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Engineering Salt and Drought Tolerance in Cotton Gossypium Hirsutum L.

Thesis Info

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Author

Muhammad Arshad

Program

PhD

Institute

Quaid-I-Azam University

City

Islamabad

Province

Islamabad.

Country

Pakistan

Thesis Completing Year

2015

Thesis Completion Status

Completed

Subject

Biotechnology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/13660/1/Muhammad_Arshad_Biotechnology_2015_QAU_17.06.2016.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726058043

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In the modern era of agricultural crop production, commercial applications of crop inputs and industrial activities have increased salt concentrations in our soil and water resources making them toxic for growing crops. During abiotic stresses, plants employ several strategies, among which salt pumping and compartmentalization into intracellular organelles (lumen of vacuole) is a major strategy. The present study involved the development of transgenic cotton cv. coker-312 tolerant to salt and drought stress. In the first part of the study, silicon carbide whisker mediated cotton callitransformationmethod was established using GUS and AVP1 gene. Fertile transgenic plants were produced from embryogenic calli of four events. PCR and southern hybridization of kanamycin resistant putative AVP1 embryogenic calli showedthe presence andstable integration of transgenes in 1-4 copies in the genome. Results of the salt tolerance study of T1AVP1transgenic cotton showed significantly greater salt tolerance at 200mM NaCl than nontransformed control plants. Moreover, transgenes segregate in a Mendelian fashion indicated the authenticity of the method. In the second part of the study, AtNHX1 gene was cloned underthe control of double CaMV 35S promoter and terminatorin plant transformation vector pGA482.Agrobacterium tumefaciensharboringAVP1 and AtNHX1 genes were used to inoculate coker-312 hypocotyls. Putative transgenic kanamycin resistant calli of different events were produced and putative regenerated transgenic plants were obtained. The Overall transformation efficiency was 20-25 %. Regenerated T0plants were grown to maturity in the containment for getting selfed T0seeds and subsequently T1 and T2 seeds. PCR and Southern analyses showed the presence and genomic integration of 1-4 copies ofthe AVP1 and AtNHX1genes. Northern analyses/cDNA analyses showed transcripts of transgenes in the transgenic plants. Transgenic plants (T2) having AVP1and AtNHX1genes verified by PCR were exposed to salt studies executed with AVP1 transgenic in both hydroponic as well as in pots while plants having AtNHX1 gene were exposed only in pots. After 4-6 weeks of germination, NaCl was applied in solutions up to 200 mM. Transgenic plants showedsignificantlyhigher accumulation of Na+, K+, total free amino acids, proline, total soluble sugars and higher nitrate reductase activities than non-transformed control plants. A water stress study was performed on AVP1 T2 transgenic cotton after two weeks and 8-10 weeks of germination in pots. Stress was applied by withholding water for upto 10 days. Results revealed that transgenic AVP1 plants have significantly high water contents and low water potentials with less square shedding, retain high number of bolls than non-transformed control plants. Photosynthesis rate was significantly higher in transgenic (AVP1/AtNHX1) cotton progenies than non-transformed control plants. Transgenic cotton also showed significantlybetter growth and yield when compared with non-transformed plants. Fiber analyses by scanning electron microscopy and HV1 indicated that fiber of transgenic plant were healthy with uniform twisting in contrast to weak and shriveled fiber of the nontransformed control plants. Transgenic plants hadhigh fiber strength, length and low mic value than non-transformed control cotton plants respectively. Transgenic germplasm developed in present study might be useful for its application in cotton breeding program aiming salt/drought tolerance in cotton cultivars.
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النتائج

النتائج

1۔ أولاً:

تقسیم قصائد وأشعار نازك الملائکۃ وبروین شاکر إلی قسمین وھما: الأشعار المتشابھۃ والأشعار المختلفۃ۔

2۔ ثانیاً:

شرح مجمل بسیط لأبیاتھما الشعریۃ، ودراسۃ مقارنۃ شعریۃ من حیث الأفکار والأسلوب والتعبیر والمعنیٰ.

3۔ ثالثاً:

أحیاناً تتشابه الشاعرتان في الأفکار والأسلوب والتعبیر، وأحیاناً أخری تختلفان عن بعضھما إختلافاً کلیاً.

4۔ رابعاً:

الشاعرتان نازك الملائکۃ وبروین شاکر کلاھما بارعتان في صناعۃ الألفاظ وإیصال الفکرۃ إلی القاريء أو السامع بشکل واضح ودقیق، وبتعبیر صادق يھز الکیان.

5۔ خامساً:

الحزن سائد في معظم قصائدھما، ولکن کان لدیھما القابلیۃ في التأقلم مع البيئۃ والظروف السیاسیۃ والإجتماعیۃ والعائلیۃ.

6۔ سادساً:

أعمال نازك الملائکۃ الأدبیۃ والشعریۃ والعلمیۃ أکثر من بروین شاکر، کما أن نازك الملائکۃ أکبر سناً، وأکثر تجربۃ، وأوسع خیالاً، وأدق تعبیراً، وأشمل وأوضح معان، وأطول قصائد من بروین شاکر.

7۔ سابعاً:

مع أن بروین شاکر کانت تقل عن نازك الملائکۃ في أمور کثیرۃ، ولکنھا کانت لا تقل عنھا شھرۃً وصدقاً وأمانۃً وعزۃً وشرفاً وحزناً وحریۃً في إختیار الأفکار الجدیدۃ المناسبۃ والمتأقلمۃ مع الجیل الجدید.

8۔ ثامناً:

دافعت الشاعرتان عن حقوق المرأۃ دفاعاً عظیماً، ولھما نفس الشعور في حمایۃ المرأۃ علی أساس أنھا أضعف قوۃً وأقل مرتبۃً من الرجال، وأنھا المظلومۃ والمعصومۃ في عالم الظلم والظلام وھو عالم الرجال.

 

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