The Development and Behaviour of Mycelial Strands in Merulius lacrymans (Wulf.) Fr.: II. Hyphal Behaviour during Strand Formation

The system of hyphal branching by Merulius lacrymans was observedin mycelium which had grown from a wood food-base on to glassslides during incubation in sterile moist chambers. A hierarchyof branches and sub-branches arose from the region of clampconnexions, or nodes, of relatively wide main hyphae. Therewas evidence that the sequence of branches occurring at nodesin basipetal succession represented the time sequence of branchdevelopment at any one node. Later-formed branches at any nodewere smaller than earlier branches, but such earlier branchesusually became smaller towards the tip as growth continued.Mycelial strands were built up by growth and branching of thigmo-tropicallysensitive ‘tendril’ hyphae in association with thewide main hyphae. Tendril hyphae were characteristically narrow,thin-walled hyphae arising both as later-formed branches fromthe nodes of the main hyphae and as the narrowed tips of earlierbranches. Although this branching behaviour could be seen amongstaerial hyphae growing over agar media, hyphae growing in contactwith or within the agar behaved differently and did not formstrands.     

Growth and mycelial strand production of Rigidoporus lignosus with various nitrogen and carbon sources

Growth and differentiation of mycelial strands in Rigidoporus lignosus have been shown to depend on suitable combinations of the pH of the media and the nature of the nitrogen and carbon sources. Amino acids as sole nitrogen sources gave rise to vegetative mycelium. At pH 4.5, growth and mycelial strand differentiation required asparagine, as the fungus failed to grow in the absence of this amino acid. However, at pH 6, differentiation of strands occurred appreciably in asparagine-deficient media, suggesting a close balance between pH and amino acid requirements. Ammonium was required for strand differentiation, while nitrate, as a sole nitrogen source, maintained the fungus undifferentiated. Of the carbohydrates tested, only glucose, fructose and mannose supported strand differentiation. Starch was found to be particularly effective in promoting growth of vegetative mycelium. Strand differentiation required more specific conditions than growth of the vegetative mycelium.     

Sporulation in Rhizopus sexualis and Some Other Fungi Following a Period of Intense Respiration

Rhizopus sexualis grown at 20° C. on liquid I per cent.malt or glucose-asparagine medium showed a peak of respiratoryactivity between 40 and 55 hours-after inoculation. Rate ofrespiration then fell until it reached a steady low level whichcoincided with maximum mycelial growth. Zygospore initiationoccurred at or just after the peak of respiration. At a low temperature (9° C.) or with high concentrationsof glucose the respiration peak was less marked and no zygosporesdeveloped. Single ‘plus’ or ‘minus’strains of the heterothallic species Mucor hiemalis and Phycomycesblakesleeanus showed a pattern of respiration and mycelial growthsimilar to that of R. sexualis but no zygospores were formed.Zygospores did not develop without a preliminary period of intenserespiration, but such a peak period could occur without beingfollowed by zygospore formation. A strain of Sordaria fimicola was grown at 25° C. on a syntheticmedium with 5.0 per cent. sucrose or glucose as source of carbon.Respiration reached a peak at approximately 4 and 5 days respectively,the actual peak value being twice as large on sucrose as onglucose. Dry weight of mycelium was greater on glucose thanon sucrose. Perithecia were formed only on the sucrose medium.Visible peri-thecial initials were first seen shortly afterthe occurrence of the respiratory peak. Mature perithecia werepresent 3 days later. The possible significance of these results is discussed.     

Rhizomorph Behaviour in Armillaria mellea (Vahl) Quel: II. Logistics of Infection

The growth of rhizomorphs of Armillaria mellea from small woodyinocula through tubes of soil was characterized by a progressivedecline in weekly growth increments. Initial growth rate wasrelated to the size of the inoculum food-base; the subsequentdecline is attributed partly to depletion of nutrient reservesin the inoculum through fungal respiration and growth, and partlyto competition for nutrients between the main growing apex ofthe rhizomorph and its subordinate branch apices. The vigourof infection of potato tubers by rhizomorphs from such smallwoody inocula increased with size of the inoculum, and decreasedwith increasing distance between inoculum and tuber. When inoculumpotential of A. mellea was low, the fungus was generally overtakenin the tuber tissues by soft-rotting bacteria, which preventedits further advance.     

A Possible Role for Urease as a Storage Protein in Aspergillus tamarii

A marked decrease in mycelial urease activity during the endogenousphase of undifferentiated Aspergillus tamariicultures was foundto be independent of preparative procedures but related to thedepletion of external nutrients. The enzyme, which was synthesizedduring the active growth stage, was produced in similar quantitieswith ammonium or urea as sole nitrogen source and at its peakrepresented c. 8·5 per cent of the total soluble proteinpool of the mycelium. It was found to show maximum activityat pH 8·20–8·65 when measured in cell-free,phosphate-buffered extracts. Isolation of urease from differentstages of the endogenous phase by affinity chromatography hasshown that the observed decrease in activity was due to breakdownof the enzyme protein in mature cultures, followed by the progressivedeactivation of residual enzyme during the autolytic stage.Since selective inhibition of 80–90 per cent of activityby acetohydroxamic acid in media containing urea as the onlynitrogen source or total repression of urease synthesis by L-histidinein ammonium-grown cultures did not interfere with normal growth,it was concluded that in A. tamarii urease fulfils the functionof a storage protein with a measure of catalytic activity. Aspergillus tamarii, urease, storage protein, nitrogen metabolism     

Gibberellin-like Substances in the Sporophores of Agaricus bisporus (Lange) Imbach

Sporophores of cultivated Agaricus bisporus (Lange) Imbach,were shown to contain a gibberellin-like substance active inthe dwarf maize (d-5), -amylase and other bioassays. Ethyl-acetateextraction followed by paper, column, and thin-layer chromatographyrevealed the presence of one major active substance. Ficin hydrolysisof dried sporophore powder, after the complete removal of freesubstances, released more gibberellin-like substances, one ofwhich appeared identical to the free compound. The free substance was predominantly in the lamellae and residualpileus tissue. The major active substance released by ficinoccurred mostly in the lamellae but also in substantial equalamounts in both stipes and pilei. No activity was found in extractsof dikaryotic vegetative mycelium on malt agar. The level ofactivity in extracts from sporophores stored at – 20 °Cfell sharply after 7 d, and then remained constant over a periodof 6 weeks. The content of gibberellin-like substances in youngand old whole sporophores showed wide variation between experiments.In most cases young 2-d tissue had higher levels than old, 11-dtissue on a fresh-weight basis. Purified sporophore extractsand authentic gibberellins had no stimulating effect on growthof sporophores or of cultured vegetative mycelium. The inhibitorsof diterpene biosynthesis, CCC, and AMO-1618 induced a smallincrease in mycelial growth rate. Ethyl-acetate extraction ofhorse-straw compost prior to inoculation with Agaricus bisporusshowed the presence of gibberellin-like activity in significantamounts.     

Development of the Anther of Annona squamosa L.

In the stamen of Annona squamosa the initial hypodermal archesporiumcomprises a uniseriate row of cells, of which subsequently alternatecells become septal initials and sporogenous cells respectively.Morphologically the tapetum has a triple origin from the parietalcell derivatives, the connective cells and the septal initials;the tapetum derived from the former two is secretory while thelast forms a periplasmodium. Cytokinesis of microspore mothercell is by successive cytoplasmic constriction. The pollen grainsremain loosely attached as tetrads. Annona squamosa L., anther development     

Cap Formation during the Elongation of Lateral Roots of Vicia faba L.

Cell proliferation was examined in the cap initials of newly-emerged0·2 and 4·0 cm long lateral roots of Vicia fabafrom an investigation of the passage of labelled cells throughmitosis following a 1-h pulse with tritiated thymidine. Celldoubling time was found to increase in this group of initialcells as the secondary roots elongated, this increase beinga result of a gradual lengthening in the duration of the mitoticcycle of both the fast and slow cycling meristematic cells andof a decrease in the size of both the growth fraction and thoseproliferating cells with a short cycle time. The rate of cellproduction by the cap initials was maximal in the newly emergedsecondary roots and showed a gradual decline with subsequentlateral elongation. This change in the rate of cell proliferationin the cap initials has been shown to be related to the initiationof the quiescent centre following lateral root emergence fromthe tissues of the primary. The number of cells making up the cap of 0·2–4·0cm long secondary roots is known to be constant and thus therate at which cells are sloughed off of the cap into the soilmust be the same as the rate of cap cell formation in theseroots, all of the cap cells being replaced every 6–9 days.The rate of cap cell formation in 0·2–4·0cm long secondary roots was used to calculate that one 11-dayold Vicia plant will have contributed between 56 000 and 85000 cap cells to the rhizosphere from its root system sincegermination.     

Glutamine requirement for aerial mycelium growth in Neurospora crassa

Five amino acids are accumulated during vegetative growth of Neurospora crassa, particularly.during the prestationary growth phase. Alanine, glutamine, glutamate, arginine and ornithine.comprised over 80% of the total amino acid pool in the mycelium. Amino acid pools of different amino acid auxotrophs were followed during the partial transformation of a mycelial mat into an aerial mycelium. The mycelial mat under starvation and in direct contact with air rapidly formed aerial mycelium, which produced thereafter a burst of conidia. During this process,glutamine and alanine in the mycelial mat were consumed more rapidly than other amino acids;in the growing aerial mycelium, glutamate and glutamine were particularly accumulated. Of the amino acids that were initially accumulated in the mycelial mat, only a high glutamine pool was required for aerial mycelium growth induced by starvation. This requirement for glutamine could not be satisfied by a mixture of the amino compounds that are synthesized via glutamine amidotransferase reactions. It is proposed that glutamine serves as a nitrogen carrier from the mycelial mat to the growing aerial mycelium.     

Biochemical Changes in Cultures of Nectria galligena during the Autolytic Phase of Growth

Some biochemical changes occurring in cultures of Nectria galligenaduring its autolytic phase of growth have been investigated.In nitrate-grown and autolysed cultures of this fungus the degreeof autolysis amounted to 57 per cent. The amount of myceliallipids decreased continuously with the age of the culture. Totalmycelial nitrogen did not substantially change within the first50 days of autolysis. The constancy in the amount of bound aminoacids released from mycelium of Nectria galligena strongly suggeststhat mycelial protein are not affected by autolysis.     

Growth behaviour and glucoamylase production by Aspergillus niger N402 and a glucoamylase overproducing transformant in recycling culture without a nitrogen source

When wild-type Aspergillus niger N402 and a glucoamylase-overproducing transformant were grown in recycling culture without a nitrogen source, hyphal tip extension and glucoamylase production still occurred, but overproduction of glucoamylase by the transformant strain stopped. The mycelium retained a low metabolic activity. Light micrographs of mycelial samples showed that some hyphae were broken at their tip and partially empty, while after continuing recycling fermentation for more than 500 h many small and empty pieces of broken mycelium could be found. A model has been developed to calculate the mycelial growth and death rates. The mycelial death rate just exceeded the mycelial growth rate and as a consequence the amount of biomass in the fermentor vessel slightly decreased. It is concluded that the cytoplasmic contents of broken mycelial threads were released into the medium and acted as a nitrogen source for the growing parts of the mycelium.     

Initiation of Procambial Strands in Axillary Buds of Dactylis glomerata L., Secale cereale L., and Lolium perenne L.

In axillary buds of Dactylis glomerata L., Secale cereale L.,and Lolium perenne L., the first two procambial strands of theprophyll and the median strand of the first normal leaf areinitiated in the bud in isolation from the vascular system ofthe parent axis. They rapidly form connections with the vascularsystem of the parent axis, presumably by downward extension,as is the case of the strands of leaf primordia on the mainaxis.     

A Unique Pattern of Mycelial Elongation of Blakeslea trispora and Its Effect on Morphological Characteristics and β-Carotene Synthesis

The mycelial morphology of Blakeslea trispora was of crucial importance in the production of β-carotene in submerged cultures of B. trispora. After the spores were inoculated, the time-course variation of mycelial morphology was closely examined under the microscope. With the addition of the non-ionic surfactant (Span 20: Sorbitan monolaurate, E493) to the culture medium, a unique pattern of mycelial elongation was observed: 1) slow formation of germ tubes from spores and 2) appearance of mycelia with very short length, which allowed a well-dispersed growth of B. trispora without significant pellet aggregation. Span 20 appears to act like a paramorphogen. Without Span 20, however, the fungal culture finally formed a big clump of mycelium owing to heavy cross-linking of long mycelia. But the short mycelium maintained in the course of cultivation seemed to be irrelevant to growth inhibition, because the final concentration of dry mycelium was much higher with Span 20 after 3-day cultivation. The 20-fold increase in specific yield of β-carotene (mg β-carotene produced per g mycelium) was achieved with this drastic change in the pattern of mycelial elongation. The reason for this result might be more effective mass transfer and/or enhanced sensitivity to environmental oxidative stress in the well-dispersed mycelial cultures of B. trispora.     

In Vitro Culture and Plant Regeneration in Vicia faba subsp. Equina (var. Spring Blaze)

Explants of stem, leaves, roots, and cotyledons from etiolatedaxenically grown Vicia faba seedlings were cultured on a rangeof media. Shoot organogenesis was only obtained with nodal stemand cotyledonary node explants when cultured on MS medium with3% sucrose, 2.0 mg 1^–1 BAP and 02 mg 1^–1 NAA. Callusproliferation accompanied shoot organogenesis from nodal stemexplants. Successive subculture of nodal stem callus resultedin proliferation of regenerative callus which contained severalshoot bud initials. The capacity for shoot regeneration fromthis callus was maintained for 9 months. Histological studiesreveal de novo formation of meristematic centres in callus andtheir further development into bud primordia. High frequencyrooting of these adventitious shoots was obtained on half-strengthMS medium with 1.5% sucrose, 0.1 mg 1^–1 NAA and 0.5 mg1^–1 kinetin. Key words: Vicia faba, adventitious shoots, axillary shoots, de novomeristem formation, organogenesis, tissue culture     

Localized, Cold-Induced Inhibition of Translocation in Mycelia and Strands of Serpula lacrimans

Thompson, W., Brownlee, C, Jennings, D. H. and Mortimer, A.M. 1987. Localized, cold-induce inhibition of translocationin mycelia and strands of Serpula lacrimans. —J. exp.Bot. 38: 889–899 The effect has been investigated of localized low temperatureon translocation of ³²P across myceliui of Serpula lacrimansusing two gas-flow detectors capable of recording radioactivitycontinuously. When the temperature of a band of mycelium wasreduced to 0 ? C, radioactivity ceased to accumulai and in factdeclined under the detector (number 2) separated from the sourceof radioactivity by tr cold-treated mycelium. In the myceliumbeneath the other detector (number 1), closest to the sourceradioactivity, the rate of accumulation of radioactivity increased.When the temperature was raised t 20 ?C, radioactivity beganto accumulate in the mycelium under detector 2 and, apart froma sma fluctuation, continued to accumulate at a uniform rate.In the mycelium under detector 1, the accumulation of radioactivitystopped for a short time but then recommenced at a rate similarto thi found at 0 ?C. In other experiments the distributionof radioactivity (¹⁴C) throughout the myceliui was measuredat the end in homogenized samples. In these experiments a bandof mycelium we subjected to 0 ?C or to 20 ?C for the whole experimentalperiod, or only after the mycelium had bee translocating radioactivityalready for 16 h. These experiments showed that the changesin the rate of accumulation of ³²P in living mycelium underthe two gas flow detectors used for in situ measurements werenot due to a reversal of the flow of translocation. The resultsare consistent with an hypothesis that a turgor-driven massflow of solution is the mechanism for translocation in thisfungus and are considered in relation to the results of similarexperiments on phloem translocation in higher plants. Key words: Serpula lacrimans, mycelium, translocation, low-temperature, phloem transport     

Winter Growth of Autumn-sown White Lupin (Lupinus albus L.) Main Apex Growth Model

The winter growth of winter white lupin (cv. Lunoble) was investigated.Over three consecutive years, 1987–1989, it was sown atdifferent times at Lusignan (France) and in 1989, at nine differentlocations with various sowing times. The production of primordia,the vernalization requirements and the final number of leaveson the main stem were related to field measurements of dailymaximum and minimum temperatures. A statistical model for the main apex growth with a system oftwo equations was developed, with a threshold level for leafprimordia production at 3°C. The number of leaf primordiaproduced by a vegetative apex (y) in terms of the cumulativesums of temperature over 3°C (x) followed the curvilinearregression y = 4.76+ 0.0268x + 0000015 6x². The upper and lowertemperature limits for vernalization were estimated as 14 and1°C respectively. The vernalization requirements of a vegetative apex (y) decreasedwhen the number of initials produced (x) increased accordingto the negative exponential regression y = exp (7.2— 0.02626.x). The two equations were used for the prediction of the finalnumber of leaves of a lupin crop. The predictive accuracy ofthe model was checked against independent data. The agreementbetween observed and predicted final leaf number was often close,but some deviations did occur with low leaf number. The modeldescribed most of the growth phenomena which occur during thephase sowing to floral initiation of the main stem of a winterlupin crop, and its possible uses are discussed Lupinus albus L, white lupin, growth, model, vernalization, primordia, apex, thermal time     

Leaf Development in Lolium temulentum L.: Progressive Changes in Soluble Polypeptide Complement and Isoenzymes

Ougham, Helen J., Jones, Thomas W. A. and Evans, Mair LL. 1987.Leaf development in Lolium temulentum L.: progressive changesin soluble polypeptide complement and isoenzymes.—J. exp.Bot. 38: 1689–1696. The spectrum of soluble polypeptides extracted from segmentsof the developing 4th leaf of Lolium temulentum simplified withincreasing distance from the leaf base. Most of the metabolicallyimportant isoenzymes analysed also exhibited gradients of activitywith respect to distance from the base, and in some cases twoor more contrasting gradients were observed for a given enzyme. Key words: Gradients, isoenzymes, leaves, Lolium temulentum,, soluble polypeptides     

Origin and Development of Axillary Buds in Jute (Corchorus capsularis)

The bud initials are laid in the usual manner in the primordialmeristem in the case of branching plants of Corchorus capsularis.The non-branching character of some varieties is due to thedifferent structural organization of the shoot apex. In thenon-branching plants the absence of bud initials is associatedwith early vacuolation of the meristematic cells. Cases occurof sporadic development of a few axillary and extra-axillarybuds.     

The Wood Parenchyma of Triplochiton scleroxylon K. Schum

Unusual features in the histology of the secondary xylem wererevealed during an investigation into growth-ring developmentin Triplochiton in Nigeria. The main observation was the presenceof two kinds of apotracheal parenchyma: (1) strands made uplongitudinally of 2 to 4 (occasionally more) cells containingstarch and, (2) empty fusiform strands, without cross-walls.Each kind of parenchyma formed ‘units’, i.e. uniseriatetangential rows of like strands between the wood-rays. The starchystrands (storage parenchyma) became more numerous towards theend of each growth ring.     

Growth morphology of Streptomyces akiyoshiensis in submerged culture: influence of pH, inoculum, and nutrients.

Most media in which the growth of shaken submerged cultures of Streptomyces akiyoshiensis was examined did not support the formation of well-dispersed mycelial suspensions. Investigation of the culture conditions promoting dispersed growth showed the pH of the culture medium to be of critical importance; an initial value of 5.5 minimized aggregation of the mycelium while supporting adequate biomass production. In cultures started at this pH, spore inocula gave better mycelial dispersal than did vegetative inocula; with spore inocula, growth morphology was also less affected by inoculum size. The composition of the nutrient solution influenced the extent of mycelial dispersal; slow growth was often associated with clumping but no clear correlation was observed between pellet formation and the ability of carbon or nitrogen sources to support rapid growth. Increasing the phosphate concentration from 0.5 to 15 mM caused a modest decrease in mycelial aggregation. Conditions promoting a well-dispersed mycelium suitable for studying the physiological control of secondary metabolism also supported the formation of 5-hydroxy-4-oxonorvaline by S. akiyoshiensis.