Variation in mycorrhizal development and growth stimulation by 20 Pisolithus isolates inoculated on to Eucalyptus grandis W. Hill ex Maiden.

Twenty isolates of Pisolithus, covering u range of hosts, basidiocarp types and geographic locations were compared on their ability to form mycorrhiza in vitro with Eucalyptus grandis and to stimulate seedling growth in vivo. There was a large variation between isolates in the rate of mycorrhizal development and this was used to categorize isolates into six types. Mycorrhiza of seedlings inoculated with type I isolates were the most advanced with a dual-layered mantle, a fully-developed Harris net and radially elongated epidermal cells. Mycorrhiza of seedlings inoculated with types II V isolates were progressively less advanced so that mycorrhiza of type V isolates had a thin single-layered mantle, the Hartig net was only partially developed and the epidermal cells were not elongated. Root lips of seedlings inoculated with type- VI isolates were non-mycorrhizal. Inoculated seedlings were transplanted directly into undrained pots containing a yellow sand deficient in phosphorus. Growth stimulation of E. grandis seedlings in the glasshouse varied greatly between Pisolithus isolates, with dry weight increases ranging from two to 45 times that of the non-inoculated control seedlings. Extent of mycorrhizal development was positively correlated to growth stimulation in the glasshouse and it is proposed that this could be used as an indicator of isolate aggressiveness and consequently the potential of an isolate to promote tree growth.

The effectiveness of ectomycorrhizal fungi in increasing the growth of Eucalyptus globulus Labill. in relation to root colonization and hyphal development in soil.

Forty-seven different isolates of ectomycorrhizal fungi, from the different genera, were screened for their effectiveness in increasing the growth of Eucalyptus globulin La hi 11. where supply of P is deficient. Plants were grown in a P-delicient sand, in pots, in a temperature- control led glasshouse. Seedlings we re harvested 6-S and K7 d after planting, and were assessed for dry matter production and mywirrhizal colonization. Selected treatments were also assessed for P concentrations in the plant and hyphal development in the soil. Dry weights of inoculated plants ranged from 50 to 350% of the dry Weights of uninoculated plants. Growth increases in response to ectorriycorrhizal inoculation corresponded with increased P uptake by the plant.'Early'colonizing fungal species (Descolea maculata, Hebeloma westraliens, Laccaria laccata and Pisolithus tinctorius) were generally more effective in increasing plant growth than'late'colonizing species (Cortinarius spp. and Hyutenmgium spp.), although there was also variation in effectiveness among isolates of the same fungal species. Plant dry weights were positively correlated (r2 = 0·79-0·84) with the length of colonized root, indicating that fungi which colonized roots extensively were The most effective in increasing plant growth. For some fungi, however, plant growth responses to inoculation were not related to colonized root length. These responses could not.be related to the development of hyphae in soil by the mycorrhizal fungi.

Ectomycorrhiza formation in Eucalyptus.: IV. Ectomycorrhizas in the sporocarps of the hypogeous fungi Mesophellia and Castorium in Eucalypt forests of Western Australia.

Mesophellia and Castorium are common hypogeous macrofungi in the karri (Eucalyptus diversicolor F. Muell.) and jarrah (Eucalyptus marginata Donn ex Sm.) forests of south-western Australia. Sporocarps of Mesophellia and Castorium develop 5-20 cm below the soil surface in close association with eucalypt roots. During differentiation of the sporocarps, eucalypt roots become trapped within the peridium where they branch profusely and form a dense ectomycorrhizal layer. Mature sporocarps of M. trabalis nom. ined. contain approximately S m of roots of 45 cm2 surface area. Anatomical studies have shown that these roots have Hartig nets penetrating to the hypodermis and are similar to the superficial eucalypt ectomycorrhizas formed in soil and litter. The association of Mesophellia and Castorium sporocarps with tree roots suggests that these are important mycorrhizal fungi in forests of southern Australia.

Effects of high soil moisture on formation of ectomycorrhizas and growth of karri (Eucalyptus diversicolor) seedlings inoculated with Descolea maculata, Pisolithus tinctorius and Laccaria laccata.

Seedlings of Eucalyptus diversicolor P. Muell. I noculated with the ectomycorrhizal fungi Descolea maculata Bougher (two isolates), Pisolithus tinctorius (Pers.) Coker & Couch and Laccaria laccata (Scop, ex Fr.) Berk. & Br. were raised under glasshouse conditions in a yellow sand at a gradient of four soil moisture levels ranging from above field capacity to near waterlogged. All fungi enhanced growth of seedlings above that of uninoculated seedlings, but in soils near saturation there was no response to inoculation. Reduced mycorrhizal formation in relation to increasing soil moisture occurred to various degrees for all fungi. This was particularly marked with Pisolithus tinctorius. In contrast, Laccaria laccata maintained a relatively high number of mycorrhizal roots at all moisture levels applied, except at the wettest soil treatment. An isolate of D. maculata from a swamp environment did not produce a greater number of mycorrhizal roots at high soil moisture than an isolate of this species from a forest environment.

Growth and phosphorus acquisition of karri (Eucalyptus diversicolor F. Muell.) seedlings inoculated with ectomycorrhizal fungi in relation to phosphorus supply.

Growth and phosphorus acquisition of pot-grown seedlings of karri (Eucalyptus diversicolor F. Muell.) were examined following inoculation with four ectomycorrhizal fungi -Descolea maculata Bougher (two isolates), Pisolithus tinctorius (Pers.) Coker & Couch, and Laccaria laccata (Scop, ex Fr.) Berk. & Br. Seedlings were raised in steam-sterilized sand to which 13 rates of phosphorus (0.100 mg P kg-1 soil) were applied. All fungi except P. tinctorius produced a plant growth response. L. laccata produced the largest growth response. Responses were greatest at low rates of application of P to soil. There was no effect of the fungi on growth at levels of P application above 28 mg P kg-1 soil. A threshold effect (no increase in growth with increasing additions of P) characteristic of non-mycorrhizal seedlings was eliminated by mycorrhizal infection. Mycorrhizal inoculation increased P content of plant tissues at sub-optimal levels of P supply. The effect of mycorrhizas on seedling P status diminished with increasing soil P. One isolate of D. maculata often had greater rates of P accumulation and produced higher concentrations of P in plant tissues than L. laccata, but did not produce greater plant biomass. Frequency of infection for all fungi was low in soils with no additional P, and greatest with the addition of 2 mg P kg-1 soil (L. laccata and D. maculata isolate A), or 4 mg P kg-1 soil (D. maculata isolate B). Infection was reduced with increasing soil P, and not evident at 36 mg P kg-1 soil or higher levels of soil P. L. laccata had higher infection frequency and mycorrhizal root length at all levels of soil P than the D. maculata isolates. Two fungi produced basidiomes. This occurred at levels of soil P application ranging from 4 to 28 mg P kg-1 soil for D. maculata (isolate B), and at 4 to 28 mg P kg-1 soil for L. laccata.

Ectomycorrhizal formation by micropropagated clones of Eucalyptus marginata inoculated with isolates of Pisolithus tinctorius.

Eucalyptus marginata Donn ex Sm. and Pisolithus tinctorius (Pers.) Cok and Couch were co-cultured to obtain ectomycorrhizal formation in vitro. One isolate of P. tinctorius formed mycorrhizas with aseptic seedlings of a juvenile clone derived from a 4-month-old seedling, and four clones derived from crowns of mature trees. A second P. tinctorius isolate formed mycorrhizas with only the clones from mature trees. Successful combinations resulted in formation of a mantle followed by a Hartig net and epidermal cell elongation. The fungal/seedlings or fungal/seedling clone combinations which did not produce ectomycorrhizal roots, were characterized by a mantle but lacked a Hartig net, and formed an abundance of polyphenols throughout the root. Genotype, maturity and fungal specificity are key factors influencing successful ectomycorrhizal formation on E. marginata by P. tinctorius in vitro.

ECTOMYCORRHIZA FORMATION IN EUCALYPTUS: III. SUPERFICIAL ECTOMYCORRHIZAS INITIATED BY HYSTERANGIUM AND CORTINARIUS SPECIES.

Cortinarius and Hysterangium species are a dominant component of the macrofungi in eucalypt forests of Western Australia, and their hyphae occupy 10 % of the soil surface area. Anatomical studies of jarrah (Eucalyptus marginata Donn ex Sm.) and karri (E. diversicolor F. Muell) roots collected in association with hyphae of Cortinarius globuliformis Boug., C. ochraceus Clel. and Hysterangium inflatum Rodway showed mycorrhizal structures with mantles one to five cells thick and Hartig nets penetrating to the hypodermis. These superficial ectomycorrhizas were similar in size to those of non-colonized feeder roots and lacked the expanded cortex and broad mantle of pyramidal ectomycorrhizas formed by Laccaria laccata (Scop, ex Fr.) Berk. & Br. Colonized host roots responded with polyphenol accumulation in the epidermis/hypodermis and lignification of the outer cortex for both the superficial and pyramidal types.

Occurrence of calcium oxalate in karri (Eucalyptus diversicolor F. Muell.) forest ecosystems of south western Australia.

The amount of calcium in Eucalyptus diversicolor (karri) ecosystems is high in comparison with other eucalypt forest ecosystems in Australia. A large proportion of this calcium is present as crystalline Ca oxalate. Whewellite (CaC2O4·H2O) accumulates in leaf tissue of the over- and understorey species. Up to 70% of the calcium in the leaves of karri is stored as whewellite, the highest concentrations occurring in the oldest leaves. Synthesis of Ca oxalate by karri may be related in part to the high levels of exchangeable soil calcium. Oxalate of plant origin is rapidly metabolized on the forest floor during decomposition of leaf litter. About 70% of the whewellite in fresh karri leaf litter is degraded during the first wet season following leaf fall. However, additional oxalate is formed in the soil and litter layer from microbial production of oxalic acid. These crystals are of the dihydrate calcium salt, weddelite (CaC2CO4· 2H2O). Microbial production of oxalate, particularly by ectomycorrhizal fungi, may be an important mechanism for mobilizing of phosphate from the strongly phosphateadsorbing soils on which these forests grow.

A light and electron microscope study of the interaction of soil bacteria with Phytophthora cinnamomi Rands.

Light- and electron-microscopic examination showed that bacteria became associated with the hyphae and asexual reproductive structures of P. cinnamomi in soil. In suppressive soils this association appears to be correlated with hyphal lysis, inhibition of zoospore production, and sporangial breakdown. One notable feature of the microbial association between P. cinnamomi and soil bacteria is the formation of extensive slime material. Many of the bacteria isolated from the fungal hyphosphere display antagonism to the growth of P. cinnamomi in vitro. The bacteria are morphologically varied and include Pseudomonas, Bacillus, and Streptomyces spp. These observations suggest that the appropriate manipulation of the antagonistic bacteria may provide a means of biological control of P. cinnamomi.