Artificial seeds is A novel method for In Vitro production of Arabidopsis thaliana Nossen (NO-0) plants

Abstract

Somatic embryos are regarded the primary plant material for producing artificial seeds, and these seeds provide for agricultural institutions genetically uniform seeds, unlike the traditional seeds that often produce plants with different traits   from the parent plant; in addition to the possibility of storing it for a long time Arabidopsis thaliana, locally known as mouse ears, is a promising model because it is self-fertile, which makes it conserved, and can produce thousands of seeds, which makes it ideal for mutation experiments. It is diploid (10=2n), and the identification of selectable traits is more straightforward because it has a relatively small genome (125 Mpb). The genome organization in Arabidopsis thaliana is simple and particularly suitable for genetic and molecular biology experiments.. This study produced Arabidopsis thaliana Nossen (NO-0) plants from the differentiation of callus tissues from its growing seedlings on MS medium (roots, stems, leaves). The B5 medium free of growth regulators successfully induced root callus, while the highest response for callus induction from stem segments was observed on B5 medium containing 0.05 mgL-1 kinetin (Kin) and 0.5 mgL -1 2-4, Dichlorophenoxy acetic acid (2,4-D). The B5 medium containing 0.05 mgL-1 (Kin) kinetin and 1.0 mg/L-1 (2,4-D)  2,4-Dichlorophenxy acetic acid (2,4-D) also affected leaf callus induction. The results of the statistical program SPSS version 16 also showed a significant difference at a probability level of 1% in the percentage of calyx creation for the plant parts used .The MS medium containing 0.5 mgL-1 Thidiazuron (TDZ) and 1.0 mgL-1  6-Benzyl adenine (BA) was effective in producing embryonic phases from transferred root callus. Meanwhile, the embryonic phases from stem and leaf callus were produced on MS medium supplemented with 0.5 mgL-1 Thidiazuron (TDZ), resulting in a group of 290 shoots. A 2% sodium alginate concentration was used in experiments to encapsulate somatic embryos, yielding the best form of artificial seeds in spherical structures (Beads) and achieving a high conversion rate of 80% when stored for 20 days at 4°C, and 60% when stored for 20 days at 25°C. There are significant differences at the 5% level in the germination of seeds stored for 20 days at a temperature of 25°C. The number of plants resulting from the artificial seeds was 36 green shoots. The experiments were carried out over a period of six months in the Tissue Culture and Genetic Applications Laboratories at the College of Education for Pure Sciences/University of Mosul.