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Abstract
SUMMARY : This investigation aimed to induce useful mutations in Atropa belladonna L. plants.
Seeds of Atropa belladonna L. were subjected to four doses of Gamma-rays (50, 80,110 and 150 Gy) and parted to three parts as follows :-
The first part was carried out at Biotechnology Lab. Botany Dept. National Research Center to improve the germination percentage in M1 seeds (irradiated seeds with 50, 80, 110 and 150 Gy) of Atropa belladonna L. by using GA3. The gibbrillic acid concentrations were 20, 40, 60, 80, 100 and 120 ppm. There was an increase in seed germination percentage by increasing gamma-ray doses until 150 Gy which gave decrease in seed germination percentage, and with regard to the effect of GA3 on seed germination percentage, there was increasing in this percentage by increasing GA3 concentration until 100 ppm. The best interaction was between 110 Gy and 100 ppm which was the highest percentage. At, M2, a new mutation in seed colour (Red seeds) for almost mutants under study will be consider as a genetic solving of germination problems, that the germination percentage reached to 100%.
The second part was to induce mutations, this part was carried out at the greenhouse of Botany Dept. N.R.C, Egypt during 2003/2004, 2004/2005 and 2005/2006 at greenhouse of Genetics Dept., Fac. of Agriculture, Zagazig Univ. The gamma ray doses were 50, 80, 110 and 150 Gy. The mutants belong any apparent morphological characters change of plant height, no. of leaves, branching, position of flowers and large leaf area were screened and isolate 25 morphological apparent genotypes. Three promising high alkaloid mutants were selected from the previous mutants M-11-1, M-11-2 and M-15-1 respectivily. These promising mutants consider as a very important for production of high alkaloid contents, they possessed twice values than the control especially M-11-1 which gave the largest value (4.01 mg/g) at M2 generation as compared with control (2.03 mg/g). High alkaloid mutants at M2 generation were sown at M3 generation and possessed stable morphological criteria by comparison M2 generation. Molecular studies on these mutants were done for identification of them by ISSR technique confirm the difference between these mutants and control on the basis of molecular aspects. Each mutant of the three mutants can be distinguish by unique molecular markers, i.e. mutant M-11-1 can be distinguished by three molecular markers with molecular sizes 1397, 1149 and 874 bp. Mutant M-11-2can be distinguished by four molecular markers with molecular sizes 1537, 1075, 839 and 510 bp. Mutant M-15-1 can be distinguished by three molecular markers with molecular sizes 1749, 817and 756 bp. These findings confirm the true genetic variation between these mutants and mother genotype, as well as, it consider as primary study for finger printing of them.
The highest of total alkaloids appeared in almost mutants than the control, they possessed twice values than the control, especially M-11-1, which gave 4.01 mg/g and 109.67 mg/plant at M2 generation by comparison, 2.03 mg/g and 53.41 mg/plant for the control.
from the above results appeared the importance of high alkaloids mutants in the genetic improvement of alkaloid contents in Atropa belladonna L., therefore, should be molecular studies on these mutants will accure for identification of them and confirm of differences between these mutants and control on the basis molecular genetic marker.
The third part was carried out at Tissue Culture Lab. Botany Dept. National Research Center to induce callus of Atropa belladonna L. This achieved by using M1 plants with several concentrations of 2, 4-D. There was increase in callus induction frequency and callus fresh weight by increasing gamma-ray doses and also there were increase in callus induction frequency and callus fresh weight by increasing the concentration of 2, 4-D until 2.0 mg/l after that there was reduction. It can be concluded that the best interaction was between all gamma-ray doses and 2.0 mg/l 2, 4-D concentration and this drew the attention to the importance of the interaction between 2.0 mg/l 2, 4-D and different doses of gamma-ray..