Yellow Leaf Syndrome Research Articles

Further evidence of the association of a phytoplasma and a virus with yellow leaf syndrome in sugarcane

Using the polymerase chain reaction (PCR), reverse‐transcriptase–PCR (RT–PCR) and double‐antibody‐sandwich enzyme‐linked immunosorbent assay (DAS–ELISA), a phytoplasma (sugarcane yellows phytoplasma, ScYP) and a virus (Sugarcane yellow leaf virus, ScYLV) were detected in sugarcane with yellow leaf syndrome (YLS) in Mauritius. Samples were collected from clones undergoing quarantine, in a variety‐collection plot and in commercial fields. A 1·25 kb DNA fragment encoding the phytoplasma 16S rRNA was consistently amplified by nested PCR. Of 134 samples with and without symptoms derived from 113 varieties, 111 were infected by either ScYLV or ScYP. The phytoplasma was detected in 63 samples by PCR. Restriction fragment‐length polymorphism (RFLP) analysis of the phytoplasma 16S rDNA amplified product indicated that sugarcane yellows phytoplasma group III, which is related to Western X phytoplasma, is present in Mauritius. ScYLV was detected by RT–PCR and ELISA. The virus was more widely distributed than the phytoplasma, and was found in 70 and 100 samples by ELISA and RT–PCR, respectively. There was a significant correlation between the presence of the phytoplasma and YLS symptoms, while such correlation was not significant for ScYLV detected by RT–PCR. ELISA was less sensitive than RT–PCR for detection of ScYLV. Forty‐one samples were coinfected with both microorganisms. Eighty‐five per cent of the samples displayed symptoms when ScYLV and SCYP coexisted, while 55 and 38% were observed when ScYP or ScYLV, respectively, was present alone. The results indicate that the presence of both organisms enhanced the syndrome.

First Report of Sugarcane yellow leaf virus (ScYLV) in Costa Rica.

In Costa Rica, sugarcane plants with symptoms similar to those described for yellow leaf syndrome (YLS) (1,2) were first observed in 1994 in research plots of imported material in the midland areas of San Carlos and Turrialba. Recently, the same symptoms have been observed in commercial plantations in Turrialba. Symptomatic plants were characterized by yellowing of the leaves and central veins, the yellowing being more intense near the leaf tips. In severe cases, veins became reddish, and necrosis developed along the leaf edges, beginning at the leaf tip and extending to the base of the leaf. Growth of stems and roots was also reduced in infected plants. Minipurifications of six plants of four different varieties were examined by immunospecific electron microscopy (ISEM) using polyclonal antibodies (1). They were: one symptomatic plant each of the varieties H782313 and H608521; two symptomatic plants of H657052, and one asymptomatic plant each of H608521 and H827318. Isometric particles of approximately 28 nm were observed in the asymptomatic H827318 plant and in all symptomatic plants, with the exception of one plant of H657052. The size and morphology of the particles was similar to those reported for Sugarcane yellow leaf virus (ScYLV) (2). The presence of ScYLV was verified by indirect enzyme-linked immunosorbent assay (ELISA) using polyclonal antibodies (1). Twenty-two of 24 symptomatic plants and five of 13 asymptomatic plants were positive for ScYLV. These findings confirm the association of ScYLV with the yellows syndrome of sugarcane observed in Costa Rica. However, as was also reported by Scaglusi and Lockhart (1), ScYLV was not detected in several symptomatic plants, and research is continuing to determine whether other pathogens are associated with this syndrome in Costa Rica.

Occurrence of sugarcane yellow leaf virus in india

Occurrence of sugarcane yellow leaf syndrome (SCYLV) has been reported from four states (U.P., Haryana, Maharashtra, Tamil Nadu) of India. The association of SCYLV was confirmed by DAC-ELISA test using polyclonal antiserum. This is the first report of the occurrence of sugarcane yellow leaf virus in India.

Diversity of phytoplasmas in northern Australian sugarcane and other grasses

Plant pathogenic phytoplasmas found in grasses in northern Australia have the potential to adversely affect sugarcane production. To begin assessment of this threat, the number of grass species with phytoplasmas, the identity of those phytoplasmas, and their relationship with disease symptoms were determined. Sugarcane with and without symptoms of yellow leaf syndrome was included in the surveys. Molecular methods were used to detect and characterize phytoplasmas in grass species exhibiting symptoms typical of phytoplasma disease. Sugarcane samples were from the Ord River Irrigation Area, Western Australia, and Samford, Queensland. Samples of other grasses were from Wyndham, Kununurra and Broome, Western Australia, and Darwin, Northern Territory. Our survey identified four new phytoplasma host species and confirmed four known previously. Counting phytoplasmas, phytoplasma variants, and mixtures of phytoplasmas and variants, these eight host species had 33 different infections. Two phytoplasmas were new, cenchrus bunchy shoot which is related to Candidatus phytoplasma australiense, and sorghum bunchy shoot which is not closely related to any described phytoplasma. Twenty‐five phytoplasma isolates were detected in sugarcane. Of these, tomato big bud phytoplasma was the most common. In most cases no clear association between phytoplasmas and symptoms could be determined. None of the phytoplasmas in Australian sugarcane, but two in other grasses, were closely related to phytoplasmas associated with white leaf and grassy shoot diseases in Asian sugarcane. This study demonstrates that diversity of phytoplasmas and grass host species in northern Australia is greater than previously thought, and that symptoms alone are not always reliable indicators of phytoplasma presence or absence. It provides the groundwork to improve future field surveys, and for initiation of transmission trials to determine whether insect vectors capable of transmitting phytoplasmas from native grasses to sugarcane are present in the region.

First Report of Sugarcane yellow leaf virus in Peru.

Two sugarcane cultivars, H 50-7209 and H 32-8560, have exhibited unusual, severe leaf yellowing for more than 18 years at Agro Industrial Paramonga S.A. (AIPSA) in Peru. In 1999, these varieties occupied about 4,600 ha (74% of the cultivated area), and almost all fields showed these symptoms. Symptoms first appear on the upper third of the leaf blades, which turns light green to light yellow in young canes up to 6 to 8 months of age. Between 10 and 16 months of age, the symptoms are visible on the spindle and first to third visible dewlap leaves. Tips and margins of older leaves become necrotic, and leaves can turn completely necrotic as the necrosis progresses down the leaves. The abaxial surface of leaf midribs is rarely bright yellow, which differs from the characteristic symptom of yellow leaf syndrome caused by the Sugarcane yellow leaf virus (ScYLV) (1). The most severe symptoms occur when the leaves of stalks that flower turn completely yellow and die. Samples from 98 plants exhibiting different types of yellowing were collected from six commercial fields of cultivars H 50-7209 and H 32-8560 and the germ plasm collection (cultivars PCG 59-1609, Trojan, CP 48-103, CP 72-2086, Q 87, and PR 908) at Paramonga. Tissue blot immunoassay was used to detect ScYLV in the midrib of the top visible dewlap leaf using antiserum provided by B. E. L. Lockhart (University of Minnesota) (2). ScYLV was detected in all 49 commercial field samples and in 35 out of 49 germ plasm samples. All six cultivars of the germ plasm collection were found to be infected, but ScYLV was detected in only a few leaves of Trojan and CP 72-2086. Eighteen cuttings from diseased stalks of cultivars H 50-7209 and H 32-8560 were grown in a greenhouse in Montpellier, France. Yellowing of the underside of the midribs and of the leaf tips appeared after 3 months in cultivar H 50-7209 but only after 9 months in cultivar H 32-8560. At 9 months, the top leaf with a visible dewlap and the four leaves immediately below it of cultivar H 50-7209 exhibited severe yellowing. Reverse transcription polymerase chain reaction with specific ScYLV primers, provided by M. S. Irey (U.S. Sugar Corp., Clewiston, FL) were used to detect ScYLV in the top visible dewlap leaf (1), and ScYLV was found in all nine samples taken from 6-month-old plants of the two cultivars. This is the first report of ScYLV in Peru.

Factors Affecting the Transmission and Spread of Sugarcane yellow leaf virus.

Sugarcane, Saccharum spp. hybrid, is widely infected in the United States and many other countries with a yellowing and stunting disease called sugarcane yellow leaf syndrome. The causal agent, Sugarcane yellow leaf virus (ScYLV), is a Polerovirus of the Luteoviridae family. In this study, it was transmitted by the sugarcane aphid, Melanaphis sacchari, and also by the corn leaf aphid, Rhopalosiphum maidis, and the rice root aphid, R. rufiabdominalis. Two other aphids that infest sugarcane in Hawaii did not transmit the virus. Some Hawaiian sugarcane cultivars are susceptible to ScYLV, while others remain virus-free in the field. The latter were not infected when inoculated with viruliferous M. sacchari. Virus-free plants of susceptible cultivars were produced through apical meristem culture and were readily reinfected by viruliferous M. sacchari. They were also quickly reinfected when planted in a field in proximity to other infected sugarcane naturally infested with M. sacchari. Sugarcane cultivars are hybrids of several Saccharum species. In a field-grown collection of Saccharum and related species, 11 to 71% of the clones of four of the species were infected with ScYLV. None of the related genus Erianthus plants were infected, but four clones were infected experimentally by aphid inoculation. A low to moderate percentage of corn, rice, and sorghum seedlings became infected when inoculated with ScYLV, but barley, oats, and wheat proved to be very susceptible. None of seven weeds common in sugarcane fields were infected with ScYLV.

Transmission, Characterization, and Serology of a Luteovirus Associated with Yellow Leaf Syndrome of Sugarcane

ABSTRACT A previously uncharacterized luteovirus was associated with one form of yellow leaf syndrome (YLS), a widespread disease of sugarcane (Saccharum sp.). The virus was named Sugarcane yellow leaf luteovirus (ScYLV), and was identified in major sugarcane-producing areas of the world. Typical disease symptoms were reproduced when ScYLV was transmitted by Melanaphis sacchari or Rhopalosiphum maidis from infected to healthy sugarcane, suggesting that this virus may be the causal agent of one form of YLS. The only known hosts of ScYLV are Saccharum and Erianthus spp. Virions of ScYLV were 24 to 29 nm in diameter in sodium phosphotungstate at pH 5.0, had a buoyant density of 1.30 g/cm(3) in Cs(2)SO(4), and contained a 5.8-kb genomic ssRNA. The capsid protein had an estimated relative molecular mass of 27 kDa and was not glycosylated. A polyclonal rabbit antiserum raised against ScYLV did not detect any of eight other luteoviruses by enzyme-linked immunosorbent assay or immunosorbent electron microscopy, but in immunoblot assays, antibodies to ScYLV detected the RPV serotype of Barley yellow dwarf luteovirus. It is concluded that ScYLV is a previously undescribed luteovirus that is biologically and serologically distinct from other members of the group and may be the causal agent of one form of YLS of sugarcane.

First Report of Virus and Phytoplasma Pathogens Associated with Yellow Leaf Syndrome of Sugarcane in Cuba.

Yellow leaf syndrome (YLS) has been seen recently in sugarcane (Saccharum sp.) in Cuba. The primary symptom is a yellow discoloration of the midrib that may spread from the midrib to the lamina in cane 6 months and older. In certain cultivars, such as CP 5243, EPC 17-395, and F31-156, a reddish coloration has been observed. In severe cases, plants are stunted and can be pulled easily. YLS was first reported from Hawaii, followed by Brazil, Florida, and Australia, where it is associated with a luteovirus: sugarcane yellow leaf virus (ScYLV). However, in South Africa, YLS is associated with a phytoplasma: sugarcane yellow leaf phytoplasma (ScYLP) (1). A survey performed in Jovellanos, Matanzas, Cuba, for ScYLV, using enzyme-linked immunosorbent assay with antiserum provided by B. E. L. Lockhart, showed that only a small percentage of canes with YLS carried the virus. A nested polymerase chain reaction (PCR) (1) was used to amplify phytoplasma 16S/23S rDNA from sugarcane leaves with YLS symptoms, also collected from Jovellanos. Restriction fragment length polymorphism analysis with HaeIII, RsaI, and AluI produced patterns similar to those of members of the aster yellows group for 260 of 277 samples tested. Sequencing of the 16S/23S intergenic rDNA PCR products, followed by BLAST (basic local alignment search tool) analysis, confirmed the high homology (97%) of these amplimers to the DNA of phytoplasmas belonging to the aster yellows I-A subgroup. This is the first report of ScYLV and ScYLP from Cuba, and it demonstrates the difficulty of determining the identity of the YLS pathogen based on symptoms alone. Reference: (1) C. P. R. Cronjé et al. Ann. Appl. Biol. 133:177, 1998.

First Report of Sugarcane Yellow Leaf Virus in the French West Indies.

Unusually severe leaf yellowing symptoms, similar to those described for yellow leaf syndrome (1), have been observed in several sugarcane clones in Guadeloupe since 1994, and since 1997 in Martinique. Leaf samples exhibiting various types of yellowing were taken from five different sugarcane clones, and analyzed by immunosorbent electron microscopy. Spherical particles, 24 to 28 nm in diameter and characteristic of luteoviruses, were found in two of five samples. The two infected samples showed yellowing on the underside of the midrib and one had a pinkish coloration on the upper side. The presence of sugarcane yellow leaf virus (ScYLV), the causal agent of sugarcane yellow leaf disease, was confirmed by reverse transcription-polymerase chain reaction (2) in these two samples and in 36 of 184 sugarcane clones bred in Guadeloupe and sent to Cirad's quarantine station in Montpellier, France. Following these observations, surveys were undertaken with a tissue blot enzyme immunoassay to analyze the distribution of ScYLV in sugarcane clones in the French West Indies. The midrib base of the first visible dewlap leaf was used to detect the presence of the virus in the phloem. In a first survey, clones of various origins worldwide were taken from germplasm collections. Two to three leaf samples per clone were analyzed from 78 clones in a collection in Guadeloupe and from 36 in a collection in Mar-tinique. Fifty of the 114 clones were infected by ScYLV, and ScYLV was detected in 21 of the 32 clones exhibiting severe leaf yellowing (score 3 or higher on a 1 to 5 scale). In a second survey, 19 leaf samples were taken from each of 53 clones from plants produced by Cirad's breeding program in Guadeloupe. The virus was detected in at least one sample for 25 of these 53 clones. ScYLV incidence in commercial fields was tested in Martinique in the variety B5992, which constitutes 57% of the cultivated area. Twenty leaves from different stools were sampled in six different fields, five of which had ScYLV-infected plants. The percentage of virus-infected stalks ranged from 0 to 90% whereas the percentage of stalks showing symptoms ranged from 50 to 100%. ScYLV appears widespread in the French West Indies, perhaps because a vector (Melanaphis sacchari) exists in Martinique and Guadeloupe. However, ScYLV was not found in all symptomatic plants, indicating that even if this luteovirus is a causal agent of leaf yellowing in the French West Indies, there may be other causal agents as well.

First Report of Yellow Leaf Syndrome of Sugarcane in Morocco.

Yellow leaf syndrome of sugarcane (Saccharum officinarum L.) has been described from a number of cane-growing countries. Symptomatic leaves were collected from the CTCS and other localities of the Gharb region. The first symptoms were observed on cv. CP 66-346 as a reddening of the adaxial leaf surface; in other cultivars, particularly cv. CP 72-1210, the older leaf midribs turned bright yellow, followed by yellowing of the blade starting from the leaf tip and spreading downward. The older leaves dried out and in severe cases the plant died. The disease is widespread in both the Gharb and Loukkos cane-growing regions. Severe symptoms were observed in the spring of 1998 following a 5-month drought and an atypical flowering of the cane (in Morocco cane has not flowered for the past 25 years). Sugarcane yellow leaf phytoplasma was identified by the amplification of a band of the expected size of approximately 1,250 bp following nested PCR (polymerase chain reaction) with universal primers in samples of CP 66-346, LCP 85-384, and clones originated from fuzz, but not in cultivars CP 74-383 and CP 72-1210. This is the first report of yellow leaf syndrome on sugarcane in Morocco.

Association of a phytoplasma with a yellow leaf syndrome of sugarcane in Africa

Summary.Evidence is presented for the association of a phytoplasma, provisionally named sugarcane yellows phytoplasma (ScYP), in sugarcane affected by a yellow leaf syndrome. The phytoplasma was consistently detected in leaves of more than 40 varieties from eight African countries. It was present in all symptomatic as well as some asymptomatic field grown cane samples but not in plants grown from true seed, and it was also observed in phloem sieve tubes by transmission electron microscopy. Phytoplasma 16S rDNA was confirmed by PCR, and restriction fragment analysis using Rsal and Haelll confirmed that PCR‐amplified products were of phytoplasma rather than of plant or of other pathogen origin. Sequences obtained from the intergenic spacer region, between the 16S and 23S rDNA genes, confirmed the identity of the phytoplasma as belonging to the western X group of phytoplasmas.