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Rapid detection of tomato spotted wilt virus by real-time RT-LAMP and in-field application
Full text linkTomato spotted wilt virus (TSWV) is considered one of the most threatening viruses worldwide for diferent economically important agricultural crops. In this scenario, it is important to perform an early detection by laboratory tests to prevent TSWV spread. A rapid and sensitive TSWV detection protocol based on real time reverse transcription loop-mediated isothermal amplifcation (RT-LAMP) assay was developed in this work, also using cost-efective and simplifed sample preparation procedure, to assess the suitability of the RT-LAMP assay in feld conditions on tomato and pepper samples. A set of six primers was designed within the nucleotide sequence region coding for the nucleocapsid protein (N) of segment S, targeting a 220-nucleotide sequence. Sensitivity, specifcity, accuracy, and in-feld application of the real-time RT-LAMP assay were evaluated. The developed real-time RT-LAMP assay proved to be one thousand and one hundred times more sensitive than end-point RT-PCR and real-time RT-PCR methods, respectively, detecting a total of 9.191 × 101 genome copies as minimum target, and no cross-reactivity were detected with other viruses belonging to Tospoviridae and Bromoviridae families used as outgroup. In addition, the in-feld application of the assay using the rapid sample preparation gave adequate and reliable results within 60 minutes, with an acceptable reaction delay when compared to canonical RNA extraction. The in-feld analyses showed an increase of TSWV-positive samples (37%) detection compared
with end-point RT-PCR and real-time RT-PCR (32% and 29%, respectively), particularly on asymptomatic samples, confrming that the real-time RT-LAMP assay can be implemented as a routine test both in-feld and laboratory conditions as a rapid and sensitive technique for TSWV detection
Estimated costs of plum pox virus and management of sharka, the disease it causes
No full textThe disease “sharka”, caused by Potyvirus plumpoxi (plum pox virus), is the most harmful viral disease affecting stone fruits. The virus spreads over long distances through illegal and insufficiently controlled exchange of infected propagative plant material. Once established in an area, the virus spreads locally through vegetative propagation of infected plant material, and naturally through aphid-vectors. Previously considered a European problem, sharka has now been reported in 54 Prunus-growing countries in all continents except Oceania, although the disease has been eradicated from the United States of America. The economic cost of the disease in the 28 years from 1995 to 2023 is estimated to be€ 2.4× 10 9, equivalent to approx. 0.17% of the stone fruit industry’s value. This includes more than over€ 2× 10 9 in direct fruit losses,€ 1.4 million from international rejection of symptomatic fruit, and over€ 100 million in eradication and disease limitation costs. Indirect costs include€ 137 million, mainly associated with ELISA analyses, and approx.€ 130 million in costs related to research and science networks. Cumulative global losses from the sharka pandemic since the decade 1910/20 probably surpass€ 13× 10 9. These outlays exclude indirect trade costs, economic losses, genetic erosion of traditional cultivars, and the costs of developing new cultivars tolerant or resistant to plum pox virus. The decline in these costs compared to the previously evaluated€ 10 billion from the 1970s to 2006 is analyzed. Four case studies (for Spain, Turkey, Chile, and Greece) illustrate different sharka scenarios and management strategies
Screening Local Sicilian Tomato Ecotypes to Evaluate the Response of Tomato Brown Rugose Fruit Virus Infection
Full text linkTomato is one of the most important crops worldwide, with a production of ≈190 million tons,
but it is constantly threatened by several viral diseases. Tomato brown rugose fruit virus (ToBRFV),
identified in 2014 on tomato plants and subsequently reported in many countries, represents one of
the major threats to tomato crops, due to production losses, different transmission modes and its rapid
spread. This work aimed to evaluate 37 local Sicilian tomato ecotypes against ToBRFV infection. After a
preliminary screening by molecular analyses for tomato mosaic virus (ToMV) and pepino mosaic virus
(PepMV), and ToBRFV detection, tomato plants were grown in a greenhouse for their morphological
characterization and for evaluating resistance and tolerance to ToBRFV. Resistance and tolerance levels
were estimated by mechanical inoculation with ToB SIC01/19 ToBRFV isolate in ten plants per ecotype
and evaluating virus accumulation by RT-qPCR and visual observation of symptoms. All ecotypes were
infected with ToBRFV, showing several symptoms with different disease severity. No tomato ecotype
showed a high level of resistance, but two ecotypes, Pop27 and Pop35, showed very moderate symptoms
and therefore a high tolerance. These Sicilian tomato ecotypes could be used in genetic breeding programs
as parental ones to obtain cultivars tolerant to ToBRFV
Perfil nutricional de granada cv. Mollar de Elche en parcelas tratadas para reducir el pardeamiento interno de la corteza
Full text linkEn la Comunidad Valenciana destaca la granada cv. Mollar de Elche. Recientemente, y cada
vez con mayor incidencia, la corteza está desarrollando un pardeamiento interno tras su frigoconservación,
principalmente en la zona de la corona. Actualmente, se desconoce si este daño
influye en el perfil nutricional del zumo.
Para estudiarlo, se seleccionaron frutos de parcelas con baja (P1) y alta incidencia de daño (P2).
Para minimizar los daños, se realizaron tratamientos con calcio y boro (Ca+B) y cobre (Cu), para
compararlo con árboles sin tratamiento (CTL). Los frutos fueron recolectados y almacenados
a 5 y 9 °C durante 4 y 8 semanas. Se exprimió por separado la corona y el pedúnculo y se
analizaron los sólidos solubles totales y el índice de color, así como los azúcares y antocianos
individuales.
En ambas parcelas, el zumo de la corona presentó más cianidina-3-O-glucósido y glucosa que
el pedúnculo. Las concentraciones de pelargolina-3-O-glucósido y fructosa fueron, también,
superiores en la corona únicamente de la P1. Al comparar las parcelas, se observó una menor
concentración de todos los metabolitos en la fruta dañada de P2, así como una coloración
menos rojiza.
El efecto de los tratamientos aplicados afectó de forma diferente al perfil nutricional del zumo,
dependiendo del grado de incidencia del pardeamiento. Los frutos de la P1 presentaron menor
concentración de los metabolitos analizados cuando se aplicó Ca+B; mientras que los mayores
valores se encontraron en los frutos tratados con Cu. En la P2 esta tendencia sólo se observó
en el antociano mayoritario (cianidina-3-O-glucósido).
Tras la frigo-conservación, el zumo de la fruta almacenada a 9 °C, que presentó menor incidencia
de daño que la almacenada a 5 °C, tuvo mayor coloración y concentración de antocianos.
Trabajo financiado por el proyecto 52203 GVA-IVIA, co-financiado por los Fondos Europeos de
Desarrollo Regional (ERDF)
Potency evaluation of different GNRH analogues on ovulation induction and reproductive performance of doe rabbit
Full text linkGonadotropin-releasing
hormone (GnRH) -supplemented
extenders have emerged as
a welfare-orientated
method to induce ovulation in the artificial insemination (AI) of
rabbits. The main factor that limits the bioavailability of GnRH analogue on intravaginal
administration is the proteolytic activity of enzymes present in rabbit seminal
plasma. The use of GnRH analogues with higher biological potency would allow us to
decrease their concentration in the seminal dose without compromising effectiveness.
The current study was designed to assess the efficacy of various GnRH analogues
concerning their ability to induce ovulation in rabbit AI. The base solution used
for experimental extenders contained an aminopeptidase inhibitor. Four experimental
groups were used, females from the Control group were induced to ovulate with an
intramuscular administration of 1 μg of buserelin, while in the other three groups females
received an intravaginal administration of 3.5 μg of buserelin (BUS), deslorelin
(DES) or fertirelin (FER) within the seminal dose. Results showed that the ovulation
frequency was similar in all groups studied. A concentration of 3.5 μg of the different
GnRH analogues tested in this study showed similar potency in inducing ovulation in
non-lactating
females, yielding comparable results in terms of pregnancy rate at birth
and prolificacy
Ionomic Concentration and Metabolomic Profile of Organically and Conventionally
Full text linkThis study evaluates mineral concentrations, biocomponents contents and fruit quality attributes in ‘Rojo Brillante’ persimmon grown under organic and conventional managements. During two seasons, the concentrations of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn), zinc (Zn), and boron (B) were determined in leaves and fruits. Weight, color, firmness, and total soluble solids (TSS) were also evaluated in fruits. Moreover, in the second season, organic acids (citric, succinic, and fumaric acids), main sugars (sucrose, glucose, and fructose), carotenoids (β-carotene, β-cryptoxanthin, violaxanthin, lutein, and zeaxanthin), phenolic compounds (gallic and ρ-coumaric), and ascorbic acid concentrations were determined in fruit flesh. The crop yield in the conventional plots was bigger than that for organic crops. Nevertheless, the highest agronomic efficiency was found in organic management. In general, the greater nutrient supply in the conventional compared to in the organic system did not result in higher concentrations of macro- and microelements in leaves and fruits. The organic fruit had higher color values and lower firmness values than the conventional fruit. The concentrations of malic acid, β-cryptoxanthin, and ascorbic acid were higher in the organic compared to in the conventional fruit, while no crop system effect was found in the other evaluated biocompounds
Extracción de semen en gallos II
No full textExtracción de semen en gallos mediante la técnica del masaje dorso-abdominal.Para la extracción de semen en gallos, se usa la técnica del masaje dorso-abdominal. En él, uno de los operarios realiza un masaje desde la base de las alas en dirección hacia la cloaca y el otro operario realiza la extracción propiamente dicha (“ordeño”) ejerciendo presión en la cloaca para lograr la eyección del semen. En este vídeo se muestra con más detalle la parte de la presión en la cloaca que realiza el operario 2. La persona a cargo de realizar el ordeño debe esperar a que se produzca cierta eversión de la cloaca, que indica que en ese momento es posible extraer el semen. Además, debe prestar atención al aspecto de la extracción porque puede aparecer material procedente de la cloaca que contaminará los eyaculados. Por otra parte, al ser una zona muy vascularizada, hay que tener cuidado con apoyar objetos que puedan provocar algún corte (como el borde los tubos, si se recoge así el eyaculado) o con ejercer una presión excesiva, ya que en esos casos podemos obtener un eyaculado contaminado con sangre
Extracción de semen en gallos I
No full textExtracción de semen en gallos: detalle del masaje dorso-abdominalPara la extracción de semen en gallos, se usa la técnica del masaje dorso-abdominal. En él, uno de los operarios realiza un masaje desde la base de las alas en dirección hacia la cloaca y el otro operario realiza la extracción propiamente dicha (“ordeño”) ejerciendo presión en la cloaca para lograr la eyección del semen. El masaje dorso-abdominal debe ser continuo desde la base de las alas hacia la cloaca. En este primer vídeo se muestra con más detalle la parte del masaje que realiza el operario 1
Influence of temperature on ‘Rojo Brillante’ persimmon drying. Quality characteristics and drying kinetics
Full text linkA physico-chemical characterization and drying kinetics study was performed for whole ‘Rojo Brillante’ persimmon subjected to hot-air drying at 35 ᵒC, 40 ᵒC and 45 ᵒC. Fruit was dried until a moisture content of 30 g water/100 g product, which is required to be considered dried persimmon. This moisture value was obtained after 12, 8 and 7 d when the drying process was applied at 35 ᵒC, 40 ᵒC and 45 ᵒC, respectively. All the drying treatments caused flesh structure changes, shrinkage and secondary epidermis formation. The final product's quality characteristics especially the texture, strongly depended on drying temperature. The fruit dried at 35 ᵒC exhibited a gelatinous internal texture and a thin secondary epidermis, similarly to that obtained during a natural drying process. The drying treatment at 40 ᵒC resulted in fruit having more rubbery flesh. However, the fruit dried at 45 ᵒC had a corky texture, with a very thick secondary epidermis that resulted in excessively tough fruit for commercialization purposes. The differences in flesh structure were revealed in the microstructural study by Cryo-FESEM. In all cases, soluble tannins content lowered to non-astringency values. Hot-air drying technology proved to be a good alternative to valorize discarded ‘Rojo Brillante’ persimmon
New Paralogs of the Heliothis virescens ABCC2 Transporter as Potential Receptors for Bt Cry1A Proteins
Full text linkThe ATP-binding cassette (ABC) transporters are a superfamily of membrane proteins. These active transporters are involved in the export of different substances such as xenobiotics. ABC transporters from subfamily C (ABCC) have also been described as functional receptors for different insecticidal proteins from Bacillus thuringiensis (Bt) in several lepidopteran species. Numerous studies have characterized the relationship between the ABCC2 transporter and Bt Cry1 proteins. Although other ABCC transporters sharing structural and functional similarities have been described, little is known of their role in the mode of action of Bt proteins. For Heliothis virescens, only the ABCC2 transporter and its interaction with Cry1A proteins have been studied to date. Here, we have searched for paralogs to the ABCC2 gene in H. virescens, and identified two new ABC transporter genes: HvABCC3 and HvABCC4. Furthermore, we have characterized their gene expression in the midgut and their protein topology, and compared them with that of ABCC2. Finally, we discuss their possible interaction with Bt proteins by performing protein docking analysis