A fast one-step reverse transcription and polymerase chain reaction (RT-PCR) amplification procedure providing highly specific complementary DNA from plant virus RNA

Sambade, A.; Martin, Sara; Olmos, Antonio; Garcia, M. L.; Cambra, M.; Grau, Oscar; Guerri, José; Moreno, Pedro

doi:10.1016/s0166-0934(00)00145-2

Cited by 11 publications

(8 citation statements)

References 6 publications

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“…Two digestive tracts were pooled and ground in Tri‐Reagent (Molecular Research Center Inc.) with a sterile Teflon pestle, and total RNA was immediately extracted according to the manufacturer's instructions. Total RNA was submitted to a modified one‐step reverse transcription and polymerase chain reaction (RT‐PCR) sequence‐independent amplification procedure (Bohlander, Espinosa, Le Beau, Rowley, & Diaz, ; Sambade et al, ; Figure ), detailed in Supplementary Information SI‐1. A negative control was included using RT‐PCR grade water instead of RNA.…”

Section: Methodsmentioning

confidence: 99%

Gut microbiota ofSpodoptera frugiperda(J.E. Smith) larvae as revealed by metatranscriptomic analysis

Rozadilla

Cabrera

Virla

et al. 2020

J Applied Entomology

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The invasive fall armyworm, Spodoptera frugiperda (J.E. Smith; Lepidoptera: Noctuidae), is considered one of the most harmful pests in corn, rice and cotton crops and has developed resistance to the most widespread methods used to control it, chemical insecticides and transgenic crops based on Bt toxins. On the other hand, gut microbiota plays an important role in the insect's fitness, and metatranscriptomic analyses of the insect host are an invaluable tool for identifying new pest control targets. Notwithstanding, there are extremely few reports on the functionally active profile of the gut microbiota in insects, especially in lepidopterans. Moreover, most studies have only described the bacterial community even though all the components of the microbiota, Bacteria, Archaea, fungi, protozoa and viruses, influence different physiological aspects of the insect host. This is the first time an unbiased and comprehensive metatranscriptomic approach has been used to describe the taxonomic and functional profile of the gut microbiota of S. frugiperda larvae, both bacterial and non‐bacterial. We identified novel and very active components which were putatively playing an important nutritional role, such as ammonia‐oxidizing Archaea and Bacteria, and a xylan‐ and hemicellulose‐degrading Actinobacteria. This analysis also allowed us to identify potential biocontrol agents.

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Section: Methodsmentioning

confidence: 99%

Gut microbiota ofSpodoptera frugiperda(J.E. Smith) larvae as revealed by metatranscriptomic analysis

Rozadilla

Cabrera

Virla

et al. 2020

J Applied Entomology

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“…Sources of antibodies. CTV-specific monoclonal antibody MAb-17G11 generated and produced at the Indian River Research and reverse transcriptase-polymerase chain reaction (RT-PCR) (Huang et al, 2004;Hung et al, 2000;Sambade et al, 2000), direct tissue blot immunoassay (DTBIA) (Garnsey et al, 1993;Lin et al, 2000Lin et al, , 2006, and in situ immunoassay (ISIA) (Lin et al, 2002(Lin et al, , 2000. The previous results indicated that CTV is located in the phloem tissues of leaf veins, stems, and petioles, and in the meristematic cells of young shoots (Brlansky and Lee 1988;Kitajima and Costa 1968;Kitajima et al, 1974;Sasaki et al, 1980;Schneider 1959;Schneider and Sasaki 1972;Zhou et al, 2002).…”

Section: Methodsmentioning

confidence: 99%

Visualization of the Distribution Pattern of Citrus Tristeza Virus in Leaves of Mexican Lime

Lin¹,

Powell²

2006

HortSci

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The distribution pattern of citrus tristeza virus (CTV) T-36 isolate in leaves of infected mexican lime [Citrus aurantifolia (Christm.) Swingle] plants was visualized using a whole-leaf-blot immunoassay (WLBIA) procedure in combination with a computer scanning imaging technique and CTV-specific monoclonal antibody 17G11 (CTV MAb 17G11). The distribution pattern of CTV T-36 in leaves varied with the age of the leaves and shoots of infected plants. In the young leaves, especially the about 5-day-old leaves and the completed expanded leaves, CTV T-36 was easily detected in most of the leaf veins, the main veins and the large and small primary veins. In the old leaves, CTV T-36 only was detected in the main veins, sometimes in a few of the large primary veins with weak signals, and seldom in the small primary veins. The distribution density and immunoassay reaction signals of CTV T-36 reacted to CTV MAb 17G11 in leaves from new shoots were much higher than that in leaves from old shoots. ELISA test results using leaves with different ages from different shoots of the same mexican lime plants infected with CTV T-36 supported the visualized-test results obtained by the WLBIA in combination with computer scanning imaging technique. This is the first reported visual analysis of the distribution pattern of CTV in leaves of infected citrus plants. The results indicate that the WLBIA in combination with computer scanning imaging technique is a useful tool for studying the distribution of plant viruses in leaves of virus-infected plants.

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“…Specimens were obtained from different environments, altitudes and food sources, namely, (1) a transgenic maize ( Zea mays ) field at 495 meters above sea level (MASL) where insecticides and fertilizers were applied (named Sf_MM; 26°49′50″S, 65°16′59.4″W), (2) Sorghum halepense at 495 MASL (Sf_MS; 26°49′50″S, 65°16′59.4″W), (3) a maize field at 2,283 MASL where no insecticides or fertilizers were used (Sf_TV; 26°55′40.75″S, 65°45′19.90″W), and (4) a colony established from larvae originally collected from the same transgenic maize field as Sf_MM, reared for 9 generations under controlled conditions on an artificial diet adapted from reference ( 8 ), without the addition of antibiotics (Sf_LR). For all samples, total RNA extracted from fifth instar larval guts (two digestive tracts per sample), was submitted to a one-step reverse transcription and PCR sequence-independent amplification procedure, modified from reference ( 9 ), as described previously ( 7 , 10 ). High-throughput pyrosequencing of the samples was performed using a Roche GS FLX (Macrogen, Inc., Republic of Korea), yielding ~1 Gb of metatranscriptomic reads with lengths of 50 to 1,600 base nucleotides (nt) (652 nt average).…”

Section: Genome Announcementmentioning

confidence: 99%

Metatranscriptomic Analysis of Larval Guts from Field-Collected and Laboratory-Reared Spodoptera frugiperda from the South American Subtropical Region

2015

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A fast one-step reverse transcription and polymerase chain reaction (RT-PCR) amplification procedure providing highly specific complementary DNA from plant virus RNA

Cited by 11 publications

References 6 publications

Gut microbiota ofSpodoptera frugiperda(J.E. Smith) larvae as revealed by metatranscriptomic analysis

Gut microbiota ofSpodoptera frugiperda(J.E. Smith) larvae as revealed by metatranscriptomic analysis

Visualization of the Distribution Pattern of Citrus Tristeza Virus in Leaves of Mexican Lime

Metatranscriptomic Analysis of Larval Guts from Field-Collected and Laboratory-Reared Spodoptera frugiperda from the South American Subtropical Region

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