Comparative transmission of, and varietal reaction to, three isolates of rice tungro virus disease

Comparative transmission by leafhoppers of three tungro isolates obtained from the Philippines, India and Malaysia, and of an infectious clone of the Philippine isolate of rice tungro bacilliform virus (RTBV) by agroinoculation, was conducted on 12 rice cultivars. The symptoms, including height of inoculated plants were recorded and the efficiency of RTBV and rice tungro spherical virus (RTSV) transmission was determined by enzyme-linked immunosorbent assay. In most cases, the reduction of height and leaf symptoms of plants infected with RTBV and/or RTSV by the three isolates were similar in any given cultivar. On cultivar ASD 7 , the Malaysian isolate showed more severe yellow orange leaf discolouration symptoms than the Indian isolate which in turn had more severe leaf discolouration than the Philippine isolate. On the other hand, cultivars ASD 7 and Ptb 18 produced the most severe yellow orange leaf discolouration when agroinoculated with an infectious RTBV clone of the Philippine isolate. There was some variation in the transmission profile of the two tungro viruses among the three isolates. However, there was no one clear set of characteristics by which one could use cultivars to distinguish isolates. The amount of viral DNA in agroinfected plants of cultivars Utri merah, Balimau putih, Utri Rajapan and ARC 11554 was low, while the amount was high in cultivars TN1, ASD7, Ptb 18 and TKM 6. There was high correlation between the amount of viral coat protein by ELISA and viral nucleic acid by DNA hybridisation on 10 agroinoculated rice cultivars; this might indicate that similar proportions of the total RTBV DNA are encapsidated in each cultivar.     

Studies on a Nigerian isolate of banana streak badnavirus: I. Purification and enzyme linked immunosorbent assay

A Nigerian isolate of banana streak badnavirus (BSV) was purified and a polyclonal antiserum was produced in mice. The antiserum titre was between 1:10 000 and 1:40 000 in enzyme linked immunosorbent assay (ELISA), and showed a good specificity to BSV antigens. Comparative tests were carried out to determine the sensitivity and reliability of BSV antigen detection by double antibody sandwich (DAS)-ELISA, triple antibody sandwich (TAS)-ELISA, antigen coated plate (ACP)-ELISA, and protein-A coated antibody sandwich (PAS)-ELISA. TAS-ELISA using rabbit polyclonal antiserum to trap BSV and mouse polyclonal antiserum to detect the virus particles, was more sensitive than ACP-ELISA and PAS-ELISA and detected BSV in plant extracts from both symptomatic and some asymptomatic plants. However, immunosorbent electron microscopy detected more BSV-infected plants from asymptomatic plant samples than did TAS-ELISA. Results of this study showed that detection of BSV antigens in sap extracts by TAS-ELISA was most efficient with symptomatic tissues which occurred most frequently in the ‘cool rainy’ season. This suggests that for more reliable BSV-indexing of field samples, tissue sampling should be done during the rainy season when most BSV-infected plants express severe symptoms.     

Effects of Priming and Endosperm Integrity on Seed Germination Rates of Tomato Genotypes: II. GERMINATION AT REDUCED WATER POTENTIAL

Seed germination rates (GR =inverse of time to germination)are sensitive to genetic, environmental, and physiological factors.We have compared the GR of tomato (Lycopersicon esculentum Mill.)seeds of cultivar T5 to those of rapidly germinating L. esculentumgenotypes PI 341988 and PI 120256 over a range of water potential(). The influence of seed priming treatments and removal ofthe endosperm/testa cap enclosing the radicle tip on germinationat reduced were also assessed. Germination time-courses atdifferent 's were analysed according to a model that identifieda base, or minimum, allowing germination of a specific percentage(g) of the seed population (_b(g)), and a ‘hydrotime constant’(_H) indicating the rate of progress toward germination per MPa.h.The distribution of _b(g) determined by probit analysis was characterizedby a mean base (_b) and the standard deviation in _b among seeds(_b). The three derived parameters, _b, _b) and _H, were sufficientto predict the time-courses of germination of intact seeds atany . A normalized time-scale for comparing germination responsesto reduced is introduced. The time to germination at any (t_g())can be normalized to be equivalent to that observed in water(t_g(0)) according to the equation t_g(0)=[l–(/_b(g))]t_g().PI 341988 seeds were more tolerant of reduced and had a morerapid GR than T5 seeds due to both a lower _b and a smaller _H.The rapid germination of PI 120256, on the other hand, couldbe attributed entirely to a smaller _H. Seed priming (6 d in–1.2 MPa polyethylene glycol 8000 solution at 20 ?C followedby drying) increased GR at all >_b(g), but did not lower theminimum allowing germination; i.e. priming reduced _H withoutlowering _b. Removing the endosperm/testa cap (cut seeds) markedlyincreased GR and lowered the mean required to inhibit germinationby 0.7 to 0.9 MPa. However, this resulted primarily from downwardadjustment in _b during the incubation of cut seeds at low inthe test solutions. The difference in _b between intact and cutseeds incubated at high was much less (0.l MPa), indicatingthat at the time of radicle protrusion, the endosperm had weakenedto the point where it constituted only a small mechanical barrier.In the intact seed, endosperm weakening and the downward adjustmentin embryo _b ceased at < –0.6 MPa, while the reductionin _H associated with priming proceeded down to at least –1.2MPa. Based on these data and on the pressure required to pushthe embryos from the seeds at various times after imbibition,it appears that the primary effect of priming was to shortenthe time required for final endosperm weakening to occur. However,as priming increased GR even in cut seeds, priming effects onthe embryo may control the rate of endosperm weakening. Key words: tomato, Lycopersicon esculentum Mill., water potential, germination rate, seed priming, genetic variation     

Some biological and genomic properties of rice tungro bacilliform badnavirus and rice tungro spherical waikavirus from Nepal

A survey of rice fields during the main growing seasons in 81 locations from 21 districts of the Southern Terai region of Nepal indicated that rice tungro was primarily restricted to the Hardinath (Janakpur) and Parwanipur (Bara) regions. The tungro incidence in Hardinath ranged from 17% to 51% and in Parwanipur from 6% to 61% causing about 89% grain yield loss in Hardinath. Both rice tungro bacilliform badnavirus (RTBV) and rice tungro spherical picornavirus (RTSV) were found in tungro isolates collected from Hardinath and Parwanipur. These isolates were transmitted by Nephotettix virescens and leaf extracts reacted to antisera against RTBV and RTSV. In a dot blot hybridisation assay, leaf extracts of 12 weed species collected from the tungro-affected area in Hardinath and Parwanipur also reacted with RTBV DNA probes. On mass inoculation of 15 popular rice cultivars most became more than 50% infected and only cv. Radha 9 had low (22.2%) infection. RTBV DNA and the coat protein region of RTSV from the Hardinath isolate were cloned and partially characterised. A comparative analyses by restriction endonuclease digestion, cross hybridisation, the polymerase chain reaction and partial sequencing indicated that the Nepalese RTBV DNA clone and the cDNA clones of the RTSV RNA were more similar to the various tungro isolates from the Indian subcontinent than to those from the Philippines.     

Evaluation of micropropagated plantain and banana (Musa spp.) for banana streak badnavirus incidence under field and screenhouse conditions in Nigeria

Between 1991 to 1996, more than 50 Musa hybrids and 10 landraces were evaluated under field and screenhouse conditions for virus symptoms resembling those caused by banana streak badnavirus (BSV). The symptoms included chlorotic streaks, leaf deformation, stunting, cigar leaf death, distortion of the peduncle, bunch or fruits, and internal pseudostem necrosis. Immunosorbent electron microscopy (ISEM) of randomly selected plants with one or more of these symptoms confirmed the presence of BSV particles in 15 tropical Musa plantain hybrids (TMPx) and five Musa landraces. Under both field and screenhouse conditions, the incidence of symptomatic plants in the hybrids was significantly higher than in the landraces. The hybrids also generally had a higher concentration of BSV antigens, as determined by enzyme-linked immunosorbent assay (ELISA). By contrast, most BSV-infected landraces were symptomless and had very low or undetectable amounts of BSV antigens. There was a significant variation in incidence of symptomatic plants between genotypes, experiments and year of observation. These results are discussed in relation to the higher natural BSV incidence observed on some Musa hybrids as compared with their parental genotypes.     

Studies on a Nigerian isolate of banana streak badnavirus: II. Effect of intraplant variation on virus accumulation and reliability of diagnosis by ELISA

Monitoring of banana streak badnavirus (BSV) antigens and symptoms in naturally BSV-infected plantain and banana (Musa spp.) plants showed a great variation in symptom expression, distribution and relative concentration of BSV between and within plants. Expression and distribution of symptoms was erratic within individual leaves as well as between different leaves of the same plant. The concentration of BSV antigens detected by triple antibody sandwich enzyme linked immunosorbent assay (TAS-ELISA) varied in different plant parts including leaf lamina, midrib and pseudostem, roots and young ‘cigar’ leaf. The concentration of BSV antigens was high in symptomatic tissues but was low or below the limits of detection in most asymptomatic tissues. During ‘hot dry’ seasons when symptoms were not fully expressed, the concentration of BSV antigens in leaf tissues declined drastically, often below the detection limit of TAS-ELISA. These results suggested that for more reliable detection of BSV antigens by TAS-ELISA, it is advisable to index plants using composite tissue samples comprising as many leaves as possible for each plant and collected during cool and/or rainy seasons when symptom expression is generally severe.     

Occurrence of papaya ringspot potyvirus and cucurbit viruses in Nepal

A survey of papaya and 10 cucurbitaceous vegetables (ashgourd, zucchini, watermelon, cucumber, pumpkin, bottlegourd, snakegourd, spongegourd, bittergourd and choyote) during 1989 and from 1992 to 1994 in more than 68 locations (both experimental plots and farmers' fields) covering 18 terai and inner-terai districts of Nepal, indicated that these crops were heavily affected with various virus-like symptoms. The most commonly observed symptoms were severe mosaic, leaf distortion, oily streaks or spots on papaya; leaf distortion, blisters and shoe stringing on zucchini; and mosaic or yellow mosaic, blisters, and leaf distortion on other cucurbits. Average incidence of plants with symptoms ranged from 75% to 100% on papaya; 85% to 100% on zucchini; 4% to 100% on cucumber; 4% to 100% on pumpkin and 10–100% on bottlegourd, choyote and watermelon. The virus isolated from papaya and zucchini was confirmed as papaya ringspot potyvirus — watermelon strain (PRSV-W). It was also detected in survey samples from ashgourd, bittergourd, snakegourd, spongegourd, zucchini, watermelon, bottlegourd and cucumber. Leaf extracts of some cucumber, choyote, pumpkin, zucchini and snakegourd samples reacted with cucumber mosaic cucumovirus (CMV) and zucchini yellow mosaic potyvirus (ZYMV) antisera. Leaf extracts of ashgourd, cucumber and pumpkin reacted with antibodies against cucurbit aphid-borne yellow luteovirus (CABW). No samples reacted with antiserum to watermelon mosaic-2 potyvirus (WMV-2) or squash mosaic potyvirus (SqMV). Some papaya and most cucurbits leaf samples cross-reacted with antibodies against Moroccan (Mor) and Algerian (Alg) isolates of WMV. The Nepalese PRSV isolate was related to but distinct from a PRSV-W type strain from France. This is the first report on the identity of ZYMV and CABW in Nepal.     

Effects of Priming and Endosperm Integrity on Seed Germination Rates of Tomato Genotypes: I. GERMINATION AT SUBOPTIMAL TEMPERATURE

The bases of differences in germination rates (GR_g = inverseof time to germination [t_g] of percentage g) among three cold/salt-toleranttomato (Lycopersicon esculentum Mill.) accessions (PI 341988,PI 120256, and PI 174263) and one cold/salt-sensitive tomatocultivar (T5) were investigated. The effects of seed priming(6 d imbibition in aerated –1.2 MPa polyethylene glycolsolution at 20 ?C followed by redrying) and of removing theendosperm/testa cap covering the radicle on the temperaturesensitivity of GR_g, and the interaction of these treatmentswith genotypes, were also examined. GR_g decreased linearly withdecreasing temperature for all genotypes and seed treatments.The minimum or base temperatures for germination (T_b) variedby 1 ?C among the tomato lines, so genotypic differences inGR_g were due to differing thermal time requirements for germination.The mean thermal time requirement for germination of T5 seeds was 22% and 19% greater than that of PI 341988 andPI 120256 seeds, respectively, but only 9% greater than thatof PI 174263 seeds. Seed priming did not lower Tb of any genotype,but significantly reduced by 24, 49, 41, and 49% in T5, PI 341988, PI 120256, and PI 174263, respectively,indicating that priming increased the rate at which the seedsprogressed towards germination when T>T_b, but did not lowerthe minimum temperature at which germination could occur. Primingincreased the GR_g of T5 seeds to equal or exceed those of control(non-primed) seeds of the cold/salt-tolerant genotypes at anyT>T_b, but the PI lines exhibited an even greater responseto priming. Times to germination within each seed lot were normallydistributed on a logarithmic scale. Priming increased the variancein tg within a seed lot when compared to control seeds. However,the variation in thermal time for germination between the 10thand 90th percentiles of the seed population (_T(10–90))was relatively unaffected by priming due to the reduction in in primed seeds. Removing the endosperm cap and testa opposite the radicle tip decreased almost 6-fold and and reduced T_b by 5 ?C in T5 and PI 341988,implicating processes in the endosperm/testa as the limitingfactors in germination at suboptimal temperatures. Key words: Lycopersicon esculentum Mill., tomato, genetic variation, seed priming, thermal time, germination rate