scholarly journals Transformation of an artificial microRNA to soybean (Glycine max (L.) Merr.) to reduce the parasitism of Meloidogyne incognita

2021 ◽  
Vol 63 (5) ◽  
pp. 60-64
Author(s):  
Vu Phong Nguyen ◽  
◽  
Thi Truc Mai Ha ◽  
Le Tram Dang ◽  
The Phuong Nguyen ◽  
...  

Effectors are specific proteins secreted by nematodes into plant cells that facilitate their parasitism to host plants. Inactivation of these effectors could reduce the parasitic ability of nematodes on plants and the damage caused by nematodes. Gene Minc14137encodes an effector unknown function that is cloned from the Meloidogyne incognita. Artificial microRNAs capable of inactivating the gene Minc14137 were synthesised and inserted into an expression vector in soybean. This construct was transformed into the soybean cotyledon node mediated by Agrobacterium tumefaciens and regenerated transgenic plants. Copy number and the expression level of miRNA in T0 transgenic plants were determined by the qPCR technique. In T1 transgenic soybean plants, the pathogenic ability of root-knot nematode is reduced by 44.6-50.5% compared to the control plants. Results show that effector MINC14137 could play an important role in the parasitism of Meloidogyne incognita.

Author(s):  
Thi Thanh Nhan Pham ◽  
Huu Quan Nguyen ◽  
Thi Ngoc Lan Nguyen ◽  
Xuan Tan Dao ◽  
Danh Thuong Sy ◽  
...  

The dehydration responsive element binding (DREB) is a plant protein subfamily expressed when soybean plants face abiotic stresses. These DREB proteins are also considered to activate the transcription of drought-resistant genes. In this study, we present the determined results of relationships between overexpression of Glycine max DREB2 (GmDREB2) with the transcription level of Glycine max pyrroline-5-carboxylate synthetase (GmP5CS) gene, proline accumulation and drought tolerant ability transgenic soybean plants as the basis for selection of transgenic lines with high drought tolerance. GmDREB2 was inserted into a plant transgenic vector and the 35S-GmDREB2-cmyc construct was transferred into the soybean plants by Agrobacterium-mediated transformation. Recombinant GmDREB2 protein with a molecular weight of approximately 20 kDa was expressed in four transgenic soybean lines in the T1 generation. The GmP5CS gene was shown to have significantly (P<0.05) increased expression in the T2 transgenic soybean lines and higher than compared to non-transgenic plants with considering both in well watered condition and stressed conditions were from 1.06 to 1.31-fold and 1.37 to 1.95-fold, respectively. The proline content of four transgenic soybean lines increased 155.81% to 187.42 % after five days in drought-stress conditions and increased from 180.52 % to 233.74 % after nine days under drought-stress conditions (P<0.05). Therefore, the overexpression of GmDREB2 resulted in increasing transcription level of P5CS gene, proline accumulation and drought-stress tolerance of the transgenic soybean plants. The GmDREB2 transformation into soybean plants was confirmed by the results of genetically modified lines in the T2 generation (T2-1, T2-6, T2-7, and T2-8) with higher drought tolerant ability than those of non-transgenic plants.


2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Di Qin ◽  
Xiao-Yi Liu ◽  
Cristina Miceli ◽  
Qi Zhang ◽  
Pi-wu Wang

Abstract Background Cry8-like from Bacillus thuringiensis (Bt) encodes an insecticidal crystal (Cry) protein. Holotrichia parallela (Coleoptera: Scarabaeoidae), commonly known as the dark black chafer, is a troublesome pest of soybean (Glycine max). To test whether cry8-like can confer resistance against H. parallela to soybean, we introduced cry8-like from the Bt strain HBF-18 into soybean cultivar Jinong 28. Results Quantitative reverse transcription-PCR analysis demonstrated that cry8-like was expressed most highly in soybean leaves. In addition, Southern blot assays revealed that one copy of the integrated fragment was present in the transformed plants. Eight independent cry8-like transgenic lines were subsequently fed on by H. parallela. Under H. parallela feeding stress, the survival rates of the non-transgenic plants were 92% lower than those of the transgenic plants. The mortality rate of H. parallela increased when the larvae fed on the roots of T1 transgenic soybean plants. Moreover, the surviving larvae were deformed, and their growth was inhibited. Conclusions Collectively, our data suggest that transgenic soybean plants expressing the cry8-like gene are more resistant to H. parallela than non-transgenic plants and that transgenic expression of the cry8-like gene may represent a promising strategy for engineering pest tolerance. The events generated in this study could thus be utilized in soybean breeding programs.


Fruits ◽  
2009 ◽  
Vol 64 (5) ◽  
pp. 295-303 ◽  
Author(s):  
Hang Ye ◽  
Wen-jun Wang ◽  
Guo-jie Liu ◽  
Li-xin Zhu ◽  
Ke-gong Jia

2021 ◽  
Vol 219 ◽  
pp. 112312
Author(s):  
Yinlong Xiao ◽  
Ying Du ◽  
Yue Xiao ◽  
Xiaohong Zhang ◽  
Jun Wu ◽  
...  

Plant Disease ◽  
2017 ◽  
Vol 101 (6) ◽  
pp. 948-956 ◽  
Author(s):  
Alois A. Bell ◽  
Robert C. Kemerait ◽  
Carlos S. Ortiz ◽  
Sandria Prom ◽  
Jose Quintana ◽  
...  

Locally severe outbreaks of Fusarium wilt of cotton (Gossypium spp.) in South Georgia raised concerns about the genotypes of the causal pathogen, Fusarium oxysporum f. sp. vasinfectum. Vegetative complementation tests and DNA sequence analysis were used to determine genetic diversity among 492 F. oxysporum f. sp. vasinfectum isolates obtained from 107 wilted plants collected from seven fields in five counties. Eight vegetative complementation groups (VCG) were found, with VCG 01117B and VCG 01121 occurring in 66% of the infected plants. The newly recognized VCG 01121 was the major VCG in Berrien County, the center of the outbreaks. All eight VCG resulted in significant increases in the percentages of wilted leaves (27 to 53%) and significant reductions in leaf weight (40 to 67%) and shoot weight (33 to 60%) after being stem punctured into Gossypium hirsutum ‘Rowden’. They caused little or no significant reductions in shoot weight and height or increases in foliar symptoms and vascular browning in a soil-infestation assay. Soil infestation with Meloidogyne incognita race 3 (root-knot nematode) alone also failed to cause significant disease. When coinoculated with M. incognita race 3, all VCG caused moderate to severe wilt. Therefore, the VCG identified in this study belong to the vascular-competent pathotype, and should pose similar threats to cotton cultivars in the presence of the root-knot nematode. Use of nematode-resistant cultivars, therefore, is probably the best approach to control the disease in Georgia.


Author(s):  
Anil Baniya ◽  
Soumi Joseph ◽  
Larry Duncan ◽  
William Crow ◽  
Tesfamariam Mengistu

AbstractSex determination is a key developmental event in all organisms. The pathway that regulates sexual fate has been well characterized at the molecular level in the model free-living nematode Caenorhabditis elegans. This study aims to gain a preliminary understanding of sex-determining pathways in a plant-parasitic nematode Meloidogyne incognita, and the extent to which the roles of the sex determination genes are conserved in a hermaphrodite species, C. elegans, and plant-parasitic nematode species, M. incognita. In this study, we targeted two sex-determining orthologues, sdc-1 and tra-1 from M. incognita using RNA interference (RNAi). RNAi was performed by soaking second-stage juveniles of M. incognita in a solution containing dsRNA of either Mi-tra-1or Mi-sdc-1 or both. To determine the effect of RNAi of the target genes, the juveniles treated with the dsRNA were inoculated onto a susceptible cultivar of cowpea grown in a nutrient pouch at 28 °C for 5 weeks. The development of the nematodes was analyzed at different time points during the growth period and compared to untreated controls. Our results showed that neither Mi-sdc-1 nor Mi-tra-1 have a significant role in regulating sexual fate in M. incognita. However, the silencing of Mi-sdc-1 significantly delayed maturity to adult females but did not affect egg production in mature females. In contrast, the downregulation of Mi-tra-1 transcript resulted in a significant reduction in egg production in both single and combinatorial RNAi-treated nematodes. Our results indicate that M. incognita may have adopted a divergent function for Mi-sdc-1 and Mi-tra-1distinct from Caenorhabditis spp. However, Mi-tra-1 might have an essential role in female fecundity in M. incognita and is a promising dsRNA target for root-knot nematode (RKN) management using host-delivered RNAi.


PLoS ONE ◽  
2013 ◽  
Vol 8 (4) ◽  
pp. e61259 ◽  
Author(s):  
Alice Teillet ◽  
Katarzyna Dybal ◽  
Brian R. Kerry ◽  
Anthony J. Miller ◽  
Rosane H. C. Curtis ◽  
...  

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