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      Whitefly-tolerant transgenic common bean ( Phaseolus vulgaris) line

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          Abstract

          Common bean ( Phaseolus vulgaris L.) is a staple food in Brazil with both nutritional and socioeconomic importance. As an orphan crop, it has not received as much research attention as the commodity crops. Crop losses are strongly related to virus diseases transmitted by the whitefly Bemisia tabaci, one of the most important agricultural pests in the world. The main method of managing whitefly-transmitted viruses has been the application of insecticides to reduce vector populations. Compared to chemical vector control, a more sustainable strategy for managing insect-borne viruses is the development of resistant/tolerant cultivars. RNA interference has been applied to develop plant lines resistant to the whitefly in other species, such as tomato, lettuce and tobacco. Still, no whitefly-resistant plant has been made commercially available to date. Common bean is a recalcitrant species to in vitro regeneration; therefore, stable genetic transformation of this plant has been achieved only at low frequencies (<1%) using particle bombardment. In the present work, two transgenic common bean lines were obtained with an intron-hairpin construct to induce post-transcriptional gene silencing against the B. tabaci vATPase (Bt- vATPase) gene, with stable expression of siRNA. Northern blot analysis revealed the presence of bands of expected size for siRNA in leaf samples of the line Bt-22.5, while in the other line (11.5), the amount of siRNA produced was significantly smaller. Bioassays were conducted with both lines, but only the line Bt-22.5 was associated with significant mortality of adult insects (97% when insects were fed on detached leaves and 59% on the whole plant). The expression of the Bt- vATPase gene was 50% lower ( p < 0.05) in insects that fed on the transgenic line Bt-22.5, when compared to non-transgenic controls. The transgenic line did not affect the virus transmission ability of the insects. Moreover, no effect was observed on the reproduction of non-target organisms, such as the black aphid Aphis craccivora, the leafminer Liriomyza sp. and the whitefly parasitoid Encarsia formosa. The results presented here serve as a basis for the development of whitefly-tolerant transgenic elite common bean cultivars, with potential to contribute to the management of the whitefly and virus diseases.

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            A plant DNA minipreparation: Version II

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              Bemisia tabaci: a statement of species status.

              Bemisia tabaci has long been considered a complex species. It rose to global prominence in the 1980s owing to the global invasion by the commonly named B biotype. Since then, the concomitant eruption of a group of plant viruses known as begomoviruses has created considerable management problems in many countries. However, an enduring set of questions remains: Is B. tabaci a complex species or a species complex, what are Bemisia biotypes, and how did all the genetic variability arise? This review considers these issues and concludes that there is now sufficient evidence to state that B. tabaci is not made up of biotypes and that the use of biotype in this context is erroneous and misleading. Instead, B. tabaci is a complex of 11 well-defined high-level groups containing at least 24 morphologically indistinguishable species.
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                Author and article information

                Contributors
                URI : https://loop.frontiersin.org/people/1940368/overview
                URI : https://loop.frontiersin.org/people/357445/overview
                URI : https://loop.frontiersin.org/people/1940261/overview
                URI : https://loop.frontiersin.org/people/1940209/overview
                URI : https://loop.frontiersin.org/people/1940271/overview
                URI : https://loop.frontiersin.org/people/742519/overview
                URI : https://loop.frontiersin.org/people/1940722/overview
                URI : https://loop.frontiersin.org/people/1253378/overview
                URI : https://loop.frontiersin.org/people/1165838/overview
                URI : https://loop.frontiersin.org/people/266046/overview
                URI : https://loop.frontiersin.org/people/1708595/overview
                Journal
                Front Plant Sci
                Front Plant Sci
                Front. Plant Sci.
                Frontiers in Plant Science
                Frontiers Media S.A.
                1664-462X
                25 August 2022
                2022
                : 13
                : 984804
                Affiliations
                [1] 1Laboratory of Entomology, Embrapa Arroz e Feijão , Santo Antônio de Goiás, Brazil
                [2] 2Laboratory of Biotechnology, Embrapa Arroz e Feijão , Santo Antônio de Goiás, Brazil
                [3] 3Escola de Agronomia, Universidade Federal de Goiás , Goiânia, Brazil
                [4] 4Laboratory of Genetic Engineering Applied to Tropical Agriculture, Embrapa Recursos Genéticos e Biotecnologia , Brasília, Brazil
                [5] 5Laboratory of Biotechnology, Embrapa Algodão , Santo Antônio de Goiás, Brazil
                Author notes

                Edited by: Maria Celeste Gonçalves-Vidigal, Universidade Estadual de Maringá, Brazil

                Reviewed by: Kelvin Kamfwa, University of Zambia, Zambia; Qijiian Song, United States Department of Agriculture, United States

                *Correspondence: Patricia Valle Pinheiro, patricia.pinheiro@ 123456embrapa.br

                This article was submitted to Plant Breeding, a section of the journal Frontiers in Plant Science

                Article
                10.3389/fpls.2022.984804
                9453422
                36092396
                88dfc74b-9789-4d76-b690-2dbe1be10a3d
                Copyright © 2022 Ferreira, de Faria, da Costa Moura, de Mendonça Zaidem, Pizetta, de Oliveira Freitas, Coelho, Silva, Barrigossi, Hoffmann, de Souza, Aragão and Pinheiro.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 02 July 2022
                : 25 July 2022
                Page count
                Figures: 7, Tables: 2, Equations: 0, References: 40, Pages: 13, Words: 9319
                Funding
                Funded by: Brazilian Agricultural Research Corporation – Embrapa
                Award ID: 20.18.03.035.00.00
                Award ID: 10.19.00.089.00.00.00
                Award ID: 20.18.04.008.00.00
                Categories
                Plant Science
                Original Research

                Plant science & Botany
                dry bean,bemisia tabaci,rna interference,insect pest management,vatpase
                Plant science & Botany
                dry bean, bemisia tabaci, rna interference, insect pest management, vatpase

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