systems has become imperative in effectively educating masses of knowledge communities while maximizing the learner’s productivity. Barring this important role, e-learning systems face major challenges such as having context-aware and reusable learning contents. Furthermore, aspects oflearner profiling and categorization for deliverance of relevant learning contents, personalization and adaptive content recommendation to learners need to be focused. Currently, learning contents are static and not machine processable.Learner profiling may not fully comprehend the implicit as well as explicit characteristics of learners with subjective consideration of academic aspects at abstract level of granularity. Learner categorization techniques lack in dynamically considering the cognitive and inclinatory attributes of learners at finer level of granularity across the learning cycle. The learning contents offered may not accord with learning capacity of learners (lack personalization) with minimal support for content adaptivity.In proposed research, Ontology based Adaptive Semantic E-learning Framework (OASEF) is presented that exploits comprehensive set of learner attributes identified for effectively profiling the learners based on discriminative ones. Machine learning based dynamic and adaptive technique named Learner Categorization based on Hybrid Artificial Intelligence Techniques (LCHAIT) has been proposed for learner categorization. A supervised mode of learning was employed on a labeled data set modeled through a LearnerOntology. It has diverse learner’s profiles with implicit and explicit attributes pertinent to learner’s perspectives of demographics, academics, inclinations and behaviors. A comparative analysis of LCHAIT with three other machine learning techniques (Fuzzy Logic, Case Based Reasoning, and Artificial Neural Networks) is also presented. The learning contents maintained in the ontologies (CourseOntology, AssessmentOntoloy and DomainOntology) were recommended by considering the learner’s category to ensure personalization by a dynamic content recommender named Knowledge based Adaptive Semantic e-Learning Recommender (KASER). The efficacy of all categorization techniques was empirically measured while categorizing the learners based on their profiles through metrics of accuracy, precision, recall, f-measure and associated costs. These empirical quantifications assert LCHAIT as a better option than contemporary techniques as exhibited by greater accuracy of performance metrics. The performance of KASER was measured through degree of correctness in recommending the relevant learning contents compared with domain experts. Overall performance of OASEF was measured while recording the learner’s results spanning three years. The comparative analysis of proposed framework exhibits visibly improved results compared to prevalent approaches. These improvements are signified to the comprehensive attribute selection, learner profiling, dynamic techniques for learner categorization and effective content recommendation while ensuring personalization and adaptivity.