Simultaneous detection of CTV, CEVd and HSVd using Arizona 861 S1 Citron and RT-PCR

Abstract

The simultaneous use of a rapid diagnostic method with the effective cleaning of infected budwood is a powerful preventative method for controlling viral diseases. The objective of this study was the multiple detection of CTV, CEVd, and HSVd, through bio-amplification in Arizona 861-S1 citron plants, combined with two incubation temperatures and multiple RT-PCR in two steps. Citron indicator plants were inoculated with the three pathogens, both individually and simultaneously, with four un-inoculated plants as controls. After four months of incubation at the two different temperatures, both the multiple and individually inoculated plants showed symptoms associated with these pathogens, which were subsequently corroborated through the use of multiple RT-PCR in two steps. This diagnostic method could be employed as an efficient, cost-effective alternative to the methods that are currently in use for detecting CTV, CEVd and HSVd, nevertheless its effectiveness should also be evaluated with tests using different isolates of these pathogens.

Caracterización del Viroide exocortis de los cítricos en diferentes condiciones de indexado

“…The use of the crop in vitro in this study proved to be useful to carry out the transmission and development of characteristic CEVd symptoms with an efficiency similar to the in vivo indexing procedures, as reported in other studies for diseases related to viroids in economically important plants Mahfouze et al, 2010;Černi et al, 2012). The presence of symptoms in the samples can simplify the molecular diagnosis of the viroids (Camps et al, 2014) since they distribute themselves homogenously in the plant tissues (Bani et al, 2015;Lin et al, 2015b) and it is difficult to determine is the sampled tissue corresponds to the (Bani et al, 2015;Lin et al, 2015b) y es difícil determinar si el tejido muestreado corresponde al tejido infectado a menos que la expresión de síntomas sirva como indicador.…”

“…El uso del cultivo in vitro en este estudio demostró ser útil para llevar a cabo la trasmisión y desarrollo de síntomas característicos de CEVd con una eficiencia similar a los procedimientos de indexado in vivo, tal como se reporta en otros estudios para enfermedades asociadas a viroides en plantas de interés económico Mahfouze et al, 2010;Černi et al, 2012). La presencia de síntomas en las muestras puede facilitar el diagnóstico molecular de los viroides (Camps et al, 2014) porque se distribuyen de manera heterogénea en los tejidos de…”

Citrus Virus and Viroid Diseases

“…Furthermore, it causes stem pitting in the grafting region and a reduction in fruit size and production [14]. This virus is transmitted by infected propagating material and aphids [2,5,[8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27]. Aphids are a significant role in transmission to remote distances.…”

“…CTV has two strains: mild isolates cause only mild or no symptoms in sensitive citrus indicator hosts and usually result in no economic loss. Severe isolates can cause decline, stem pitting, or both and may vary in intensity [14,19].…”

“…The significant symptoms that are caused by this viroid are bark scaling and cracking, general decline, decreased size and stunting, and leaf epinasty and cracks in the petiole (Figure 4) [10,19]. This viroid can infect various kinds of citrus cultivars like P. trifoliata and its hybrids, rangpur lime (Citrus limonia), lemon (C. limon), and citron (C. medica) [10].…”