African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12368

Review

Advances on the application of non-coding RNA in crop improvement

Diriba Guta Dekeba
  • Diriba Guta Dekeba
  • Ethiopian Institute of Agriculture Research, National Agricultural Biotechnology Research Center, Holeta, Oromia, Ethiopia.
  • Google Scholar


  •  Received: 14 April 2021
  •  Accepted: 26 July 2021
  •  Published: 30 November 2021

 ABSTRACT

Thanks to committed plant breeding researchers over the few decades, many problems associated with food supply and qualities have been improved. Food security is an exceptionally serious worldwide issue via world climate change, the increase in human population, and the use of plants for bioethanol production in current years. Improved tolerance to abiotic and biotic stress, resistance to herbicides, improved yield, and plants with wonderful nutritional value are essential goals of crop improvement. RNAi applications are the modern innovation that can assist in the solution for these issues. A natural protection mechanism against invading viruses, nucleic acids, and transposons non-coding RNAs (ncRNAs), are identified as effector molecules in RNA-mediated gene silencing and used in the genetic modification of crops. These ncRNAs are concerned with the regulation of growth, development, and response to stress at the transcriptional and translational levels. Improving crop yields is the final purpose of molecular plant breeding. ncRNAs, along with transfer RNAs (tRNAs) and ribosomal RNAs (rRNAs), as well as small non-coding RNAs (sncRNAs) and the long non-coding RNAs (lncRNAs), have been recognized as essential regulators of gene expression in plants in plant immunity and adaptation to abiotic and abiotic stages biotic stress.

 

Key words: Crop improvement, long RNA, ncRNA, RNA interference, small RNA.


 INTRODUCTION

The world population is growing rapidly; however, world food security is still threatened in recent years via climate change, the increase in human population and the use of plants for bioethanol production in recent years (FAO, 2020). In addition, the future capacity to meet the world's food security needs has come to be unstable as the area of  arable   and   cultivated  land  continues  to  decrease. These issues and threats have led scientists to look for options to increase crop productivity. Proponents of new technologies, which include recombinant DNA, have a promise in current green transformation, with genetically engineered plants which include transgenes achieving focused traits (Wheeler and von Braun, 2013; Sang and Chanseok, 2016; Bader et al., 2020; Mezzetti et al., 2020).

 

Non-coding RNAs refer to transcripts that no longer code for proteins; however play vital regulatory roles within the cell, which precludes the possibility of the production of exogenous protein products. Non-coding RNA can be a gene silencing phenomenon that involves sequence-specific gene legislation encapsulated via double-stranded RNA, resulting in inhibition of protein production (Zheng and Qu, 2015; Brant and Budak, 2018; Yan et al., 2020, 2021).

 

Some classes of regulatory ncRNAs especially regulate a single goal gene, while others modulate more than one gene at the genome-wide level via various molecular mechanisms. These cumulative consequences advise that regulatory ncRNAs may want to be possible goals for molecular plant breeding (Zhou and Luo, 2013).

 

Non-coding RNAs are divided into functionally vital RNAs such as transfer RNA (tRNA), ribosomal RNA (rRNA), small non-coding RNAs, including Micro-RNAs (miRNAs), small interfering RNAs (siRNAs), piwi-interacting RNAs (piRNAs), and the long non-coding RNAs (lncRNAs). Two lncRNAs that have been substantially studied are the X-inactivation-specific transcript (XIST) and the HOX-antisense-intergenic RNA (HOTAIR) (Renyi and Jiankang, 2014; Santosh et al., 2014; Brant and Budak, 2018).

 

Post-transcriptional gene silencing (PTGS) is mentioned in many organisms; fungi, animals, and plants (Zhou and Luo, 2013; Brant and Budak, 2018). These products are the result of non-coding dsRNA, the DICER or Dicer-like enzyme, which performs this cleavage. Small non-coding RNA consists of an RNA-induced silencing complex (RISC) and Argonaute proteins (AGOs). Drosha and Pasha are part of the microprocessor protein complex. Drosha and Dicer are RNase III enzymes, Pasha is a dsRNA-binding protein, while Argonauts are RNase H enzymes (Wilson and Doundna, 2013; Kamthan et al., 2015; Brant and Budak, 2018).

 

The goals of this review had been to discuss non-coding RNA and its use in plant improvement. The RNA interferences in the adaptation of plants to the environment serve the tolerance to abiotic stress and the plant protection towards biotic stress. The RNA interference increases the yield potential of plants, such as manipulating plant improvement and improving nutrition. They modulate a broad range of gene regulatory networks via regulating a specific subset of downstream genes that are closely associated to agricultural traits such as seed maturation, flower development, pathogen resistance, and other abiotic stress resistance.

 

Natures of non-coding RNAs (ncRNAs)

 

Ribosomal RNA (rRNA)

 

Ribosomal RNA is the  most  abundant  small  non-coding RNA molecule that is made up of 4-10% cellular RNA. They are translation machines in so far as they translate the genetic information into a corresponding polypeptide chain in a mRNA template-directed manner (Rodnina and Winter Meyer, 2011). The mRNA sequence starts with the begin codon of AUG and is followed by termination codon signaling for subsequent folding into its functional state (Crick, 1970). The ribosomes in eukaryotes are more complex than in prokaryotic ones; in general, ribosomes have three exclusive binding sites; A (aminoacyl) site, the P (peptidyl) site and the E (parent) site in the interface between the subunits. The mRNA that binds to the 30S subunit can regularly move one codon at a time during peptide elongation (Crick, 1970).

 

The ribosomal RNA is about 60% by weight rRNA and 40% by weight protein and additionally includes two most important rRNAs and 50 or more proteins. The 60S subunit incorporates (three rRNAs 5S, 5.8S, 28S and about forty proteins) and the 40S subunit contains (an18S rRNA and about 30 proteins). The LSU rRNA acts as a ribozyme and catalyzes the formation of peptide bonds (Anita et al., 2013).

 

Transfer RNA (tRNA)

 

Transfer RNA is the link between mRNA and the peptide sequence in transfer RNA (tRNA). There is an enzyme that can understand both tRNA and an anticodon, which is complementary to the mRNA codon associated to this amino acid, and couples it to a highly standardized free energy complex known as aminoacyl-tRNA at the rate of ATP hydrolysis (Sharp at el., 1985). The mRNA binds to the 40s ribosomal subunit, followed by using the binding of the initiator tRNA loaded with formylated methionine to the P site in a reaction step that is considerably accelerated by the three initiation factors. As soon as the mRNA and the initiator tRNA are successfully bound, the large 60S subunit binds to form an 80s initiation complex, releasing the eIF factors. With the mRNA in the correct reading frame, the initiator tRNA in the P-site and the empty A-site programmed with the first interior codon of the protein to be synthesized. The ribosome has now left the initiation phase and has entered the peptide elongation phase (Valle et al., 2003).

 

Short non-coding RNAs

 

Micro RNA (miRNA)

 

Micro-RNA is the party of the small non-coding RNA (20-22nt) with partially double-stranded stem loop structures, which regulates negative gene expression in both animals and plants. It is transcribed by the RNA polymerase II enzyme and, after transcription, cleaved by the Dicer-like 1 (DCL1) enzyme and transported into the cytoplasm and incorporated into the Argonaut (AGO) protein. Mature miRNA is produced in various processes and incorporated into the protein RISC (RNA-induced Stille complex). Mature single miRNA that incorporates the RISC protein binds with other complementary mRNA sequences to produce the protein (Khraiwesh et al., 2012).

 

Small interfering RNA (siRNA)

 

Small interfering RNAs are the parts of small non-coding RNA that have the function of facilitating post-transcriptional gene silencing (PTGS) by breaking down mRNA. Small interfering RNA is generated from RNA double strands that are nearly perfectly complementary and cleaved by DCL2, DCL3 and DCL4 to produce siRNA duplexes 22nt, 24nt and 21nt, respectively (Sang and Chanseok, 2016). The other cofactors of siRNA are the RNA-dependent RNA polymerases 2 and 6 (RDR2, RDR6), SUPPRESSOR OF GENE SILENCING 3 (SGS3) and the plant-specific DNA-dependent RNA polymerases IV and V (Bologna and Voinnet, 2014). After the siRNAs have been cleaved by DCL, they are loaded into AGRONAUTS (AGOs) and companies with a complexing gene silence machinery called RISC (RNA Induced Silence Complex). The RISC recognizes the complementary sequence of mRNA and AGO-loaded small RNA (also known as 'guide RNA'), then RISC suppresses the target mRNA via a cleavage or transcription gene silencing (TGS) mechanism. Small, interfering RNA is naturally very important in the organism to protect it from various natural enemies. For example, RNA interference (RNAi) was seen as a natural defense mechanism that used exogenous siRNAs to protect organisms from viruses, but it soon became clear that endogenous siRNAs (endo-siRNAs) also play a role in regulating genomic functions (Kamthan et al., 2015).

 

Piwi-interacting RNAs (piRNA)

 

Piwi-interacting RNAs are small non-coding RNAs that have 20-31nt. The name, piRNA (Piwi-interacting RNA), reflects the fact that piRNAs bind to Piwi proteins under physiological conditions. Piwi-interacting RNAs were first discovered in Drosophila as repeat-associated siRNAs (rasiRNA), which show complementarities to a variety of transposable and repetitive elements (Minna et al., 2011).

 

The main function of these RNA molecules involves chromatin regulation and suppression of transposon activity in germ-line and somatic cells. piRNAs are antisense to expressed transposons target and cleave the transposon in complexes with  piwi  proteins.  This cleavage generates additional piRNAs which target and cleave additional transposons. This cycle continues to produce an abundance of piRNAs and augment transposon silencing (Lindsay et al., 2013).

 

Long non-coding RNA (lncRNA)

 

LncRNAs are transcripts generally longer than 200nt having little or no potential of encoding proteins. Most of the lncRNAs have similar characteristics within the mRNA, transcribed by RNA polymerase II. In addition to RNA Pol II-derived lncRNAs, there are other classes of lncRNAs transcribed by two plant-specific DNA-dependent RNA polymerases, RNA Pol IV and RNA Pol V, which play critical roles in transcriptional gene silencing mediated by RNA-dependent DNA methylation (RdDM) (Sang and Chanseok, 2016).

 

LncRNA can be divided based on genomic location; sense lncRNAs overlap with one or more exons of a transcript on the same strand; antisense lncRNAs overlap with one or more exons of a transcript on the opposite strand; intronic lncRNAs derive from an intron within another transcript; and intergenic lncRNAs occur in the interval between two genes on the same strand (Ma et al., 2013).

 

Although only a small number of lncRNAs are identified and functionally investigated until now, plant lncRNAs play important roles as regulators in complex gene regulatory networks involved in plant development and stress management (Zhang et al., 2014). The human genome has only 3% of the coding region but more than 85% of the genome is actively transcribed and the biggest challenge is the understanding of the functional role for these transcripts (Hangauer et al., 2013).

 

The lncRNA regulates the expression levels of target genes ranging from transcription to translation. Some examples of their functions are lncRNA, called LDMAR (for long-day specific male-fertility-associated RNA), which regulates the photoperiod-sensitive male sterility of the rice variety whose pollen is completely sterile in a long-day condition. Genome-wide investigation on rice also identified a set of lncRNAs that are specifically expressed during the reproduction stage (Zhang et al., 2014). On the Flowering Locus C (FLC) in Arabidopsis, an antisense lncRNA, COOLAIR, and a sense lncRNA, COLD AIR, are produced during vernalization to form an epigenetic switch for silencing the expression of the FLC gene to promote flowering (Fatica and Bozzoni, 2014, Renyi and Jiankang, 2014).


 APPLICATION OF RNAI IN CROP IMPROVEMENT

Crop yield improvement is the ultimate goal of molecular crop breeding. There are multiple physiological traits influencing crop yield, and several studies have found ncRNAs and their target genes to be involved in those traits. Some physiological traits which influence crop yield are plant manipulation, abiotic stress, biotic stress, and improvement in fruit quality, quantity, and nutritional value.

 

Plant architecture

 

Biomass

 

RNA interferences are one application that can improve the yield of crops and fruit vegetation via the manipulation of primary agronomic traits. It can increase the biomass of the crops and fruits by manipulation of plant height, prolonged vegetative segment and delayed flowering time, a number of branches or branching type, and measurement of the plants.

 

RNA interference knocks down the OsDWARF4 gene in rice to limit the plant's erect leaf and growing the photosynthesis through reducing leaves. The yield of such plants is improved or increased via dense planting conditions (Feldmann, 2006). Over-expression of the maize MIR156, Corngrass1 (Cg1) gene (Chuck et al., 2011), and red clover (Trifolium pratense L.) (Zheng et al., 2016) motives prolong the vegetative parts of maize and delayed flowering. This means that the reproductive stage is delayed, the vegetative parts are increased in size, and at the identical time, biomass of plants become increasing.

 

Moderate and low ranges of miR156 expression had 58–101% more biomass production than wild-type controls because of increases in tiller numbers in change grass plants. Over-expression of rice miR156 should improve the yield of solubilized sugar as properly as forage digestibility (Xie et al., 2012, Johnson, 2017), suspending the flowering time in the Arabidopsis (Roussin et al., 2020), and it can manipulate the white and lily flowering colors (Yamagishi and Sakai, 2020).

 

Grain yield

 

Improvements of the plants by way of special technology or techniques are mainly for increasing the yield of the crops. RNA interferences are one of the technologies which are used to improve the crop's yield by using manipulating traits. One of the functions of RNA interferences is the manipulation of the gene, which affects the characteristics of the crop whether to be increasing or reducing the crop yield.

 

Regulatory ncRNAs influence the reproductive stage. It is a very important aspect that affects crop yield and is used for genomic association. According to Ding et al. (2019), lncRNA, called LDMAR (for LONG-DAYSPECIFIC MALE-FERTILITY-ASSOCIATED RNA), regulates the photoperiod sensitive male sterility of rice with absolutely sterile pollen in a long-day condition. Genome-wide investigation on rice additionally identified a set of lncRNAs that are particularly expressed during the reproduction stage (Zhang et al., 2014).

 

The RNA mediates suppression of GA 20-oxidase (OsGA20ox2) gene which resulted in semi-dwarf plant life from a taller rice variety QX1. It increases panicle length, the quantity of seeds per panicle, and greater seed weight. OsSPL14 (Souamosa promoter binding protein-like14) is the goal of Osa-miR156 in rice to increase the yield of the rice grain, reduced tiller number, and accelerated grain yield (Wang et al., 2012; Jiao et al., 2010). Overexpression of Osa?miR1873 also resulted in some defects in yield traits, which include grain numbers and seed putting rate in rice (Zhou et al., 2020). Overexpression of the OsAGO17 gene was once also found to be involved in increased grain size, weight, and promote stem development in rice (Zhong et al., 2020).

 

Fruit improvement

 

The application of transgenic protects crops from different pests that can affect the plant products. Genetic engineering can be applied to improve the composition and quality of the harvested organs to reduce post-harvest deterioration of fruits or increase agronomic quality and nutritional value. Fruits are consumed when fresh because they are very sensitive to pests and are perishable. RNA interference is the technology that can overcome this problem (Meli et al., 2010).

 

Enhanced nutritional value and edibility

 

RNA silencing-based technology has been used in improving the nutritional value of crops. By down-regulating key genes in plant metabolic pathways usage of RNAi constructs, transgenic plants may also accumulate more favorable metabolites or produce fewer undesirable ingredients. Crops have exclusive nutritional values; some of them are allergens to human being, some pollutants to the environments, and some allergens to the different crops. But there are plants which have very essential nutritional values that are consumed by means of human beings or animals. RNA interference is to manipulate all these nutrients through adding the essential one and removing the unwanted ones.

 

Tomato (Lycopersicon esculentum) is one of the most economically essential fruit plants throughout the world rich in antioxidants, minerals, fibers, and vitamins. RNAi has been utilized in the development of tomato fruit with an improved level of carotenoids and flavonoids which are especially beneficial for human health (Kamthan et al., 2015).

 

RNAi approach has additionally been used in apple to improve the fruit quality via enhancing self-life and reducing the quantity of a major apple allergen, metabolites, and accumulation of sugars in the fruits through sorbitol synthesis, which impacts fruit starch accumulation (Romer et al., 2020) and sugar-acid stability (Teo et al., 2006).

 

Some examples of nutrition adding plants through RNAi are high lysine maize for the expression of zein proteins (Li and Song, 2020; Choudhary et al., 2021), and silencing of carotenoid β-hydroxylases which increases the β-carotene content of maize (Berman et al., 2017).

 

RNAi has also been used to down-regulate the starch-branching enzyme resulting in high- amylose wheat, which has an amazing potential to improve human health (Man and Hong, 2013). Corn with increased essential amino acids (Hasan and Rima, 2021), improved soybean by using oil quality (Yang et al., 2018), and cotton with improved fatty acid composition have been developed with the use of RNAi. Overexpression of GmPDAT genes elevated seed size and oil content (Gao et al., 2020), whereas RNAi strains had reduced seed size and oil content of soybean (Liu et al., 2020). Suppression of three carotenoid-cleavage dioxygenase genes, OsCCD1, 4a, and 4b, increases carotenoid content in rice (Ko et al., 2018).

 

Enhanced shelf life

 

The fundamental issues of fruits are post-harvest deterioration and spoilage. It is a main financial loss. This may be because of some issues like; technique of harvesting, transports, and storage. Therefore, an increase in the shelf life of vegetables and fruits through delayed ripening involves any other critical agronomic trait that is being addressed through non-coding RNA technology. Initiation of ripening in climacteric fruits like tomato is characterized through a climacteric burst of ethylene, resulting in the regulation of the expression of ripening-specific genes. Manipulating the gene which is accountable to produce ethylene and decrease the production of ethylene was once increasing the shelf life of tomato (Osorio et al., 2011). RNA interference decreases the expression of 1-aminocyclopropane-1-carboxylate (ACC) oxidase, a gene of the ethylene biosynthesis pathway in tomato, and inhibited the ethylene production at the ripening time so that the fruit can survive a long time.

 

RNA silencing used to be first genetically modified in the Flavr Savr tomato, bringing about the antisense of transcript polygalacturonase (PG) which is suppressed to PG expression. The polygalacturonase is accountable for cell wall degradation during tomato ripening. Suppression of polygalacturonase delayed the natural softening of tomatoes and allowed tomatoes to ripen on the vine for long and resulting in a greater flavorful fruit (Renyi and Jiankang, 2014). The increase in shelf life of tomatoes used to be observed after increasing abscisic acid (ABA) content. Manipulation of β?carotene stages results in an enchantment that is no longer only limited to the shelf life of tomatoes, but also their nutritional value (Diretto et al., 2020).

 

Tomato fruits are considered to be climacteric and require the gaseous hormone ethylene to ripen. In every case, the change of the expression of the mi156 gene in tomatoes led to an increase in yield and shelf life (Zhang et al., 2011). Definitely, tomato has emerged as the pre-eminent experimental model for studying fleshy fruit, which includes the developmental control of ripening, ethylene synthesis and perception. Overexpression of Pti4, Pti5, and Pti6 genes (Wang et al., 2021), and SlGRAS4 gene (Liu et al., 2021a) in tomato are very crucial to accelerated fruit ripening and elevated the complete carotenoid content.

 

Non-coding RNA is especially primary within the increased shelf life of vegetables and fruits. Tobacco (Moreno et al., 2020), and tomatoes (Arefin et al., 2020), superoxide dismutase (SOD) genes expressed at some stage in the ripening of apple fruit (Lv et al., 2020), cold storage responses genes in the peach (Antonella et al., 2020), blueberry cultivars with higher fruit firmness and longer shelf lifestyles (Liu et al., 2021), preserved post-harvest shelf life and quality of banana fruit (Yumbya et al., 2021) have been developed using RNAi. RNAi method concentrated on suppression of more than one homolog would be much effective than the knockdown of a single homolog (Gupta et al., 2013).

 

Seedless fruit development (Parthenocarpy)

 

Parthenocarpy or seedless fruit improvement is a technique of fruit production from seedless crops by way of ovaries without pollination and fertilization. Parthenocarpy or seedlessness is a highly preferred agronomic trait, particularly in safe to eat fruit crops. It produces high yields in harsh environmental conditions, as no pollination or fertilization is required. Customers constantly want fresh fruit, while it is possible to produce the fruit at any time of the year. It produces excessive yields in harsh environmental conditions, as no pollination or fertilization is required. Customers continually want clean fruit, while it is possible to produce the fruit at any time of the year.

 

In some cases, fruits produce hard seed which makes it difficult to overcome the dormancy of the seed, whereas others produce seeds with the difficult tests, as the absence of seeds can also be a positive trait for both direct clean consumption (e.g. grape, citrus, and banana) and industrial approaches (e.g. frozen eggplants, and tomato sauce) (Meli et al., 2010).

 

RNA interferers are one of the most essential primary technologies for overcoming the issues of vegetation with parthenocarpy. Some plants can move forward through RNAi innovation like; manipulation of auxin response factors8 (ARF8), which is the goal of miR167 to affect product from parthenocarpic fruits in each arabidopsis and tomatoes (Molesini, et al., 2012). Suppression of the orthologous genes of Pad-1 caused parthenocarpic fruit improvement in Solanaceae plants; hence, it is a very powerful tool in improving Solanaceae fruit production (Matsuo et al., 2020).

 

The overexpression of the PbGA20ox2 gene modified the GA biosynthetic pathway and improved GA4 synthesis, which promoted fruit set and parthenocarpic fruit improvement in pear fruits (Pyrus Bretschneider Rehd.) (Wang et al., 2020a). The PpIAA19 gene used to be concerned in the regulation of lateral root number, stem elongation, parthenocarpy, and fruit structure of tomatoes (Ding et al., 2019) and grapevines (Wang et al., 2020a). The overexpression of the MaTPD1A gene in banana plants produces seedless fruits in contrast to wild-type plants (Hu et al., 2020).

 

Biotic stress resistance

 

Phyto-pathogens are disease causing agents that can affect crops and crop products. The disease may affect the crop at different stages, seedling, vegetative, and at productive stages. RNAi strategies have been employed to improve the small RNA-mediated crop improvement defense mechanism in crop plants against various biotic stresses including an attack by viruses, bacteria, fungi, nematodes, and insects (Park and Shin, 2015).

 

Virus resistance

 

Viruses are pathogens that can influence crop products. RNA interference is one of the technologies which can overcome their effect. The functions of RNA interferences are protection mechanism of virus that invades nucleic acid molecules. Transgenic plant technology has been used to express genes encoding viral coat proteins in transgenic plants and can be resistant to viruses containing the coat proteins. The virus resistance was primarily based on the recognition of coat proteins through the transgenic plant cells and therefore on the plant immune response against the coat proteins. However, for understanding the application of genetic transformation technology, the plants can express the coat protein gene transcript and damage the protein coat; however, it cannot produce the protein that acts against the protein coat itself. Transgenic plants that are resistant to viruses have been produced by way of sense transgene-triggered RNAi against the viral RNAs (Ding, 2010; Renyi and Jian-Kang, 2014; Bahadur et al., 2015; Uslu and Wassenegger, 2020).

 

RNAi has shown a way to keep virus-resistant traits in many crops. The transgenic plant can produce the viral sequences that match coat proteins, replication-associated proteins, ATPases, or promoter areas in the viral genomes that can derive transgenic siRNAs to goal viral RNAs for degradation upon infection. Plants reportedly consisting of virus resistance included some suggested crops like papaya (Kertbundit et al., 2007), cassava (Vanitharani et al., 2004; Vanderschuren et al., 2012) and potato (Sajid et al., 2019). Some examples are potato resistance to spindle tuber viroid (PSTVd) and cassava resistance to African Cassava Mosaic Virus (ACMV) (Renyi and Jiankang, 2014). The virus prevention strategies are based totally on the ecological implication that increased carbon dioxide concentrations minimize the accumulation of the cucumber mosaic virus in Nicotiana tabacum through the viral suppressor of the RNAi (VSR) 2b protein of CMV (Guo et al., 2021).

 

Bacterial resistance

 

Bacterial diseases are extremely hard to manage due to the high rate of production of the spread. The multiplication rates of microorganism are very excessive in contrast to plant production, due to which, it can effortlessly manage plant development. RNA interference application technologies are used in various facilities to resist bacterial diseases. Some plants that can withstand RNA interference are; In Arabidopsis, it was reported that miR393 is caused through a bacterial pattern-associated molecular pattern (PAMP) peptide flg22 and negatively regulates the F-box auxin receptors TIR1, AFB2, and AFB3. This suppression of auxin signaling contributed positively to the predicament of bacterial infection (Pseudomonas syringae) (Li et al., 2010; Zhang et al., 2011a). The other application RNAi mediated the suppression of two genes from Agrobacterium tumefaciens that are involved in the formation of crown gall tumors (iaaM and ipt), which could appreciably minimize tumor production in Arabidopsis (Kamthan et al., 2015).

 

Overexpression of the rice chorismate mutase (OsCM) gene modified the downstream pathway of the aromatic amino acids, whereby the stress of bacterial leaf rot (BLB) was weakened through changing stress-sensitive genes and hormonal accumulation (Jan et al., 2020). Some of the well-reported plants that showed bacteria can be resisted through manipulating RNAi genes include tomatoes (Bento et al., 2020), citrus (Yu and Killiny, 2020), soybean (Tian et al., 2020), and Arabidopsis (Guo et al., 2020).

 

Fungal resistance

 

Genetic engineering based on RNA silencing has made a primary contribution to improving crops, that is, resistance to pathogens. Fungus is a pathogen that can affect plant production and quality. RNAi has been confirmed to be an essential strategy for producing tolerance in various crops (Kamthan et al., 2015).

 

RNA interference is done in response to fungal attack in wheat Blumeria graminis f. Sp. tritici (Bgt) through the use of the 24 miRNAs gene that causes devastating diseases of wheat powdery mildew (Xin et al., 2010). Transgenic overexpression of rice plants through OSA-miR7695 (Campo et al., 2013), miR160a or miR398b (Li et al., 2014), miR169 (Li et al., 2017) and OsamiR167d (Zhao et al., 2020) has been negatively regulated as a response to the blast fungus Magnaportheoryzae, and through overexpression of these genes, an improved resistance against rice blast infections was achieved.

 

Treating of the Fusarium species with various dsRNAs that concentrated on the genes was destructive to the fungus and leads to determination of an extended growth in in vitro cultures (Koch et al., 2018). In the reduction of the Arabidopsis thaliana line, it was observed that the activity of miR396 confers resistance to necrotrophic and hemibiotrophic fungal pathogens using artificial miRNA goal mimetics (Soto-Suárez et al., 2017). Overexpression of the BnaNPR1 gene in oilseed rape played a high-quality role in the resistance of Brassica napus to Sclerotinia sclerotiorum, which confers resistance to this pathogen (Wang et al., 2020b).

 

Insect and resistance

 

Defoliation of plants or sucking out their sap insects can cause great damage to plants by slowing, weakening, and sometimes killing their increase (Bahadur et al., 2015). The use of RNAi has been confirmed to be an essential approach for developing tolerance in various plants. Insect-resistant plants produce the dsRNA that attacks the insects, and when insects ingest, the gene expresses itself (Huvenne and Smagghe, 2010).

 

This technology is intended to protect insects from various kinds of plant production; among others, the western corn rootworm Diabrotica virgifera, the cotton bollworm Helicoverpa armigera, and the tobacco hawk Manduca sexta (Renyi and Jian-kang, 2014). Transgenic tobacco lines expressing dsRNA of v-ATPase and leading to whitefly mortality rate (Thakur et al., 2014) transcript level of goal genes in Bemisia tabaci at more than 70% mortality was observed (Raza et al., 2016; Shelby et al., 2020).

 

The knockdown of CsKrh1 mediated through RNA interference substantially decreased the transcription of vitellogenesis (Vg) within C. suppressalis, which is extremely essential in the suppression of rice pests (Tang et al., 2020). For pest management in the field through topical application or spraying, dsRNA shows a very high potential, with soybean aphids Aphis glycines (Yan et al., 2020a) mortality of up to 81.67% and with soybean Nezara viridula mortality of up to 90% recorded (Sharma et al., 2021).

 

Abiotic stress tolerance

 

Plant growth in the field can be exposed to several abiotic stresses, such as: drought, salt, heat and cold. RNA silencing coding is one of the technologies that can overcome this problem, the abiotic stress of plant products. By regulating the endogenous level of regulatory ncRNAs in response to abiotic stress, they regulate the expression of their target genes, which are closely involved in specific or multiple stresses.

 

Drought and salinity tolerance

 

Lack of water or drought and salinity are the abiotic stresses that are the main environmental stresses that restricts crop productivity. RNA silencing coding has been used successfully to enhance drought and salt tolerance cultures. RNA interference is used in oilseed rape to supply the AtHPR1 promoter, which is resistant to seed break-off during drought-induced flowering, besides affecting yield in drought stress. Transgenic rice plants that are drought stress-tolerant have been developed such as the receptor for activated C-kinase1 (RACK1) (Li et al., 2009), knockdown of a RING finger E3 ligase gene OsDSG1, and also silencing OsDIS1 for the drought-induced SINA protein through Oryza sativa (Park et al., 2010).

 

A wide variety of miRNAs has been recognized in Brassica that responds to drought, high salinity, and stress at high temperatures. Of 126 newly recognized miRNAs, miR164, miR160, and miR156 were experimentally approved to goal NAC domain-containing proteins, ARF17-like, and SPL2-like proteins, respectively (Bhardwaj et al., 2014).

 

However, the RNA interference has also regulated the gene to respond in an identical way to the salinity tolerance plants. The overexpression of GmNFYA3 in Arabidopsis led to an increased sensitivity to salinity stress, and exogenous ABA (Ni et al., 2013) as well as transgenic creeping bentgrass (Agrostis stolonifera) plants that overexpressed a rice miR319 gene (OsamiR319) also confirmed an increased tolerance to drought and salinity related with increased leaf wax content and water retention; however, there was decreased sodium intake (Zhou and Luo, 2013). In tomato, overexpression of the miR169c gene led to a reduction in stoma openings, the transpiration rate, and the loss of leaf water, which improved the drought tolerance in transgenic plants compared to wild-type controls (Zhang et al., 2011a). Accordingly, a knockdown of the OsTBP2.2 gene was generated in rice in order to increase rice sensitivity to drought stress (Zhang et al., 2020).

 

Cold and heat stress tolerance

 

Cold and heat are the abiotic types of stress that can influence plant production. With the variable conditions, the plants are pale; also, the yield and quality of plant products decrease and additionally affects the financial loss. Transgenic plants that use RNA silencing coding can overcome the problem of this stress. It has been suggested that miR319 expression changes in response to cold stress in Arabidopsis, rice, and sugar cane (Lv et al., 2020).

 

The overexpression of the Osa miR319 gene led to increased cold stress tolerance (4C) after cold acclimatization (12C) of plants (Yang et al., 2013). In rice, the overexpression of OsPCF5 and OsTCP21 led to the production of the cold-resistant transgenic plant (Yang et al., 2013).

 

Guan et al. (2013) observed a new kind of plant thermotolerance mechanism, in particular to protect the reproductive organs. It includes the induction of miR398 to downregulate its goal genes CSD (copper / zinc superoxide dismutase), CSD1 and CSD2, and CCS (a gene that codes for copper chaperones for CSD1 and CSD2). They observed that csd1, csd2 and ccsmutanten showed a higher heat stress tolerance than wild-type plants, combined with an increased accumulation of heat stress transcription factors and heat shock proteins as well as much less damage to plants (Guan et al., 2013). Corn (Yu et al., 2018) and cassava (Suksamran et al., 2020) responds to abiotic stress such as heat, cold, salt, and drought.


 CONCLUSION

One of the basic requirements of a human being is food. However, food insecurity and malnutrition are currently among the most serious concerns for human health, causing the loss of countless lives in developing countries. Therefore, there is need for an innovative technology to improve upon our crop production methods and practices. RNA silencing gene is an advanced application that can solve the agricultural problem in a short period. In crop improvement, it has a wider application in the production of transgenic plants, which have improved yield, increased nutritional qualities with improved taste, texture or appearance, and health benefits. It also enables us to produce plants that have reduced dependence on fertilizers, pesticides, and other agrochemicals, and plants which have reduced the vulnerability of crops to environmental stresses. RNA interference applications are innovative technologies that can contribute to the solution to these problems. It describes several mechanistically related pathways which are involved in controlling and regulating gene expression. RNA silencing pathways are associated with the regulatory activity of non-coding RNAs that function as factors involved in inactivating homologous sequences, promoting endonuclease activity, translational arrest, and/or chromatic or DNA modification. It also has some limitations which include identification of appropriate target genes, off-target effects, and the application of RNAi is more sophisticated and needs highly skilled human power.


 CONFLICT OF INTERESTS

The author has not declared any conflict of interests.



 REFERENCES

Anita QG, Sofia N, Helena S (2013). Non-Coding RNAs: Multi- Tasking Molecules in the Cell. International Journal of Molecular Sciences 14:16010-16039.
Crossref

 

Antonella M, Muller CT, Bruno L, McGregor L, Ferrante A, Ceverista CAA, Spadafora ND (2020). Fruit volatile profiling through GC× GC-ToF-MS and gene expression analyses reveal differences amongst peach cultivars in their response to cold storage. Scientific Reports (Nature Publisher Group) 10(1).
Crossref

 

Arefin P, Amin R, Sadiq MZA, Dey SS, Boby F, Parvaz MS (2020). A Study of Effectiveness of Natural Coatings on the Shelf Life Extension of Tomatoes by the Observation of TomloxC Gene Expression. Bioscience Biotechnology Research Asia 17(3):459-465.
Crossref

 

Bader AS, Hawley BR, Wilczynska A, Bushell M (2020). The roles of RNA in DNA double strand break repair. British Journal of Cancer 122(5):613-623.
Crossref

 

Bahadur B, Rajam MV, Sahijram L, Krishnamurthy KV (2015). Induced mutations and crop improvement. Plant Biology and Biotechnology 1(23):593-617.
Crossref

 

Bento FM, Marques RN, Campana FB, Demétrio CG, Leandro RA, Parra JRP, Figueira A (2020). Gene silencing by RNAi via oral delivery of dsRNA by bacteria in the South American tomato pinworm, Tuta absoluta. Pest Management Science 76(1):287-295.
Crossref

 

Berman J, Zorrilla-López U, Sandmann G, Capell T, Christou P, Zhu C (2017). The silencing of carotenoid β-hydroxylases by RNA interference in different maize genetic backgrounds increases the β-carotene content of the endosperm. International Journal of Molecular Sciences 18(12):2515.
Crossref

 

Bhardwaj AR, Joshi G, Pandey R, Kukreja B, Goel S, Jagannath A, Agarwal M (2014). A genome-wide perspective of miRNA ome in response to high temperature, salinity, and drought stresses in Brassica juncea (Czern) L. Plos One 9(3):92456.
Crossref

 

Bologna NG, Voinnet O (2014). The diversity, biogenesis, and activities of endogenous silencing small RNAs in Arabidopsis. Annual Review of Plant Biology 65:473-503
Crossref

 

Brant EJ, Budak H (2018). Plant small non-coding RNAs and their roles in biotic stresses. Frontiers in Plant Science 9:1038.
Crossref

 

Campo S, Peris-Peris C, Sire C, Moreno AB, Donaire L, Zytnicki M (2013). Identification of a novel microRNA (miRNA) from rice that targets an alternatively spliced transcript of the Nramp6 (Natural resistance-associated macrophage protein6) gene involved in pathogen resistance. New Phytologist 199(1):212-227.
Crossref

 

Choudhary M, Grover K, Singh M (2021). Maize's significance in the Indian food situation to mitigate malnutrition. Cereal Chemistry 98(2):212-221.
Crossref

 

Chuck GS, Tobias C, Sun L, Kraemer F, Li C, Dibble D (2011). Overexpression of the maize corn grass1 microRNA prevents flowering, improves digestibility, and increases the starch content of switchgrass. Proceedings of the National Academy of Sciences, 108(42):17550-17555.
Crossref

 

Crick F (1970). Central dogma of molecular biology. Nature 227:561-563.
Crossref

 

Ding SW (2010). RNA based antiviral immunity. Nature Reviews Immunology 10:632-644.
Crossref

 

Ding Y, Zeng W, Wang X, Wang Y, Niu L, Pan L, Wang Z (2019). Over- expression of Peach PpIAA19 in Tomato Alters Plant Growth, Parthenocarpy and Fruit Shape. Journal of Plant Growth Regulation 38(1):103-112.
Crossref

 

Diretto G, Frusciante S, Fabbri C, Schauer N, Busta L, Wang Z, Giuliano G (2020). Manipulation of β?carotene levels in tomato fruits results in increased ABA content and extended shelf life. Plant Biotechnology Journal 18(5):1185-1199.
Crossref

 

Fatica A, Bozzoni I (2014). Long non-coding RNAs: new players in cell differentiation and development. Nature Reviews Genetics 15:7-21.
Crossref

 

Feldmann KA (2006). Steroid regulation improves crop yield. Nature Biotechnology 24:46-47.
Crossref

 

Gao L, Chen W, Xu X, Zhang J, Singh TK, Liu S, Liu Q (2020). Engineering Trienoic Fatty Acids into Cottonseed Oil Improves Low-Temperature Seed Germination, Plant Photosynthesis and Cotton Fiber Quality. Plant and Cell Physiology 61(7):1335-1347.
Crossref

 

Guan Q, Lu X, Zeng H, Zhang Y, Zhu J (2013). Heat stress induction of miR398 triggers a regulatory loop that is critical for thermo-tolerance in Arabidopsis. Plant Journal 74(5):840-851.
Crossref

 

Guo H, Ge P, Tong J, Zhang Y, Peng X, Zhao Z, Sun Y (2021). Elevated Carbon Dioxide Levels Decreases Cucumber Mosaic Virus Accumulation in Correlation with Greater Accumulation of rgs-CaM, an Inhibitor of a Viral Suppressor of RNAi. Plants 10(1):59.
Crossref

 

Guo W, Chen W, Zhang Z, Guo N, Liu L, Ma Y, Dai H (2020). The hawthorn CpLRR-RLK1 gene targeted by ACLSV-derived vsiRNA positively regulates resistance to bacteria disease. Plant Science 300:110641.
Crossref

 

Gupta A, Pal RK, Rajama M (2013). Delayed ripening and improved fruit processing quality in tomatoes by RNAi-mediated silencing of three homologs of 1- aminopropane-1-carboxylate synthase gene. Journal of Plant Physiology 170(11):987-995.
Crossref

 

Hangauer MJ, Vaughn IW, McManus MT (2013). Pervasive transcription of the human genome produces thousands of previously unidentified long intergenic noncoding RNAs. PLoS genetics 9(6):e1003569.
Crossref

 

Hasan MM, Rima R (2021). Genetic engineering to improve essential and conditionally essential amino acids in maize: transporter engineering as a reference. Transgenic Research 30(2):207-220.
Crossref

 

Hu C, Sheng O, Dong T, Yang Q, Dou T, Li C, Bi F (2020). Overexpression of MaTPD1A impairs fruit and pollen development by modulating some regulators in Musa itinerant. BMC Plant Biology 20(1):1-12.
Crossref

 

Huvenne H, Smagghe G (2010). Mechanisms of dsRNA uptake in insects and potential of RNAi for pest control: a review. Journal of Insect Physiology 56(3):227-235.
Crossref

 

Jan R, Khan MA, Asaf S, Lee IJ, Bae JS, Kim KM (2020). Overexpression of OsCM alleviates BLB stress via phytohormonal accumulation and transcriptional modulation of defense-related genes in Oryza sativa. Scientific reports 10(1):1-15.
Crossref

 

Jiao Y, Wang Y, Xue D, Wang J, Yan M, Liu G, Dong G, Zeng D, Lu Z, Zhu X, Qian Q, Li J (2010). The regulation of OsSPL14 by OsmiR156 defines the ideal plant architecture in rice. Nature Genetics 42(6):541-544.
Crossref

 

Johnson CR (2017). Reproduction and confinement of miR156 transgenic switchgrass (Panicum virgatum L.).

View

 

Kamthan A, Chaudhuri A, Kamthan M, Datta A (2015). Small RNAs in plants: recent development and application for crop improvement. Front. Plant Science 6:208-210.
Crossref

 

Kertbundit S, Pongtanom N, Ruanjan P, Chantasingh D, Tanwanchai A, Panyim S, Ju?í?ek M (2007). Resistance of transgenic papaya plants to Papaya ring spot virus. Biologia Plantarum 51(2):333-339.
Crossref

 

Khraiwesh B, Zhu JK, Zhu J (2012). Role of miRNAs and siRNAs in biotic and abiotic stress responses of plants. Biochimica et Biophysica Acta 1819(2):137-148.
Crossref

 

Ko MR, Song MH, Kim JK, Baek SA, You MK, Lim SH, Ha SH (2018). RNAi-mediated suppression of three carotenoid-cleavage dioxygenase genes, OsCCD1, 4a, and 4b, increases carotenoid content in rice. Journal of Experimental Botany 69(21):5105-5116.
Crossref

 

Koch A, Stein E, Kogel KH (2018). RNA-based disease control as a complementary measure to fight Fusarium fungi through silencing of the azole target Cytochrome P450 Lanosterol C-14 α-Demethylase. European Journal of Plant Pathology 152(4):1003- 1010.
Crossref

 

Li C, Song R (2020). The regulation of zein biosynthesis in maize endosperm. Theoretical and Applied Genetics 133(5):1443-1453.
Crossref

 

Li DH, Hui LIU, Yang YL, Zhen PP, Liang JS (2009). Down-regulated expression of RACK1 gene by RNA interference enhances drought tolerance in rice. Rice Science 16(1):14-20.
Crossref

 

Li Y, Lu YG, Shi Y, Wu L, Xu YJ, Huang F, Wang WM (2014). Multiple rice microRNAs are involved in immunity against the blast fungus Magnaporthe oryzae. Plant Physiology 164(2):1077-1092.
Crossref

 

Li Y, Zhang Q, Zhang J, Wu L, Qi Y, Zhou JM (2010). Identification of microRNAs involved in pathogen-associated molecular pattern-triggered plant innate immunity. Plant Physiology 152(4):2222-2231.
Crossref

 

Li Y, Zhao SL, Li JL, Hu XH, Wang H, Cao XL, Wang WM (2017). Osa-miR169 negatively regulates rice immunity against the blast fungus Magnaporthe oryzae. Frontiers in Plant Science 8:2.
Crossref

 

Lindsay MA, Griffiths-Jones S, Sato K, Siomi MC (2013). Piwi-interacting RNAs: biological functions and biogenesis. Essays in Biochemistry 54:39-52.
Crossref

 

Liu JY, Zhang YW, Han X, Zuo JF, Zhang Z, Shang H, Zhang YM (2020). An evolutionary population structure model reveals pleiotropic effects of GmPDAT for traits related to seed size and oil content in soybean. Journal of Experimental Botany 71(22):6988-7002.
Crossref

 

Liu Y, Shi Y, Su D, Lu W, Li Z (2021). SlGRAS4 accelerates fruit ripening by regulating ethylene biosynthesis genes and SlMADS1 in tomatoes. Horticulture Research 8(1):1-11.
Crossref

 

Liu Y, Wang Y, Pei J, Li Y, Sun H (2021a). Genome-wide identification and characterization of the COMT gene family during the development of blueberry fruit. BMC Plant Biology 21(1):1-16.
Crossref

 

Lv J, Zhang J, Han X, Bai L, Xu D, Ding S, Li J (2020). Genome-wide identification of superoxide dismutase (SOD) genes and their expression profiles under 1- methyl cyclopropane (1-MCP) treatment during the ripening of apple fruit. Scientia Horticulturae 271:109471.
Crossref

 

Ma L, Bajic VB, Zhang Z (2013). On the classification of long non-coding RNAs. RNA Biology 10(6):925-933.
Crossref

 

Man Z, Hong L (2013). MicroRNA-mediated gene regulation: potential applications for plant genetic engineering. Plant Molecular Biology 83(1-2):59-75.
Crossref

 

Matsuo S, Miyatake K, Endo M, Urashimo S, Kawanishi T, Negoro S, Fukuoka H (2020). Loss of function of the Pad-1 aminotransferase gene, which is involved in auxin homeostasis, induces parthenocarpy in Solanaceae plants. Proceedings of the National Academy of Sciences 117(23):12784-12790.
Crossref

 

Meli VS, Ghosh S, Prabha TN, Chakraborty N, Chakraborty S, Datta A (2010). Enhancement of fruit shelf life by suppressing N-glycan processing enzymes. Proceedings of the National Academy of Sciences of the United States of America 107(6):2413-2418.
Crossref

 

Mezzetti B, Smagghe G, Arpaia S, Christiaens O, Dietz-Pfeilstetter A, Jones H, Sweet J (2020). RNAi: What is its position in agriculture? Journal of Pest Science 93:1125-1130.
Crossref

 

Minna UK, Michael TYL, Christopher KG (2011). Non-coding RNAs as regulators of gene expression and epigenetics. Cardiovascular Research 90(3):430-440.
Crossref

 

Molesini B, Youry P, Tiziana P (2012). Fruit improvement using intragenesis and artificial microRNA. Trends in Biotechnology 30:2.
Crossref

 

Moreno JC, Mi J, Agrawal S, Kössler S, Ture?ková V, Tarkowská D, Schöttler MA (2020). Expression of a carotenogenic gene allows faster biomass production by redesigning plant architecture and improving photosynthetic efficiency in tobacco. The Plant Journal 103(6):1967-1984.
Crossref

 

Ni Z, Hu Z, Jiang Q, Zhang H (2013). GmNFYA3, a target gene of miR169, is a positive regulator of plant tolerance to drought stress. Plant Molecular Biology 82(1-2):113-129.
Crossref

 

Osorio S, Alba R, Damasceno CM, Lopez-Casado G, Lohse M, Zanor MI (2011). Systems biology of tomato fruit development: combined tran- script, protein, and metabolite analysis of tomato transcription factor and ethylene receptor (Nr) mutants reveals novel regulatory interactions. Plant Physiology 157(1):405-425.
Crossref

 

Park GG, Park JJ, Yoon J, Yu SN, An G (2010). A RING finger E3 ligase gene, Oryza sativa Delayed Seed Germination 1 (OsDSG1), controls seed germination and stress responses in rice. Plant Molecular Biology 74(45):467-478.
Crossref

 

Park JH, Shin C (2015). The role of plant small RNAs in NB-LRR regulation. Briefings in Functional Genomics 14:268-274.
Crossref

 

Raza A, Malik HJ, Shafiq M, Amin I, Scheffler JA, Scheffler BE, et al (2016). RNA Interference based Approach to Down Regulate Osmoregulators of Whitefly (Bemisia tabaci): Potential Technology for the Control of Whitefly. PLoS ONE 11(4):e0153883.
Crossref

 

Renyi L, Jiankang Z (2014). Non-coding RNAs as potent tools for crop improvement. National Science Review 1(2):186-189.
Crossref

 

Rodnina MV, Wintermeyer W (2011). The ribosome as a molecular machine: the mechanism of tRNA-mRNA movement in translocation. Biochemical Society Transactions 39(2):658-662.
Crossref

 

Romer E, Chebib S, Bergmann KC, Plate K, Becker S, Ludwig C, Schwab W (2020). Tiered approach for the identification of Mal d 1 reduced, well tolerated apple genotypes. Scientific Reports 10(1):1-13.
Crossref

 

Roussin-Léveillée C, Silva-Martins G, Moffett P (2020). ARGONAUTE5 represses age- dependent induction of flowering through physical and functional interaction with miR156 in Arabidopsis. Plant and Cell Physiology 61(5):957-966.
Crossref

 

Sajid IA, Tabassum B, Yousaf I, Khan A, Adeyinka OS, Shahid N, Husnain T (2019). In vivo gene silencing of potato virus X by small interference RNAs in transgenic potato. Potato Research 1:13.
Crossref

 

Sang YS, Chanseok S (2016). Regulatory non-coding RNAs in plants: potential gene resources for the improvement of agricultural traits. Plant Biotechnology Reports 10(2):35-47.
Crossref

 

Santosh B, Akhil V, Pramod KY (2014). Non-coding RNAs: biological functions and applications. Cell Biochemistry & Function 33(1):14-22.
Crossref

 

Sharma R, Christiaens O, Taning CN, Smagghe G (2021). RNAi?mediated mortality in southern green stinkbug Nezara viridula by oral delivery of dsRNA. Pest Management Science 77(1):77-84.
Crossref

 

Sharp SJ, Schaack J, Cooley L, Burke DJ, Soll D (1985). Structure and Transcription of Eukaryotic tRNA Genes. CRC Critical Reviews in Biochemistry 19(2):107-144.
Crossref

 

Shelby EA, Moss JB, Andreason SA, Simmons AM, Moore AJ, Moore PJ (2020). Debugging: Strategies and Considerations for Efficient RNAi-Mediated Control of the Whitefly Bemisia tabaci. Insects 11(11):723.
Crossref

 

Soto-Suárez M, Baldrich P, Weigel D, Rubio-Somoza I, San Segundo B (2017). The Arabidopsis miR396 mediates pathogen-associated molecular pattern-triggered immune responses against fungal pathogens. Scientific reports 7(1):1-14.
Crossref

 

Suksamran R, Saithong T, Thammarongtham C, Kalapanulak S (2020). Genomic and transcriptomic analysis identified novel putative cassava lncRNAs involved in cold and drought stress. Genes 11(4):366.
Crossref

 

Tang Y, He H, Qu X, Cai Y, Ding W, Qiu L, Li Y (2020). RNA interference? mediated knockdown of the transcription factor Krüppel homologue 1 suppresses vitellogenesis in Chilo suppressalis. Insect Molecular Biology 29(2):183-192.
Crossref

 

Teo G, Suzuki Y, Uratsu SL, Lampinen B, Ormonde N, Hu WK, Dandekar AM (2006). Silencing leaf sorbitol synthesis alters long-distance partitioning and apple fruit quality. Proceedings of the National Academy of Sciences 103(49):18842-18847.
Crossref

 

Thakur N, Upadhyay SK, Verma PC, Chandrashekar K, Tuli R, Singh PK (2014) Enhanced Whitefly Resistance in Transgenic Tobacco Plants Expressing Double Stranded RNA of v-ATPase A Gene. PLoS ONE 9(3):e87235.
Crossref

 

Tian SN, Liu DD, Zhong CL, Xu HY, Yang S, Fang Y, Liu JZ (2020). Silencing GmFLS2 enhances the susceptibility of soybean to bacterial pathogen through attenuating the activation of GmMAPK signaling pathway. Plant Science 292:110386.
Crossref

 

Uslu VV, Wassenegger M (2020). Critical view on RNA silencing-mediated virus resistance using exogenously applied RNA. Current Opinion in Virology 42:18-24.
Crossref

 

Valle M, Zavialov A, Li W, Stagg SM, Sengupta J, Nielsen RC, Nissen P, Harvey SC, Ehrenberg M, Frank J (2003). Incorporation of aminoacyl-tRNA into the ribosome as seen by cryo-electron microscopy. Nature Structural Biology 10(11):899-906.
Crossref

 

Vanderschuren H, Moreno I, Anjanappa RB, Zainuddin IM, Gruissem W (2012). Exploiting the combination of natural and genetically engineered resistance to cassava mosaic and cassava brown streak viruses impacting cassava production in Africa. PloS one 7(9):e45277.
Crossref

 

Vanitharani R, Chellappan P, Pita JS, Fauquet CM (2004). Differential roles of AC2 and AC4 of cassava geminiviruses in mediating synergism and suppression of posttranscriptional gene silencing. Journal of Virology 78(17):9487-9498.
Crossref

 

Wang C, Wang W, Abdelrahman M, Jiu S, Zheng T, Gong P, Fang J (2020a). Study on miRNAs-Mediated Seed and Stone-Hardening Regulatory Networks and the Mechanism of miRNAs' Manipulating Gibberellin-Induced Seedless Berries in Grapevine (Vitis vinifera L.).
Crossref

 

Wang Z, Zhang WH, Ma LY, Li X, Zhao FY, Tan XL (2020b). Overexpression of Brassica napus NPR1 enhances resistance to Sclerotinia sclerotiorum in oilseed rape. Physiological and Molecular Plant Pathology 110:101460.
Crossref

 

Wang S,Wu K, Yuan Q, Liu X, Liu Z, Lin X, (2012). Control of grain size, shape and quality by OsSPL16 in rice. Nature Genetics 44:950-954.
Crossref

 

Wang Y, Feng G, Zhang Z, Liu Y, Ma Y, Wang Y, Niu X (2021). Overexpression of Pti4, Pti5, and Pti6 in tomato promote plant defense and fruit ripening. Plant Science 302:110702.
Crossref

 

Wheeler T, von Braun J (2013). Climate change impacts on global food security. Science 341:508-513.
Crossref

 

Wilson RC, Doudna JA (2013). Molecular mechanisms of RNA interference. Annual Review of Biophysics 42:217-239.
Crossref

 

Xie K, Shen J, Hou X, Yao J, Li X, Xiao J, Xiong L (2012). Gradual increase of miR156 regulates temporal expression changes of numerous genes during leaf development in rice. Plant Physiology 158(3):1382-1394
Crossref

 

Xin M, Wang Y, Yao Y, Xie C, Peng H, Ni Z (2010). Diverse set of microRNAs are responsive to powdery mildew infection and heat stress in wheat (Triticum aestivum L.). BMC Plant Biology 10(1):123-134.
Crossref

 

Yamagishi M, Sakai M (2020). The MicroRNA828/MYB12 Module Mediates Bicolor Pattern Development in Asiatic Hybrid Lily (Lilium spp.) Flowers. Frontiers in Plant Science 11.
Crossref

 

Yan S, Qian J, Cai C, Ma Z, Li J, Yin M, Shen J (2020). Spray method application of transdermal dsRNA delivery system for efficient gene silencing and pest control on soybean aphid Aphis glycines. Journal of Pest Science 93(1):449-459.
Crossref

 

Yan S, Ren B, Zeng B, Shen J (2020a). Improving RNAi efficiency for pest control in crop species. BioTechniques 68(5):283-290.
Crossref

 

Yan S, Ren BY, Shen J (2021). Nanoparticle?mediated double?stranded RNA delivery system: A promising approach for sustainable pest management. Insect Science 28(1):21-34.
Crossref

 

Yang C, Li D, Mao D, Liu X, Li C, Li X (2013). Over expression of microRNA319 impacts leaf morphogenesis and leads to enhanced cold tolerance in rice (Oryza sativa L.). Plant, Cell & Environment 36(12):2207-2218.
Crossref

 

Yang J, Xing G, Niu L, He H, Guo D, Du Q, Yang X (2018). Improved oil quality in transgenic soybean seeds by RNAi-mediated knockdown of GmFAD2-1B. Transgenic Research 27(2):155-166.
Crossref

 

Yu X, Killiny N (2020). RNA interference-mediated control of Asian citrus psyllid, the vector of the huanglongbing bacterial pathogen. Tropical Plant Pathology 45:298-305.
Crossref

 

Yu X, Meng X, Liu Y, Li N, Zhang A, Wang TJ, Jiang L, Pang J, Zhao X, Qi X, Zhang M (2018). The chromatin remodeler ZmCHB101 impacts expression of osmotic stress-responsive genes in maize. Plant Molecular Biology 97(4):451-65.
Crossref

 

Yumbya P, Ambuko J, Hutchinson M, Owino W, Juma J, Machuka E, Mutuku JM (2021). Transcriptome analysis to elucidate hexanal's mode of action in preserving the post-harvest shelf life and quality of banana fruits (Musa acuminata). Journal of Agriculture and Food Research 100114.
Crossref

 

Zhang W, Gao S, Zhou X, Chellappan P, Chen Z, Zhou X, Zhang X, Fromuth N, Coutino G, Coffey M, Jin H (2011). Bacteria responsive microRNAs regulate plant innate immunity by modulating plant hormone networks. Plant Molecular Biology 75:93-105.
Crossref

 

Zhang X, Zou Z, Zhang J, Zhang Y, Han Q, Hu T, (2011a). Over expression of sly- miR156a in tomato results in multiple vegetative and reproductive trait alterations and partial phenocopy of the sft mutant. FEBS Letters 585:435-439.
Crossref

 

Zhang Y, Zhao L, Xiao H, Chew J, Xiang J, Qian K, Fan X (2020). Knockdown of a Novel Gene OsTBP2.2 Increases Sensitivity to Drought Stress in Rice. Genes 11(6):629.
Crossref

 

Zhang YC, Liao JY, Li ZY, Yu Y, Zhang JP, Li QF, Qu LH, Shu W, Chen YQ (2014). Genome-wide screening and functional analysis identify a large number of long noncoding RNAs involved in the sexual reproduction of rice. Genome Biology 15:5-12.
Crossref

 

Zhao ZX, Feng Q, Cao XL, Zhu Y, Wang H, Chandran V, Wang WM (2020). Osa?miR167d facilitates infection of Magnaporthe oryzae in rice. Journal of Integrative Plant Biology 62(5):702-715.
Crossref

 

Zheng LL, Qu LH (2015). Application of microRNA gene resources in the improvement of agronomic traits in rice. Plant Biotechnology Journal 13:329-336.
Crossref

 

Zheng Q, Liu J, Goff BM, Dinkins RD, Zhu H (2016). Genetic manipulation of miR156 for improvement of biomass production and forage quality in red clover. Crop Science 56(3):1199-1205.
Crossref

 

Zhong J, He W, Peng Z, Zhang H, Li F, Yao J (2020). A putative AGO protein, OsAGO17, positively regulates grain size and grain weight through OsmiR397b in rice. Plant Biotechnology Journal 18(4):916-928.
Crossref

 

Zhou M, Luo H (2013). MicroRNA-mediated gene regulation: potential applications for plant genetic engineering. Plant Molecular Biology 83:59-75.
Crossref

 

Zhou SX, Zhu Y, Wang LF, Zheng YP, Chen JF, Li TT, Wang WM (2020). Osa?miR1873 fine?tunes rice immunity against Magnaporthe oryzae and yield traits. Journal of integrative plant biology 62(8):12213-1226.
Crossref

 




          */?>