Exames electrono-microscópicos de tecido foliar ou radicular de milho (Zea maysL.) afetado pelo enfezamento do milho, tanto a forma vermelha como a pálida, e de órgãos da cigarrinha vectora, Dalbulus maidisDe L. & W., portadora, demonstraram a ocorrência de corpúsculos pleomórficos, do tipo micoplasma, consistentemente associados a essa anomalia. Não se notou diferença na morfologia entre os microrganismos do tipo micoplasma associados às duas formas do enfezamento da planta nem tampouco entre estes e os presentes no inseto vector. Verificou-se também que na planta esses corpúsculos do tipo micoplasma ocorriam somente nos vasos crivados, e que no inseto eles se achavam presentes em diversos tipos de tecidos de diferentes órgãos - tubo digestivo, músculo, túbulos de Malphigi, epiderme, gânglio nervoso, glândula salivar, tecido adiposo. Nas células do inseto, os corpúsculos ocorriam dispersos no citoplasma ou em cavidades limitadas por membrana.

Corn stunt in São Paulo State is usually of minor importance during normal summer crops but might induce significant losses if corn is planted late in the season. It is transmitted by leaf - hopper Dalbulus maidisDe L. & W. and two different forms of this disease have been recognized the chlorotic, similar to the Rio Grande type of the U.S., and the red, resembling the Mesa Central type of Mexico. Electron microscopic examination of tissues from both affected plants or leafhopper reared on diseased plants, demonstrated the presence of pleomorphic, mycoplasma-like bodies, consistently associated with the corn stunt, which probably represent the causal agent the latter. No significant difference could be observed in the morphology of these mycoplasma-like bodies associated with the two disease types or with the insect vector. In the affected plants, these corpuscles were only found in the sieve tubes; in the insect body they were detected in several types of tissues of different organs - intestine, Malphigian tubules, muscle, nervous ganglion, salivary gland, fat body, epidermis. In the insect cells, the pleomorphic bodies occurred either scattered in the cytoplasm or in cytoplasmic cavities bounded by membrane.

Derivados de tomateiro, que incorporam resistência ao vírus de mosaico Y (PVY) e mosaico comum (TMV), foram obtidos do cruzamento entre a var. Ângela, resistente às estirpes locais do vírus de mosaico Y, e T 15-1-6-1, que é altamente resistente às estirpes locais do TMV, por meio de inoculação com dois vírus, retrocruzamentos para Ângela e seleção de gerações avançadas do híbrido. Tipos selecionados do tomateiro são homozigotos para os gens Tm-2ªe rt. Suas características comerciais são semelhantes às da var. Ângela. Espera-se que os novos tipos de tomateiro reduzam perdas causadas pela infecção com PVY e/ou TMV, evitem efeitos sinérgicos entre dois vírus em combinação, eliminem a infecção do tipo "internal browning" e também transmissão do TMV pelas sementes.

A tomato variety named Angela, previously described, incorporates resistance to local strains of the potato virus Y group that occur in tomato plantings in São Paulo. This variety is well accepted by tomato growers and is being used as a basis for incorporation of other types of virus disease resistance. Derivatives incorporating resistance to potato Y and tobacco mosaic viruses were obtained from crosses between Ângela and T 15-1-6-1, a tomato type that is highly resistant to tobacco mosaic virus, by inoculation and selection of advance generations of the hybrid or by backcrossing, combined with selfing. The newly selected tomato types are homozygous for the recessive gene rt that controls resistance to potato Y virus and for Tm-2a that conditions tobacco mosaic virus resistance. Their commercial qualities are similar to those of the Ângela parent plant. The new tomato types, resistant to TMV and potato virus Y, are still being tested in a large scale before being released, but it is expected that they will reduce considerably the losses caused by infection with either of the two viruses, avoid injury from the synergistic effect of virus combinations in the same plant, eliminate internal browning, and seed transmission of TMV.

Estudos citoquímicos, ao nível do microscópio óptico, foram efetuados para determinar a natureza química de inclusões intranucleares, de aspecto fibroso, induzidas pelo vírus do mosaico amarelo do salsão na maioria de suas hospedeiras. Os testes citoquímicos foram conduzidos em material foliar fresco ou fixado (aldeído glutárico, Carnoy 3:1 ou Bouin), tendo sido feitas as seguintes reações: hematoxilina férrica (testemunha); Sudan IV e Azul do Nilo (lipídios); iodo-iodeto (amido); Feulgen, Azur B e verde-de-metila-pironina (ácidos nucleicos); ninidrina-Schiff e "Fast Green", este último em soluções ácida e alcalina (proteínas). Os testes com verde-de-metila-pironina, Azur B e ninidrina-Schiff foram combinados com digestão enzimática pela ribonuclease ou pepsina. Os dados obtidos sugerem que, dentro da sensibilidade dos testes realizados, a inclusão contenha proteína, mas não tenha amido, lipídios ou ácidos nucleicos. Isso permite supor, portanto, que essas inclusões não sejam formadas de partículas de vírus.

Cytochemical studies at optical microscopic level were made to determine the chemical nature of intranuclear inclusions with fibrous aspect which were induced by the celery yellow mosaic virus (CYMV) in most of its hosts. The cytochemical tests were carried on fresh as well on fixed foliar material, fixation being in glutaric aldeid, Carnoy 3:1 or Bouin. The following reactions were tried: ferric haematoxylin (control); Sudan IV and Nile blue (for lipids); iodine ioduret (for starch); Feulgen, azur B and methyl green-pyronin (for nucleic acids); ninhydrin-Schiff and fast-green, the latter in acid and in alcaline solution (for protein). The tests with methyl green-pyronin, azur B and ninhydrin-Schiff were combined with enzimatic digestion with RNase or pepsin. The results suggest that, within the sensibility of the tests, the inclusion contains protein but does not contain starch, lipids or nucleic acids. This permit to suppose, therefore, such inclusions not to be formed of virus particles.

Fragmentos da epiderme inferior de fôlhas de trigo com sintomas típicos da anomalia conhecida como "espiga branca", examinados ao microscópio convencional, mostraram a constante ocorrência de inclusões intracelulares, de dimensões avantajadas e de aspecto fibrilar e enovelado. Freqüentemente, células adjacentes aquelas que continham inclusões apresentavam formações aciculares, em seu interior. Seções ultrafinas dessas folhas, examinadas ao microscópio electrô-nico, revelaram que tais inclusões eram de localização citoplasmática e formadas por uma massa de partículas filamentosas de 7-10 m¼ em diâmetro e de comprimento indeterminado, idênticas àquelas encontradas em preparações "leaf dip". Várias alterações celulares, como hipertrofia do nucléolo, degeneração dos cloroplastos e vesicularização do retículo endoplasmático, puderam também ser notadas. Ocasionalmente, formações cristalinas foram notadas na periferia e mesmo no interior do núcleo. A semelhança das inclusões citoplasmáticas e das partículas que as compõem, com aquelas descritas, associadas à infecção do vírus da "hoja blanca" do arroz, e também o fato de o trigo desenvolver sintomas do tipo espiga branca quando inoculado experimentalmente com o vírus da "hoja blanca", parecem reforçar a sugestão de que a espiga branca do trigo teria etiologia virosa e que o agente causal seria do grupo do vírus da "hoja blanca" do arroz. A constante associação das inclusões celulares com a condição de espiga branca, em trigo, constitui mais um elemento para sua rápida diagnose.

Light microscopic examination of epidermal strips from leaves of wheat (Triticum sativumL.) showing typical symptoms of the white spike (WS) disease, demonstrated the constant occurrence of huge, fibrous and whorled intracellular inclusions. Occasionally cells adjacent to those having inclusions contained needle-like formations. The whorled inclusions appeared in ultrathin section profiles as loose aggregates of threadlike elements, 7-10 m¼ in diameter and of indeterminable length, within the cytoplasm. They seem to represent filamentous particles found in leaf dip preparations from affected plants, in situ. Nucleolus hypertrophy, chloroplast degeneration and changes in endoplasmic reticulum were frequently noticed in affected tissues. Occasionally small crystalline inclusions were found nearby or within the nucleus. The presence of these fibrous, whorled inclusions associated to WS condition, reinforces the suggested viral etiology, due to the similarity of these inclusions with virus aggregates found in tissues infected with several other viruses of the beet yellows virus group. Besides symptomatology, the particle and inclusion morphology of rice "hoja blanca" virus are very similar to WS of wheat, suggesting that both diseases are induced by viruses of the same complex. The ease with which the inclusions are detected in epidermal strips, constitutes a quick and secure method to diagnose the WS disease.

Exames electrono-microscópicos de secções ultrafinas de tecidos foliares de cenoura (Daucus carota L.) e coentro (Coriandrum sativumL.) infetados pelo vírus do mosaico da cenoura (VMC) revelaram a ocorrência de inclusões em forma de bandas densas no citoplasma. Exames mais detalhados dessas bandas revelaram serem elas constituídas pela justaposição de lamelas delgadas (10-15m¼ de espessura), as quais parecem eventualmente desprender-se e formar outros tipos de inclusão, como as de perfil circular e em forma de catavento, comumente associadas à infecção provocada por vírus do grupo Y da batata. Ocasionalmente, partículas do VMC foram observadas nas proximidades dessas inclusões. As dimensões das bandas densas atingiram até 0,l-0,2¼ x 3-4¼, visíveis mesmo em exames ao microscópio convencional. Além da ocorrência dessas inclusões, pôde também ser observada a presença de grande número de vesículas no citoplasma, aparentemente associadas a uma atividade do complexo de Golgi.

A virus, inducing mosaic and malformation of carrot leaves (Daucus carota L.), was found in Piedade, SP. It was termed carrot mosaic virus (CMV), and is both mechanically and aphid transmitted. Morphologically it belongs to the potato virus Y group (15m¼ x 740m¼). Electron microscopic examination of thin sections of leaf tissues from carrot or coriander (Coriandrum sativum L.) infected with CMV demonstrated the presence of cytoplasmic inclusions, usually in the form of dense bands. They could reach sizes up to 0.2¼ x 4¼ and were detectable even under light microscopic examination. These bands, at higher magnification, showed to be built up of thin lamella (10-15m¼ thick), closely apposed. Occasionally individual or group of lamella appeared to loose from the bands, producing other type of profile, such as pin wheels and rings. CMV particles were sometimes observed in the vicinity of these inclusions, often laying parallel to their surface. These inclusions are identical to those observed in tissues infected with other components of the potato Y virus group, and their detection seems to corroborate the previous inclusion of CMV in this group, basead on particle morphology. Besides these cytoplasmic inclusions, some cells contained a large number of vesicles in the cytoplasm, apparently associated with Golgi complex activity.

Foi verificado haver correlação entre a manifestação de sintomas nas plantas indicadoras intercaladas no batatal para semente e a disseminação, nêle, do vírus do enrolamento das fôlhas. Médias de 12 ensaios realizados em vários pontos do Estado de São Paulo deram os seguintes valores: 49,6% em Datura stramonium, 34,0% em Physalis sp. e 28,5% em tomateiro Santa Cruz. Somente 0,9% das plantas de batata mostraram sintomas de infecção primária pelo vírus, embora a infecção no campo tenha realmente sido de 23,6%. É sugerida a introdução do uso de plantas indicadoras na legislação que regulamenta a produção de batata-semente certificada como método auxiliar para estimativa do teor do vírus do enrolamento nos tubérculos produzidos.

Leaf roll infection on the interplanted indicator plants, jimson weed, tomato, and a non-identified Physalis species is correlated to spread of the disease in the seed potato field. Result from twelve tests carried out at several localities gave the following infection averages: jimson weed, 49.6%; Physalissp., 34.0%; tomato plants, 28.5%. Potato plants showed an average of 0.9% primary leaf roll, but tuber tests indicated that field spread had been 23.6%. Potato virus Y infected 45.1% of the tobacco plants, 37.3% of Physalissp., and 18.1% of the tomato plants. Potato plants were not infected with this virus because the Ycomplex present was the one that infects tomato and pepper plants and very seldom potato plants in the field. The viruses of tomato curly top and Brazilian tobacco streak infected jimson weed at the average rates of 16.1% and 2.7%, respectively. Tomato plants showed an average of 4.1% of curly top, but no infection with the Brazilian tobacco streak virus. Potato plants were not affected by these viruses. Tomato spotted wilt virus infected several of the indicator species and also the potato plants, but there was no perpetuation of the virus through the tubers. Interplanting indicator plants in the certified seed potato fields is suggested as a means to estimate the extent of leaf roll spread in the crop.

A comparação entre as produções de 50 pares de plantas sadias e afetadas, de nove variedades de batata, mostra que a redução na produção causada pela infecção secundária do vírus do enrolamento da fôlha foi, em média, de 60,8% no pêso total dos tubérculos produzidos e de 75,5% no pêso dos tubérculos do tipo graúdo ("especial" mais "primeira"). As perdas na produção total variaram de 44,6% (variedade Patrones) a 73,4% (variedade Aquila), e na de tubérculos graúdos, de 49,8% (Delta A) a 86,0% (Gunda).

A comparison of 50 pairs of field healthy and leaf roll infected potato plants of nine varieties indicated that the total yield reduction was on the average 60.7 per cent. Yield reduction of the two largest and most marketable potato sizes ("especial" and "primeira") reached 75.6 per cent. Aquila and Gunda had the highest yield losses (73.4 and 72.0 per cent); Delta A and Patrones had the lowest (47.6 and 44.6 per cent). Reduction in the yield of the larger tuber sizes was generally greater than total losses.

Partículas alongadas de 15 mµ de diâmetro e 650 mµ de comprimento normal foram consistentemente detectadas em exames electrono--microscópicos de preparações "leaf dip" de 6 espécies diferentes de plantas infetadas pelo vírus latente da couve (Brassica oleracea L. var. acephala) (VLC). Tais partículas são consideradas como sendo de um vírus que poderia ser incluído no grupo do vírus S da batata, na classificação dos vírus alongados de Brandes. Exames de secções ultrafinas de tecidos foliar e radicular de couve infetada pelo VLC, demonstraram a ocorrência do VLC, formando agregados mais ou menos organizados no citoplasma. Não se notou alterações citológicas sensíveis como conseqüência da infecção, o que está de acordo com a ausência de sintomas externos em couve infetada pelo VLC.

Electron microscopic examination of leaf dip preparations made from cole (Brassica oleraceaL. var. acephala), Chenopodium amaranticolorCoste & Reyn., radish (Raphanus sativusL.), mustard(Brassica albaBoiss. Rabenh.), bean (Phaseolus vulgaris L.)and Gomphrena globosaL. plants infected with cole latente virus (CLV) demonstrated consistently the presence of slender rods. These particles, considered as CLV, were about 15 mµ wide and 650 mµ in normal length (NL). The NL was determined in a series of comparative measurements using turnip mosaic virus (NL = 750 mµ) as a standard. Morphological characteristics of CLV permit its inclusion in the potato virus S group, in Brandes' system of classification of elongated plant viruses. Thin section profiles of root and leaf tissues, from CLV-infected cole, showed the frequent occurrence of fibrillar aggregates in the cytoplasm. The particles component of these aggregates were identified as CLV in situ due to the constant association with the infection and their mophological similarities with the particles found in leaf dip preparations. CLV-particles appeared with their longer axis laying side--by-side within these aggregates, often forming a spiral arrangement. Vesiculated areas were occasionally seen in the cytoplasm, with virions in or around them. Virus particles were found in most of the examined cells, except within the tracheids and sieve tubes in the leaf, and within the meristematic zone, in th root. Besides these abnormalities, no other remarkable changes in normal cell structure could be noticed, in agreement with the lack of external symptoms.