scholarly journals An Improved Method for the Extraction of Nucleic Acids from Plant Tissue without Grinding to Detect Plant Viruses and Viroids

Plants ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2683
Author(s):  
Tatsuji Hataya

Gene amplification techniques such as polymerase chain reaction (PCR) are widely used for the diagnosis of plant diseases caused by viruses and viroids. It is preferable that sample preparation methods for PCR or reverse transcription (RT) PCR are rapid, straightforward, and inexpensive. We previously reported a method for the extraction of nucleic acids without mechanical tissue grinding using a buffer containing potassium ethyl xanthogenate (PEX) to detect viroid RNAs. In the present report, the previous PEX method was improved and simplified. In the simplified PEX (SPEX) method, the process of PEX buffer treatment for plant cell wall disruption is improved to one step of incubation at 80 °C for 10 min, instead of three steps that took more than 26 min at 65 °C in the previous method. Total nucleic acids could be extracted from fresh, frozen, or dried leaves of a cultivar or wild species of tobacco, tomato, citron, hop plants, and pericarps of persimmon fruits by the SPEX method. Several RNA viruses and viroids were successfully detected from the extracted nucleic acids together with an internal mRNA by RT-PCR. The SPEX method may be useful for detecting not only viruses and viroids, but also other plant pathogens.

Author(s):  
J. R. Hully ◽  
K. R. Luehrsen ◽  
K. Aoyagi ◽  
C. Shoemaker ◽  
R. Abramson

The development of PCR technology has greatly accelerated medical research at the genetic and molecular levels. Until recently, the inherent sensitivity of this technique has been limited to isolated preparations of nucleic acids which lack or at best have limited morphological information. With the obvious exception of cell lines, traditional PCR or reverse transcription-PCR (RT-PCR) cannot identify the cellular source of the amplified product. In contrast, in situ hybridization (ISH) by definition, defines the anatomical location of a gene and/or it’s product. However, this technique lacks the sensitivity of PCR and cannot routinely detect less than 10 to 20 copies per cell. Consequently, the localization of rare transcripts, latent viral infections, foreign or altered genes cannot be identified by this technique. In situ PCR or in situ RT-PCR is a combination of the two techniques, exploiting the sensitivity of PCR and the anatomical definition provided by ISH. Since it’s initial description considerable advances have been made in the application of in situ PCR, improvements in protocols, and the development of hardware dedicated to in situ PCR using conventional microscope slides. Our understanding of the importance of viral latency or viral burden in regards to HIV, HPV, and KSHV infections has benefited from this technique, enabling detection of single viral copies in cells or tissue otherwise thought to be normal. Clearly, this technique will be useful tool in pathobiology especially carcinogenesis, gene therapy and manipulations, the study of rare gene transcripts, and forensics.


2021 ◽  
Vol 83 (8) ◽  
Author(s):  
F. Al Basir ◽  
Y. N. Kyrychko ◽  
K. B. Blyuss ◽  
S. Ray

AbstractMany plant diseases are caused by plant viruses that are often transmitted to plants by vectors. For instance, the cassava mosaic disease, which is spread by whiteflies, has a significant negative effect on plant growth and development. Since only mature whiteflies can contribute to the spread of the cassava mosaic virus, and the maturation time is non-negligible compared to whitefly lifetime, it is important to consider the effects this maturation time can have on the dynamics. In this paper, we propose a mathematical model for dynamics of cassava mosaic disease that includes immature and mature vectors and explicitly includes a time delay representing vector maturation time. A special feature of our plant epidemic model is that vector recruitment is negatively related to the delayed ratio between vector density and plant density. We identify conditions of biological feasibility and stability of different steady states in terms of system parameters and the time delay. Numerical stability analyses and simulations are performed to explore the role of various parameters, and to illustrate the behaviour of the model in different dynamical regimes. We show that the maturation delay may stabilise epidemiological dynamics that would otherwise be cyclic.


2020 ◽  
Vol 5 (1) ◽  
pp. 404-440 ◽  
Author(s):  
Mehrdad Alizadeh ◽  
Yalda Vasebi ◽  
Naser Safaie

AbstractThe purpose of this article was to give a comprehensive review of the published research works on biological control of different fungal, bacterial, and nematode plant diseases in Iran from 1992 to 2018. Plant pathogens cause economical loss in many agricultural products in Iran. In an attempt to prevent these serious losses, chemical control measures have usually been applied to reduce diseases in farms, gardens, and greenhouses. In recent decades, using the biological control against plant diseases has been considered as a beneficial and alternative method to chemical control due to its potential in integrated plant disease management as well as the increasing yield in an eco-friendly manner. Based on the reported studies, various species of Trichoderma, Pseudomonas, and Bacillus were the most common biocontrol agents with the ability to control the wide range of plant pathogens in Iran from lab to the greenhouse and field conditions.


2013 ◽  
Vol 19 (3) ◽  
pp. 220-225 ◽  
Author(s):  
Siwon Lee ◽  
Eun-Ha Kang ◽  
Yong-Gil Shin ◽  
Su-Heon Lee
Keyword(s):  

2021 ◽  
Author(s):  
Quang D. Tran ◽  
Eric Galiana ◽  
Philippe Thomen ◽  
Céline Cohen ◽  
François Orange ◽  
...  

Phytophthora species cause diseases in a large variety of plants and represent a serious agricultural threat, leading, every year, to multibillion dollar losses. Infection occurs when these biflagellated zoospores move across the soil at their characteristic high speed and reach the roots of a host plant. Despite the relevance of zoospore spreading in the epidemics of plant diseases, it is not known how these zoospores swim and steer with two opposite beating flagella. Here, combining experiments and modeling, we show how these two flagella contribute to generate thrust when beating together, and identify the mastigonemes-attached anterior flagellum as the main source of thrust. Furthermore, we find that steering involves a complex active process, in which the posterior flagellum is stopped, while the anterior flagellum keeps on beating, as the zoospore reorients its body. Our study is a fundamental step towards a better understanding of the spreading of plant pathogens’ motile forms, and shows that the motility pattern of these biflagellated zoospores represents a distinct eukaryotic version of the celebrated “run-and-tumble” motility class exhibited by peritrichous bacteria.


Nanomaterials ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 2393
Author(s):  
Xiuping Wang ◽  
Fei Peng ◽  
Caihong Cheng ◽  
Lina Chen ◽  
Xuejuan Shi ◽  
...  

Plant pathogens constantly develop resistance to antimicrobial agents, and this poses great challenges to plant protection. Therefore, there is a pressing need to search for new antimicrobials. The combined use of antimicrobial agents with different antifungal mechanisms has been recognized as a promising approach to manage plant diseases. Graphene oxide (GO) is a newly emerging and highly promising antimicrobial agent against various plant pathogens in agricultural science. In this study, the inhibitory activity of GO combined with fungicides (Mancozeb, Cyproconazol and Difenoconazole) against Fusarium graminearum was investigated in vivo and in vitro. The results revealed that the combination of GO and fungicides has significant synergistic inhibitory effects on the mycelial growth, mycelial biomass and spore germination of F. graminearum relative to single fungicides. The magnitude of synergy was found to depend on the ratio of GO and fungicide in the composite. In field tests, GO–fungicides could significantly reduce the disease incidence and disease severity, exhibiting a significantly improved control efficacy on F. graminearum. The strong synergistic activity of GO with existing fungicides demonstrates the great application potential of GO in pest management.


2021 ◽  
Vol 10 (15) ◽  
pp. e296101522465
Author(s):  
Erika Valente de Medeiros ◽  
Lucas Figueira da Silva ◽  
Jenifer Sthephanie Araújo da Silva ◽  
Diogo Paes da Costa ◽  
Carlos Alberto Fragoso de Souza ◽  
...  

A better understanding of the use of biochar with Trichoderma spp. (TRI), considered the most studied tool for biological control, would increase our ability to set priorities. However, no studies exist using the two inputs on plant disease management. Here, we hypothesized that biochar and TRI would be used for the management of soilborne plant pathogens, mainly due to changes in soil properties and its interactions. To test this hypothesis, this review assesses papers that used biochar and TRI against plant diseases and we summarize the handling mechanisms for each input. Biochar acts by mechanisms: induction to plant resistance, sorption of allelopathic and fungitoxic compounds, increase of beneficial microorganisms, changes the soil properties that promote health and nutrient availability. Trichoderma as biocontrol agents by different mechanisms: mycoparasitism, enzyme and secondary metabolic production, plant promoter agent, natural decomposition agent, and biological agent of bioremediation. Overall, our findings expand our knowledge about the reuse of wastes transformed in biochar combined with Trichoderma has potential perspective to formulate products as alternative management tool of plant disease caused by soilborne fungal pathogen and add important information that can be suitable for development of strategy for use in the global health concept.


2018 ◽  
Vol 56 (1) ◽  
pp. 581-610 ◽  
Author(s):  
Cristina Rosa ◽  
Yen-Wen Kuo ◽  
Hada Wuriyanghan ◽  
Bryce W. Falk

The origin of RNA interference (RNAi), the cell sentinel system widely shared among eukaryotes that recognizes RNAs and specifically degrades or prevents their translation in cells, is suggested to predate the last eukaryote common ancestor ( 138 ). Of particular relevance to plant pathology is that in plants, but also in some fungi, insects, and lower eukaryotes, RNAi is a primary and effective antiviral defense, and recent studies have revealed that small RNAs (sRNAs) involved in RNAi play important roles in other plant diseases, including those caused by cellular plant pathogens. Because of this, and because RNAi can be manipulated to interfere with the expression of endogenous genes in an intra- or interspecific manner, RNAi has been used as a tool in studies of gene function but also for plant protection. Here, we review the discovery of RNAi, canonical mechanisms, experimental and translational applications, and new RNA-based technologies of importance to plant pathology.


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