Barbara Greco - Academia.edu (original) (raw)

Papers by Barbara Greco

Spanish Journal of Agricultural Research, 2013

ABSTRACT

Pest Management Science, 2013

The whitefly Bemisia tabaci (Gennadius) is a complex of cryptic species, some of which, namely th... more The whitefly Bemisia tabaci (Gennadius) is a complex of cryptic species, some of which, namely the Mediterranean (MED) and the Middle East-Asia Minor 1 (MEAM1), are highly invasive and injurious crop pests worldwide and able to displace local genotypes. Invasiveness of B. tabaci may depend on the phenotype of inherited bacterial endosymbionts. Here, the B. tabaci genetic diversity variation that has occurred in recent years in southern Italy was examined. Whitefly was genotyped by restriction fragment length polymorphism analysis of polymerase-chain-reaction-amplified fragments (PCR-RFLP) of the COI gene and molecular identification of endosymbionts. Possible factors leading to the observed genetic diversity were examined. Q1 and Q2 mitochondrial types of MED, the only species found, coexisted in the field, while MEAM1 disappeared. A large spreading of Q2 (70% of individuals) was observed for the first time in Italy. Q2 showed a significant female-biased sex ratio and largely outnumbered Q1 on solanaceous hosts, in greenhouses and on insecticide-treated plants. Q1, with an even sex ratio, slightly prevailed on non-solanaceous hosts, especially on wild and untreated plants. Endosymbiont composition was associated with the mitochondrial type. Hamiltonella and Rickettsia were found at near fixation in Q1 and Q2 respectively; Arsenophonus, Cardinium and Wolbachia were found in both types, although at different frequencies. Q2 invasion seems to have been favoured by the agroecological conditions of southern Italy and by the female-biased sex ratio. Endosymbionts may have a role in Q2 invasiveness, acting as sex-ratio manipulators (e.g. Rickettsia) and possibly by benefiting the host fitness.

ABSTRACT Yucca aloifolia L. (Spanish bayonet), family Asparagaceae, is the type species of the ge... more ABSTRACT Yucca aloifolia L. (Spanish bayonet), family Asparagaceae, is the type species of the genus Yucca. It is native to Mexico and the West Indies and is appreciated worldwide as an ornamental plant. In 2013, during a survey for viruses in ornamental plants in the Campania region of southern Italy, symptoms consisting of bright chlorotic spots and ring spots 1 to 3 mm in diameter with some necrotic streaks were observed on leaves of two plants of Y. aloifolia growing in a nursery located in the Pignataro Maggiore municipality, Caserta Province. Cucumber mosaic virus (CMV) infection was suspected because the symptoms resembled those caused by CMV in Yucca flaccida (1). A range of herbal plant indicators was inoculated with sap extracts of symptomatic Y. aloifolia plants and developed symptoms indicative of CMV. Furthermore, 30 nm isometric virus particles were observed in the same Y. aloifolia sap extracts by transmission electron microscopy. The identity of the virus was confirmed by positive reaction in ELISA tests with CMV polyclonal antisera (Bioreba) conducted on sap extracts of symptomatic Y. aloifolia plants and systemically infected symptomatic hosts (i.e., Nicotiana tabacum, N. glutinosa, Cucumber sativus cv. Marketer, Solanum lycopersicum cv. San Marzano). The presence of CMV in the two naturally infected Y. aloifolia and other mechanically inoculated plants was further verified by reverse transcription (RT)-PCR. Total RNAs were extracted with the E.Z.N.A. Plant RNA Kit (Omega Bio-Tek), according to the manufacturer's instructions. RT-PCR was carried out with the ImProm-II Reverse Transcription System first-strand synthesis reaction (Promega) using the primer pair CMV1 and CMV2 (2). These primers amplify part of the CP gene and part of the 3′-noncoding region of CMV RNA3 and were designed to produce amplicons of different sizes to distinguish CMV isolates belonging to subgroups I or II (3). RT-PCR products were obtained from both naturally infected Y. aloifolia and mechanically inoculated plants as well as from PAE1 isolate of CMV (2), used as positive control, but not from healthy plants. Based on the length of the amplicons obtained (487 bp), the CMV isolate from Y. aloifolia (named YAL) belonged to subgroup I (3). The amplified RT-PCR products were purified with QIAquick PCR Purification Kit (Qiagen), cloned in the pGEMT vector (Promega), and three independent clones were sequenced at MWG (Ebersberg, Germany). Sequences obtained from the two CMV-infected Y. aloifolia plants were identical. This sequence was deposited at GenBank (Accession No. HG965199). Multiple alignments of the YAL sequence with Quick Links Add to favorites E-mail to a colleague Alert me when new articles cite this article Download to citation manager Related articles found in APS Journals (Accession No. HG965199). Multiple alignments of the YAL sequence with sequences of other CMV isolates using MEGA5 software revealed highest percentage of identity (98.9%) with the isolates Z (AB369269) and SO (AF103992) from Korea and Japan, respectively. Moreover, the YAL isolate was identified as belonging to subgroup IA, based on the presence of only one HpaII restriction site in the 487-bp sequence, as previously proposed (2). Although CMV seems to not be a major threat currently for the production of Y. aloifolia, because the farming of this plant is performed using vegetative propagation, particular attention should be given to the presence of the virus in donor mother plants in order to avoid the dispersion of infected plants that could serve as sources for aphid transmission to other susceptible plant species. To our knowledge, this is the first report of CMV infection of Y. aloifolia in the world. References: (1) I. Bouwen et al. Neth. J. Plant Pathol. 84:175, 1978. (2) G. Parrella and D. Sorrentino. J. Phytopathol. 157:762, 2009. (3) Z. Singh et al. Plant Dis. 79:713, 1995.

American Journal of Plant Sciences, 2013

Spanish Journal of Agricultural Research, 2013

ABSTRACT

Pest Management Science, 2013

The whitefly Bemisia tabaci (Gennadius) is a complex of cryptic species, some of which, namely th... more The whitefly Bemisia tabaci (Gennadius) is a complex of cryptic species, some of which, namely the Mediterranean (MED) and the Middle East-Asia Minor 1 (MEAM1), are highly invasive and injurious crop pests worldwide and able to displace local genotypes. Invasiveness of B. tabaci may depend on the phenotype of inherited bacterial endosymbionts. Here, the B. tabaci genetic diversity variation that has occurred in recent years in southern Italy was examined. Whitefly was genotyped by restriction fragment length polymorphism analysis of polymerase-chain-reaction-amplified fragments (PCR-RFLP) of the COI gene and molecular identification of endosymbionts. Possible factors leading to the observed genetic diversity were examined. Q1 and Q2 mitochondrial types of MED, the only species found, coexisted in the field, while MEAM1 disappeared. A large spreading of Q2 (70% of individuals) was observed for the first time in Italy. Q2 showed a significant female-biased sex ratio and largely outnumbered Q1 on solanaceous hosts, in greenhouses and on insecticide-treated plants. Q1, with an even sex ratio, slightly prevailed on non-solanaceous hosts, especially on wild and untreated plants. Endosymbiont composition was associated with the mitochondrial type. Hamiltonella and Rickettsia were found at near fixation in Q1 and Q2 respectively; Arsenophonus, Cardinium and Wolbachia were found in both types, although at different frequencies. Q2 invasion seems to have been favoured by the agroecological conditions of southern Italy and by the female-biased sex ratio. Endosymbionts may have a role in Q2 invasiveness, acting as sex-ratio manipulators (e.g. Rickettsia) and possibly by benefiting the host fitness.

ABSTRACT Yucca aloifolia L. (Spanish bayonet), family Asparagaceae, is the type species of the ge... more ABSTRACT Yucca aloifolia L. (Spanish bayonet), family Asparagaceae, is the type species of the genus Yucca. It is native to Mexico and the West Indies and is appreciated worldwide as an ornamental plant. In 2013, during a survey for viruses in ornamental plants in the Campania region of southern Italy, symptoms consisting of bright chlorotic spots and ring spots 1 to 3 mm in diameter with some necrotic streaks were observed on leaves of two plants of Y. aloifolia growing in a nursery located in the Pignataro Maggiore municipality, Caserta Province. Cucumber mosaic virus (CMV) infection was suspected because the symptoms resembled those caused by CMV in Yucca flaccida (1). A range of herbal plant indicators was inoculated with sap extracts of symptomatic Y. aloifolia plants and developed symptoms indicative of CMV. Furthermore, 30 nm isometric virus particles were observed in the same Y. aloifolia sap extracts by transmission electron microscopy. The identity of the virus was confirmed by positive reaction in ELISA tests with CMV polyclonal antisera (Bioreba) conducted on sap extracts of symptomatic Y. aloifolia plants and systemically infected symptomatic hosts (i.e., Nicotiana tabacum, N. glutinosa, Cucumber sativus cv. Marketer, Solanum lycopersicum cv. San Marzano). The presence of CMV in the two naturally infected Y. aloifolia and other mechanically inoculated plants was further verified by reverse transcription (RT)-PCR. Total RNAs were extracted with the E.Z.N.A. Plant RNA Kit (Omega Bio-Tek), according to the manufacturer's instructions. RT-PCR was carried out with the ImProm-II Reverse Transcription System first-strand synthesis reaction (Promega) using the primer pair CMV1 and CMV2 (2). These primers amplify part of the CP gene and part of the 3′-noncoding region of CMV RNA3 and were designed to produce amplicons of different sizes to distinguish CMV isolates belonging to subgroups I or II (3). RT-PCR products were obtained from both naturally infected Y. aloifolia and mechanically inoculated plants as well as from PAE1 isolate of CMV (2), used as positive control, but not from healthy plants. Based on the length of the amplicons obtained (487 bp), the CMV isolate from Y. aloifolia (named YAL) belonged to subgroup I (3). The amplified RT-PCR products were purified with QIAquick PCR Purification Kit (Qiagen), cloned in the pGEMT vector (Promega), and three independent clones were sequenced at MWG (Ebersberg, Germany). Sequences obtained from the two CMV-infected Y. aloifolia plants were identical. This sequence was deposited at GenBank (Accession No. HG965199). Multiple alignments of the YAL sequence with Quick Links Add to favorites E-mail to a colleague Alert me when new articles cite this article Download to citation manager Related articles found in APS Journals (Accession No. HG965199). Multiple alignments of the YAL sequence with sequences of other CMV isolates using MEGA5 software revealed highest percentage of identity (98.9%) with the isolates Z (AB369269) and SO (AF103992) from Korea and Japan, respectively. Moreover, the YAL isolate was identified as belonging to subgroup IA, based on the presence of only one HpaII restriction site in the 487-bp sequence, as previously proposed (2). Although CMV seems to not be a major threat currently for the production of Y. aloifolia, because the farming of this plant is performed using vegetative propagation, particular attention should be given to the presence of the virus in donor mother plants in order to avoid the dispersion of infected plants that could serve as sources for aphid transmission to other susceptible plant species. To our knowledge, this is the first report of CMV infection of Y. aloifolia in the world. References: (1) I. Bouwen et al. Neth. J. Plant Pathol. 84:175, 1978. (2) G. Parrella and D. Sorrentino. J. Phytopathol. 157:762, 2009. (3) Z. Singh et al. Plant Dis. 79:713, 1995.

American Journal of Plant Sciences, 2013