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A potyvirus (proposed name of Zea mosaic virus [ZeMV]) isolated from maize in Israel was analyzed by serology, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of capsid proteins, symptomatology, and sequencing. Parts of the nuclear inclusion b, coat protein, and 3' regions were sequenced; the amino acid sequence of ZeMV capsid was determined by time-of-flight mass spectrometry (TOFMS). The results of these analyses were compared with those of similar analyses of the following potyviruses: Maize dwarf mosaic virus (MDMV), Sugarcane mosaic virus strain MDB (SCMV-MDB), Johnsongrass mosaic virus(JGMV), Sorghum mosaic virus (SrMV), and an isolate of MDMV from Israel. Indirect enzyme-linked immunosorbent assay using ZeMV antiserum detected only ZeMV, and reciprocal tests using MDMV, JGMV, or SrMV antisera failed to detect ZeMV. ZeMV cross-reacted weakly when SCMV-MDB antiserum was used. The mass of ZeMV capsid was determined to be 36,810 Da by SDS-PAGE and 34,216 Da by TOFMS. The ZeMV systemically infected johnsongrass (Sorghum halepense), but did not infect oat (Avena sativa), pearl millet (Pennisetum glaucum), barley (Hordeum vulgare), or rye (Secale cereale). Necrosis was caused in 19 sorghum lines by SrMV, in 15 by ZeMV, in 14 by MDMV, and in 5 by JGMV and SCMV-MDB. The nucleic acid and amino acid sequences of ZeMV clearly showed that it is not a strain of JGMV, MDMV, SCMV, or SrMV.

Mosaic diseases of sugarcane caused by various viruses have been reported in most sugarcane planting countries and threaten global sugar production. There is a lack of extensive, systematic investigation of mosaic diseases and their causal viruses in China. In this study, a total of 901 leaf samples showing mosaic symptoms were collected from commercial fields in eight provincial regions in China and tested for sorghum mosaic virus (SrMV), sugarcane mosaic virus (SCMV), sugarcane streak mosaic virus (SCSMV), and maize yellow mosaic virus (MaYMV) using RT-PCR with four specific primer pairs. Of 901 tested samples, 38.5% (347/901) of samples were infected with one of the four viruses alone. Infection by two or more viruses was seen for 42.6% (384/901) of samples. The highest incidence of virus-causing sugarcane mosaic disease was SrMV (70.1%), followed by SCMV (33.4%) and SCSMV (30.3%), and the lowest incidence was seen for MaYMV (5.1%). Three viruses (SrMV, SCMV, and SCSMV) were found in eight sugarcane-planting provinces, whereas MaYMV was only found in Fujian, Guangxi, and Sichuan provinces. Mixed infections of the three main viruses, particularly for SrMV + SCMV and SrMV + SCSMV, were commonly found in the sugarcane samples. Our systematic determination of the occurrence and distribution of four RNA viruses associated with sugarcane mosaic diseases can provide evidence to guide the development of strategies for the prevention and control of sugarcane mosaic diseases in China.

Here, we describe the full-length genome sequence of a novel potyvirus, tentatively named “Miscanthus sinensis mosaic virus” (MsiMV), isolated from Miscanthus sinensis (silver grass) held in a post-entry quarantine facility after being imported into Western Australia, Australia. The MsiMV genome is 9604 nucleotides (nt) in length, encoding a 3071-amino-acid (aa) polyprotein with conserved sequence motifs. The MsiMV genome is most closely related to that of sorghum mosaic virus (SrMV), with 74% nt and 78.5% aa sequence identity to the SrMV polyprotein region. Phylogenetic analysis based on the polyprotein grouped MsiMV with SrMV, sugarcane mosaic virus (SCMV), and maize dwarf mosaic virus (MDMV). This is the first report of a novel monopartite ssRNA virus in Miscanthus sinensis related to members of the genus Potyvirus in the family Potyviridae.

Maize dwarf mosaic (MDM) is one of the most important virus diseases of maize worldwide. Caused by the potyviruses maize dwarf mosaic virus (MDMV) or sugarcane mosaic virus (SCMV), MDM can cause up to 90% yield loss in susceptible hybrids. One of the most effective management strategies for MDM is growing potyvirus-resistant corn varieties. However, yield impacts associated with MDM and the corresponding efficacy of genetic resistance present in modern United States commercial hybrid lines is uncharacterized. In this study, we evaluated the disease response of 78 commercial hybrids to MDMV and SCMV and quantified yield losses associated with infection over multiple trials. We determined that while 97% of the hybrids tested were resistant to MDMV, 100% were susceptible to SCMV, with mean disease incidence per line averaging between 45 and 78% across six trial years. Despite only one hybrid displaying visible mosaic symptoms when inoculated with MDMV, MDMV reduced average yields by approximately 5% across all hybrids compared with the mock-inoculated treatment. The yield impact of SCMV was more severe, reducing average yields by 10% across replicated experiments. These results indicate that while most commercial hybrids are resistant to MDMV, possibly due to the presence of the major Scmv1 resistance locus on chromosome 6, additional potyvirus resistance genes are needed to manage SCMV-induced MDM. Pyramiding resistance loci, such as Scmv2 on chromosome 3 or Scmv3 on chromosome 10 in addition to Scmv1, could be an effective strategy to mitigate the yield impact of MDM disease.

Sugarcane mosaic virus (SCMV), sugarcane streak mosaic virus (SCSMV), and sorghum mosaic virus (SrMV) are the causative pathogens of mosaic disease. This study aimed to identify mosaic virus infection and its impact on photosynthetic and antioxidant gene expression in eight commercial sugarcane cultivars grown on sugarcane plantations in East Java, Indonesia. The disease incidence and severity were observed in symptomatic leave samples, and then the virus was identified. A high incidence and severity of mosaic symptoms were observed in the PS881 and NX04 cultivars compared with the other cultivars. RT-PCR analysis detected SCSMV infection in all cultivars; double infections with SCSMV and SCMV in the PS881, PS882, and Cening cultivars; and triple infections with SCSMV, SCMV, and SrMV in the PS881 cultivar. Ascorbate peroxidase (Apx) expression was upregulated in all virus-infected cultivars and significantly increased in the triple-infected PS881 cultivar. However, catalase (Cat) expression was only slightly increased in the PS881 cultivar. The chlorophyll content was reduced, and the PsaA gene was downregulated in all cultivars. The expression of PsaA, RbcS, and Sps was significantly suppressed in the triple-infected PS881 cultivar. Moreover, the downregulation of both the RbcS and Pepc genes was concomitant with that of their protein levels.

In February 2014, a severe disease on maize (Zea mays L.) broke out in the fields of central and southwestern Taiwan and caused yield losses in sweet corn production. Chlorotic spots first appeared at the base of infected leaves and later developed into systemic mottling. Diffused necrotic patches were also found on leaves or husks of the diseased plants. Moreover, severe rosetting and stunting accompanied by abnormalities in ear production were observed on mature plants. Eighteen leaf samples from symptomatic plants were collected and submitted to our Plant Diagnostic Clinic for virus diagnosis. All of the samples were first tested by reverse transcriptase (RT)-PCR to detect Maize stripe virus (MSpV) and by indirect ELISA to detect Maize dwarf mosaic virus (MDMV) or Sugarcane mosaic virus (SCMV), which were endemic to this area (1). Only 2 out of 18 samples were positive for MDMV, SCMV, or mixed infection of both viruses. Sap inoculation tests conducted on seedlings of sweet corn cv. Honey 236 indicated that the MDMV- and SCMV-negative samples still had an unknown pathogen causing original symptoms in the receptor plants. The isolate from Yunlin county reacted only with the antibody to Maize chlorotic mottle virus (MCMV) (AC Diagnostics, Fayetteville, AR) in ELISA. For further identification, the MCMV-specific primers (forward: MCMVg3514F-GGGAACAACCTGCTCCA; reverse MCMVg4014R-GGACACGGAGTACGAGA) were designed from the nucleotide sequence of MCMV coat protein (CP) gene. In RT-PCR using the AccuPower RT/PCR PreMix kit (Bioneer, Daejeon, Korea), an expected 500-bp DNA fragment was observed. This PCR product was cloned and its nucleotide sequence was determined by Mission Biotech Co., Taipei, Taiwan. BLAST analysis of the CP gene of the MCMV-Yunlin revealed the maximum nucleotide identities (99%) with Chinese Sichuan isolates (GenBank Accession No. JQ984270) and 98% identities to four Chinese Yunnan isolates (GU138674, JQ982468, JQ982469, and KF010583) and one Kenya isolate (JX286709), compared with 97% to Kansas isolate (X14736) and 96% to Nebraska isolate (EU358605). Subsequently, the complete nucleotide sequence of the viral genome (KJ782300) was determined from five overlapping DNA fragments obtained from independent RT-PCR amplification. The virus isolate was infectious to sweet corn cultivars Bai-long-wang, Devotion, SC-34, SC2015, and Zheng-zi-mi, on which similar symptoms were developed after mechanical inoculation. During the spring of 2014, a total of 224 sweet corn samples were collected from the epidemic areas of Taichung, Yunlin, Chiayi, and Kaohsiung counties. Samples (n= 161) reacted positive for MCMV in ELISA and/or RT-PCR. In the field survey, more than 20 adult thrips might be observed on an MCMV-infected plant. Two species of Frankliniella were found on maize plants: F. williamsi Hood and F. intonsa Trybom. Maize thrips (F. williamsi), an occasional pest of maize occurring during winter and spring in Taiwan, was characterized by its abdominal sternite II on which 1 or 2 discal setae of equal length with posteromarginal setae were borne (2). Samples with 1, 5, 10, and 30 F. williamsi collected in the field were tested by RT-PCR; MCMV was detectable not only in the pooled crushed bodies but also in a single maize thrips. This is the first report of MCMV occurrence on maize in Taiwan and of the virus transmitted by maize thrips.

Sugarcane mosaic is the most widespread virus disease affecting sugarcane production. We have established a collection of seven prominent sugarcane mosaic potyvirus (SCMV) strains currently causing disease in sugarcane throughout the world and originally found in sugarcane in the United States. This collection includes SCMV strains A, B, D, and E, and the sorghum mosaic virus (SrMV) strains SCH, SCI, and SCM. These viruses were propagated on Sorghum bicolor cv. Rio and purified. Cloned cDNAs representing 2.0 kb of the 3' termini, obtained after a reversetranscriptase-polymerase chain reaction (RT-PCR) on purified virions using an oligo(dT) primer and degenerate primers with sequences located in the NIb gene, have been sequenced for each of these strains. A comparative analysis of the deduced amino acid sequences in the NIb and coat protein genes and of the nucleotide sequences in the 3'-untranslated region, among these seven viruses and among six other members of the SCMV subgroup, confirms that there are at least four, but suggests that there are five, distinct viruses in the SCMV complex. Based on these seven new sequences and on the available sequence data for six other members of the SCMV subgroup, we have developed group-specific primers for use in RT-PCR-based restriction fragment length polymorphism analysis for rapid discrimination between strains of SCMV and SrMV. This is the first assay for differentiating strains of SCMV and SrMV that does not require interpretation of symptoms on differential hosts.

Sugarcane mosaic virus (SCMV) (genus, Potyvirus; family, Potyviridae) is widespread, deleterious, and the most damaging pathogen of sugarcane (Saccharum officinarum L. and Saccharum spp.) that causes a substantial barrier to producing high sugarcane earnings. Sugarcane mosaic disease (SCMD) is caused by a single or compound infection of SCMV disseminated by several aphid vectors in a non-persistent manner. SCMV has flexuous filamentous particle of 700–750 nm long, which encapsidated in a positive-sense, single-stranded RNA molecule of 9575 nucleotides. RNA interference (RNAi)-mediated antiviral innate immunity is an evolutionarily conserved key biological process in eukaryotes and has evolved as an antiviral defense system to interfere with viral genomes for controlling infections in plants. The current study aims to analyze sugarcane (Saccharum officinarum L. and Saccharum spp.) locus-derived microRNAs (sof-miRNAs/ssp-miRNAs) with predicted potential for targeting the SCMV +ssRNA-encoded mRNAs, using a predictive approach that involves five algorithms. The ultimate goal of this research is to mobilize the in silico- predicted endogenous sof-miRNAs/ssp-miRNAs to experimentally trigger the catalytic RNAi pathway and generate sugarcane cultivars to evaluate the potential antiviral resistance surveillance ability and capacity for SCMV. Experimentally validated mature sugarcane (S. officinarum, 2n = 8X = 80) and (S. spp., 2n = 100–120) sof-miRNA/ssp-miRNA sequences (n = 28) were downloaded from the miRBase database and aligned with the SCMV genome (KY548506). Among the 28 targeted mature locus-derived sof-miRNAs/ssp-miRNAs evaluated, one sugarcane miRNA homolog, sof-miR159c, was identified to have a predicted miRNA binding site, at nucleotide position 3847 of the SCMV genome targeting CI ORF. To verify the accuracy of the target prediction accuracy and to determine whether the sugarcane sof-miRNA/ssp-miRNA could bind the predicted SCMV mRNA target(s), we constructed an integrated Circos plot. A genome-wide in silico-predicted miRNA-mediated target gene regulatory network was implicated to validate interactions necessary to warrant in vivo analysis. The current work provides valuable computational evidence for the generation of SCMV-resistant sugarcane cultivars.

Rapid identification of viruses causing sugarcane mosaic by direct sequencing of RT-PCR products from crude extracts: A method for large scale virus surveys

Plant virus diseases are serious constraints to the production and productivity of a wide range of crops among which maize chlorotic mottle virus (MCMV) is a serious problem in maize production and productivity. The virus is primarily originated in South America (Peru) and later distributed into many countries of all the continents except Australia and Antarctica. It is transmitted from location to location, and from plant to plant mechanically, through seed, insect vectors, and/or soil. The virus has many host ranges of agricultural crops (Maize, Sorghum, Sugarcane and Finger millet) and wild species (Dinebra retroflexa, Setaria verticillata, Cyperus assimilis, Digitaria ternta and Oplismenus hirtellus). MCMV alone causes 10-15% crop loss and up to 100% loss when it co-infects maize plants with other maize viruses such as maize dwarf mosaic virus, Sugar cane mosaic virus or Wheat streak mosaic virus. No any single disease control measure alone is effective in controlling the MCMV. However, an integrated virus disease management options (the use of disease-resistant crop varieties, proper field sanitation, removal of infection sources, use of virus-free seeds and chemical pesticides to indirectly control insect vectors) play a critical role in controlling the virus. Since the current status of the disease is wide spreading and on increasing trend, intensive MCMV employment, combined with integrated disease management, requires ongoing practice in countries where the virus is prevalent and in those countries that have not yet reported MCMV. Keywords : Integrated management; Insect vector; plant residue; Soil transmission; Zea mays DOI: 10.7176/ALST/87-03 Publication date: April 30 th 2021

The maize production environments in sub-Saharan Africa (SSA) are conducive to the development of many disease-causing organisms, which have infected the crop, causing serious reduction in maize production and productivity. Some maize diseases infect maize in all SSA ecologies, while some others are specific to some environments and absent from others. The prominent maize diseases in SSA changed over time, and presently, the most devastating are gray leaf spot (GLS), incited by Cercospora zeae-maydis; northern corn leaf blight, incited by Exserohilum turcicum; southern corn rust (Puccinia polysora); maize streak virus (MSV) disease, transmitted by the leafhopper, Cicadulina mbila; downy mildew, by Peronosclerospora sorghi; ear rots, incited by several fungi, including Aspergillus sp. and Fusarium sp.; and the recently discovered disease that is spreading fast across SSA, the maize lethal necrosis (MLN), a disease incited by the synergistic effect of maize chlorotic mottle virus (MCMV; Tombusviridae: Machlomovirus) and several other potyvirus, such as maize dwarf mosaic virus (MDMV), sugarcane mosaic virus (SCMV), and wheat streak mosaic virus (WSMV). There are several methods used for disease control, but the most effective and economic method is breeding for host plant resistance. In this chapter, four case studies of this breeding approach are presented and their effectiveness in SSA clearly highlighted.

Maize is a staple food in Zambia and contributes immensely to food security for smallholder farmers. Disease outbreaks such as Maize Lethal Necrosis Disease (MLND) can be a key constraint to maize production. This disease is caused by synergistic co-infection with Maize Chlorotic Mottle Virus (MCMV) and any virus from the family Potyviridae, particularly, Sugarcane Mosaic Virus (SCMV), Maize Dwarf Mosaic Virus (MDMV) or Wheat Streak Mosaic Virus (WSMV). In 2011, an outbreak of MLND affecting almost all of the currently grown commercial varieties posed a challenge to maize production in Kenya and it has since been reported in DR Congo, Ethiopia, Kenya, Rwanda, Tanzania, and Uganda causing yield losses of up to 100%. Despite MLND having been reported in some neighboring countries, there is no information on the status of the disease in Zambia. Additionally, there is a lot of grain and seed trade between Zambia and other countries among which MLND has been reported. The aim of this study was to establish: (a) the status of MCMV; (b) agricultural practices used by farmers and (c) insect vectors associated with MLND. A survey was conducted in nine (9) provinces of Zambia during 2014/2015 and 2015/ 2016 cropping seasons. Farmers’ maize fields were sampled at every five to ten-kilometer interval and tested using rapid diagnostic kits capable of detecting MCMV. Four hundred and nineteen samples collected all tested negative for MCMV. Zambian Agricultural Research Institute (ZARI), with all stakeholders in the maize value chain should continue implementing measures aimed at preventing the introduction of MLND in Zambia. Key words: Survey, MLND, Losses, food security

Procedures were developed for the in vitro elimination of Sugarcane mosaic virus (SCMV), Sorghum mosaic virus (SrMV), Sugarcane streak mosaic virus (SCSMV), Sugarcane yellow leaf virus (SCYLV) and Fiji disease virus (FDV) from infected sugarcane. In vitro shoot regeneration, elongation and virus elimination through meristem tissue culture originating from both apical and axillary shoots were compared. The average rates of regeneration and elongation from apical meristem tissues were 91 and 66%, respectively, with the virus-free rate among elongated shoots ranging from 61–92%. Mature axillary buds were cultivated in vitro to produce axillary shoots, from which meristem tissues were excised and cultured. These meristem tissues regenerated (77–100%) and elongated (55–88%) in culture medium at approximately the same rate as the apical meristems. The average virus elimination rate was 90% among elongated shoots derived from mature axillary buds. All five viruses can be eliminated by meristem tissue culture from both apical and axillary shoots using a standardized procedure. The overall average efficiency of virus-free plant production was 45 and 58% from apical and axillary shoots, respectively. There were no significant differences for shoot induction or virus elimination when the meristems were harvested from either the apical or the axillary shoots. This is the first report of SrMV or SCSMV elimination from sugarcane, as well as elimination of any mixed virus infections. This new method of harvesting meristems from axillary buds greatly expands the amount of material available for therapeutic treatments and thereby increases the probability of eliminating viruses from infected sugarcane.

The amino acid sequence of the coat protein of the Johnsongrass (JG) strain of sugarcane mosaic virus (SCMV) has been determined by protein sequencing techniques. The protein contains 303 amino acid residues corresponding to a molecular weight of 33,510 and when compared to the coat proteins of other potyviruses that have been characterized (263–267 residues) is found to have additional residues at its N-terminus. The N-terminus is acetylated as shown by fast atom bombardment mass spectrometry. Partial amino acid sequences of the coat proteins of the other three Australian SCMV strains, sugarcane (SC), Queensland blue couch grass (BC) and sabi grass (Sabi) have also been obtained. The sequence data and the comparative tryptic peptide HPLC profiles showed that the JG coat protein was substantially different from those of the other three SCMV strains, the sequence homology being around 66 per cent. This is in marked contrast to the high sequence homology between SC, BC and Sabi strains (95–100 per cent) but similar to that (51–62 per cent) found between coat proteins of distinct members of the potyvirus group. On the basis of these structural findings and other information on major differences in serological, biological and biochemical properties we believe that the present JG strain should not be considered a strain of SCMV but should be regarded as an independent member of the potyvirus group. The name “Johnsongrass mosaic virus” is proposed for this new member.

Eastern gamagrass (EGG) (Tripsacum dactyloides) is a high-quality, high-production native forage grass with potential as a high-protein, perennial grain crop. We used enzyme-linked immunosorbent assay to test EGG samples for virus infection. Over a 2-yr period, we found five instances of maize dwarf mosaic virus (MDMV) and a high incidence (12 and 72% in 3-mo-old and 5-yr-old plantings, respectively) of sugarcane mosaic virus strain MDMV-B (SCMV-MDMV-B) in experimental breeding nurseries in Kansas. Incidence of SCMV-MDMV-B was higher in older breeding nurseries. SCMV-MDMV-B, but not MDMV, was found in three of 42 natural EGG stands (.)