Abstract. Binary vectors are widely used in Agrobacterium gene transfer in plants.Although new plant expression vectors have been designed, pBI121-based vectors are still more common. The availability of fewer numbers of restriction enzyme sites is the most important drawback in pBI121. In this article, two new vectors-based on pB121, pBI121 5+1 , and pBI121 GUS-6 -were introduced. The construction of new vectors was confirmed by PCR and digestion pattern analysis. Furthermore, the reporter gene (gus) was cloned in these vectors. The T-DNA transformation ability of the new vectors and pBI121 (as the control sample) to tobacco via Agrobacterium tumefaciens strain LBA4404 was evaluated. Transgenic plants were regenerated with BAP and NAA in a selective medium. Thereafter, plant DNA was extracted and successful gene transformation was confirmed by PCR analysis. The GUS assay also confirmed gene expression in transgenic leaves. The results indicate that the vectors are totally efficient in plant transformation.

Clustered regularly interspaced short palindromic repeats (CRISPRs) technique is the most effective and novelist technique for genome editing. CRISPR mechanism has been widely developed for gene editing, gene silencing, high-specific regulation of the transcription, and reducing off-target effects through double-strand breaks (DSBs) in the genomic DNA and then modifying nucleotide sequences of the target gene in diverse plant and animal species. However, the application may be restricted by a high rate of off-target effects. So, there are many studies on designing precise single-guide RNAs (sgRNAs) to minimize off-target effects. Thus, the high-efficiency design of a specific sgRNA is critical. First, in the chapter, the sgRNA origin and different types of gRNA will be outlined. Then, the off-target effect will be described. Next, the remarkable characteristics of the sgRNA will be highlighted to improve precise gene editing. Finally, some popular in silico tools will be introduced for designing sgRNA.

Nanoparticles recently play remarkable roles in modern agriculture and biotechnology due to specific exclusively physicochemical and biological characteristics of the particles. In recent years, nanoparticles are been using as fertilizers and elicitors to improve crops. Nutritionists are constantly seeking natural antioxidants without side effects to using for healthcare and treatment. Asparagus officinalis L. as medicinal plant treated by iron (0, 10, 50 and 100 mg/L) and selenium (0, 0.5,and 1 mg/L) nanoparticles as nano elicitors. Then the antioxidant capacity of A. officinalis L. was detected and measured by α, α-diphenyl-β-picrylhydrazyl (DPPH) assay, for assessment of the antioxidant activity. The iron nanoparticlesconcentration significantly increases the antioxidant activity of both male and female asparagus stem, as well as selenium nanoparticles. When combined iron and selenium used as nano elicitors then cause the antioxidant activity significantly decreases. But the integration of two nano elicitors (iron and selenium) decreased antioxidant capacity while the use of nano-selenium could enhance antioxidant capacity. The application of nano elicitor increased antioxidant capacity in the female stem than male.Nanoparticles recently play remarkable roles in modern agriculture and biotechnology due to specific exclusively physicochemical and biological characteristics of the particles. In recent years, nanoparticles are been using as fertilizers and elicitors to improve crops. Nutritionists are constantly seeking natural antioxidants without side effects to using for healthcare and treatment. Asparagus officinalis L. as medicinal plant treated by iron (0, 10, 50 and 100 mg/L) and selenium (0, 0.5,and 1 mg/L) nanoparticles as nano elicitors. Then the antioxidant capacity of A. officinalis L. was detected and measured by α, α-diphenyl-β-picrylhydrazyl (DPPH) assay, for assessment of the antioxidant activity. The iron nanoparticlesconcentration significantly increases the antioxidant activity of both male and female asparagus stem, as well as selenium nanoparticles. When combined iron and selenium used as nano elicitors then cause the antioxidant activity significantly decreases. But the integration of two nano elicitors (iron and selenium) decreased antioxidant capacity while the use of nano-selenium could enhance antioxidant capacity. The application of nano elicitor increased antioxidant capacity in the female stem than male.