Gene Silencing Efficiency and INF-β Induction Effects of Splicing miRNA 155-Based Artificial miRNA with Pre-miRNA Stem-Loop Structures

2011 ◽  
Vol 50 (1-2) ◽  
pp. 112-121
Author(s):  
Onsam Sin ◽  
Prudence Mabiala ◽  
Ye Liu ◽  
Ying Sun ◽  
Tao Hu ◽  
...  
2004 ◽  
Vol 171 (4S) ◽  
pp. 256-257
Author(s):  
Kazunori Haga ◽  
Ataru Sazawa ◽  
Toru Harabayashi ◽  
Nobuo Shinohara ◽  
Minoru Nomoto ◽  
...  

2021 ◽  
Vol 12 ◽  
Author(s):  
Yijuan Ding ◽  
Yangui Chen ◽  
Baoqin Yan ◽  
Hongmei Liao ◽  
Mengquan Dong ◽  
...  

Sclerotinia sclerotiorum is a devastating necrotrophic fungal pathogen and has a substantial economic impact on crop production worldwide. Magnaporthe appressoria-specific (MAS) proteins have been suggested to be involved in the appressorium formation in Magnaporthe oryzae. Sscnd1, an MAS homolog gene, is highly induced at the early infection stage of S. sclerotiorum. Knock-down the expression of Sscnd1 gene severely reduced the virulence of S. sclerotiorum on intact rapeseed leaves, and their virulence was partially restored on wounded leaves. The Sscnd1 gene-silenced strains exhibited a defect in compound appressorium formation and cell integrity. The instantaneous silencing of Sscnd1 by tobacco rattle virus (TRV)-mediated host-induced gene silencing (HIGS) resulted in a significant reduction in disease development in tobacco. Three transgenic HIGS Arabidopsis lines displayed high levels of resistance to S. sclerotiorum and decreased Sscnd1 expression. Production of specific Sscnd1 siRNA in transgenic HIGS Arabidopsis lines was confirmed by stem-loop qRT-PCR. This study revealed that the compound appressorium-related gene Sscnd1 is required for cell integrity and full virulence in S. sclerotiorum and that Sclerotinia stem rot can be controlled by expressing the silencing constructs of Sscnd1 in host plants.


2019 ◽  
Vol 1 (1) ◽  
Author(s):  
Yan Li ◽  
John Martin Jerome Jeyakumar ◽  
Qin Feng ◽  
Zhi-Xue Zhao ◽  
Jing Fan ◽  
...  

AbstractMicroRNAs (miRNAs) are a class of small (20–24 nucleotides (nt) long) non-coding RNAs. One mature miRNA can be transcribed from one or more gene loci known as miRNA genes (MIRs). The transcript of a MIR forms a stem-loop structure that is processed into a 20–24-nt miRNA-5p/−3p duplex by RNase III family endoribonucleases such as Dicer-like1 (DCL1). In turn, the overhang ends of the duplex are methylated by HUA ENHANCER 1 (HEN1), generating stabilized mature miRNAs. The mature miRNAs are loaded onto ARGONAUTE (AGO) proteins, forming a miRNA-induced gene silencing complex (miRISC). Then, the miRISC binds to target sites with sequences complementary to the miRNAs, leading to either cleavage or translational inhibition of the target mRNAs, or methylation of the target sequences, resulting in post-transcriptional and transcriptional gene silencing, respectively. In the past decade, more than 700 miRNAs have been identified in rice, a subset of which have been found to be responsive to the rice blast fungus, Magnaporthe oryzae, or its elicitors. Moreover, members of 10 miRNA families have been found to positively or negatively regulate rice defense against M. oryzae, namely miR160, miR164, miR166, miR167, miR169, miR319, miR396, miR398, miR444 and miR7695. This review summarizes the identification and functional characterization of the miRNAs, which respond to M. oryzae or its elicitors and describes the current understanding of the complicated but well-organized network in the context of rice-M. oryzae interaction.


2014 ◽  
Vol 9 (4) ◽  
pp. 939-949 ◽  
Author(s):  
Jian-Feng Li ◽  
Dandan Zhang ◽  
Jen Sheen

2019 ◽  
Vol 20 (22) ◽  
pp. 5620 ◽  
Author(s):  
Dandan Zhang ◽  
Nannan Zhang ◽  
Wenzhong Shen ◽  
Jian-Feng Li

Plant genome sequences are presently deciphered at a staggering speed, due to the rapid advancement of high-throughput sequencing technologies. However, functional genomics significantly lag behind due to technical obstacles related to functional redundancy and mutant lethality. Artificial microRNA (amiRNA) technology is a specific, reversible, and multiplex gene silencing tool that has been frequently used in generating constitutive or conditional mutants for gene functional interrogation. The routine approach to construct amiRNA precursors involves multiple polymerase chain reactions (PCRs) that can increase both time and labor expenses, as well as the chance to introduce sequence errors. Here, we report a simplified method to clone and express amiRNAs in Arabidopsis and rice based on the engineered Arabidopsis miR319a or rice miR528 precursor, which harbor restriction sites to facilitate one-step cloning of a single PCR product. Stem-loop reverse-transcriptase quantitative PCR (RT-qPCR) and functional assays validated that amiRNAs can be accurately processed from these modified precursors and work efficiently in plant protoplasts. In addition, Arabidopsis transgenic plants overexpressing the modified miR319a precursor or its derived amiRNA could exhibit strong gene silencing phenotypes, as expected. The simplified amiRNA cloning strategy will be broadly useful for functional genomic studies in Arabidopsis and rice, and maybe other dicotyledon and monocotyledon species as well.


2021 ◽  
Author(s):  
Shipra Saxena ◽  
Sneha Yogindran ◽  
Manmohan Arya ◽  
Yogita Sharma ◽  
Chandra Pal Singh

Insects as pests destroy annually an estimated 18–20% of the crop production worldwide. Caterpillars, the larval stage of moths, are the major pests of agricultural products owing to their voracious feeding habits. In the past few decades, the potent methods of insect control, such as insecticides and Bt toxins, have been constrained as a result of health hazards, environmental issues, and development of resistance, after their prolonged application. Thus, there is need to find alternative options to improve plant protection strategies. Recently, RNA interference (RNAi), the post-transcriptional gene-silencing mechanism, has emerged as one of such a novel, sustainable, and environment friendly approaches for insect management and crop protection. RNAi technology relies on selection of a vital insect pest target gene and its expression as a double stranded RNA or stem-loop RNA molecule, which is recognized by the host RNAi machinery and processed into small interfering RNAs (siRNAs) or microRNAs (miRNAs). The siRNA/miRNA along with the RNA-induced silencing complex (RISC) binds to the complimentary mRNA and induce gene silencing at post-transcriptional level. With effective target-gene selection and transgenic plants expressing these precursor RNA molecules, insect pests of various crops have been efficiently managed. In this chapter, we discuss the basic mechanism of RNAi and its application in controlling lepidopteran pests of important crop plants.


PLoS ONE ◽  
2015 ◽  
Vol 10 (7) ◽  
pp. e0132899
Author(s):  
Tim Thijs ◽  
Katleen Broos ◽  
Stefaan J. Soenen ◽  
Aline Vandenbulcke ◽  
Karen Vanhoorelbeke ◽  
...  

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