Abstract
Ectomycorrhizas generally improve seedling mineral nutrition and growth, so I hypothesized that decline of the Florida native pine variety Pinus elliottii var. densa Little & Dorman is related to deficiency of appropriate ectomycorrhizal (ECM) fungi in the pine's native flatwoods. At Archbold Biological Station I examined how quickly ECM fungi colonize P. elliottii var. densa seedlings and I compared the effect of local absence versus presence of adult pines on ECM colonization and pine seedling performance. Under controlled greenhouse conditions, I investigated how a wide range of ECM colonization and spread of extraradical mycelium throughout a large volume of relatively infertile, flatwoods soil enhance the mineral nutrition and growth of pine seedlings. In a field bioassay, I transplanted two-month-old pine seedlings to three flatwoods sites with low (4 pines/400 square m), medium (9 pines/400 square m), and high (19 pines/400 square m) adult pine densities. I subsequently excavated seedlings every two weeks for four-and-a-half months and determined their ECM colonization, response to shade, and response to surrounding grass density. Across all sites, pine seedlings in high shade had a higher mean chlorophyll concentration and lower stem dry weight than in full sun. Competition with grass reduced seedling survival and stem dry weight. Initial colonization was rapid and not different among sites, with 5.4 % of roots colonized 15 days after transplant. Pine seedlings had midpoint means of 29.5 %, 18.1 % and 21.3 % ECM root tips in low, medium and high adult pine density sites, respectively, suggesting that pine seedlings establishing in flatwoods encounter sufficient ECM fungi to support their growth, regardless of adult pine density. In a field experiment, I determined in the presence versus absence of adult pines if pine seedlings had higher ECM colonization and consequent improved survival, mineral nutrition, and growth. Within and beyond pine stands, I transplanted seedlings into intact or drilled, hyphae in-growth pipes buried in the ground. I placed autoclaved or fresh ECM root inoculum in two sets of intact pipes, and autoclaved inoculum in drilled pipes into which mycorrhizal hyphae could extend from the surrounding vegetation. Seven-and-a-half months after transplant, ECM hyphae had penetrated the drilled pipes and colonized pine seedlings, but roots from the surrounding vegetation also penetrated pipes. Extraneous roots reduced the survival of seedlings both within and beyond pine stands, but extraneous roots reduced seedling growth only beyond pine stands. Because percentage ECM root tips was higher in the presence (53 %) than in the absence (38.8%) of adult pines, pine stands might benefit the competitive ability of seedlings by increased ECM colonization and possibly by common mycorrhizal networks connecting seedlings to adults. Because beneficial effects of ECM in the field were small, I also examined ECM effects on pine seedlings in a greenhouse experiment. I manipulated ECM fungus colonization and the volume of flatwoods soil to which extraradical mycelium had access. In a small volume of soil (220 mL), fresh ECM root inoculum promoted the mycorrhizal colonization of seedlings versus those receiving autoclaved roots, but seedling growth and uptake of Mg, Ca, and Zn was lower with fresh than with autoclaved root inoculum. Growth and mineral nutrient uptake likely was enhanced by a pulse of nutrients from autoclaved roots, but for inoculated plants may have been reduced because of nutrient retention by saprotrophic microorganisms degrading fresh ECM roots and because of mineral nutrient retention by ECM fungi. Ectomycorrhizal seedlings with extraradical mycelium access to a large soil volume had higher mean chlorophyll concentration than those in a small soil volume. Weekly disturbance of the extraradical mycelium, however, reduced foliar contents of Mn, K, P, N, and Zn by one-third to one-half, and reduced needle dry weight of seedlings by one-third, demonstrating the importance of extraradical mycelium accessing a large volume of soil when it is nutrient-poor. My research demonstrates that ECM fungi are widespread in flatwoods and rapidly colonize pine seedlings. ECM fungus inocula are greater in the presence than in the absence of adult pines, and ECM or seedlings' connections to a common mycorrhizal network improve seedlings' belowground competitive ability. ECM especially enhance seedling mineral nutrition and growth when undisturbed, extraradical mycelium extends throughout a large volume of soil. Populations of Pinus elliottii var. densa might best regenerate in flatwoods if seedlings recruit near adult pines and where there is little competition for light, water, and mineral nutrients.