Physiologic specialization of Colletotrichum corchori,Ikata and Yoshida,the causal organism of anthracnose of jute (Corchorus capsularis L.)

Summary Four varieties of jute,Corchorus capsularis, among fifteen screened were found to differentiate six races ofColletotrichum corchori. As regard virulence only two races, CS-1 and CS-2 among the six were of real importance.Besides recognising six races on four differential jute varieties, these were also differentiated from their cultural and morphological studies on laboratory medium as six cultural races.     

Preparation and regeneration of protoplasts of three fungi of Boletaceae

Protoplasts of three fungi of Boletaceae,Suillus luteus, S. grevillei, andBoletinus cavipes, were prepared with yields of 45, 8.0, and 1.8×10⁷/g fresh mycelia under the optimal conditions, respectively. Nucleate protoplasts accounted for 42% of the whole preparation ofS. luteus and 32% of that ofS. grevillei, and 21% of the nucleate protoplasts ofS. luteus and 35% of those ofS. grevillei possesed two nuclei. Regeneration efficiency of protoplasts was 0.4% forS. luteus and 0.05% forS. grevillei. The regeneration ofB. cavipes protoplasts was also confirmed. Optimal conditions for regeneration were determined. Addition of gellan gum instead of agar to the medium and activated charcoal treatment of agar medium increased the regeneration efficiency significantly.     

Synthesis of sterigmatocystin on a chemically defined medium by species ofAspergillus andChaetomium

Sterigmatocystin (ST) is a secondary metabolite and a principal mycotoxin known to be produced by over 30 species of filamentous fungi. It is also one of the late intermediates in aflatoxin biosynthesis. We have tested the ability of 7 species ofAspergillus, including 4 strains ofA. versicolor, one species ofBipolaris, and two species ofChaetomium, to produce ST on a sucrose-salts-phenylalanine defined medium as well as on three complex substrates. Highest ST production in our survey was by a strain ofA. versicolor grown on wheat, whereas, the highest ST production on defined medium was byC. cellulolyticum. To our knowledge, this is the first report of ST production byC. cellulolyticum on any substrate. In precursor feeding studies, resting cultures of wild typeA. nidulans andA. versicolor were unable to biotransform O-methylsterigmatocystin (OMST), the last known intermediate in aflatoxin biosynthesis. These results suggest that ST is the end product of polyketide metabolism in the strains tested.     

Use of the egg parasiteEdovum puttleri [Hym.: Eulophidae] in an IPM system developed for Colorado potato beetle [Col.: Chrysomelidae] control on potatoes,Beltsville, Maryland

Edovum puttleri Grissell in conjunction with the fungal pathogenBeauveria bassiana (Balsamo) Vuillimen, a β-exotoxin ofBacillus thuringiensis varthuringiensis Berliner, and the insect growth regulator triflumuron (Alsystin 4F) were evaluated in a pest management system to control Colorado potato beetle,Leptinotarsa decemlineata (Say). Soil treatments ofB. bassiana had no significant impact on emerging Colorado potato beetle adults. Foliar applications of the β-exotoxin and triflumuron significantly reduced 1 ^st larval generation. Releases ofE. puttleri resulted in 56 % egg mass parasitization, significant reduction of 2 ^nd generation larvae, and significantly higher potato yields.Leptinotarsa texana (Schaeffer) was successfully used as an alternate host for production ofE. puttleri.      

Observations on gymnoascaceae. IX. A new species of pseudoarachniotus from soil-burial beds

Summary A new species ofPseudoarachniotus P. marginosporus, is described and illustrated. The developmental morphology of this new ascomycete is discussed. On a standard medium such as YpSs,P. marginosporus can be distinguished from other species of the genus by colonial coloration which varies from ochre-rust to very pale yellow at different stages of maturation. The colony ofP. roseus is red, red-orange or orange-yellow;P. citrinus is lemon yellow to brownish yellow in age;P. reticulatus is red orange andP. hyalinosporus is yellowish and usually with a great deal of green pigmentation in the mycelium and in the medium. Furthermore ascospores ofP. marginosporus are oblate, small and thin-walled, measuring 2.2–3.0×4.0–4.4 µ, with a minute elevated rim around the long axis, while ascospores ofP. reticulatus are globose and reticulate, and those ofP. roseus andP. citrinus are large and thick-walled, measuring about 4.4×7.7 µ. Ascospores ofP. hyalinosporus are similar in size and shape to those ofP. marginosporus, but lack a rim or other markings.     

A rapid identification method for aflatoxin-producing strains ofAspergillus flavus andA. parasiticus by ammonia vapor

The colony reverse of aflatoxin (AF)-producing strains ofAspergillus flavus andA. parasiticus turned pink when their cultures were exposed to ammonia vapor. The color change was visible for colonies grown on media suitable for AF production such as potato dextrose, coconut, and yeast extract sucrose agars after 2 d incubation at 25°C. Of the 120 strains ofA. flavus, A. parasiticus, and two related species inA. flavus group:A. oryzae andA. sojae tested in this study, only the AF-producing strains ofA. flavus andA. parasiticus showed the pink pigmentation. The color change occurred immediately after the colony was contacted with ammonia vapor. This method was useful for rapid screening the AF-producing strains ofA. flavus andA. parasiticus.     

Effect of carbohydrates on the production of thaxtomin A by <Emphasis Type="Italic">Streptomyces acidiscabies</Emphasis>

Several Streptomyces species cause plant diseases, including S. scabies, S. acidiscabies and S. turgidiscabies, which produce common scab of potato and similar diseases of root crops. These species produce thaxtomins, dipeptide phytotoxins that are responsible for disease symptoms. Thaxtomins are produced in vivo on diseased potato tissue and in vitro in oat-based culture media, but the regulation of thaxtomin biosynthesis is not understood. S. acidiscabies was grown in a variety of media to assess the impact of medium components on thaxtomin A (ThxA) production. ThxA biosynthesis was not correlated with bacterial biomass, nor was it stimulated by α-solanine or α-chaconine, the two most prevalent potato glycoalkaloids. ThxA production was stimulated by oat bran broth, even after exhaustive extraction, suggesting that specific carbohydrates may influence ThxA biosynthesis. Oat bran contains high levels of xylans and glucans, and both of these carbohydrates, as well as xylans from wheat and tamarind, stimulated ThxA production, but not to the same extent as oat bran. Starches and simple sugars did not induce ThxA production. The data indicate that complex carbohydrates may act as environmental signals to plant pathogenic Streptomyces, allowing production of thaxtomin and enabling bacteria to colonize its host.     

Glandular trichomes ofSolanum berthaultii alter host preference of the Colorado potato Beetle,Leptinotarsa decemlineata

Choice and no-choice studies were conducted to determine how the glandular trichomes of the wild potato,Solanum berthaultii Hawkes, affect host preference of the Colorado potato beetle,Leptinotarsa decemlineata (Say). Given a feeding choice betweenS. tuberosum andS. berthaultii, larvae and adults preferred the foliage ofS. tuberosum, but adults were more discriminating. When foliage ofS. berthaultii was appressed toS. tuberosum leaflets, fewer adults fed on the appressed leaflets. When given a choice between ‘trichome-intact’ and ‘trichome-removed’S. berthaultii foliage, adults preferred to feed on the latter. The preference for ‘trichome-removed’ foliage and the percent of adults initiating feeding, increased with the degree of trichome removal. These studies provide evidence that the resistance ofS. berthaultii is associated with feeding deterrents localized in the glandular trichomes, thatS. berthaultii possesses more than one mechanism of resistance to the Colorado potato beetle, and that the expression of resistance is dependent on the developmental stage of the insect.     

Fungal diseases of the respiratory tract

The proportion ofCandida and non-Candida species in the clinical material from patients. with respiratory-tract diseases was determined.C. albicans was isolated in 102 cases. An additional 89 strains of yeasts, isolated in association with respiratory diseases, belonged to 10 non-albicans Candida spp. andCryptococcus spp. The prevailing species, which occurred in 47 cases, wasC. parapsilosis. C. tropicalis, C. glabrata, andC. guilliermondii were isolated in 12, 10, and 9 cases, respectively. Four strains ofC. krusei and three strains ofC. lusitaniae and one strain each ofC. freyschussii, C. robusta, C. zeylanoides, andCryptococcus neoformans were also isolated.     

The effect of magnesium and phosphate on mycological fat formation from sweet potatoes

In an attempt to promote mycological fat formation from sweet potatoes, the sweet potato medium was supplemented with magnesium sulphate or sodium phosphate. The sweet potato medium itself was used either as such or after hydrolysis with acid or with enzyme. Two local fungi were used, namely,Aspergillus oryzae andA. terreus. Addition of magnesium or phosphate enhanced carbohydrate absorption from the external medium. This occurred to a remarkable extent in enzyme-hydrolysed sweet potato medium and when both factors were present. The presence of magnesium or phosphate suppressed fungal growth in acid-hydrolysed media particularly in case ofA. terreus. In untreated media either factor promoted growth ofA. oryzae but suppressed that ofA. terreus.     

Behavioral ecology ofMyiopharus doryphorae (riley) andM. aberrans (townsend), tachinid parasitoids of the colorado potato beetle

The behavior ofMyiopharus doryphorae andM. aberrans, North American tachinid parasitoids of the Colorado potato beetle, was recorded under field and laboratory conditions throughout three growing seasons in western Massachusetts. Eight common behaviors associated with resting, searching, feeding, and larviposition were distinguished, which together accounted for nearly all daytime activity of the females of both tachinids. Several of these behaviors, and in particular larviposition, were closely related to temperature but differed between species. A sequence of five defensive behaviors by the different larval stages of the Colorado potato beetle prevented larviposition in 49% of resisted attempts and perhaps one-fourth of total larviposition attempts byMyiopharus species, yet both parasitoids were highly successful in allocating their progeny during most of the summer. Second- and third-instar beetle larvae were least effective in resisting larviposition. Females of bothMyiopharus species actively guarded recently parasitized hosts from otherMyiopharus females for a period of several minutes after larviposition during the last month of the growing season when second- and third-instar Colorado potato beetle larvae were most scarce. Laboratory studies based on the field observation that femaleM. aberrans doggedly pursued circum-diapausing adult beetles led to the first recorded account ofM. aberrans larvipositing in adult hosts. Flies gained access to a beetle’s vulnerable abdominal dorsum at the instant it lifted its elytra to initiate flight. The late-season switch ofM. aberrans to adult Colorado potato beetles contributed to a seasonal sequence of larviposition-related behaviors concordant with prevailing host densities, which should lend complementarity toM. doryphorae andM. aberrans as biological controls of pest populations.     

Fungicide inhibition of aflatoxins,diacetoxyscirpenol and zearalenone production

The effect of fungicides on the production of aflatoxin byAspergillus flavus IMI 89717, diacetoxyscirpenol and zearalenone byFusarium graminearum was studied. In a yeast extract - sucrose medium, dicloran, iprodione and vinclozolin fungicides significantly inhibited mycelial growth ofA. flavus at 250 ppm and significantly decreased aflatoxin production at 100, 250 and 500 ppm, respectively. In potato — dextrose broth, these fungicides diminished the mycelial growth ofF. graminearum and production of diacetoxyscirpenol and zearalenone at 100 ppm. Sensitivity of toxigenic mycelia to fungicides increased approximately five-fold in a yeast extract-starch medium with an appreciable reduction in sugar uptake andα-amylase activity.     

A novel J8 domain gene,IbJ8, inIpomoea batatas (L). Lam.

Sweet potato cDNAs that encode IbJ8, the smallest known J-domain protein, were isolated and characterized. This genome has at least two copies ofIbJ8, which is expressed preferentially in the leaves, flowers, petioles, and stems. Spatial and temporal patterns were studied at different developmental stages, and expression was greater in younger leaves than in older ones. Moreover, expression in roots that arose from single-leaf cuttings was lower at 15, 20, and 30 d than at 40 d, and then the signal was undetectable at 60 d after planting. These results suggest thatIbJ8 expression may be related to the organ age or developmental stage.     

Antibiose Manifestée Par Une Souche De Byssochlamys Nivea Westling, 1909. I. Conditions D'apparition

The juice of heat processed strawberries, contaminated by a strain ofByssochlamys nivea, inhibited the growth ofEscherichia coli.The antibiotic property of the isolated strain, growing in a liquid medium, appears only after several days and disappears at a later period.     

Biological control of the root rot and wilt diseases ofLens culinaris medic

Summary Biological control visualizes a control of disease with the help of living organisms. In case of root disease, organisms occur in soil which suppress the activity of disease fungi. In the present investigation antagonists have been isolated and used in the control of root rot and wilt disease ofLens culinaris caused byS. rolfsii andF. oxysporum respectively.Isolation of antagonistic micro-organisms were done from the rhizosphere soil of Bombay 20 and local varieties and from field soil. Antagonists were screened against certain test organisms and pathogens. The selected antagonists were used for the control of the diseases of lentil crop by inoculating the antagonists in the soil and thereby to see the effect of decreased plant disease in sterilized and unsterilized soil.Amendments in the form of different organic materials were also given to control the diseases in natural soil.The most effective results were obtained withT. viride, Streptomyces gougeroti Streptomyces species 12 and 13, bacterial isolates no. 16 and 18 in case ofF. oxysporum f.lentis inoculated pots, where there was no disease. In case ofS. rolfsii inoculated pots not a single antagonist was able to eradicate the disease completely.Portion of the Ph. D. thesis accepted by the University of Saugor, Sagar, India.     

Rates of growth and increase ofMyzus persicae on virus-infected potatoes according to type of virus-vector relationship

Growth, reproduction and survival (=performance) of the aphidMyzus persicae Sulzer was measured on virus-free and virus-infected potato plants The principle objective was to evaluate if various viral infections affected aphid performance differently, and if so, whether any order in the performance response of the aphid was discernible according to the type of virus-vector relationship. Three viruses varying in their dependency onM. persicae as a vector were used. Plants infected with potato leafroll virus (PLRV), a circulative virus highly dependent uponM. persicae for dispersal and transmission, were superior hosts as determined by the significantly greater mean relative growth rate (MRGR) and intrinsic rate of increase (r_m) ofM. persicae compared with those of aphids reared on other plants. Plants infected with potato virus Y, a noncirculative virus less dependent uponM. persicae for dispersal than PLRV, were intermediate in their quality based upon intermediate MRGR and r_m values. Plants infected with potato virus X, a nonvectored virus independent ofM. persicae, were least suitable hosts along with the group of virus-free plants according to the lower MRGR and r_m values.     

Purification of antibiotics produced byLentinus squarrosulus and preliminary characterization of a compound active againstRigidoporus lignosus

Purification and study of the antibiotic substances produced byLentinus squarrosulus have been carried out. The substances, excreted in the culture medium, were extracted withn-butanol. The butanolic extract inhibited growth ofRigidopurus lignosus, the agent of white rot ofHevea brasiliensis, and also ofMucor ramannianus, Rhodotorula mucilaginosa, Saccharomyces cerevisiae, andBacillus subtilis. The antibiotic fractions were purified by silicic acid column chromatography, then by reversed-phase (C18) high performance liquid chromatography (HPLC) followed by preparative adsorption thin-layer chromatography on silica gel. Two purified compounds were obtained: Ls1, which was active againstB. subtilis, and Ls2, which in addition was also active againstR. lignosus, M. ramannianus, and yeasts. Only the Ls2 compound is analyzed in this work. It was characterized by chemical reactions, ultraviolet spectroscopy, infrared spectroscopy, and proton nuclear magnetic resonance spectroscopy, which indicated the hydrophilic character of the molecule and the presence of alcoholic functions as well as a glycosidic moiety. The properties differed from those of already known antibiotics produced by different species ofLentinus.     

An allele of dihydroflavonol 4-reductase associated with the ability to produce red anthocyanin pigments in potato (Solanum tuberosum L.)

The potato R locus is necessary for the production of red pelargonidin-based anthocyanin pigments in any tissue of the plant, including tuber skin and flower petals. The production of pelargonidins in plants requires the activity of dihydroflavonol 4-reductase (DFR) to catalyze the reduction of dihydrokaempferol into leucopelargonidin. To test the hypothesis that potato R encodes DFR, portions of both dfr alleles were sequenced from a diploid potato clone known to be heterozygous Rr. Sequence comparison revealed a polymorphic BamHI restriction site. The presence or absence of this site was monitored in three diploid populations that segregated for R, as well as in a wide range of tetraploid breeding clones and cultivars, by amplifying a fragment of dfr and digesting the products with BamHI. An identically sized dfr restriction fragment lacking the BamHI site was present in all potato clones that produced red anthocyanin pigments, while the same fragment was absent in many potato clones with white tuber skin and flowers. An independent RFLP test using DraI to detect sequence polymorphism was performed on a subset of the potato clones. This test also revealed dfr-derived bands that were present in all red-colored potatoes and absent in several white clones. The presence of shared restriction fragments in all red-colored potatoes provides strong evidence that R does indeed code for DFR. The data are also consistent with a 48 year-old hypothesis by Dodds and Long, that R was selected just once during the domestication of potato. A cDNA clone corresponding to the red allele of dfr was sequenced and compared to two other alleles. The red allele is predicted to encode a 382 amino acid protein that differs at ten amino acid positions from the gene products of the two alternative alleles. Several of these differences map in a region known to influence DFR substrate specificity in Gerbera.Communicated by J. Dvorak     

Rapid propagation of Phalaenopsis from floral stalk-derived leaves

Summary An efficient and rapid in vitro method was developed for regeneration of Phalaenopsis using leaf segments derived in vitro from flower stalk nodes. Leaf segments of four cultivars Tinny Sunshine ‘Annie’, ‘Taisuco Hatarot’, Teipei Gold ‘Golden Star’, Tinny Galaxy ‘Annie’ cultured on Murashige and Skoog medium supplemented with N ⁶-benzyladenine (BA; 88,8 μM) and α-naphthaleneacetic acid (NAA; 5,4 μM) produced an average of 10–13 protocorm-like bodies (PLBs) after 12 wk. PLB proliferation was achieved on a modified Hyponex medium (1 gl⁻¹ 6.5N−4.5P−19K+20N−20P−20K+2gl⁻¹ peplone +3% (w/v) potato homogenate +0.05% activated 1 gl⁻¹ charcoal) and an optimal number of 13–18 PLBs developed from single PLB sections of different cultivars. Plantlet development was also achieved on a modified Hyponex medium. By repeated subculture of PLBs on a proliferation medium, and culturing them in the plantlet regeneration medium, plantlets could be produced continuously. Approximately 6 mo, were required from the initiation of culture to the production of plantlets for transplant to community pots.     

An account of some physiological studies in two species of phytophthora

Summary The paper deals with some physiological studies of two typical strains ofPhytophthora palmivora Butl. andP. parasitica Dast. var.macrospora Ashby causing severe fruit rots ofAchras sapota L. andAnona squamosa L. respectively. The effect of temperature on the germination of sporangia had a marked effect. Maximum percentage of indirect germination (by formation of zoospores) occurred at 20° C, whereas direct germination of sporangia (by formation of germ tube) was at 30° C, which further continued upto 37° C. Maximum indirect germination at 20° C shows the favourable cool temperature for the epidemic of the diseases (Fruit rots) in nature. The best substrate (medium) for germination of sporangia was found to be tap water. Next to this was the host decoction or extract. Glucose solution also accelerated sporangial germination. The effect of two dyes viz., Malachite green and crystal violet was also studied in relation to growth and sporulation ofPhytophthora isolates. Their addition to the medium in various fractional dilutions had a profound influence in the rate of growth and sporulation. An interesting observation noted was that growth of the isolates was inversely porportional to the various concentrations of the dyes, under study. An attempt was also made to study the influence of various vitamins. In all, six vitamins were included in the study. Out of these, thiamine and riboflavin were found to be the best sources promoting good growth and sporulation of both the species ofPhytophthora under study.Forms a part of Senior author's M. Sc. (Agric.) Thesis, University of Poona, Poona (India).Respectively, Ex-Junior Research Fellow, I.C.A.R., New Delhi; Professor of Plant Pathology and Principa, College of Agriculture, Junagad (Gujarat); Plant Pathologist, Wheat Rust Research Station, Mahabaleshwar, Poona, India.