Specific and multiple‐target gene silencing reveals function diversity of BnaA2 . NIP5 ;1 and BnaA3 . NIP5 ;1 in Brassica napus

2021 ◽  
Author(s):  
Mingliang He ◽  
Cheng Zhang ◽  
Liuyang Chu ◽  
Sheliang Wang ◽  
Lei Shi ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Gene Silencing ◽  
Target Gene ◽  
Multiple Target ◽  
Multiple Target Gene

Artificial microRNA (amiRNA) induced gene silencing in alfalfa (Medicago sativa)

Botany ◽  
2013 ◽  
Vol 91 (2) ◽  
pp. 117-122 ◽  
Author(s):  
Julian C. Verdonk ◽  
Michael L. Sullivan
Keyword(s):  
Gene Silencing ◽  
Medicago Sativa ◽  
Target Gene ◽  
Target Genes ◽  
Mirna Precursor ◽  
Crop Species ◽  
Forage Crop ◽  
Ribonucleic Acids ◽  
Endogenous Gene ◽  
Medicago Sativa L

Gene silencing is a powerful technique that allows the study of the function of specific genes by selectively reducing their transcription. Several different approaches can be used, however they all have in common the artificial generation of single stranded small ribonucleic acids (RNAs) that are utilized by the endogenous gene silencing machinery of the organism. Artificial microRNAs (amiRNA) can be used to very specifically target genes for silencing because only a short sequence of 21 nucleotides of the gene of interest is used. Gene silencing via amiRNA has been developed for Arabidopsis thaliana (L.) Heynh. and rice using endogenous microRNA (miRNA) precursors and has been shown to also work effectively in other dicot species using the arabidopsis miRNA precursor. Here, we demonstrate that the arabidopsis miR319 precursor can be used to silence genes in the important forage crop species alfalfa (Medicago sativa L.) by silencing the expression of a transgenic beta-glucuronidase (GUSPlus) target gene.

scholarly journals Development and Application of a Green Fluorescent Protein Sentinel System for Identification of RNA Interference in Blastomyces dermatitidis Illuminates the Role of Septin in Morphogenesis and Sporulation

2007 ◽  
Vol 6 (8) ◽  
pp. 1299-1309 ◽  
Author(s):  
T. Krajaejun ◽  
G. M. Gauthier ◽  
C. A. Rappleye ◽  
T. D. Sullivan ◽  
B. S. Klein
Keyword(s):  
Green Fluorescent Protein ◽  
Rna Interference ◽  
Gene Silencing ◽  
Rapid Detection ◽  
Target Gene ◽  
Fluorescent Protein ◽  
Yeast Cells ◽  
Blastomyces Dermatitidis ◽  
Dimorphic Fungus ◽  
Green Fluorescent

ABSTRACT A high-throughput strategy for testing gene function would accelerate progress in our understanding of disease pathogenesis for the dimorphic fungus Blastomyces dermatitidis, whose genome is being completed. We developed a green fluorescent protein (GFP) sentinel system of gene silencing to rapidly study genes of unknown function. Using Gateway technology to efficiently generate RNA interference plasmids, we cloned a target gene, “X,” next to GFP to create one hairpin to knock down the expression of both genes so that diminished GFP reports target gene expression. To test this approach in B. dermatitidis, we first used LACZ and the virulence gene BAD1 as targets. The level of GFP reliably reported interference of their expression, leading to rapid detection of gene-silenced transformants. We next investigated a previously unstudied gene encoding septin and explored its possible role in morphogenesis and sporulation. A CDC11 septin homolog in B. dermatitidis localized to the neck of budding yeast cells. CDC11-silenced transformants identified with the sentinel system grew slowly as flat or rough colonies on agar. Microscopically, they formed ballooned, distorted yeast cells that failed to bud, and they sporulated poorly as mold. Hence, this GFP sentinel system enables rapid detection of gene silencing and has revealed a pronounced role for septin in morphogenesis, budding, and sporulation of B. dermatitidis.

scholarly journals RNA interference as a gene silencing tool to controlTuta absolutain tomato (Solanum lycopersicum)

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e2673 ◽  
Author(s):  
Roberto A. Camargo ◽  
Guilherme O. Barbosa ◽  
Isabella Presotto Possignolo ◽  
Lazaro E. P. Peres ◽  
Eric Lam ◽  
...  
Keyword(s):  
Rna Interference ◽  
Gene Silencing ◽  
Target Gene ◽  
Target Genes ◽  
Larval Mortality ◽  
Tuta Absoluta ◽  
Specific Gene ◽  
Gene Transcript ◽  
Pest Species ◽  
In Planta

RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. We selected two target genes (Vacuolar ATPase-AandArginine kinase) based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet forT. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on “in planta-induced transient gene silencing” (PITGS), a well-established method for silencing plant genes, used here for the first time to deliverin planta-transcribed dsRNA to target insect genes.Tuta absolutalarvae that fed on leaves containing dsRNA of the target genes showed an ∼60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic ‘Micro-Tom’ tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage byT. absolutain these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.

Effect of diet delivered various concentrations of double-stranded RNA in silencing a midgut and a non-midgut gene of Helicoverpa armigera

2013 ◽  
Vol 103 (5) ◽  
pp. 555-563 ◽  
Author(s):  
R. Asokan ◽  
G. Sharath Chandra ◽  
M. Manamohan ◽  
N.K. Krishna Kumar
Keyword(s):  
Gene Silencing ◽  
Juvenile Hormone ◽  
Pest Management ◽  
Helicoverpa Armigera ◽  
Target Gene ◽  
Specific Gene ◽  
Upstream Gene ◽  
Double Stranded Rna ◽  
Helicoverpa Armígera ◽  
Rnai Response

AbstractRibonucleic acid interference (RNAi) is a sequence-specific gene silencing mechanism induced by double-stranded RNA (dsRNA). Recently, RNAi has gained popularity as a reverse genetics tool owing to its tremendous potential in insect pest management, which includes Helicoverpa armigera. However, its efficiency is mainly governed by dsRNA concentration, frequency of application, target gene, etc. Therefore, to obtain a robust RNAi response in H. armigera, we evaluated various concentrations of dsRNA and its frequency of applications delivered through diet in silencing a midgut gene, chymotrypsin and a non-midgut gene, juvenile hormone acid methyl transferase (jhamt) of H. armigera. The extent of target gene silencing was determined by employing reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Our study revealed four significant findings: (i) single application of dsRNA elicited a delayed and transient silencing, while multiple applications resulted in early and persistent silencing of the above genes; (ii) silencing of the non-midgut gene (jhamt) through diet delivered dsRNA revealed prevalence of systemic silencing probably due to communication of silencing signals in this pest; (iii) the extent of silencing of chymotrypsin was positively correlated with dsRNA concentration and was negatively correlated with jhamt; (iv) interestingly, over-expression (15–18 folds) of an upstream gene, farnesyl diphosphate synthase (fpps), in juvenile hormone (JH) biosynthetic pathway at higher concentrations of jhamt dsRNA was the plausible reason for lesser silencing of jhamt. This study provides an insight into RNAi response of target genes, which is essential for RNAi design and implementation as a pest management strategy.

scholarly journals RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum)

2016 ◽  
Author(s):  
Roberto A Camargo ◽  
Guilherme O Barbosa ◽  
Isabella Presotto Possignolo ◽  
Lazaro E. P. Peres ◽  
Eric Lam ◽  
...  
Keyword(s):  
Rna Interference ◽  
Gene Silencing ◽  
Target Gene ◽  
Target Genes ◽  
Larval Mortality ◽  
Tuta Absoluta ◽  
Specific Gene ◽  
Gene Transcript ◽  
Pest Species ◽  
In Planta

RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. We selected two target genes [Vacuolar ATPase-A and Arginine kinase] based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet for T. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on “in planta-induced transient gene silencing” (PITGS), a well-established method for silencing plant genes, used here for the first time to deliver in planta-transcribed dsRNA to target insect genes. Tuta absoluta larvae that fed on leaves containing dsRNA of the target genes showed an ~60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic ‘Micro-Tom’ tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage by T. absoluta in these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.

scholarly journals Gene Silencing: The Mechanism to Down Regulate the Target Gene

2018 ◽  
Vol 9 (6) ◽  
pp. 682-690
Author(s):  
Shreya ◽  
◽  
Ajeet Kumar Pandey ◽  
H. R. Bhandari ◽  
◽  
...  
Keyword(s):  
Gene Silencing ◽  
Target Gene

scholarly journals Transformation of plants with target gene encoding glutathione S-transferase to induce stress resistance

2020 ◽  
Author(s):  
E. V. Mikhaylova ◽  
M. Yu. Shein ◽  
V. Yu. Alekseev ◽  
E. A. Baimukhametova ◽  
Kh. G. Musin ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Nicotiana Tabacum ◽  
Transgenic Plants ◽  
Stress Resistance ◽  
Target Gene ◽  
Induce Stress ◽  
Eruca Sativa ◽  
Glutathione S Transferase ◽  
Gene Encoding

Transgenic plants Nicotiana tabacum, Brassica napus, B. rapa, Eruca sativa with target gene AtGSTF11 were created. Their stress resistance is being studied.

scholarly journals Gene silencing of BnaA09.ZEP and BnaC09.ZEP confers orange color in Brassica napus flowers

2020 ◽  
Vol 104 (4) ◽  
pp. 932-949
Author(s):  
Yingjun Liu ◽  
Shenhua Ye ◽  
Gaigai Yuan ◽  
Xiaowei Ma ◽  
Shuangping Heng ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Gene Silencing ◽  
Orange Color
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