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- W129821843 abstract "Sugarcane bacilliform virus (SCBV) is an economically important pathogen of sugarcane in Australia which limits access to foreign sugarcane germplasm. Although SCBV is present in the major cane growing regions worldwide, very little is known about its variability, virulence and the yield losses resulting from infection. The limited information on SCBV has resulted in quarantine measures being introduced to protect the Australian sugarcane industry, with a major consequence being restricted access to imported sugarcane germplasm for breeding programs. Foreign sugarcane germplasm plays an important role in breeding of new commercial varieties for the Australian sugar industry and is essential for the long term productivity, profitability and sustainability of the sugar industry. This study was aimed at characterising Australian isolates of SCBV to enable the development of reliable and robust molecular and/or antibody-based diagnostic tests which could be used to not only assess the impact of SCBV on the Australian sugarcane industry, but could also be used to screen imported sugarcane germplasm for the virus. SCBV virions (SCBV-IM) were purified from the sugarcane accession Ireng Maleng and the dsDNA genome was cloned and sequenced. The genome of SCBV-IM comprised 7687 bp with an organisation typical of other badnaviruses. When the entire nucleotide sequence of SCBV-IM was compared to that of the Moroccan SCBV isolate (SCBV-Mo), less than 75% similarity was present. Within the coding regions, ORF I, ORF II and ORF III had 83%, 71% and 73% nucleotide similarity to SCBV-Mo, respectively. At the amino acid level, ORFs I, II and III from SCBV-IM showed 91%, 84% and 85% similarity to the equivalent regions in SCBV-Mo, respectively. To further investigate the level of sequence variability within Australian SCBV isolates, virions were purified from three further sugarcane accessions and a 220 bp fragment of the reverse transcriptase-coding region was amplified. Five clones from each sub-population were selected and sequenced. Analysis of these sequences revealed considerable variability in the virus population with variability within one plant as great as it was between isolates. However, since the use of specific primers could also be selecting for a sub-population of SCBV sequences, it was possible that the variability may actually be greater than that reported. These results indicated that SCBV isolates are complex and variable and may represent a continuum of genetic variability.High molecular weight DNA species larger than the SCBV 7.6 kbp unit-length genome were found in DNA extracted from purified SCBV-IM virions. We confirmed that these high molecular weight nucleic acids were virus-specific and open circular in conformation. Using field inversion gel electrophoresis (FIGE), the SCBV-IM DNA was separated into four discrete bands with sizes ranging from between 1 to 4 genome copies. The DNA was shown to comprise overlapped individual genome-length molecules and not covalently-bonded continuous DNA strands. We presume that these DNA molecules are concatamers formed during replication as a result of a terminal overlap on the sense strand. The presence of these concatamers within virions may explain the observation of particles with lengths corresponding to one, two or three times the modal length of 130 nm.Four SCBV-infected Saccharum officinarum plants were examined for the presence of integrated viral DNA. Southern blot analysis of viral DNA and total DNA extracted from the same plant source were compared with, or without, restriction digestion. The resulting restriction patterns from viral and total DNA were almost identical suggesting that there were no integrated SCBV sequences in the sugarcane cultivars tested. Although larger-than-single-genome copy bands were detected in both the viral and the total DNA samples, this was probably due to the presence of genomic concatamers. SCBV integration studies using Southern analyses were further complicated by high sequence variability which precluded the restriction digestion of all viral DNA species. As such, some of the SCBV DNA species remain as concatamers which appear as larger-than-unit-length SCBV products. An antiserum derived from a mixture of purified SCBV isolates has been used routinely in the past to screen for SCBV infection, but the heterogeneity reported for badnaviruses has cast doubt on the ability of this antiserum to detect all SCBV isolates. We attempted to determine whether antiserum generated against proteins other than the viral capsid could be used to detect SCBV infections, thus improving the reliability and robustness of SCBV diagnosis. The complete coding regions of SCBV ORF I and ORF II were bacterially expressed and used as antigens for antiserum production. Both ORF I and II proteins were found to be highly immunogenic and generated high-titre antisera, designated AS-I and AS-II, respectively. The diagnostic utility of both antisera to detect SCBV in six different infected sugarcane plants was tested using both immunosorbent electron microscopy (ISEM) and western blots. The currently used SCBV antiserum (AS-V), generated against a mixture of purified SCBV isolates, was included for comparison. In western analyses, neither AS-I nor AS-V was able to conclusively detect SCBV in any of the six infected plants due to reactivity with numerous non-specific proteins. In contrast, AS-II reacted specifically with a protein of the expected size (~13.5 kDa) in 2/6 infected plants. When compared using ISEM, AS-V, AS-I and AS-II trapped virions from 6/6, 6/6 and 2/6 SCBV-infected plants, respectively. However, the number of virions trapped using AS-V was approximately 30-fold more than that trapped using either AS-I or AS-II. These results highlight the variability between SCBV isolates and suggest that ISEM with antisera raised against mixtures of viral proteins may be a useful tool for the detection of viral isolates." @default.
- W129821843 created "2016-06-24" @default.
- W129821843 creator A5040176405 @default.
- W129821843 date "2003-01-01" @default.
- W129821843 modified "2023-09-26" @default.
- W129821843 title "Characterisation of an Australian isolate of sugarcane bacilliform virus" @default.
- W129821843 cites W1149578017 @default.
- W129821843 cites W118023291 @default.
- W129821843 cites W129486756 @default.
- W129821843 cites W1411046992 @default.
- W129821843 cites W14367026 @default.
- W129821843 cites W1485513336 @default.
- W129821843 cites W149574725 @default.
- W129821843 cites W1560207482 @default.
- W129821843 cites W1560669833 @default.
- W129821843 cites W1591726902 @default.
- W129821843 cites W1762283362 @default.
- W129821843 cites W1816016449 @default.
- W129821843 cites W1957831660 @default.
- W129821843 cites W1968618897 @default.
- W129821843 cites W1970298427 @default.
- W129821843 cites W1987265965 @default.
- W129821843 cites W1991173380 @default.
- W129821843 cites W1996062853 @default.
- W129821843 cites W1998471485 @default.
- W129821843 cites W2002152077 @default.
- W129821843 cites W2013321754 @default.
- W129821843 cites W2025175490 @default.
- W129821843 cites W2027246680 @default.
- W129821843 cites W2027882209 @default.
- W129821843 cites W2029267120 @default.
- W129821843 cites W2030425354 @default.
- W129821843 cites W2035128586 @default.
- W129821843 cites W2035315789 @default.
- W129821843 cites W2038676266 @default.
- W129821843 cites W2040752239 @default.
- W129821843 cites W2041265591 @default.
- W129821843 cites W2041414302 @default.
- W129821843 cites W2042799128 @default.
- W129821843 cites W2049635419 @default.
- W129821843 cites W2051681993 @default.
- W129821843 cites W2051994618 @default.
- W129821843 cites W2057150035 @default.
- W129821843 cites W2061115429 @default.
- W129821843 cites W2064160521 @default.
- W129821843 cites W2071095693 @default.
- W129821843 cites W2072962648 @default.
- W129821843 cites W2077131281 @default.
- W129821843 cites W2089898829 @default.
- W129821843 cites W2092172777 @default.
- W129821843 cites W2093291221 @default.
- W129821843 cites W2094147473 @default.
- W129821843 cites W2119698111 @default.
- W129821843 cites W2120678660 @default.
- W129821843 cites W2124129179 @default.
- W129821843 cites W2129376543 @default.
- W129821843 cites W2135737180 @default.
- W129821843 cites W2149638143 @default.
- W129821843 cites W2155241571 @default.
- W129821843 cites W2157576377 @default.
- W129821843 cites W2157744927 @default.
- W129821843 cites W2159329209 @default.
- W129821843 cites W2171510852 @default.
- W129821843 cites W2284448514 @default.
- W129821843 cites W2318014962 @default.
- W129821843 cites W2492241659 @default.
- W129821843 cites W2502637633 @default.
- W129821843 cites W2558766452 @default.
- W129821843 cites W2615352060 @default.
- W129821843 cites W2624838652 @default.
- W129821843 cites W263105778 @default.
- W129821843 cites W2780574853 @default.
- W129821843 cites W2785522896 @default.
- W129821843 cites W2896212243 @default.
- W129821843 cites W2896291633 @default.
- W129821843 cites W3034743378 @default.
- W129821843 cites W3184793796 @default.
- W129821843 cites W410953118 @default.
- W129821843 cites W563991718 @default.
- W129821843 cites W621196644 @default.
- W129821843 cites W644654585 @default.
- W129821843 cites W8138582 @default.
- W129821843 cites W818485975 @default.
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