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1

Xiang, Dao Hui, Guang Bin Yang, Song Liang, Yan Feng Wang, and Qiang Qin. "Study on Milling Force of High Volume Fraction SiCp/Al Composites with Ultrasonic Longitudinal and Torsional Vibration High Speed Milling." Advanced Materials Research 910 (March 2014): 114–17. http://dx.doi.org/10.4028/www.scientific.net/amr.910.114.

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High volume fraction SiCp/Al composites were milled in ultrasonic longitudinal and torsional vibration high speed milling and high speed milling in this experiment, study on effects of different milling parameters (milling depth ap, feed engagement fz, the milling speed vz) on milling force. The results shows that in the same cutting parameters ,the three milling force of ultrasonic longitudinal and torsional vibration high speed milling are smaller than that of high speed milling, and milling forces of two milling method increase with the add of the milling depth and feed engagement ,but they aren't fold increase. Milling forces of two milling method decrease with the add of the the minlling speed,and the changes become gently when the the minlling speed run up to 170m/min, with the further increase of the milling speed,ultrasonic vibration high speed milling will translate to high speed milling, this is because the torsional vibration is submerged.
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2

Martins, Ana Luiza. "Biblioteca Mindlin: a alegria da pesquisa." Revista BBM, no. 1 (September 11, 2018): 159–67. http://dx.doi.org/10.11606/issn.2595-5802.v1i1p159-167.

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A autora rememora as lembranças de sua experiência no convívio com Guita e José Mindlin na biblioteca pessoal do casal, hoje, Biblioteca Brasiliana Guita e José Minlin – BBM-USP. Relembra detalhes dos aspectos físicos do espaço e menciona algumas obras e textos que eram encontrados lá e a relação de Mindlin com eles. Recorda que a biblioteca era visitada por pesquisadores acadêmicos, mas também por jornalistas, escritores, bibliófilos e estudiosos do livro e que, dessas visitas, no início, eram realizadas pesquisas informais, que, posteriormente, foram sistematizadas e organizadas com regularidade, das quais resultaram obras fundamentais para a cultura nacional.
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3

Dai, Liang, Kristiaan Pelckmans, and Er-Wei Bai. "Identifiability and convergence analysis of the MINLIP estimator." Automatica 51 (January 2015): 104–10. http://dx.doi.org/10.1016/j.automatica.2014.10.091.

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4

Pelckmans, K., L. Dai, and E. Bai. "On the Convergence Analysis of the MINLIP Estimator*." IFAC Proceedings Volumes 45, no. 16 (July 2012): 482–87. http://dx.doi.org/10.3182/20120711-3-be-2027.00212.

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5

Jaisankar, Parasuraman, Bikash Pal, and Venkatachalam Sesha Giri. "MICROWAVE ASSISTED McFADYEN–STEVENS AND HUANG–MINLON REACTIONS." Synthetic Communications 32, no. 16 (January 2002): 2569–73. http://dx.doi.org/10.1081/scc-120005941.

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6

Pelckmans, Kristiaan. "MINLIP for the identification of monotone Wiener systems." Automatica 47, no. 10 (October 2011): 2298–305. http://dx.doi.org/10.1016/j.automatica.2011.08.026.

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7

Jaisankar, Parasuraman, Bikash Pal, and Venkatachalam Sesha Giri. "ChemInform Abstract: Microwave Assisted McFadyen-Stevens and Huang-Minlon Reactions." ChemInform 33, no. 47 (May 18, 2010): no. http://dx.doi.org/10.1002/chin.200247030.

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8

Berger, Ulrich, Alison Jones, and Monika Seisenberger. "Program extraction applied to monadic parsing." Journal of Logic and Computation 29, no. 4 (June 6, 2019): 487–518. http://dx.doi.org/10.1093/logcom/exv078.

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Abstract This article outlines a proof-theoretic approach to developing correct and terminating monadic parsers. Using modified realizability, we extract formally verified and terminating programs from formal proofs. By extracting both primitive parsers and parser combinators, it is ensured that all complex parsers built from these are also correct, complete and terminating for any input. We demonstrate the viability of our approach by means of two case studies: we extract (i) a small arithmetic calculator and (ii) a non-deterministic natural language parser. The work is being carried out in the interactive proof system Minlog.
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9

Spiteller, Peter, Jovan Jovanovic, and Michael Spiteller. "NMR analysis of (1S, 1aR, 6aR)-2’, 3’, 6, 6a-tetrahydrospiro cycloprop a indene-1(1aH), 1’- 1H indene." Journal of the Serbian Chemical Society 70, no. 10 (2005): 1133–36. http://dx.doi.org/10.2298/jsc0510133s.

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The aldol condensation product of 1H-indan-1-one, (2E)-2-(2,3-dihydro-1H-in den-1-ylidene)-2,3-dihydro-1H-inden-1-one, subjected to Huang?Minlon reduction conditions was shown, via 1D and 2D NMR analysis, to be a mixture of (1S,1aR,6aR)-2?,3?,6,6a-tetrahydro-spiro cycloprop a indene-1(1aH),1? 1H indene and its 1R,1aS,6aS enantiomer and not 2,3,1?,3?-tetrahydro- 1,2? -biindenylidene as originally expected. The full NMR assignment, the coupling constants in the proton NMR, and the couplings in the HMBC and NOESY of the title compound are summarized in the Table.
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10

Chodounská, Hana, and Alexander Kasal. "Allopregnanolone Derivatives: Synthesis of 3α-Hydroxy-7a-homo-5α-pregnan-20-one." Collection of Czechoslovak Chemical Communications 65, no. 7 (2000): 1156–62. http://dx.doi.org/10.1135/cccc20001156.

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3α-Hydroxy-7a-homo-5α-pregnan-20-one and 3α-hydroxy-7a-homo-5α-pregnane-7,20-dione were prepared from (20R)-3α-(methoxymethoxy)-7-oxo-5α-pregnan-20-yl benzoate. Homo- logation of the B ring was carried out by the Demyanov rearrangement of the corresponding cyanohydrin. Alkaline hydrolysis of the protecting group and oxidation of the formed 20-hydroxy group and deprotection of the 3-hydroxy group were performed either with (20R)-3α-(methoxymethoxy)-7-oxo-7a-homo-5α-pregnan-20-ol derivative or with the product of the Huang Minlon deoxygenation at carbon 7.
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11

Kasal, Alexander, and Miloš Buděšínský. "Partial Modifications of Hydroxy Groups in 5-Methyl-5β-estr-9-ene Derivatives." Collection of Czechoslovak Chemical Communications 61, no. 2 (1996): 276–87. http://dx.doi.org/10.1135/cccc19960276.

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In compounds of the Westphalen type the substituent in position 6 has been removed: the 6β-hydroxyl in 3β,17β-dibenzoyloxy-5-methyl-5β-estr-9-en-6β-ol (1) by oxidation to ketone 2 and Huang Minlon reduction, the 6β-chlorine atom in 3β-acetoxy-6β-chloro-5-methyl-5β-estr-9-en-17-one (13) by reduction with tributyltin hydride. Androstenedione, testosterone and methyltestosterone analogues 4, 5, and 16, respectively, were prepared from 3β,17β-dihydroxy-5-methyl-5β-estr-9-ene (3) by partial oxidation or partial acylation followed by oxidation. Alternatively, these derivatives were prepared from compound 13 without the need of partial transformations.
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12

MIYAMOTO, KENJI, and HELMUT SCHWICHTENBERG. "Program extraction in exact real arithmetic." Mathematical Structures in Computer Science 25, no. 8 (November 10, 2014): 1692–704. http://dx.doi.org/10.1017/s0960129513000327.

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The importance of an abstract approach to a computation theory over general data types has been stressed by Tucker in many of his papers. Berger and Seisenberger recently elaborated the idea for extraction out of proofs involving (only) abstract reals. They considered a proof involving coinduction of the proposition that any two reals in [−1, 1] have their average in the same interval, and informally extract a Haskell program from this proof, which works with stream representations of reals. Here we formalize the proof, and machine extract its computational content using the Minlog proof assistant. This required an extension of this system to also take coinduction into account.
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13

Rosen, Oleg M., Ali A. Abbyasov, and John C. Tipper. "MINLITH—an experience-based algorithm for estimating the likely mineralogical compositions of sedimentary rocks from bulk chemical analyses." Computers & Geosciences 30, no. 6 (July 2004): 647–61. http://dx.doi.org/10.1016/j.cageo.2004.03.011.

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14

Prager, Susanne. "The Coming of the "Future King": Burmese Minlaung Expectations Before and During the Second World War." Journal of Burma Studies 8, no. 1 (2003): 1–32. http://dx.doi.org/10.1353/jbs.2003.0000.

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15

Hernest, Mircea-Dan. "Synthesis of Moduli of Uniform Continuity by the Monotone Dialectica Interpretation in the Proof-system MinLog." Electronic Notes in Theoretical Computer Science 174, no. 5 (June 2007): 141–49. http://dx.doi.org/10.1016/j.entcs.2007.01.023.

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16

Ghosh, Pranab, and Prasanta Chakraborty. "ChemInform Abstract: Reaction of 2-Bromo-3-keto Triterpenoids: Deoxygenation and Simultaneous Debromination by Huang-Minlon Reduction Condition." ChemInform 44, no. 46 (October 24, 2013): no. http://dx.doi.org/10.1002/chin.201346187.

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17

Krokus, Christian. "The Sufi and the Friar: A Mystical Account of Two Men in the Abode of Islam by Minlib Dallh." Journal of Ecumenical Studies 54, no. 3 (2019): 470–71. http://dx.doi.org/10.1353/ecu.2019.0025.

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18

O'MAHONY, ANTHONY. "THE SUFI AND THE FRIAR: A MYSTICAL ENCOUNTER OF TWO MEN OF GOD IN THE ABODE OF ISLAM by Minlib Dallh, State University of New York Press, Albany, 2017, pp. xi + 201, $20.95, pbk." New Blackfriars 101, no. 1091 (December 11, 2019): 103–5. http://dx.doi.org/10.1111/nbfr.12529.

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19

Mufarrih, AM, Moh Nasir Hariyanto, and Nanang Qosim. "Analisa Kekasaran Permukaan Titanium Grade 2 pada Proses Frais." Jurnal Pendidikan Teknik Mesin Undiksha 8, no. 2 (August 1, 2020): 53. http://dx.doi.org/10.23887/jptm.v8i2.27766.

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Titanium Grade 2 termasuk jenis bahan yang sering dipergunakan di industri, utamanya pada bahan untuk implan biomedis. Titanium Grade 2 mempunyai sifat perbandingan kekakuan terhadap berat yang baik, tahan terhadap korosi dan memiliki sifat biokompatibel yang baik di dalam tubuh. Namun memiliki konduktifitas panas yang rendah, sehingga perlu memilih perameter pemesinan yang tepat untuk menghasilkan nilai kekasaran permukaan yang baik. Penelitian ini bertujuan untuk mengetahui karakteristik Titanium Grade 2 yaitu kekasaran permukaan hasil pemesinan frais. Desain penelitian menggunakan metode Taguchi L9, dengan 2 faktor dan 3 level. Parameter pemesinan yang digunakan ialah putaran spindel 500; 700; 900 rpm dan kecepatan pemakanan 25; 50; 75 mm/menit. Variabel respon yang diteliti ialah kekasaran permukaan. Proses frais dilakukan menggunakan Mesin CNC Dahlih. Kekasaran permukaan diukur menggunakan Mitutoyo surface roughess tester. Analisis data menggunakan analisis ANOVA. Hasil penelitian menunjukan bahwa ada pengaruh variasi parameter pemesinan terhadap respon kekasaran permukaan. Variabel putaran spindel mempunyai p-value sebesar 0,039 dan variabel gerak makan memiliki p-value sebesar 0,025. Hal ini menunjukkan bahwa kedua variabel bebas tersebut memiliki pengaruh yang signifikan terhadap respon kekasaran permukaan. Kekasaran permukaan terendah dapat dicapai dengan pengaturan putaran spindel sebesar 700 rpm dan kecepatan pemakanan sebesar 25 mm/menit. Kata kunci: titanium grade 2, kekasaran permukaan, frais, anova Daftar RujukanBagno, A., & Di Bello, C. (2004). Surface treatments and roughness properties of Ti-based biomaterials. Journal of Materials Science: Materials in Medicine. https://doi.org/10.1023/B:JMSM.0000042679.28493.7fBruce, 2011. (2013). Analisis Kekasaran Permukaan Dan Getaran Pada Pemesinan Bubut Menggunakan Pahat Putar Modular (Modular Rotary Tools) Untuk Material Titanium 6Al-4V Eli. Journal of Chemical Information and Modeling. https://doi.org/10.1017/CBO9781107415324.004Davim, J. P. (2011). Machining of hard materials. Machining of Hard Materials. https://doi.org/10.1007/978-1-84996-450-0Ganguli, S., & Kapoor, S. G. (2016). Improving the performance of milling of titanium alloys using the atomization-based cutting fluid application system. Journal of Manufacturing Processes. https://doi.org/10.1016/j.jmapro.2016.05.011Karkalos, N. E., Galanis, N. I., & Markopoulos, A. P. (2016). Surface roughness prediction for the milling of Ti-6Al-4V ELI alloy with the use of statistical and soft computing techniques. Measurement: Journal of the International Measurement Confederation. https://doi.org/10.1016/j.measurement.2016.04.039Kiswanto, G., Mandala, A., Azmi, M., & Ko, T. J. (2020). The effects of cutting parameters to the surface roughness in high speed cutting of micro-milling titanium alloy ti-6al-4v. Key Engineering Materials, 846 KEM, 133–138. https://doi.org/10.4028/www.scientific.net/KEM.846.133Mufarrih, A., Istiqlaliyah, H., & Ilha, M. M. (2019). Optimization of Roundness, MRR and Surface Roughness on Turning Process using Taguchi-GRA. In Journal of Physics: Conference Series. https://doi.org/10.1088/1742-6596/1179/1/012099Nithyanandam, J., Das, S. L., & Palanikumar, K. (2015). Inluence of Cutting Parameters in Machining of Titanium Alloy. Indian Journal of Science and Technology, 8(8), 556–562. https://doi.org/10.17485/ijst/2015/v8i/71291Oshida, Y. (2012). Bioscience and Bioengineering of Titanium Materials: Second Edition. Bioscience and Bioengineering of Titanium Materials: Second Edition. https://doi.org/10.1016/C2011-0-07805-5Setyowidodo, I., Sutanto, S., Mufarrih, A., & Sholehah, I. M. (2020). Exhaust temperature and peltier element optimization of thermoelectric generator output. In IOP Conference Series: Materials Science and Engineering. https://doi.org/10.1088/1757-899X/850/1/012007Shucai, Y., Chunsheng, H., & Minli, Z. (2019). A prediction model for titanium alloy surface roughness when milling with micro-textured ball-end cutters at different workpiece inclination angles. International Journal of Advanced Manufacturing Technology. https://doi.org/10.1007/s00170-018-2852-6Soepangkat, B. O. P., Pramujati, B., Effendi, M. K., Norcahyo, R., & Mufarrih, A. M. (2019). Multi-objective Optimization in Drilling Kevlar Fiber Reinforced Polymer Using Grey Fuzzy Analysis and Backpropagation Neural Network–Genetic Algorithm (BPNN–GA) Approaches. International Journal of Precision Engineering and Manufacturing. https://doi.org/10.1007/s12541-019-00017-zTapiero, H., Townsend, D. M., & Tew, K. D. (2003). Trace elements in human physiology and pathology. Copper. Biomedicine and Pharmacotherapy. https://doi.org/10.1016/S0753-3322(03)00012-XThepsonthi, T., & Özel, T. (2012). Multi-objective process optimization for micro-end milling of Ti-6Al-4V titanium alloy. International Journal of Advanced Manufacturing Technology. https://doi.org/10.1007/s00170-012-3980-zWennerberg, A., & Albrektsson, T. (2009). Effects of titanium surface topography on bone integration: A systematic review. Clinical Oral Implants Research. https://doi.org/10.1111/j.1600-0501.2009.01775.x
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20

Chattopadhyay, Sarmishtha, Sajal Kumar Banerjee, and Alok Kumar Mitra. "The Huang—Minlon Modification of Wolff—Kishner Reduction in Rapid and Simple Way Using Microwave Technology." ChemInform 34, no. 26 (July 1, 2003). http://dx.doi.org/10.1002/chin.200326055.

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21

Gerhardy, Philipp, and Ulrich Kohlenbach. "Extracting Herbrand Disjunctions by Functional Interpretation." BRICS Report Series 10, no. 32 (October 6, 2003). http://dx.doi.org/10.7146/brics.v10i32.21800.

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Carrying out a suggestion by Kreisel, we adapt Gödel's functional interpretation to ordinary first-order predicate logic(PL) and thus devise an algorithm to extract Herbrand terms from PL-proofs. The extraction is carried out in an extension of PL to higher types. The algorithm consists of two main steps: first we extract a functional realizer, next we compute the beta-normal-form of the realizer from which the Herbrand terms can be read off. Even though the extraction is carried out in the extended language, the terms are ordinary PL-terms. In contrast to approaches to Herbrand's theorem based on cut elimination or epsilon-elimination this extraction technique is, except for the normalization step, of low polynomial complexity, fully modular and furthermore allows an analysis of the structure of the Herbrand terms, in the spirit of Kreisel, already prior to the normalization step. It is expected that the implementation of functional interpretation in Schwichtenberg's MINLOG system can be adapted to yield an efficient Herbrand-term extraction tool.
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22

Monika, Pardeep Singh, and Satish Chand. "Computer vision-based framework for pedestrian movement direction recognition." Journal of Intelligent & Fuzzy Systems, February 21, 2023, 1–13. http://dx.doi.org/10.3233/jifs-223283.

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Pedestrians are the most critical and vulnerable moving objects on roads and public areas. Learning pedestrian movement in these areas can be helpful for their safety. To improve pedestrian safety and enable driver assistance in autonomous driver assistance systems, recognition of the pedestrian direction of motion plays an important role. Pedestrian movement direction recognition in real world monitoring and ADAS systems are challenging due to the unavailability of large annotated data. Even if labeled data is available, partial occlusion, body pose, illumination and the untrimmed nature of videos poses another problem. In this paper, we propose a framework that considers the origin and end point of the pedestrian trajectory named origin-end-point incremental clustering (OEIC). The proposed framework searches for strong spatial linkage by finding neighboring lines for every OE (origin-end) lines around the circular area of the end points. It adopts entropy and Q measure for parameter selection of radius and minimum lines for clustering. To obtain origin and end point coordinates, we perform pedestrian detection using the deep learning technique YOLOv5, followed by tracking the detected pedestrian across the frame using our proposed pedestrian tracking algorithm. We test our framework on the publicly available pedestrian movement direction recognition dataset and compare it with DBSCAN and Trajectory clustering model for its efficacy. The results show that the OEIC framework provides efficient clusters with optimal radius and minlines.
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23

Wiesnet, Franziskus, and Nils Köpp. "Limits of real numbers in the binary signed digit representation." Logical Methods in Computer Science Volume 18, Issue 3 (August 19, 2022). http://dx.doi.org/10.46298/lmcs-18(3:24)2022.

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We extract verified algorithms for exact real number computation from constructive proofs. To this end we use a coinductive representation of reals as streams of binary signed digits. The main objective of this paper is the formalisation of a constructive proof that real numbers are closed with respect to limits. All the proofs of the main theorem and the first application are implemented in the Minlog proof system and the extracted terms are further translated into Haskell. We compare two approaches. The first approach is a direct proof. In the second approach we make use of the representation of reals by a Cauchy-sequence of rationals. Utilizing translations between the two represenation and using the completeness of the Cauchy-reals, the proof is very short. In both cases we use Minlog's program extraction mechanism to automatically extract a formally verified program that transforms a converging sequence of reals, i.e.~a sequence of streams of binary signed digits together with a modulus of convergence, into the binary signed digit representation of its limit. The correctness of the extracted terms follows directly from the soundness theorem of program extraction. As a first application we use the extracted algorithms together with Heron's method to construct an algorithm that computes square roots with respect to the binary signed digit representation. In a second application we use the convergence theorem to show that the signed digit representation of real numbers is closed under multiplication.
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24

Bougard, Kayleigh, Hans Jacob Maree, Gerhard Pietersen, Julia Christine Meitz-Hopkins, and Rachelle Bester. "First report of Coguvirus eburi infecting pear (Pyrus communis) in South Africa." Plant Disease, August 29, 2021. http://dx.doi.org/10.1094/pdis-08-21-1630-pdn.

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Coguvirus eburi is a member of the genus Coguvirus in the family Phenuviridae (Khun et al., 2020). The species Coguvirus eburi was established to include citrus virus A (CiVA), which is a negative-sense, single-stranded RNA virus that was first found infecting sweet orange in southern Italy via high-throughput sequencing (HTS) (Navarro et al., 2018). This virus was also found to infect pome fruits in France, such as pear (Svanella-Dumas et al., 2019). More recently CiVA infections have been associated with impietratura disease in citrus (Beris et al. 2021). In the summer of 2021, leaf samples were collected from a pear tree (Pyrus communis cv. Bosc, B175) in the Koue Bokkeveld, South Africa as part of a virus survey. Sample B175 displayed no visual disease symptoms. One gram of leaf petioles was used for total RNA extraction, using a modified CTAB extraction protocol (Ruiz-García et al. 2019). Ribo-depleted RNA was prepared (Ribo-Zero Plant kit) and a sequencing library constructed (Illumina TruSeq Stranded Total RNA). The RNA library was paired-end (2 × 100 bp) sequenced on an Illumina HiSeqX instrument (Macrogen, South Korea). A total of 47,750,152 reads were obtained. Raw data was trimmed for quality with Trimmomatic (SLIDINGWINDOW:3:20, MINLEN:20) (Bolger et al. 2014). De novo assembly performed with CLC Genomics Workbench 11.0.1 (Qiagen) (Default parameters) using high quality reads yielded 75250 contigs. BLASTn analysis identified two viral contigs with high nucleotide (nt) identity to apple stem pitting virus (ASPV) and CiVA. The CiVA contig was 9400 nts and on closer examination, a concatemer of CiVA RNA1 and RNA2. The concatenation occurred due to the characteristic near-identical nucleotides shared at the 5’ and 3’ ends of RNA1 and RNA2 of these negative-stranded RNA viruses (Navarro et al., 2018). After splitting and curation, the RNA1 contig was 6664 nts and the RNA2 contig 2686 nts. A total of 51397 and 34820 reads were used to construct these contigs resulting in an average depth of coverage of 761 and 1281 for RNA1 and RNA2, respectively. The contigs had the highest nt identity to the complete CiVA GenBank accessions MT720885.1 (95.53%) and MW148460.1 (96.03%), spanning 99.6% and 98.1 % of the genomes of RNA1 and RNA2, respectively. These contigs were submitted as partial genomes to GenBank as accessions MZ463039 and MZ463040. Reverse transcription polymerase chain reaction (RT-PCR) was used to validate the presence of CiVA in sample B175. Two RT-PCR assays, directed at RNA1 and RNA2 respectively (Bester et al. (2021)) were used to generate amplicons. Amplicon sequences were confirmed with bi-directional Sanger sequencing. Twenty-one additional samples from the same orchard as B175 as well as other samples from the Koue Bokkeveld and Elgin areas, including cultivars Abate (10 samples), Forelle (10 samples), Early Bon Chretien (3 samples), Packham’s Triumph (12 samples) and Rosemarie (3 samples), were all surveyed for CiVA using the same RT-PCR assays as mentioned above. Thirty-six of the 59 samples tested were positive for CiVA, which further confirms the presence and wide-spread distribution of this virus in the limited survey conducted in pears in South Africa. However, no association with any disease symptoms or specific cultivar were identified. This is the first report of CiVA infecting pear in South Africa. This study therefore contributed to investigating the distribution of this virus and will assist the South African plant material certification scheme to assess the incidence of CiVA in South Africa.
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Bester, Rachelle, Shadelene Unisha Demas, and Hans Jacob Maree. "First report of Tomato spotted wilt orthotospovirus infecting agapanthus (Agapanthus praecox) in South Africa." Plant Disease, February 1, 2023. http://dx.doi.org/10.1094/pdis-12-22-2765-pdn.

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Agapanthus praecox Willd. is an ornamental flowering plant that is indigenous to southern Africa and was reported to be a host of tomato spotted wilt orthotospovirus (TSWV) in Australia in 2000 (Wilson et al. 2000). Tomato spotted wilt orthotospovirus (TSWV) belonging to the genus Orthotospovirus of the family Tospoviridae is a single-stranded negative sense RNA virus known to cause disease symptoms in many crops and ornamental plant species. This virus is in the top 10 of most economically important plant viruses worldwide (Rybicki 2015; Scholthof et al. 2011). In May 2021, leaf material from three agapanthus (Agapanthus praecox) plants displaying chlorotic mottling, and yellow lesions (Supplementary material 1A) was collected in Mbombela, South Africa. One gram of symptomatic leaf material was used for total RNA extraction from each of the three samples using a CTAB extraction protocol (Ruiz-García et al. 2019). The three RNA extracts were pooled, and a sequencing library was constructed using the Ion Total RNA-Seq Kit v2.0 and RiboMinus™ Plant Kit for RNA-Seq (ThermoFisher Scientific) (Central Analytical Facility (CAF), Stellenbosch University). The RNA library was sequenced on an Ion Torrent Proton Instrument (CAF). A total of 34,392,939 single-end reads were obtained. Data was trimmed for quality with Trimmomatic (CROP:250, MINLEN:50). De novo assembly was performed on the remaining 32,281,645 trimmed reads (average readlength: 100 nt, range: 50-250 nt) using SPAdes 3.13.0 and resulted in 4,788 contigs. BLASTn analysis identified viral contigs longer than 1,000 nucleotides (nts) with high nucleotide (nt) identity to TSWV (6 contigs), as well as to the newly discovered viruses, agapanthus tungro virus (AgTV) (1 contig), and agapanthus velarivirus (AgVV) (4 contigs) (Read et al 2021). Read mapping was performed against the relevant reference sequence with the highest nt identity to the contigs. For TSWV, 4995, 21221 and 14574 reads mapped to segment L (KY250488), M (KY250489) and S (KY250490) of isolate LK-1, respectively resulting in 99.97%, 100.00% and 99.97% genome coverage of the reference accessions. The nt identity between the reference accessions and the consensus sequences generated (OP921761-OP921763) were 97.26%, 97.64% and 97.82% for segment L, M and S. The presence of TSWV was confirmed in the HTS sample using an RT-PCR assay (primers L1 and L2) targeting the L segment of TSWV (Mumford et al. 1994). In July 2022, additional leaf samples displaying symptoms of chlorotic mottling, streaking, and ringspots were collected from 31 symptomatic and 3 asymptomatic agapanthus plants in public gardens in Stellenbosch, South Africa. Using the above-mentioned RT-PCR assay, 13 of the symptomatic samples tested positive for TSWV. All six plants displaying ring spot symptoms (Supplementary material 1B) were infected with TSWV. However, plants that displayed yellow streaking (five samples) and chlorotic mottling (two samples) (Supplementary material 1C-D) were also positive for TSWV which could be due to the presence of other viruses, plant growth stage, infection time or just variable symptom expression in a single host species as reported previously (Sherwood et al. 2003). The 275 bp RT-PCR amplicons of the HTS sample and three additional positive samples were validated with bidirectional Sanger sequencing (CAF) and had 96% identity to accession KY250488. The pairwise nt identity between amplicons was 98.55-100%. This is the first report of TSWV infecting agapanthus in South Africa. This study contributes information towards the distribution and incidence of TSWV and highlights the need for nurseries to screen plant material before propagation.
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