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Home > Cloning and Over-Expression of a Germin-Like Protein Gene for its Functional Analysis

Cloning and Over-Expression of a Germin-Like Protein Gene for its Functional Analysis

Thesis Info

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Author

Yasmin, Tayyaba

Program

PhD

Institute

Pir Mehr Ali Shah Arid Agriculture University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2009

Thesis Completion Status

Completed

Subject

Natural Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/handle/123456789/443

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725740292

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Germin and germin-like proteins (GLPs) constitute a large and highly diverse family of developmentally regulated proteins showing a wide range of distribution from Myxomycetes to flowering plants. These proteins have been cited in various plant organs like roots, leaves, nectar glands and seeds etc. Germin-like proteins also seem to be involved in many stress related processes, but their definite role in these processes remains obscure. The present study was conducted to elucidate the function of germin-like protein gene (OsRGLP1) of rice by over-expressing OsRGLP1 cDNA under the transcriptional control of strong promoter in tobacco. The cDNA of OsRGLP1, 958 bp was successfully cloned in pTZ57R/T cloning vector. OsRGLP1 cDNA was inserted in p1301 in sense direction down stream of CaMV35S promoter and transformed into Agrobacterium strain EHA101. Transgenic tobacco plants were produced through Agrobacterium mediated transformation and confirmed through hygromycin resistance and PCR amplification of OsRGLP1. Transgenic plants were assessed for growth, morphology, oxalate oxidase and superoxide dismutase activities. Morphologically no difference was observed between T0 transgenic and control plants except that leaves of some mature transgenic plants showed necrotic lesions. Transgenic plants were evaluated for the presence of H2O2 which is reported to be involved in necrosis and cell death. Interestingly, higher levels of H2O2 were observed in transgenic plants. Seeds of T0 /T1 transgenic plants were similar in shape, size and coat color to the control plants when harvested, whereas, seed coat of transgenic seeds turned lighter in color after one year storage at room temperature. Germination rate and percentage in transgenic T1 seeds was significantly lower than control seeds. Segregation analysis indicated a 1:1 ratio for hygromycin phosphotransferase gene in T1 seeds. There was no difference in fresh/dry weights, plant heights and shoot lengths, number of leaves and nodes of transgenic and control plants. Number of roots in transgenic plants was lower while roots were longer than control plant roots. No oxalate oxidase activity either of native or transgene origin could be observed in transgenic plants revealing that OsRGLP1 may not posses this activity. Significantly higher levels of heat resistant superoxide dismutase activity were observed in transgenic plants as compared with control. This added activity was insensitive to KCN and sensitive to H2O2 suggesting being due to FeSOD activity. Present study indicates for the first time that OsRGLP1 may posses SOD activity along with a striking observation that this activity is FeSOD type instead of MnSOD, if at all observed. Upstream regulatory region of OsRGLP1 was also cloned, sequenced and analyzed to gain insight into the regulatory aspects. The promoter region contained many regulatory elements related to diverse functions, most intriguing were the elements related to biotic and abiotic stress which emphasizes the possible involvement of OsRGLP1 in these processes. These findings may suggest that OsRGLP1 gene product may act as FeSOD responsible for dismutation of stress generated reactive oxygen species into H2O2 which in turn may play important roles like defense against pathogen and root elongation.
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المبحث السابع: قصيدة بديع حقي

المبحث السابع:قصيدة بديع حقي

 حصلت الشاعرۃ علی قصیدۃ حرۃ منشورہ للشاعر بدیع حقی وذلک قبل صدور قصیدتھا وقصیدۃ بدر السیاب، وهذا مقطع من قصیدۃ بدیع حقي:

 

أي نسمۃ۔
حلوۃ الخفق علیلۃ۔
تمسح الأوراق في لین ورحمۃ۔
تھرق الرعشۃ في طیبات نغمۃ
وأنا في الغاب أبکي۔
أملاً ضاع وحلماً ومواعید ظلیلۃ۔
والمنى قد ھربت من صفرۃ الغصن النحیلۃ۔
فا نمحى النور وھام الظلّ یحکي

بعض وسواسي وأوھامي البخیلۃ[1]۔

 وذکرت نازک عن الباحث الدکتور أحمد مطلوب بأنہ أورد في کتابہ "النقد الحدیث في العراق"، وقد حصل علی أقدم قصیدۃ في الشعر الحر المعنونۃ ’’بعد موتي‘‘ المنشورۃ في جریدۃ العراق سنۃ 1921م تحت عنوان ’’النظم الطلیق‘‘ ولکن لم یذکر الشاعر اسمُہ بل رمز لاسمہ (ب۔ن)، ویدل ذلک علی أن ھناک محاولات ومباحثات حول الشعر الحر ، ودارت مناقشات بین النقاد والدارسین حول الأسبقیۃ الزمنیۃ في نظم الشعر الحر: أي القصیدتين سبقت الأخری، ھل کانت قصیدۃ ’’الکولیرا‘‘ لنازک أم قصیدۃ ’’ھل کان حباً؟‘‘ لبدر شاکر السیاب، وإلی من ترجع الریادۃ (ریادۃ الشعر الحر)؟

 وقد اعترفت نازک، بأن المسألۃ لیست مصادفۃ، إذا ھو مشروع ثقافي وإبداع کامل المحکم بالوعي والإرادۃ[2]. وتقول أیضاً بأن الریادۃ لم تکن لھؤلاء الشعراء الذین نظموا قصیدۃ واحدۃ او اثنین وعادوا إلی أسلوب الشطرین۔

وتعتقد نازک بأنّھا لو لم تبدأ ھي حرکۃ الشعر الحر، لبدأھا بدر شاکر السیاب، ولو لم یبدأھا بدر السیاب لبدأھا شاعر عربي آخر غیرہ، لأن في تلک السنین أصبح العصر یقبل الشکل الجدید ویرحب بکل ما ھو حدیث. ویتضح من المناقشات السابقۃ أن نازک الملائکۃ ھي ’’ رائدۃ الشعر الحر ‘‘ ۔

 

 



بدھ مت کا تصور امن اور عصری صورت حال

The religion of Buddhism attributed to Siddhartha Gautama (563-482 B.C). Afterwards he came to be known by the name of "Buddha"(mean: awakened). The religion he founded stressed on universal compassion and monasticism. His concept of compassion flowing freely towards all creatures or all living being. That is why, Buddha has forbidden all kind of wars, battles and conflicts as they cause ferocity or bloodshed. When Emperor Asoka (273-233 B.C.) accepted Buddhism as a true believer, then he set up good examples of piety, sympathy and compassion. If we look all around the modern age of Buddhism, they put off the peace living teachings of Gautama. The obvious example of that conduct is oppressive attacks on Rohingya Muslims. Since the £2012, (when the conflicts started between the Rohingya Muslims and the extremist Buddhist) innumerable Muslims are killed, mosques, houses and villages are being set on fire and destroyed.

Estimation of Covariance Matrix and Inference About Regression Coefficients and Autocorrelation in Heteroskedastic Regression Models

The inference about the parameters of regressionmodel and autocorrelation is a challenging task in the presence of heteroskedasticity. In this scenario the heteroskedastic consistent covariance matrix estimators (HCCMEs) and variance ratio (VR) tests are widely used methods to obtain valid inference about regression parameters and autocorrelation respectively. For this purpose, several methods have been suggested in literature. But these methods generally work well when the sample size is sufficiently large and heteroskedasticity level is not very high. In this thesis, we have studied these methods for linear regression model in small samples with high level of heteroskedasticity. In first part of this thesis, we suggest a new heteroskedastic consistent (HC) covariance matrix estimator which takes into account the effect of leverage observations and has better approximation of its true asymptotic distribution. We point out that the basic motivation behind this new modified HC estimator is to provide an estimator which does not require any user specified values. In terms of bias and mean squared error (MSE) a Monte Carlo simulation study provided evidence that this new estimator has better small sample properties over some existing estimators. Real life example also evaluated the finite sample behavior in comparison to those existing estimators. In the second part we suggest heteroskedastic consistent covariance matrix estimator, HC6d, which is based on deviance measure. We have studied the finite sample behavior of the test statistic based on this new HC estimator. We compare its performance with other HC estimators namely HC1, HC3 and HC4m, which are also used in case of leverage observations. Extensive simulation are used to study the effect of various levels of heteroskedasticity on the performance of the quasi tests based on HC estimators. Results showed that the test statistic based on new suggested estimator has better asymptotic approximation and less size distortion in small samples especially when high level heteroskedasticity is present in the data. It has been known that the autocorrelation test do not perform well in presence of heteroskedasticity and variance break case. The remedies to this have been suggested by Jeong and Kang (2012), Shim et al. (2006) and Kim (2006). In the last part of the thesis, we suggested three weighted variance ratio test to test the autocorrelation in presence of heteroskedasticity and variance break. We show through monte carlo simulation that new suggested tests perform well in small samples and are comparable with other tests in terms of size and much better in terms of power properties and also when lag length is misspecified.