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MORAD MOKHTAR MOKHTAR MOHAMED

Thesis: Master

Department: Genetics

Factuality: Agriculture

University: CAIRO

Date : 2017

Biotechnological Studies on Egyptian Date Palm

ABSTRACT

In recent years, date palm has been subjected to intensive genome sequencing studies. The advances in bioinformatics have provided the scientists with tools to develop useful SSR markers that help the breeder accelerating their breeding programs. A total of 172,075 SSR motifs was identified in date palm genome sequence with a frequency of 450.97 SSRs per Mbp. Out of these, 130,014 SSRs (75.6%) were located within the intergenic regions. While, only 42,061 SSRs (24.4%) were located within the genic regions. A number of 111,403 of SSR primer pairs were designed, that represent 292.39 SSR primers per Mb. Out of the 111,403 only 31,380 SSR primers were developed in the genic regions, while 80,023 primers were developed in the intergenic regions. A number of 250,507 SNPs were recognized in 84,172 SSR flanking regions, which represent 75.55% of the total SSR flanking regions. Out of 12,274 genes only 463 genes comprising 896 SSR primers were mapped onto 111 pathways using KEGG data base. The most abundant enzymes were identified in the pathway related to the biosynthesis of antibiotics. Validation of the designed SSR primers was conducted using in silico and in vitro PCR. We tested 1031 SSR primers using both publicly available date palm genome sequences as templates in the in silico PCR reactions. When using the date palm genome PDK30 sequence, all the 1031 SSR primer pairs successfully found complementary sequences. However, only 903 SSR primers could successfully hit within the ATBV01 genome. For in vitro validation, 31 SSR primers among those used in the in silico PCR were synthesized and tested for their ability to detect polymorphism among six Egyptian date palm cultivars. All tested primers have successfully amplified products, but only 16 primers detected polymorphic amplicons among the studied date palm cultivars.

Key words: Date palm, SSR, Intergenic regions, Genic regions, SNPs, pathways, KEGG, In vitro PCR, In silico PCR.


NERMEIN MOHAMED ALI

Thesis: Master

Department: Genetics

Factuality: Agriculture

University: Ain Shams

Date : 2017

Regeneration and Transformation in Sweet Potato

ABSTRACT

Nermein Mohamed Ali: Regeneration and Transformation in Sweet potato. Unpublished M.Sc. Thesis, Department of Genetics, Faculty of Agriculture, Ain Shams University, 2017. Genetic transformation is considered as one of the most favourable options for improvement of crop traits. In this study the regeneration frequency and transformation system were established on the Egyptian sweet potato (Ipomoea batatas (L.) Lam.) cv. Abees and Mabruka. The effect of different hormone combinations and type of explant on shoot regeneration was evaluated. The regeneration percentages from Abees and Mabruka cv. 26.3 and 13.3%, respectively were obtained on Murashige and Skoog MS basal salt mixture + 1.0 mg/l BA + 30.0 g/l

sucrose + 2.2 g/l Phytagel with Abees cv. and the same media was used for cv. Mabruka with only cytokinin type different as 5.0 Kin and shoots were rooted on MS medium + 30 g/l sucrose and 2.2 g/l Phytagel. The

Agrobacterium-mediated and microprojectile bombardement transformation system were successfully introducing the reporter GUS and selectable bar marker genes in the sweet potato explants under pressure of 900& 1100 psi and microcarrier travel distance (6 & 9 cm). Incorporation and expression of the GUS and bar genes into sweet potato plants were confirmed using polymerase chain reaction (PCR) and GUS histochemical assay. Several factors were found to be important for regeneration and transformation in sweet potato. The most effective factors were plant genotype and the type of explants. Co-cultivation time and optical density of the Agrobacterium suspension were also critical for sweet potato transformation. This work is an attempt to open the door for further genetic improvement of sweet potato using important agronomic traits.

Key words:

Sweet potato (Ipomoea batatas (L.) Lam.), Agrobacterium tumefaciens, microprojectile bombardement, microcarrier, regeneration.


 

Sally Mohamed Hassan

Thesis: Master

Department: Genetics

Factuality: Agriculture

University: Ain Shams

Date : 2017

Insertion of Chitinase Gene to Attenuate Early Blight Disease in Some Potato Virus Resistant Lines

ABSTRACT

Potato (Solanum tuberosum L.) an agro-economically important food crop in the world, is sensitive to many fungal pathogens including Alternaria solani, the causal agent of early blight disease. Chitinase is cell wall degrading enzyme which has been shown to have high antifungal activity against a wide range of phytopathogenic fungi. In the present study, plasmid pRI 201-AN binary vector, containing the kanamycine selectable marker in plant and the chitinase gene was used in potato transformation. Leaves of Desriee cultivar, PVY5 and PVY15 lines were transformed with the pRI 201-AN construct via the Agrobacterium tumefaciens delivery system. Transformed leaves were incubated for 5 days in dark on callus induction media which contained MS with 5 mg/l 2-4, D, 1 mg/l cefatoxine (200mg/ml) and 1 mg/l kanamycine (25mg/ml). After that callus was transferred to regeneration media which contained MS with 1 mg/l IAA, 1 mg/l BA, 10 mg/l GA3, 1 mg/l cefatoxine (200mg/ml) and 250 µg/l kanamycine (25mg/ml) and their expression at the transcriptional level was confirmed by polymerase chain reaction (PCR) by usingchitinase and vector primers. After the transformed plants were evaluated, the positive transgenic plants were detected by using forward and reverse primer of kanamycine.

Keywords: Chitinase; Potato; Alternaria solani; early blight disease;Agrobacterium tumefaciens.


 

 

 

 

 

 

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