A Promising Approach of Hi- Tech Plant Breeding Int J Biol Sci. Department of Horticultural Science, Kyungpook National University, Daegu 7. Korea. 2. Institute of Horticultural Sciences, University of Agriculture, Faisalabad 3. Pakistan. 3. Department of Plant Science, and Plant Genomics and Breeding Institute, Seoul National University, Seoul 1. Korea. How. to cite this article. Younis A, Siddique MI, Kim CK, Lim KB. RNA Interference (RNAi) Induced Gene Silencing: A Promising Approach of Hi- Tech Plant Breeding. Int J Biol Sci 2. Available from. http: //www. Abstract. RNA interference (RNAi) is a promising gene regulatory approach in functional genomics that has significant impact on crop improvement which permits down- regulation in gene expression with greater precise manner without affecting the expression of other genes. RNAi mechanism is expedited by small molecules of interfering RNA to suppress a gene of interest effectively. RNAi has also been exploited in plants for resistance against pathogens, insect/pest, nematodes, and virus that cause significant economic losses. Keeping beside the significance in the genome integrity maintenance as well as growth and development, RNAi induced gene syntheses are vital in plant stress management. Modifying the genes by the interference of small RNAs is one of the ways through which plants react to the environmental stresses. Hence, investigating the role of small RNAs in regulating gene expression assists the researchers to explore the potentiality of small RNAs in abiotic and biotic stress management. This novel approach opens new avenues for crop improvement by developing disease resistant, abiotic or biotic stress tolerant, and high yielding elite varieties. Keywords: abiotic stress, biotic stress, crop improvement, functional genomics, post transcriptional gene silencing, si. RNA. Introduction. RNA interference (RNAi) is a biological mechanism which leads to post transcriptional gene silencing (PTGS) trigger by double stranded RNA (ds. RNA) molecules to prevent the expression of specific genes . RNAi mechanism has the potential in identification and functional assessment of thousands genes within any genome that is responsible for crop improvement. This promising approach also imparts its effective and efficient role to knock down the expression of any particular gene through short interfering RNA molecules in any target cell and moreover to assess the changes that occur in signaling pathways. Recently, RNAi has become a powerful and more reliable technique to inhibit the expression of targeted genes and also determine gene loss- of- function phenotype which leads to gene functional analysis when no mutant alleles are unavailable . RNAi technique was first time applied on Petunia hybrida L. New pattern of flower color in transgenic Petunia was observed due to overexpression of chs. A gene that encodes major enzymes in anthocyanin biosynthesis pathway . Several methods to induce RNAi, RNAi vectors, in vitro dicing and synthetic molecules are reported . Introduction of short pieces of double stranded RNA (ds. RNA) and small interfering RNA (si.
Climate: Jatropha is a highly adaptable species and is especially tolerant of severe heat. This plant thrives in warmer weather. The plant is susceptible to freeze damage but can tolerate a light frost of. Alkemade et al., Biodiversity Scenarios: Projections of 21st Century Change in Biodiversity and Associated Ecosystem Services, Convention on Biological Diversity, Montreal, Canada. Agrobacterium tumefaciens has the ability to transfer a portion of its DNA, T-DNA (transferred DNA), to the genome of a host plant ( Sheng and Citovsky, 1996). Gene transfer from Agrobacterium to plant cells. Disease Treatment and Prevention. The benefits for the treatment or prevention of disease or infection that may accrue from either dietary or topical administration of Moringa preparations (e.g. Plant biotechnology refers to the development of technologies based on biological systems to improve agricultural practices. The hallmark of green biotechnology is the genetic modification of crops in order to. Biotechnology Research International Volume 2011 (2011), Article ID 576286, 11 pages http://dx.doi.org/10.4061/2011/576286. RNA) into the cytosol, initiate the pathway culminating targeted degradation of the specific cellular m. RNA (Figure 1). During RNAi mechanism, silencing initiate with enzyme Dicer and ds. RNA is processed to convert the silencing trigger to ~2. RNAs (si. RNAs). The antisense strand of si. RNA become specific to endonuclease- protein complex, RNA- induced silencing complex (RISC), which then targets the homologous RNA and degrade it at specific site that results in the knock- down of protein expression (Figure 2) . Plant pathologists and plant biotechnologists have adopted different approaches to develop pathogen resistant genotypes but in last decade RNAi- induced gene silencing emerged as an effective tool to engineer pathogen resistant plants . This approach proved to be effective to create resistance against some diseases of economic importance caused by bacteria, fungi and viruses . Double stranded RNA (ds. RNA) act as an igniter in RNA interference and activate the homologous m. RNAs to inhibit its translation and transcription to silence the susceptible genes . This RNAi approach has opened new avenues in the development of eco- friendly techniques for plant improvement as specific genes are suppressed which cause stress and expression of novel genes for disease resistance. Figure 1. Model of Dicer and RNA- induced silencing complex (RISC) A. Dicer binding and cleaving ds. RNA (*Cleavage into precisely sized fragments is determined by the fact that one of the active sites in each Dicer protein is defective. Different colors show two separate molecules of Dicer). Reprinted by permission from Macmillan Publishers Ltd: Nature, 4. Click on the image to enlarge.) Figure 2. Diagrammatic representation of mechanism of RNAi. Silencing triggers in the form of double- stranded RNA presented in the cell as synthetic RNAs, replicating viruses or transcribed from nuclear genes. These are recognized and processed into small interfering RNAs by Dicer. The duplex si. RNAs are passed to RISC (RNA- induced silencing complex), and the complex becomes activated by unwinding of the duplex. Activated RISC complexes can regulate gene expression at many levels. Reprinted by permission from Macmillan Publishers Ltd: Nature, 4. Click on the image to enlarge.)Host gene silencing - hair pin RNAi (HGS- hp. RNAi) is reported as more stable gene silencing method in plants . This method can be employed to increase fungal and bacterial disease resistance by changing the gene expression against pathogens through genetic engineering in the host plant. Flagellin (a bacterial component) can stimulate the expression of specific mi. RNA to increase disease resistance signaling pathway in Arabidopsis . Over- expression of a gene At. FAAH in Arabidopsis, responsible for fatty acid (N- acylethanolamines) metabolism can alter phyto- hormone signals through intersecting with plant defense pathways to increase resistance against bacterial pathogens . In rice, RNAi can knockdown Os. SSI2 (Os. SSI2- kd), meant for fatty acid desaturase activity that cause increase resistance against bacterial pathogen (Xanthomonas oryzae pv. Oryzae) of leaf blight and blast fungus (Magnaporthe grisea) . Gene silencing can be obtained by host- induced gene (Avra. Triticum aestivum) and barley (Hordeum vulgare) through a transient gene expression resistant to RNAi because of silent point mutations. This suggests that the transfer of RNA from host plant to fungal pathogen Blumeria graminis, leads to RNAi- based plant protection against these pathogens . RNA silencing hosts target protein translation and process the virus- mediated ds. RNA, which results by pathogen replication into vsi. RNAs (virus- mediated si. RNAs). The vsi. RNAs then target and suppress gene expression and protein translation in the virus genes. For stabilization of the defense system, virus encodes 'viral suppressor of RNA silencing protein', that has been identified and isolated from various plant virus . Plants can also control viral diseases by RNAi and reveal resistance when having proper anti- sense or hairpin RNAi constructs . Although classical breeders have been developed various insect/pest resistant cultivars, however, this approach is tedious and time consuming as complexity increases with some added traits. The practice of using pesticides to control pests has become a common approach around the world, but having dramatic health and environmental effects its use seems to be very limited in coming years. Among transgenic approaches to control specific insect/pest, Bt- based toxins proved effective and replaced chemical insecticides in many crops. Most of the commercially used biotechnological approaches to control insect/pests on crops are subjected to expression of Bt insecticidal proteins which help in the permeabilization of gut epithelial cell's membrane in susceptible insects . However, this approach is limited for some specific crops to manage some specific pests, and there is also a threat that some insects can develop resistance against Bt . After the successful induction of transgene- encoded RNAi in plants . They further revealed the herbivorous insect RNAi efficiency can be stimulated by ingestion of transgenic ds. RNA producing plants that is gene- specific and proved effective against cotton bollworms (Helicoverpa armigera) damage. A gene 'CYP6. AE1. Helicoverpa armigera . This identified gene expressed in the insect midgut was correlated with larva growth when food contains gossypol. Therefore, after feeding on plant material exhibiting ds. RNA specific to gene 'CYP6. AE1. 4', the effect of the transcript decreased in midgut and larva growth also retarded . The gene silencing of 'glutathione- S- transferase' (GST1) can trigger RNAi process when herbivorous insects feed on plant material expressing ds. RNA . When ds. RNA was injected in whitefly body cavity, RNAi was induced that knocked- down the genes expression to 7. RNAi- mediated death of whitefly via oral dissemination of ds. RNA, targets ADP/ATP, translocase, a- tubulin, ribosomal protein L9 actin ortholog and v- ATPase. A genes responsible for insects mortality . They further revealed that transgenic corn plants which are engineered for WCR ds. RNAs expression exhibited a substantial reduction in insect damage that suggests RNAi pathway is effective and can be exploited further to control coleopteran insects . This mechanism was also practiced in Acyrthosiphon pisum for silencing of a gene C0. Later on, Pitino et al. The delivery of ds. RNAs from plant to nematode occurs by the ingestion process of plant cytoplasm and after its injection into the nematode body accelerate the RNAi that results in inactivation of targeted genes through ds. RNA . Nevertheless, to target host genes destined for interaction, efficient regulation of ds.
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