Essam A. Zaki

Cloning and sequencing of an envelope-like gene in Gossypium.

Abstract. Eukaryotic genomes harbor mobile genetic elements known as long terminal repeat (LTR) retrotransposons. LTR retrotransposons are closely related to the infectious and endogenous retroviruses. The viral envelope (env) gene of the retroviruses, which is responsible for their infective properties, distinguishes them from the LTR retrotransposons. Here, we report the cloning and sequencing of an envelope-like gene in Gossypium, implying that enveloped retroviruses are not limited to animals.

DNA Sequences of RAPD Fragments in the Egyptian cotton Gossypium barbadense

Random Amplified Polymorphic DNAs (RAPDs) is a DNA polymorphism assay based on the amplification of random DNA segments with single primers of arbitrary nucleotide sequence. Despite the fact that the RAPD technique has become a very powerful tool and has found use in numerous applications, yet, the nature of molecular variation(s) uncovered by the RAPD technique is still unclear. The aim of the following study, therefore, was to investigate the molecular nature of RAPD DNA fragments in four Gossypium barbadense cultivars. Five RAPD DNA fragments, generated by improved RAPD-PCR technique, and representing polymorphic and nonpolymorphic bands were analyzed at the molecular level using DNA sequence analysis. Nonpolymorphic RAPD DNA fragments showed homologies to previously characterized plant structural genes. Comparative nucleotide sequence analysis of two comigrating nonpolymorphic fragments revealed that these two DNA sequences are highly similar to each other, indicating that similarity of fragment size is a good predicator of homology. Polymorphic RAPD DNA fragments, on the other hand, showed homologies to middle and high-repetitive DNA sequences. These results promote the initiative to integrate these RAPD markers in cotton breeding applications, and DNA fingerprinting. (African Journal of Biotechnology: 2003 2(5): 129-132)

Molecular distribution of gypsy-like retrotransposons in cotton Gossypium Spp.

PCR primers specific for conserved domains of the reverse transcriptase (RT) genes of gypsy-like retrotransposons amplified their corresponding gene in two Gossypium barbadense cultivars. Analysis with the FASTA software showed a high DNA sequence homology to pine, gypsy LTR-retrotransposon. Using the PCR product as a hybridization probe, gypsy-like retrotransposons were detected in wild type species of Gossypium, suggesting that gypsy-like retrotransposons are present in the Gossypium genome. This supports the view that gypsy-like retrotransposons are major components of plant genomes. Our results suggest gypsy-like retrotransposons have played a fundamental role in the shaping and evolution of the Gossypium genome.

Sequence heterogeneity of the envelope-like domain in cultivated allotetraploid Gossypium species and their diploid progenitors

Retroviralenvelope (env)-like sequences in 2 cultivated allotetraploid cottons and their diploid progenitors have been identified and characterized in this study. DNA sequence analysis reveals that these sequences are heterogeneous. The observed sequence diversity, however, seems to preserve coding information. This is evidenced by the detection of the transmembrane domain (TM), which is the most conserved feature of the divergent retroviral env genes. The high ratio of synonymous to nonsynonymous changes suggests that these sequences are evolving under purifying selection. Phylogenetic analysis shows thatGossypium sequences closely cluster with a lineage of plant endogenous retroviruses that have anenv-like gene. These results provide evidence for the antiquity and the wide diversity ofenv-like sequences in theGossypium genome.

Phylogenetic and molecular evolutionary analyses of gypsy group retrotransposon families in the Egyptian cotton Gossypium barbadense

Gypsy group retrotransposons in the Egyptian cotton, Gossypium barbadense, was examined by phylogenetic and molecular evolutionary analyses. DNA sequences of gypsy group retrotransposons in two G. barbadense cultivars revealed that these sequences are heterogeneous and represent two distinct families. Sequence variation between these families seems to preserve coding information of the reverse transcriptase domain. The high ratio of synonymous to nonsynonymous changes indicates that the reverse transcriptase domain of these families is evolving under purifying selection. Our phylogenetic analysis revealed that the closest relatives of cotton retroelements are found in other plants gypsy group retrotransposons. Cotton retroelements-encoded transcripts were detected in their related respective young seedlings using RNA slot-blot hybridization, suggesting their transcriptional activity. The wide distribution of gypsy group retrotransposons and the detection of their encoded transcripts illustrate their active role in the Gossypium genome. Key words : Evolution, Gossypium, gypsy, retrotransposons, reverse transcriptase, substitution rates, transcription. African Journal of Biotechnology Vol.2(9) 2003: 271-275

Isolation, characterization, and phylogenetic analysis of copia-like retrotransposons in the Egyptian cotton Gossypium barbadense and its progenitors

We have used the polymerase chain reaction to analyze copia-like retrotransposons in the Egyptian cotton and its progenitors. All three cotton species studied contain reverse transcriptase fragments from copia-like retrotransposons. Sequence analysis of these reverse transcriptase fragments reveals that each is different from the others, with predicted amino acid diversities between 9 and 94%. The detection of stop codons and insertions/deletions in the derived amino acid sequences of the Gossypium RT clones, suggests that these clones represent defective retrotransposons. The presence of these sequences in G. barbadense progenitors, however, suggests the presence of active retrotransposons capable of producing new functional copies at an appropriate rate to compensate for the mutational loss of old ones. Phylogenetic analysis provided strong bootstrap support for a monophyletic origin of plant copia-like retrotransposons, yet showed high diversity within all species. Our results suggest that both vertical transmission of copia-like retrotransposons within G. barbadense lineages, and horizontal transmission between G. barbadense and its progenitors have played major roles in the evolution of copia-like retrotransposons in Gossypium. Keywords: Genome structure, Gossypium, repetitive DNA, polyploidy, sequence diversity, retrotransposons. (African Journal of Biotechnology: 2003 2(6): 165-168)