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Assessment of Molecular Diversity in Rice Germ Plasm in Nuclear, Mitochondrial and Chloroplast Genomes

Thesis Info

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Author

Shah, Shahid Masood

Program

PhD

Institute

Pakistan Institute of Engineering and Applied Sciences

City

Islamabad

Province

Islamabad

Country

Pakistan

Thesis Completing Year

2013

Thesis Completion Status

Completed

Subject

Biotechnology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/2501/1/3083S.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725543715

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Rice has twenty three species but out of these only two species Oryza sativa L and Oryza glaberrima are being cultivated around the world. Asian cultivated rice (O. sativa L.) grasps an exclusive place among domesticated crop species. It is the first fully sequenced crop genome and a critical staple food. Rice along with its wild relatives is being grown on diverse series of agroecological environments and climates. Pakistan is famous for special Basmati rice varieties mainly grown in the Punjab province. Along with Basmati rice, Indica varieties are also grown throughout Pakistan and being exported to different countries. The comparison of genetic diversity of rice varieties grown in Pakistan along with other countries has been given little attention. Information and genetic characterization of rice accessions would be useful in determining present trends in rice breeding and in assessing alternative for improving cultivars. In the present study microsatellite and SNP markers were used to characterize rice accessions from different countries along with Pakistan to provide information about their genetic diversity, which may help for maintenance or expansion of diversity in future breeding programs and to investigate rice adulteration for export purposes. The first study was carried out to evaluate the genetic diversity within a diverse collection of rice (O. sativa L.) accessions and to determine differences in the patterns of diversity within the aromatic and non-aromatic rice varieties. Forty rice accessions were evaluated by using 24 microsatellite markers distributed over the whole rice genome. A total of 66 alleles were detected at 24 SSR loci and the number of alleles per marker ranged from 2 to 4, with an average of 2.75. Polymorphism Information Content (PIC) value ranged from 0.0476 (RM315) to 0.5993 (RM252), with an average of 0.3785 per marker. The average genetic diversity over all SSR loci for the 40 genotypes was 0.4477, ranging from 0.0488 to 0.6638. Major allele frequency ranged from 0.4250 (RM252) to 0.9750 (RM315), with an average of 0.6472. The dendrogram based on the cluster analysis by microsatellite polymorphism grouped 40 rice cultivars into three groups, effectively differentiating Basmati cultivars from non-basmati cultivars. xiii Understanding genetic diversity and population structure in rice populations are of great importance and a prerequisite for genetic characterization and crop improvement. Ninety five accessions from sixteen countries were characterized by one hundred and fifty nine SSR markers. SSR makers were well distributed on the nuclear, mitochondrial and chloroplast genome. The overall results revealed that Indian rice accessions were more divergent than International Rice Research Institute (IRRI) Philippines and Pakistan. Cluster-analysis based on genetic differences coefficients clearly separated all the genotypes into three major groups but was unable to separate the accessions based on their geographical area. The population structure analyses showed that most accessions exhibit some degree of admixture, with many individuals within a population sharing the same introgressed segment due to artificial selection. The model-based structure analysis revealed the presence of three subpopulations. This was basically consistent with clustering based on genetic distance. SSRs proved to be an efficient tool in assessing the genetic diversity of rice genotypes. The same ninety-five accessions of rice were also analyzed in order to profile SSR variation in mitochondrial and chloroplast genomes as well, by using 62 pairs of SSR primers. Out of 42 microsatellite loci for mitochondria only 12 SSR markers showed polymorphism whereas in case of chloroplast 6 out of 20 were found polymorphic. The average gene diversity for both mitochondrial and chloroplast was 0.32 oscillating between 0.041 to 0.620. The PIC value ranged from 0.040 to 0.543 with an average of 0.282. While allelic richness ranged from 2-4 alleles with an average of 2.779 alleles. Mononucleotide repeats stood first (50% polymorphic) for detecting polymorphism for organelle genome followed by tri- (25%), tetra- (14.29%) and dinucleotide (12.5%). Cluster and population analysis revealed two groups of accessions but principal coordinate analysis disagreed with this grouping pattern. The overall results revealed the monophyletic origin of rice. Single nucleotide polymorphism is the base pair change most frequently present in the eukaryotes and are responsible for major diversity. Multiplex single nucleotide polymorphism has been acknowledged as having great potential to find out genetic xiv diversity and structure of diverse rice accessions. One hundred and five accessions of rice were analyzed by RiceOPA2.1 from diverse environment. Twenty accessions representing all five groups of rice were used as reference. Overall call percentage was 97.7%. A total of 746 alleles were detected in this study. The lowest polymorphism information content (PIC) was 0.0706 for SNP id3002191 as compared to the highest PIC value 0.5222 for SNP ud4000438 with an average of 0.3353. The average genic diversity (H) over all SNPs loci for the 105 genotypes was 0.4196, ranging from 0.0733 to 0.5919. Neighbor Joining and Unweighted Pair Group Method with Arithmetic mean cluster analysis clearly separated all the genotypes in two major clusters. Structure analysis based on admixture model and principal coordinate analysis produced the same results. The results revealed that the Indian accessions are more diverse followed by IRRI and Pakistan. This comprehensive and comparative study based on microsatellites and SNPs provides a clear image of genetic diversity of Pakistani rice accessions in comparison with those of other countries accessions. A narrow genetic base has been reported for rice cultivars in Pakistan as compared to that in other countries. So there is a strong need to introduce diverse material in breeding program keeping in view preference of the farmers’ community. The overall results derived from analyses of genetic diversity could be used for designing effective breeding programs aimed at broadening the genetic bases of commercially grown varieties particularly in Pakistan. These results could also be useful for monitoring purity, genotype identification and for plant variety protection.
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پروفیسر ایم ایم شریف

پروفیسر ایم۔ایم شریف
پروفیسر ایم۔ایم شریف اگرچہ اصلاً پنجابی تھے لیکن درحقیقت وہ تھے علی گڑھ کاسرمایۂ گراں پایہ۔ وہ یہاں طالبِ علم رہے، پھر فلسفہ کے پروفیسر اورصدر شعبہ ہوئے۔یونیورسٹی کے پرووائس چانسلر بھی بنے اوراپنے علم وفضل، اخلاق وعادات اورہمدردی ومحبت کے باعث یونیورسٹی کی سوسائٹی میں نہایت مقبول اور ہردلعزیز ہوکر رہے،تقسیم کے بعد علی گڑھ کی یہ دولت پاکستان منتقل ہوگئی تواب اُن کے تحقیقی اورتصنیفی جوہر کھلے، چنانچہ ادب، فلسفہ اوراسلامیات پر انھوں نے انگریزی اور اُردو میں اتنی فکرانگیز اورمحققانہ کتابیں اورمقالات لکھے جوایک مصنف کے فخر کرنے اور اس کو علم وادب کی دنیا میں زندہ جاوید بنانے کے لیے کافی ہیں۔ اسی بناء پر اُن کو پاکستان میں علمی اعزاز بھی بڑے سے بڑاحاصل ہوا،ادارۂ ثقافتِ اسلامی کے ڈائرکٹر ہوئے،پاکستان فلاسفکل کانگریس کے صدر چُنے گئے،پنجاب یونیوسٹی میں فیکلٹی آف آرٹس کے ڈین مقرر ہوئے۔ان کاقلم بڑا شگفتہ نگار اورباغ وبہار تھا۔ طبعاً اپنے نام کی طرح نہایت شریف اوراعلیٰ اخلاق وکردار کے انسان تھے۔راقم الحروف کی پہلی ملاقات اُس وقت ہوئی تھی جبکہ مسلم یونی ورسٹی کی انجمن تمدن وتاریخِ اسلامی کی دعوت پر۴۰ء میں راقم نے یونیورسٹی کے یونین ہال میں عمر میں پہلی مرتبہ ’’مسلمانوں کے زوال کے اسباب‘‘ پرتقریر کی تھی اورپروفیسر محمد شریف مرحوم نے اُس جلسہ کی صدارت کی تھی ۔تقسیم کو داد دیجیے کہ موصوف کاانتقال ہوگیااوراس کی اطلاع ہم کوکئی ماہ کے بعد ابھی پچھلے دنوں لاہور کے ماہنامہ ثقافت کے چار مہینوں کے یکجائی نمبرسے ہوئی، اناﷲ وانا الیہ راجعون۔ اﷲ تعالیٰ مرحوم کومغفرت وبخشش کی نعمتوں سے نوازے ۔
[جولائی ۱۹۶۶ء]

اسم محمد ﷺ اور رسول اللہ کی جلالت قدر کا تحقیقی اور تجزیاتی مطالعہ

Regarding Nabuwat, our Holy Prophet (S.A.W.W) has attained excellence/perfection. His name is foremost (S.A.W.W) among all prophet hood is towards both the worlds. He will be brought before all prophets most respectfully on the Day of Judgment. His teachings are not cancelled as compared to the prophets before him (S.A.W.W). His (S.A.W.W) virtues and excellences are uncountable and unlimited. Below is described his (S.A.W.W) magnificence and dignity in respect of his name; Muhammad (S.A.W.W).

Tectonic Evolution of the Margala Hills and a Part of the South Eastern Hazara Ranges, Pakistan

The Margala Hills and Hazara Ranges are the south-western continuation of the HazaraKashmir Syntaxis in North Pakistan, constituting an east-northeast trending fold-thrust belt of Jurassic to Eocene platform sediments, partially overlying Late Precambrian basement. The current field investigations have revealed that at least three macro-scale fold-thrust nappes, which are structurally linked to a regional basal detachment located at base-Hazara Slate, fabricate the structural grain of the Margala Hills and southeast Hazara Ranges. From north to south these nappes include the Hazara fold-thrust nappe which is exposed along Hazara Thrust. Below the Hazara nappe lays the Garamthun nappe which is exposed at surface along Garamthun Fault. Southeast of the Garamthun Fault the hanging wall sequence of the Main Boundary Thrust constitutes the third fold-thrust nappe of the area and constitute the foothills of Margala Hills. All these fold-thrust nappes are characterised by east-northeast trending major anticlinal folds detached at the level of Jurassic rocks. The stratigraphic set up of these fold-thrust nappes implies a gradual south younging of structuration of the study area. The lithostratigraphic offset along the mapped faults is very little and does not support large scale horizontal movements with the thrust faulting alone. The large scale fold-thrust belt allochthony is shared by fold nappe mechanism which is also believed to be responsible for the thickening of the sedimentary pile. Mesoscopic and macroscopic folds data indicate the presence of at least three distinct lines of folding including northeast-southwest followed by southeast and north-south trends. The northwest-southeast compression is the prominent grain of the southeast Hazara and Margala Hills. Mesoscopic fault planes can be categorised into three distinct sets. The most prominent is the top to the southeast followed by top to the west-northwest and northeast. The top the southeast set is consistent with the movement direction on the macroscopic scale faults (Hazara Thrust, Garamthun Fault and Main Boundary Thrust) and the pitch data is suggestive of a dominant thrust slip motion in addition to left lateral strike slip adjustments. The second set that is the top to the west-northwest can be considered as the back thrusting phenomenon associated with the top to the southeast major displacement. The third set and a few other less prominent fault planes are generally oriented discordant to set one and two and indicate the transpressional related adjustments of the region. Most of dips in the hanging vii wall of Main Boundary Thrust are found to be overturned to the northwest and the axial planes of the folds mostly dip southeast. This back folding is most prominent and consistent along the hanging wall of Main Boundary Thrust and gradually defuse in the northwest direction. The synthesis of the structural data indicate that in the whole Miocene southward directed major shortening created a southwards propagating, flat-ramp thrust fault, cutting up section. This basal thrust carried the Margala Hills Mesozoic and part of its Murree cover towards the south above thicker Murree molasse and underlying Northern Potwar Deformed Zone Mesozoic nappes. Increased shortening took place by tight folding of the hanging wall. This could be the main reason of folds found in the Margala Hills. Late (?) Pliocene major, crustal or lithospheric scale folding created the Hazara Kashmir Syntaxis and created north-south to northeast-southwest axis folds in the Margala Hills. This event is responsible in the first instance for the east or west verging structures in the whole area, for the northwards or northwestwards overturned folds of the Margala Hills; and for the major crustal thickening and subsequent very important uplift of the whole area. The same event could have created strike slip faulting. Repeated south verging thrusting in Quaternary resulted in the southwards movement of the Salt Range above Pleistocene sediments, and flat, mostly south verging thrust faults cutting the older structures. This last event may have generated deeper ramps in the Northern Potwar Deformed Zone and may have formed folds and uplifts in the Northern Potwar deformed Zone and beneath the frontal part of Margala Hills. Strike slip faulting may accompany this event, together with gravitational collapse-induced normal faulting.