Detection ofCitrus tristeza virus(CTV) from Satsuma Owari mandarins(Citris unshiu)by direct tissue blot immunoassay (DTBIA), DAS‐ELISA, and biological indexing

2008 ◽  
Vol 36 (4) ◽  
pp. 239-246 ◽  
Author(s):  
S. Korkmaz ◽  
B. Cevik ◽  
S. Onder ◽  
K. Koc ◽  
O. Bozan
Keyword(s):  
Biological Indexing ◽  
Tissue Blot Immunoassay ◽  
Tristeza Virus ◽  
Das Elisa

scholarly journals Resistance to barley yellow dwarf luteovirus in Aegilops species

1994 ◽  
Vol 74 (3) ◽  
pp. 631-634 ◽  
Author(s):  
K. M. Makkouk ◽  
W. Ghulam ◽  
A. Comeau
Keyword(s):  
Triticum Aestivum ◽  
Agricultural Research ◽  
Sandwich Elisa ◽  
Yellow Dwarf ◽  
Aegilops Species ◽  
Elite Group ◽  
Barley Yellow Dwarf ◽  
Tissue Blot Immunoassay ◽  
Das Elisa ◽  
Barley Yellow Dwarf Luteovirus

One thousand and ninety-seven Aegilops accessions were evaluated for their reaction to a PAV serotype of barley yellow dwarf luteovirus (BYDV). The accessions tested belong to the species bicornis, biuncialis, caudata, crassa, columnaris, comosa, cylindrica, kotschyi, longissima, mutica, neglecta (= triaristata 4 ×), ovata, peregrina, searsii, sharonensis, speltoides, tauschii (= squarrosa), triuncialis, umbellulata, uniaristata, vavilovii and ventricosa. The first evaluation of virus levels in the different accessions was conducted at International Center for Agricultural Research in the Dry Areas (ICARDA), Aleppo, Syria, using double antibody sandwich ELISA (DAS-ELISA). Accession reaction ranged from highly resistant to highly susceptible. Thirty-eight Aegilops accessions resistant at ICARDA, were evaluated at Sainte-Foy, Quebec, Canada, by tissue-blot immunoassay. Diversity of response to BYDV infection was again observed in this elite group. Seven accessions belonging to the species biuncialis, caudata, neglecta and triuncialis were highly BYDV resistant at both locations; five of these originated from Bulgaria. Key words: Introgression, interspecific, Triticum aestivum, BYDV, ELISA, immunoassay, tissue blot

scholarly journals Prereaction of Citrus tristeza virus (CTV) Specific Antibodies and Labeled Secondary Antibodies Increases Speed of Direct Tissue Blot Immunoassay for CTV

Plant Disease ◽  
2006 ◽  
Vol 90 (5) ◽  
pp. 675-679 ◽  
Author(s):  
Youjian Lin ◽  
Phyllis A. Rundell ◽  
Lianhui Xie ◽  
Charles A. Powell
Keyword(s):  
Citrus Tristeza Virus ◽  
Plant Viruses ◽  
Specific Antibodies ◽  
Rabbit Igg ◽  
Tissue Blot Immunoassay ◽  
Polymerase Chain ◽  
Bacteria And Fungi ◽  
Secondary Antibodies ◽  
Tristeza Virus ◽  
Citrus Tristeza

An improved direct tissue blot immunoassay (DTBIA) procedure for detection of Citrus tristeza virus (CTV) within 1 h is described. Prints of fresh young stems of citrus plants that were infected or not infected with CTV were made by gently and evenly pressing the fresh-cut surface of the stems onto a nitrocellulose membrane. The tissue blots were air-dried for 5 min, incubated with prereaction solutions of CTV-specific antibodies and labeled secondary antibodies, goat anti-mouse Ig (H+L)-alkaline phosphatase conjugate or goat anti-rabbit IgG alkaline phos-phatase conjugate, for up to 20 min, rinsed with PBST buffer for 5 min, and immersed into an NBT-BCIP substrate solution for 15 to 20 min. Then the blots were rinsed in water for a few seconds to stop the reactions, and the results were observed and recorded under a light microscope. All samples from greenhouse plants that were infected with CTV decline inducing isolate T-36 were positive to CTV-specific polyclonal antibody 1212 (PCA 1212) and monoclonal antibodies 17G11 (MAb 17G11) and MCA13 (MAb MCA13), whereas samples from greenhouse plants infected with non-decline-inducing isolate T-30 were positive to PCA 1212 and MAb 17G11, but not to MAb MCA13. The noninfected greenhouse plants were negative to all of the antibodies. The improved DTBIA was at least as reliable as other immunological procedures and almost as reliable as polymerase chain reaction for detecting CTV in field trees. The improved DTBIA enables the detection of CTV within 1 h by having a prereaction of CTV-specific antibodies and labeled secondary antibodies in solutions before they are applied to the tissue blots. This DTBIA procedure may be useful in detecting other plant viruses and other pathogens such as bacteria and fungi.

Biological indexing and genetic analysis of Citrus tristeza virus in Pakistan

2017 ◽  
Vol 83 (6) ◽  
pp. 382-389 ◽  
Author(s):  
Sagheer Atta ◽  
Mengji Cao ◽  
Ummad ud din Umar ◽  
Yan Zhou ◽  
Fangyun Yang ◽  
...  
Keyword(s):  
Genetic Analysis ◽  
Citrus Tristeza Virus ◽  
Biological Indexing ◽  
Tristeza Virus ◽  
Citrus Tristeza

scholarly journals Incidence of Citrus Tristeza Virus and its Vector Toxoptera citricida in Different Parts of Assam and Nagaland, India

2021 ◽  
Vol 10 (9) ◽  
pp. 68-78
Author(s):  
Maongkar T. Changkiri Pulin Patgiri ◽  
Palash Deb Nath Rokozeno ◽  
Otto S. Awomi
Keyword(s):  
Citrus Tristeza Virus ◽  
Field Survey ◽  
Disease Incidence ◽  
Citrus Limon ◽  
Toxoptera Citricida ◽  
Aphid Vectors ◽  
Tristeza Virus ◽  
Das Elisa ◽  
Different Parts ◽  
Citrus Tristeza

A field survey was conducted in 2018 to study the incidence of citrus tristeza virus (CTV) and its aphid vectors, in different citrus growing regions of the states of Assam and Nagaland, India. Leaf samples of Assam lemon (Citrus limon) and Khasi mandarin (Citrus reticulata) were collected from four districts of Assam (Jorhat, Tinsukia, Sivasagar and Golaghat) and two districts of Nagaland (Mokokchung and Wokha). Citrus leaf samples were collected from a total of 190 citrus plants and were used for detection of CTV infection through Double Antibody Sandwich-Enzyme linked Immuno-Sorbent Assay (DAS-ELISA). According to the results, 75 per cent CTV disease incidence was detected in surveyed areas of Assam and 24.55 per cent CTV disease incidence was detected in surveyed areas of Nagaland. District wise, the highest CTV disease incidence (96.67 %) was detected in Tinsukia district of Assam and the lowest (21.43 %) was detected in Mokokchung district of Nagaland. Aphid samples were also collected during the survey and the presence of the vector Toxoptera citricida, in all the locations was determined.

scholarly journals Keefektifan Eliminasi Penyakit Sistemik (Huanglongbing dan Citrus Tristeza Virus) pada Jeruk dengan Embriogenesis Somatik

2016 ◽  
Vol 23 (2) ◽  
pp. 107
Author(s):  
Sri Widyaningsih ◽  
Farida Yulianti ◽  
Nirmala Frianti Devy
Keyword(s):  
Citrus Tristeza Virus ◽  
Tristeza Virus ◽  
Das Elisa ◽  
Citrus Tristeza

<p>Perbanyakan tanaman dengan teknik embriogenesis somatik diduga mampu mengeliminasi penyakit sistemik pada tanaman jeruk. Namun tingkat keefektifan eliminasi penyakit sistemik tersebut sangat bergantung pada eksplan dan status penyakit pada pohon induk. Tujuan penelitian ialah mengetahui keefektifan perbanyakan dengan embriogenesis somatik dalam membersihkan penyakit sistemik pada jeruk (huanglongbing dan citrus tristeza virus). Penelitian dilaksanakan di Laboratorium Terpadu dan Screenhouse Balai Penelitian Tanaman Jeruk dan Buah Subtropika (Balitjestro) Tlekung, Kota Batu. Penelitian dilaksanakan pada Bulan Januari 2010 sampai dengan Desember 2011. Pengujian status penyakit sistemik dilakukan terhadap hasil perbanyakan melalui teknik embriogenesis somatik yang berasal dari tanaman induk terinfeksi dan bebas penyakit sistemik huanglongbing (HLB) dan citrus tristeza virus (CTV). Analisis penyakit HLB menggunakan metode pengujian PCR, sedangkan analisis penyakit CTV menggunakan metode DAS-ELISA. Pengujian dilakukan pada empat stadia pertumbuhan (kalus, embrio, planlet, dan semai) hasil perbanyakan. Hasil penelitian menunjukkan bahwa teknik embriogenesis somatik efektif mengeliminasi penyakit HLB, namun kurang efektif untuk penyakit CTV. Hal ini ditunjukkan oleh hasil pengujian yang menunjukkan bahwa semua varietas dan semua stadia yang diuji bebas dari HLB, namun untuk CTV hanya terjadi pada varietas keprok Kinnow, siam Kintamani, dan nipis. Pada varietas JC (Japanche citroen) fase embrio, 40% dari sampel yang diuji masih terinfeksi CTV. Oleh karena itu, pengambilan nuselus sebagai sumber eksplan pada perbanyakan tanaman jeruk dengan embriogenesis somatik perlu dilakukan pada tanaman yang bebas dari penyakit sistemik.</p>

scholarly journals First Report of Citrus tristeza virus in Citrus Orchards in Bosnia and Herzegovina

Plant Disease ◽  
2013 ◽  
Vol 97 (12) ◽  
pp. 1665-1665
Author(s):  
D. Delić ◽  
M. Afechtal ◽  
K. Djelouah ◽  
B. Lolić ◽  
A. Karačić
Keyword(s):  
Coat Protein ◽  
Bosnia And Herzegovina ◽  
Coat Protein Gene ◽  
Citrus Tristeza Virus ◽  
Protein Gene ◽  
Rt Pcr ◽  
First Report ◽  
Citrus Orchards ◽  
Tristeza Virus ◽  
Das Elisa

The citrus growing area in Bosnia and Herzegovina (B&H) is limited to the confluence of the river Neretva, which is close to the Adriatic coastal region. Approximately 6 ha are grown in the country. Mandarins (Citrus reticulata Blanco) and lemons (Citrus limon L.) grafted on trifoliate orange (Poncirus trifoliata (L.) Raf.) are the most cultivated species. In June 2012, 25 samples were collected from individual trees from three locations in Herzegovina district of B&H (Mostar, Čapljina, and Ljubuški). Samples of different Citrus spp. (C. reticulata Blanco, C. aurantium L., C. limon L., C. sinensis (L.) Osbeck, P. trifoliata (L.) Raf., and Fortunella margarita Lour) and varieties were collected from infield plants, commercial citrus orchards, and a nursery. Out of 25, 10 citrus trees exhibited leaves chlorosis, whereas as all others were apparently symptomless. Double antibody sandwich (DAS)-ELISA test, using commercial kit from the DSMZ, Germany (product code AS-0988), was carried out to confirm the presence of Citrus tristeza virus (CTV). In addition, further analyses were performed using reverse transcription (RT)-PCR targeting the coat protein gene (2). CTV was detected in 8 out of the 25 tested samples with DAS-ELISA, whereas CTV was detected in 14 samples by RT-PCR. Being grafted on P. trifoliata rootstock, no typical CTV symptoms in the field were observed on the CTV-infected trees; interestingly, the lab analyses evidenced the CTV presence in all inspected locations of the Herzegovina district. To our knowledge, this is the first report of CTV in Bosnia and Herzegovina; nevertheless, the virus presence is also reported from neighboring countries Croatia (1) and Montenegro (3). The PCR products of four samples were additionally analyzed by sequencing. The preliminary results by sequencing of the coat protein gene of four selected CTV isolates (Accessions HF947341, HF947342, HF947343, and HF947347) showed 99% nucleotide identity with the CTV resistance breaking isolates from Montenegro (FR871866) and Croatia (EU579422). Although a very small number of samples were tested in this study, CTV appears to be widely distributed in the citrus orchards of the country. This could be related to the traditional use of tolerant P. trifoliata rootstock that prevents the development of the tristeza decline as well as to the virus isolates present in the region, which appear not to cause another economically devastating CTV disease such as stem pitting. Further research will be dedicated to the biological properties of the genetic variability of these identified CTV isolates and the assessment of potential aphid vectors. References: (1) S. Černi et al. Plant Dis. 89:342, 2005. (2) M. E. Hilf et al. Options Méditerranéennes B 65:89, 2009. (3) T. Papic et al. Plant Dis. 89:434, 2005.

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