Expression and functional characterisation of a soluble form of Tomato yellow leaf curl virus coat protein

2014 ◽  
Vol 70 (10) ◽  
pp. 1624-1631 ◽  
Author(s):  
Lan-Lan Wang ◽  
Xue-Mei Wei ◽  
Xiao-Dong Ye ◽  
Hong-Xing Xu ◽  
Xue-Ping Zhou ◽  
...  
2017 ◽  
Author(s):  
Manal Tashkandi ◽  
Zahir Ali ◽  
Fatimah Aljedaani ◽  
Ashwag Shami ◽  
Magdy M. Mahfouz

AbstractCRISPR/Cas systems confer molecular immunity against phages and conjugative plasmids in prokaryotes. Recently, CRISPR/Cas9 systems have been used to confer interference against eukaryotic viruses. Here, we engineered Nicotiana benthamiana and tomato (Solanum lycopersicum) plants with the CRISPR/Cas9 system to confer immunity against the Tomato yellow leaf curl virus (TYLCV). Targeting the TYLCV genome with Cas9-single guide RNA at the sequences encoding the coat protein (CP) or replicase (Rep) resulted in efficient virus interference, as evidenced by low accumulation of the TYLCV DNA genome in the transgenic plants. The CRISPR/Cas9-based immunity remained active across multiple generations in the N. benthamiana and tomato plants. Together, our results confirmed the efficiency of the CRISPR/Cas9 system for stable engineering of TYLCV resistance in N. benthamiana and tomato, and opens the possibilities of engineering virus resistance against single and multiple infectious viruses in other crops.


Plant Disease ◽  
2002 ◽  
Vol 86 (12) ◽  
pp. 1404-1404 ◽  
Author(s):  
H. Delatte ◽  
B. Reynaud ◽  
J. M. Lett ◽  
M. Peterschmitt ◽  
M. Granier ◽  
...  

In April 2001, reduced leaf size, leaf curling, yellowing symptoms, and reduced yield were observed in tomato plants in the southwestern (Toliary, Morondava, Miandrivazo) and northern (Antsiranana) regions of Madagascar. Symptoms were similar to those caused by Tomato yellow leaf curl virus (TYLCV, genus Begomovirus, family Geminiviridae). Large populations of Bemisia tabaci (Gennadius) were observed colonizing tomato, other crops, and weeds. Leaf samples were collected from tomato plants from 14 sites located in northern, central, and southern Madagascar. Two plant samples collected near Antsiranana, one sample near Morondava, and one sample near Toliary were positive in triple-antibody sandwich enzyme-linked immunosorbent assay using a begomovirus-specific antibody purchased from ADGEN (Nellies Gates, Auchincruive, Scotland, UK). A 500-bp product was amplified and cloned (2) from two leaf samples collected near Toliary and one near Morondava using a pair of degenerate primers that are expected to amplify a region of the A component of begomoviruses between the intergenic conserved nonanucleotide sequence and the first 200 nucleotides of the coat protein ORF. The sequences corresponding to the two Toliary samples (GenBank Accession Nos. AJ422123 and AJ422124) and the Morondava sample (GenBank No. AJ422125) showed the most significant alignments (NCBI, BLAST) with begomoviruses, Tobacco leaf curl virus from Zimbabwe (GenBank Accession No. AF 350330) and Tomato leaf curl virus from Tanzania (GenBank Accession No. U73498) with 76 to 77% nucleotide identity (Clustal method, MegAlign, DNASTAR, London) and South African cassava mosaic viruses (SACMV GenBank Accession Nos. AJ422132 and AF155806) and East African cassava mosaic viruses from Malawi (GenBank Accession Nos. AJ006459 and AJ006460) with 74 to 75.5% nucleotide identity. The low nucleotide identity suggests that the begomovirus isolated from tomato in Madagascar is a new species. Since the core region of the coat protein gene is a molecular marker for provisional classification of begomoviruses (1), this region was amplified for the Morondava isolate with degenerate primers. The 519nt core fragment obtained showed the most significant alignments with SACMV (GenBank Accession No. AF329227), Cassava geminivirus from Mozambique (GenBank Accession No. AF329240), and with TYLCV (GenBank Accession Nos. AB014346 and AF105975) with 81 to 82% nucleotide identity. According to the current taxonomic criteria (4), the begomovirus from Madagascar is a new one that is related to begomoviruses from the southern part of Africa and to TYLCV and is provisionally named Tomato yellow leaf curl Morondava virus (TYLCMV). Tomato yellow leaf curl disease was previously described in Madagascar by Reckhaus (3) who presumed that it was caused by TYLCV. Although symptoms in the tomato plant from which TYLCMV was isolated were similar to those induced by TYLCV, TYLCV was not detected in our samples. References: (1) J. K. Brown et al. Arch. Virol. 146:1581, 2001 (2) M. Peterschmitt et al. Plant Dis. 83:303, 1999. (3) P. Reckhaus, Maladies et ravageurs des cultures maraîchères: A l'exemple de Madagascar. GTZ, Weikersem, 1997. (4) M. H. V. van Regenmortel et al. Virus Taxonomy. Seventh Rep. Int. Comm. Taxon. Viruses. Academic Press, San Diego, 2000.


EPPO Bulletin ◽  
2002 ◽  
Vol 32 (1) ◽  
pp. 31-35
Author(s):  
A. F. Arsenio ◽  
E. Neto ◽  
N. Ramos ◽  
S. Mangerico ◽  
E. Fortunato ◽  
...  

2020 ◽  
pp. 30-34
Author(s):  
С.Ф. Гавриш ◽  
Т.А. Редичкина ◽  
А.В. Буц ◽  
Г.М. Артемьева

Дана информация об изучении коллекции гибридов F1томата (Solanum lycopersicum L.) зарубежной селекции различных фирм-оригинаторов, рекомендованных производителями семян как толерантные к вирусу желтой курчавости листьев томата. Все гибриды обладали комплексом хозяйственно ценных признаков и набором генов устойчивости к основным заболеваниям томата, в том числе к новому для юга России опасному патогену с максимальным потенциальным риском – вирусу желтой курчавости листьев томата (Tomato yellow leaf curl virus — TYLCV). Исследования проведены в 2017-2018 годах в лаборатории пасленовых культур ООО «НИИСОК» и в лаборатории молекулярной диагностики растений ООО «Семеновод». Всего было протестировано 34 гибрида F1 томата. Гибриды оценивали по совокупности хозяйственно ценных признаков, также проводили молекулярно-генетический анализ на наличие и аллельное состояние основных генов устойчивости: к вирусу табачной мозаики (Tm2а), фузариозному увяданию (I2), вертициллезному увяданию (Ve), к кладоспориозу (Cf9), нематодам (Mi1.2), вирусу бронзовости томата (Sw5), вирусу желтой курчавости листьев томата (Ty3a). Установлено, что все проанализированные гибриды томата с заявленной оригинаторами семян устойчивостью к вирусу желтой курчавости листьев были гетерозиготны по гену Ty3a. На основании проведенных исследований и с учетом требований рынка разработаны модели гибридов F1 томата юга России. Перспективный гибрид томата должен обладать индетерминантным типом роста с укороченными междоузлиями (4,5-5 см) а также хорошей облиственностью. Плоды томата должны быть с красной равномерной окраской без зеленого пятна у плодоножки, с плоскоокруглой или округлой формой плода и со средней массой 220-270 г. Для повышения транспортабельности томатов необходимо, чтобы плоды отличались высокой прочностью и характеризовались хорошей лежкостью. Урожайность гибрида томата должна быть более 30 кг/м2, а товарность - не менее 85%. Гибрид томата должен обладать следующим набором генов устойчивости в гетерозиготном состоянии: Ty3a, Mi1.2, Cf-9, а также в гомозиготном состоянии: Tm2a, I2, Ve. The article provides information on the study of the collection of F1 tomato hybrids (Solanum lycopersicumL.) of foreign breeding from various firms-originators recommended for cultivation in regions with a strong spread of tomato yellow leaf curl virus. All hybrids had a complex of economically valuable traits and a set of genes for resistance to the main diseases of tomato, including a new dangerous pathogen for the South of Russia with a maximum potential risk — the tomato yellow leaf curl virus (TYLCV). The studies were carried out in 2017-2018 in the Solanaceae Laboratory of LLC NIISOK and in the Molecular Diagnostics Laboratory of Plants of LLC Semenovod. A total of 34 F1 tomato hybrids were tested. The hybrids were assessed by a set of economically valuable traits. Molecular genetic analysis was also carried out for the presence and allelic state of the main resistance genes: Tomato mosaic virus (Tm2a), Fusarium wilt (I2), Werticillium wilt (Ve), Cladosporium fulvum (Cf9), Nematodes (Mi1.2), Tomato spotted wilt virus (Sw5), Tomato yellow leaf curl virus (Ty3a). It was found that all the analyzed tomato hybrids with the declared by seed originators resistance to yellow leaf curl virus were heterozygous for the Ty3a gene. Based on the conducted research and taking into account the market requirements, models of F1 tomato hybrids for protected ground for the South of Russia have been developed. A promising tomato hybrid should have an indeterminate growth type with shortened internodes (4.5-5 cm) and good foliage. Tomato fruits should have a uniform red color without green shoulders, with a flat-round or round shape of the fruit and with an average weight of 220-270 g. To increase the transportability of tomatoes, it is necessary that the fruits are highly firm and characterized by good shelf life. The yield of tomato hybrid should be more than 30 kg/m2, and marketability should be at least 85%. The tomato hybrid should have the following set of resistance genes in a heterozygous state: Ty3a, Mi1.2, Cf-9, and also in a homozygous state: Tm2a, I2, Ve.


2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Adi Moshe ◽  
Eduard Belausov ◽  
Annette Niehl ◽  
Manfred Heinlein ◽  
Henryk Czosnek ◽  
...  

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Nathalie Becker ◽  
Loup Rimbaud ◽  
Frédéric Chiroleu ◽  
Bernard Reynaud ◽  
Gaël Thébaud ◽  
...  

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