Fungal Assciations: I. Synergistic Relation between Rhizoctonia solani Kuhn and Fusarium solani Snyder and Hansen in causing a Potato Tuber Rot

Rhizoctonia solani, Fusarium solani, and Phoma foveata werechosen for the study of disease caused by these fungi in differentcombinations in potato tubers. An initial Rhizoctonia infection,when followed by a Fusarium infection, gave an extensive rottingwith external pimple-like formations in some cases. This typeof rotting could not be brought about by individual infectionswith either of the two fungi, or jointly by them when Fusariumwas inoculated first. Microscopic observations of infected matureand young potato tubers showed that Rhizoctonia grew intracellularlywhen infected alone, whereas it grew inter- as well as intra-cellularlyin the successive double infection. Fusarium formed more haustorium-likestructures when inoculated alone that when it followed Rhizoctonia.The length of these structures in the double infection was greaterin mature than in young tubers. Atmospheric humidity affectedthe amount of rotting, the shape and colour of the rot, andthe morphology of the fungus in the tissue.     

The Galls on Forsythia: Inoculation Studies

Two fungi, Fusarium lateritium and Phomopsis dominici and twobacteria (S3) and (G1) resembling Agrobacterium radiobactervar. tumefaciens and Corynebacterium fascians isolated fromgalls on Forsythia intermedia failed to produce galls upon reinoculation.Similarly, cultures of C. fascians, Pseudomonas savastanoi andP. savastanoi var. fraxini failed to induce galls. Both A. radiobactervar. tumefaciens and tumerous carrot tissue infected with A.radiobacter var. rhizogenes produced tumerous tissue in inoculatedplants but only the latter induced an overgrowth resemblinga natural gall on Forsythia.     

Variation in 14C Partitioning in the Tips of Growing Stolons of Potato (Solanum tuberosum L.)

¹⁴C partitioning was examined in growing stolons of field-grownpotato (Solanum tuberosum L.) cv. Maris Piper. Considerablevariation was evident on single plants and on a fresh weightbasis many stolon tips, which showed no signs of sub-apicalswelling, had higher specific activities (cpm g^–1 f. wt)of ¹⁴C in both ethanol soluble and insoluble forms than larger,visibly tuberized stolons. Furthermore, many tips of low freshweight had a higher insoluble to soluble ¹⁴C ratio than visiblytuberized stolons suggesting greater efficiency of conversionof soluble ¹⁴C to insoluble ¹⁴C in the smaller stolons. Theresults suggest that the onset of visible ‘tuberization’,namely the sub-apical swelling of the stolon, is preceded byincreased soluble carbon accumulation at the stolon tip togetherwith an increase in the conversion of soluble to insoluble formsof carbon. Tuberization, ¹⁴C, stolon tip     

Effect of wounding or infection by Phytophthora infestans on the contents of terpenoids in potato tubers

Inoculation of the potato cut surfaces with an incompatiblerace of Phytophthora infestans induced an accumulation of rishitin,but only a trace occurred when infected by a compatible race.When the tuber was cut, a large amount of steroid-glycoalkaloids(solanine) accumulated in the cut surface tissue, although onlya trace was found in intact tissue. Only a small amount of solanine,if any, was contained in the wound periderm tissue. Most ofthe solanine seemed to be distributed in tissue neighbouringthe newly formed meristematic tissue zone. Distribution of solanineas a function of distance from the cut surface was exponential.Infection of the cut surface by an incompatible race of Phytophthorainfestans reduced the accumulation of solanine. The higher theconcentration of zoospore used, the less the solanine content.It has been reported that the higher the concentration of zoosporesused for inoculation of the cut surface, the less the numberof renewed meristematic cells in the wound tissue. In experimentsusing fresh and aged tubers, a good correlation between thenumber of renewed meristematic cells and solanine content wasfound. The accumulation of solanine in the wound tissue andits reduction due to infection by an incompatible race may berelated to renewed meristematic cells formation and its reductioncaused by the infection. No drastic change in carotenoids or sterol contents was found2 days after cutting or inoculation, when the tubers were cut,or cut and then infected by the incompatible race. ¹ Studies on the phytoalexin (No. 10). (6) in References constitutes"Studies on the phytoalexin No. 9". ² Present adress: Plant Pathology Laboratory, Faculty of Agriculture,Nagoya University, Chikusa-ku, Nagoya, 464, Japan. (Received April 1, 1972; )     

CHANGE OF B-TYPE HAEM CONTENT IN RELATION TO PEROXIDASE BIOSYNTHESIS IN INJURED SWEET POTATO ROOTS

The methods of quantitative analysis of b-type haem in plantswere investigated. With an improved method developed was determinedthe haem content in the supernatant, mitochondrial, and microsomalfractions of sweet potato tissue. The activities of peroxidase,catalase, and cytochrome oxidase, as well as the contents ofb-type haem and acid-insoluble nitrogen in the cellular fractionswere determined at different incubation times after cuttingof sweet potato tissue. Peroxidase and catalase increased withtime in each celluler fraction, following a short lag phase.In the mitochondrial fraction, b-type haem, cytochrome oxidase,and acid insoluble nitrogen increased linearly with time. Inthe microsomal and supernatant fraction, b-type haem increasedwith time following a short lag phase. The increase in haemcontent of the supernatant fraction appeared to be associatedwith peroxidase formation. Time course analysis showed that ⁵⁹Fe-incorporation into b-typehaem of the supernatant fraction increased with time and thatincorporation was markedly inhibited by blasticidin S. The incorporationof ⁵⁹Fe into mitochondrial haem did not increase with time andwas not inhibited by blasticidin S. Blasticidin S inhibited⁵⁹Fe-incorporation into microsomal haem. Time course analysisof b-type haem content, ⁵⁹Fe-incorporation into b-type haem,and peroxidase activity suggest that in the injured tissue haemis synthesized from low molecular weight compounds and is incorporatedinto peroxidase as the haem moiety. ¹ This paper constitutes Part 57 of the Phytopathological Chemistryof Sweet Potato with Black Rot. ² Present address: Institute for Plant Virus Research, Chiba.     

Types of Rot, Rate of Rotting, and Analysis of Pectic Substances in Apples Rotted by Fungi

The rate at which the fungi grew through apples was determinedin various ways and used to estimate their rate of linear advance.Five fungi were studied;they were Sclerotinia fructigena (firm-browncoloured rot, rapid growth through apples), Botrytis cinerea(soft, light-brown coloured rot, rapid growth through apples),Psyrenochaeta furfuracea (firm to soft rot, variable in colourbut generaly dark, slow growth through apples), Penicilliumexpansion A (soft, white rot, slow growth through apples) andPenicillium expansum B (soft, white rot, medium rate of growththrough apples). S. fructigena which had the highest rate oflinear advance which was about three times that of P. furfuraceawhich had the lowest. Methods for extracting different types of pectic substancesfrom sound and rotted tissues are described, and details aregiven of a rapid and reasonably accurate colorimetric methodof determining the anhydrogalacturonic acid content of theseextracts. The firm-rot fungi reduced the water-insoluble pecticsubstances by 10–20 per cent., but the soft-rot fungicaused much larger changes, up to 70 per cent. being degraded,The firm-rot and soft-rot fungi had different effects on thepectic substances insoluble in dilute acid but soluble in dilutealkali. The soft-rot fungi had little effect on these substances,or reduced their concentration, whereas the firm-rot fungi causedsubstantial increases compared with sound tissue. These resultsare considered in terms of pectic enzyme activity. Analysisof extracts by paper chromatography showed that galacturonicacid, absent from sound tissue, was present in each type ofrotted tissue. Di- and tri-galacturonic acids were present inrots caused by P. expansum, and these rots probably also containedproducts from the break-down of other polysaccharides.     

Fruit Softening II. Precursor Incorporation into Pectin by Pear Tissue 6

The incorporation of radioactivity from precursors of methylester and galacturonosyl residues into pectin was investigatedusing tissue slices cut from ripening pear fruits. Incorporationfrom ¹⁴CH₃ methionine into methyl ester of water soluble pectinincreased 10 fold in 4 d at 18 °C and declined in laterstages of ripening. Activity from [³H]inositol could not bedetected in gaJacturonic acid released enzymically from solublepolysaccharides. When l³H]glucose was used as a precursor, activitycould be detected in galacturonic acid released from both thesoluble and insoluble polysaccharide fractions. Methionine wasa more efficient precursor of methyl ester groups than S-adenosylmethionine or S-methyl methionine; incorporation from all threeprecursors was inhibited under nitrogen. Radioactively labelledmethyl ester did not decline during a 225 min ‘chase’following a 15 min ‘pulse’ of [¹⁴CH₃]methionine;the total pectin content of slices increased by 20% during this4 h incubation.     

Ultrastructure of the Haustorial Apparatus of Dimargaris cristalligena

Infection of the reference host, Cokeromyces recurvatus by Dimargariscristalligena was studied by means of light and electron microscopy.Morphologically, the infection apparatus comprised an appressorium,a septate neck region with a collar and a nucleate lobed regionseparated from an extrahaustorial membrane by a boundary layercontinuous with the wall of the parasite. An electron-opaquepenetration jacket was formed during initial stages of infectionand also in cases of apparent ‘self-parasitism’.Reactions of the host to infection are described, and cytologicalfindings are related to current knowledge of host infectionby haustorium-producing parasitic fungi. Dimargaris cristalligena, Cokeromyces recurvatus, ultrastructure, haustorium, Dimargari-taceae, mycoparasitism, host-parasite-interface     

Host-Parasite Relations in Loose Smut of Wheat: II. The Distribution of 14C-Labelled Assimilates

The effect of infection by Ustilago nuda on the distributionof ¹⁴C-assimilates in spring wheat plants at several stagesof growth was examined Only small differences were detectedbetween healthy and infected plants at the early stages, butthere were marked differences between the older plants The amountof assimilate exported by the fed leaf appeared to be dependenton the demand for assimilates in all plants, and was greatestduring a period of rapid growth of host or parasite, such asthe sporulation of the fungus in the infected ear The distributionof exported assimilates from the fed leaf of healthy plantsvaried with the position of the fed leaf, its stage of development,and the age of the plant, the pattern was very similar to thatfound in other plant diseases This pattern was altered by thepresence of the pathogen, which acted as an alternative siteof accumulation Totally different sites received ¹⁴C-labelledassimilates in infected plants, in some cases assimilates weretranslocated over unusually long distances, for example fromthe main shoot flag leaf to infected tiller ears. The differencein distribution correlated well with the differences in growthcaused by the pathogen. In addition, the ratio between the radioactivityin alcohol-insoluble and that in alcohol-soluble compounds wasaffected by the infection, in young infected plants more radioactivitywas present in the insoluble fraction and in older infectedplants in the soluble fraction     

Mixed Crystals of Threonine and Allothreonine

Study of the soluble proteins of sweet potato (Ipomoea batatas L. cv. Norin 1) roots showed that the major protein had an apparent molecular weight of 25,000, and accounted for 60 ~ 70% of the total soluble protein extracted from fresh tissue. The 25-kDa protein exists in two forms, which can be resolved into two bands by nondenaturing polyacrylamide gel electrophoresis. Immunodiffusion and crossed immunoelectrophoresis showed that these forms are immunologically identical. This protein was identified as the antigenic component A of sweet potato root.¹⁾ It was degraded to proteins of lower molecular weight (9,500 to 20,000) if the tissue was cut or infected by Ceratocystis fimbriata. As almost none of this 25-kDa protein was detected in roots stored for one year at 10 ~ 12°C, it is probably the storage protein of these roots. Another major protein was identified as β-amylase by immunodiffusion and immunoelectrophoresis. The amount of β-amylase did not change appreciably after cutting or infection, but it was present in only trace amounts in the roots stored for one year, Cutting, infection, or storage of root tissue resulted in the production of new isozymes of peroxidase, acid phosphatase, and esterase. Increases in some other proteins in cut and in diseased tissues were detected by gel electrophoresis.     

The Structure and Development of Trifolium subterraneum L. Root Nodules: II. IN PLANTS GROWN AT SUB-OPTIMAL ROOT TEMPERATURES

Cold root temperature affected infection thread proliferation,cell invasion, and release of Rhizobium and the subsequent developmentof this infection in Trifotium subterraneum. These events werealso modified by both host cultivar and bacterial strain. At7 °C bacteroid development was only substantial with strainTA1, with either sparsely or abundantly nodulating lines ofthe host. At 11 °C strain SU297 also readily formed effective,bacteroid-filled nodules with both lines. Strain 0403 formeda few bacteroids with the abundant line only at 7 °C andreadily formed bacteroids with the sparse line only at 19 °C.At 15 °C 0403 nodules were effective on abundant lines,but mostly ineffective on sparse lines. The development of Rhizobium rods into bacteroicis and theirsubsequent degeneration wa slower at low temperatures with bothstrains. Low root temperatures favoured the deposition of starchthroughout the nodule. At higher temperatures, when bacteroidswere more active in nitrogen fixation, starch was mostly confinedto a narrow band of the youngest bacteroid filled cells andto the zone of bacteroid degeneration.     

Cytochrome Oxidase and Mitochondrial Protein in Extracts of Leaves of Nicotiana glutinosa L. Infected with Tobacco Mosaic Virus

Leaves of N. glutinosa were inoculated with tobacco mosaic virus,and infection allowed to develop in situ or in discs cut fromthe leaves and floated on water or culture solution. Extracts,made from the leaves and discs when their respiration was greaterthan that of comparable uninfected tissue, contained amountsof protein N, chlorophyll, glycollic oxidase, and mitochondrialprotein N, similar to those in extracts from uninfected tissue,but slightly less cytochrome oxidase and considerably more polyphenoloxidase.No evidence was obtained for the view that the virus-inducedrespiration reflects an increase in the amount of mitochondrialmaterial.     

The Analysis of the Alcohol-insoluble Nitrogen of Plants by Quantitative Procedures based on Paper Chromatography

Part I. The amino-acid composition of egg albumin, zein, edestin,cucumber (Cucumis) seed globulin, and squash (Cucurbita) seedglobulin has been determined by a quantitative paper chromatographicmethod. The chromatographic analyses account completely forthe amino-acid nitrogen of the protein hydrolysate as determinedby the Kjeldahl method. More complete analyses than hithertoof cucumber and squash seed globulin have been obtained. Theseanalyses can now be regarded as complete since histidine andtryptophane, which have not been determined chromatographically,have previously been reported. The two cucurbit seed globulinsanalysed closely resemble edestin, and evident differences betweenthese three on the one hand and zein on the other are referredto in the text. It is evident that the paper chromatographicmethod is applicable to the quantitative amino-acid analysisof the protein-N fraction of the plants as well as to the analysisof pure proteins. Part II. The analysis of the alcohol-soluble and insoluble fractions(hydro-lysate) of potato tubers of the variety ‘Sebago’has been carried out. Attention is drawn to differences whichexist between the samples and varieties which have been investigated,namely ‘King Edward’, ‘Sebago’, and‘Katahdin’. These differences relate particularlyto the relative proportion of soluble to insoluble nitrogen,to the amount of amides present in the soluble fraction, andto the ratio of asparagine to glutamine. There is no correlation between the relative proportions ofamino-acids as they exist free in the tuber and as they occurin the alcohol-insoluble (protein) fraction. Certain compounds(e.g. -aminobutyric acid) are present free in the tissue, butthey do not occur in the protein. Also the amides far exceedtheir possible occurrence combined in the protein. Proline especiallyis much more abundant in the combined than in the free state,and the ratios between the various free amino-acids are quitedifferent from those which apply to the protein. The latterdata suggest that the soluble compounds which occur free arenot directly combined as such to form protein. The utility of the quantitative procedure based on partitionchromatography on paper in the analysis of the alcohol-insoluble(protein) fraction of plants is, therefore, established.     

Interaction between rhizobia and potato tissues10

Several cultivars of Solanum tuberosum L., the potato, weregrown on tissue culture media and their roots inoculated withstrains of rhizobia known to infect legumes at root junctionsor between epidermal cells. Infection incidence and severityshowed considerable cultivar/bacterial strain interaction. Bacteriaspread through intercellular spaces and invaded cells in a non-structuredway: some infections penetrated to the root xylem. There wasno evidence that potato root cells produced nod-inducing factorsand nitrogenase activity was not detected. In some host/rhizobialcombinations outgrowths were formed on roots. These varied fromloose infected callus tissue at the junctions of lateral roots,to modified lateral roots of limited growth which showed somecolonization by rhizobia. Key words: Solarium tuberosum L, potato, glycoprotein, immunogold labelling, rhizobia, flavonoids     

Effects of crop plants on abundance of Pochonia chlamydosporia and other fungal parasites of root‐knot and potato cyst nematodes

The effects of a host plant on reproduction/abundance of fungal populations in relation to soil nutrients released by plants in the rhizosphere were studied. Abundance in the soil and potato rhizosphere of the fungi Paecilomyces lilacinus, Monographella cucumerina (CABI 380408) and Pochonia chlamydosporia var. chlamydosporia (Pc280, potato cyst nematode biotype) and P. chlamydosporia var. catenulata (Pc392, root‐knot nematode biotype) were assessed. The different ability of break crops (oilseed rape, sugarbeet and wheat) in the potato rotation to support Pa. lilacinus, Pochonia isolates Pc280 and Pc392 and abundance of the latter two isolates in soil and rhizosphere of potato plants infected with Meloidogyne incognita were also studied. Potato chits and crop seedlings were planted into boiling tubes containing 5000 chlamydospores or conidia g^?1 in acid washed sand (pH 6) and kept in a growth chamber at 20°C, and 16 h of light for up to 9 weeks. The abundance of the fungi in sand (fallow) differed significantly between fungal species, being in general less abundant in the absence than in the presence of the plant, although there was no interaction between plant species and fungal isolate. There was evidence of a different response to Me. incognita for Pc392 than for Pc280 but there was no significant effect of the presence of the nematode on the rate of increase of the fungus.     

Hexose Accumulation and Turgor-Sensitive Starch Synthesis in Discs Derived from Source versus Sink Potato Tubers

The net total uptake (sum of soluble and insoluble components)of the hexoses, D-glucose and D-fructose, into sink potato (Solanumtuberosum L.) storage parenchyma was biphasic with respect tosubstrate concentration. Analysis of radioactive products revealedthat the biphasic kinetics were composed of a linear, solublecomponent superimposed on saturating starch synthesis. In contrast,in source tuber tissue, there was negligible conversion of D-glucoseto starch and the shape of the kinetic was the result of a biphasicsoluble component. The uptake of D-fructose into source tissuewas linear with respect to substrate concentration. Uptake ofthe non-metabolizable glucose analogue, 3-oxymethyl-D-glucose(3-OMG), into both sink and source tissue, demonstrated biphasickinetics, indicating the presence of a carrier for glucose.The data demonstrate that in sink potato tubers, metabolismgreatly influences apparent uptake kinetics, the kinetics ofstarch synthesis masking the kinetics of hexose transport atthe plasmalemma. Uptake of L-glucose was linear with respectto substrate concentration, an observation consistent with thissugar not being recognized by a carrier. As in the case of sucrose, in sink tuber tissue the conversionof D-glucose and D-fructose to starch was sensitive to turgor,showing a marked optimum in external osmotica containing 300mol m^–3 mannitol. The mechanisms controlling turgor-sensitivestarch synthesis in the potato tuber would, therefore, appearto be common to all three sugars. Key words: Hexose (transport), partitioning, Solanum (source, sink tubers), starch synthesis     

The Nature of the Resistance of Oats to the Take-all Fungus: II. INHIBITION OF GROWTH AND RESPIRATION OF OPHIOBOLUS GRAMINIS SACC. AND OTHER FUNGI BY A CONSTITUENT OF OAT SAP

Growth of Ophiobohu gramimt and of O. gramims var. avenae isinhibited by concentrations of 3·3-4·0µg./ml.,and respiration by concentrations of 55µg./nil. of a partiallypurified substance from oat-leaf sap. The two varieties appearto be equally sensitive. The filtrate of boiled sap is inhibitorybut here dilution of the sap permits better growth of isolatesof var. avenae. Sap from oat roots is inhibitory to O.graminisonly, and fractionation of the sap shows that the inhibitorcan be masked by a growth stimulant. Inhibition of growth andrespiration can be reduced by glutathione and ascorbic acid,particularly if the inhibitor and reducing agent are previouslyincubated together for a few hours, suggesting that the inhibitoris inactivated on reduction. The capacity of var. avenae toovercome inhibition in the favourable medium provided by thecrude sap more readily than can the type variety is suggestedas the cause of the slight differential activity of the filtrateof leaf sap and the full differential activity of the root sap.Susceptibility of oats to var. avenae would thus be due to conditionsenabling the fungus to overcome toxicity rather than to an absenceof toxicity. Activity of the inhibitor against growth and respiration ofa number of fungi and a few other organisms has been tested.Bacteria and oat and barley roots are not affected but abouthalf of the fungi tested are inhibited although none is as sensitiveas O. gramims. No members of the fungi imperfecti tested aresensitive.     

Day and Night Temperature Control of Floral Induction in Stylosanthes guianensis var. guianensis cv. Schofield

Floral initiation in seedlings of Stylosanthes guianensis var.guianensis cv. Schofield grown at a photoperiod marginal forflowering (12–11.75 h) was promoted by a combination oflow day (25 °C) and low night (16 or 21 °C) temperatures,and completely inhibited by a 35 °C day temperature. Additionally,earliness of floral initiation under naturally decreasing daylengthwas negatively related to temperature regime over the range35/30 to 20/15 °C (day/night). Stylosanthes guianensis var, guianensis, flowering, temperature, photoperiod, short-day plant     

Observations on the Biology of Fomes annosus, with Particular Reference to East Anglian Pine Plantations: II. Spore Production, Stump Infection, and Saprophytic Activity in Stumps

F. annosus produces conidia abundantly in culture but rarelyunder natural conditions. Sporophorea are formed on stumps anddying trees throughout the year provided that the humidity issufficiently high. Spore discharge in the plantations may occurat any time of year, but slowa down or ceases during very dryweather. It takes place at low temperatures, ceasing only whensporophores freeze. In the laboratory, F. annosus basidiospores germinate in relativehumidity exceeding 92 per cent. on freshly cut, unsterilizedpine wood. Viability is lost rapidly in light at temperaturesabove 15°C. Under humid conditions F. annosus spores will infect freshlycut pine stumps. There is evidence that spores of F. annosus washed down intosoil remain viable for at least a short period and that stumpscovered with soil are infected from this source. F. annosusspores can infect stumps for only a few weeks after felling,colonization after a longer interval probably beingprevdntedby competing fungi. In stumps inoculated with a mixed suspensionof F. annosus and Peniophora gigantea spores, Fomes at firstcolonized the wood but was soon replaced by P. gigantea. Otheraspects of competition are discussed. The incidence of natural infection in stumps and the factorsaffecting it are discussed Growth rates of F. annosus on malt agar and in lengths of pineroot are given. The fungus grows in stump roots at about 1 m.per year, and so closely approaches adjacent trees within ayear of thinning the plantation: In large stumps, F. annosusmay survive 15 or even 30 years after felling. Data are givenwhich suggest that the infective capacity of stumps containingF. annosus is greater in alkaline than in acid mils, while replacementby other fungi is slower. Competition of F. annosus with other fungi during colonizationof stumps is discussed. The parasite can grow along roots alreadyoccupied by certain fungi but not along roots containing others.The probable course of succession in stumps rotted by F. annousis described.     

氮素水平对花生氮素代谢及相关酶活性的影响

 在大田高产条件下研究了氮素水平对花生(Arachis hypogaea)可溶性蛋白质、游离氨基酸含量及氮代谢相关酶活性的影响, 结果表明, 适当提高氮素水平既能增加花生各器官中可溶性蛋白质和游离氨基酸的含量, 又能提高硝酸还原酶、谷氨酰胺合成酶和谷氨酸脱氢酶等氮素同化酶的活性, 使其达到同步增加; 氮素水平过高虽能提高硝酸还原酶和籽仁蛋白质含量, 但谷氨酰胺合成酶(GS)和谷氨酸脱氢酶(GDH)的活性下降; N素施肥水平不改变花生植株各器官中可溶性蛋白质、游离氨基酸含量以及硝酸还原酶(NR)、谷氨酰胺合成酶、谷氨酸脱氢酶活性的变化趋势, 但适量施N (A2和A3处理)使花生各营养器官中GS、GDH活性提高; 氮素水平对花生各叶片和籽仁中GS、GDH活性的高低影响较大, 但对茎和根中GDH活性大小的影响较小。