Characterization of the carbohydrase productions of an ectomycorrhizal fungus, Tricholoma matsutake

To evaluate the potential of the production of the ectomycorrhizal fungus Tricholoma matsutake to produce carbohydrases, (1) the distribution of carbohydrase activities among the different strains (18 strains) was investigated and (2) the abilities of T. matsutake and saprophytic fungi to produce β-glucosidase were compared. The results showed that the carbohydrase productions patterns of T. matsutake still resemble one another. Moreover, this fungus exhibited markedly higher β-glucosidase than did the saprophytic mushrooms. Tricholoma matsutake showed weak production of α-amylase and α-glucosidase in a static cultur filtrate. On the other hand, glucoamylase activity was not observed. Surprisingly, we discovered that β-glucosidase demonstrated strong activity. This finding suggests that this fungus has saprotrophic abilities. The carbohydrase production systems in T. matsutake were characterized from our experimental results. Also, we point out some weak points in the carbohydrase production systems of T. matsutake.     

Tricholoma matsutake Y1 strain associated with Pinus densiflora shows a gradient of in vitro ectomycorrhizal specificity with Pinaceae and oak hosts

Tricholoma matsutake produces commercially valuable yet uncultivable matsutake mushrooms during an ectomycorrhizal association with coniferous trees. In the Far East, most matsutake are harvested in managed Pinus densiflora forests. To determine whether T. matsutake has host plant specificity, we synthesized mycorrhiza in vitro between T. matsutake Y1 that originated from a P. densiflora forest and various Pinaceae and oak hosts. The strain Y1 formed a continuous Hartig net, a sign of ectomycorrhization, in the lateral roots of Pinus sylvestris, Pinus koraiensis, Pinus parviflora var. pentaphylla, Picea glehnii, Picea abies, and Tsuga diversifolia seedlings in vitro, which resembled those formed with the natural host Pinus densiflora. The strain conferred a discontinuous Hartig net with Pinus thunbergii, Picea yezoensis, Abies veitchii, and Larix kaempferi. However, no such development by this strain was observed on the roots of Quercus serrata, unlike T. bakamatsutake B1, a false matsutake that is symbiotic with oak trees. The data suggest that T. matsutake can be associated with diverse conifers but may establish ectomycorrhizal relationships only with specific host plant species.     

四种外生菌根真菌对油松幼苗的抗猝倒病和促生作用

为了筛选拮抗油松猝倒病菌及对油松苗木具有促生作用的外生菌根（ectomycorrhizal,ECM）真菌,通过对盆栽油松幼苗进行人工接种,测定了ECM真菌对油松根的侵染率、对猝倒病的防治效果以及对油松株高和地径的影响。结果表明,供试ECM真菌中,绒粘盖牛肝菌Suillus tomentosus、灰环粘盖牛肝菌Suillus laricinus、灰鹅膏菌Amaniata vaginata和血红铆钉菇Gomphidius viscidus可在盆栽条件下与油松合成菌根。灰鹅膏菌对油松猝倒病防治效果最好,为43%     

Tricholoma matsutake in a natural Pinus densiflora forest: correspondence between above- and below-ground genets, association with multiple host trees and alteration of existing ectomycorrhizal communities

Tricholoma matsutake (matsutake) is an ectomycorrhizal (ECM) fungus that produces economically important mushrooms in Japan. Here, we use microsatellite markers to identify genets of matsutake sporocarps and below-ground ECM tips, as well as associated host genotypes of Pinus densiflora. We also studied ECM fungal community structure inside, beneath and outside the matsutake fairy rings, using morphological and internal transcribed spacer (ITS) polymorphism analysis. Based on sporocarp samples, one to four genets were found within each fairy ring, and no genetic differentiation among six sites was detected. Matsutake ECM tips were only found beneath fairy rings and corresponded with the genotypes of the above-ground sporocarps. We detected nine below-ground matsutake genets, all of which colonized multiple pine trees (three to seven trees per genet). The ECM fungal community beneath fairy rings was species-poor and significantly differed from those inside and outside the fairy rings. We conclude that matsutake genets occasionally establish from basidiospores and expand on the root systems of multiple host trees. Although matsutake mycelia suppress other ECM fungi during expansion, most of them may recover after the passage of the fairy rings.     

Survival of inoculated Laccaria bicolor in competition with native ectomycorrhizal fungi and effects on the growth of outplanted Douglasfir seedlings

The survival, development and mycorrhizal efficiency of a selected strain of Laccaria bicolor along with naturally occurring ectomycorrhizal fungi in a young plantation of Douglas fir was examined. Symbionts were identified and their respective colonization abilities were determined. Eight species of symbiotic fungi, which may have originated in adjacent coniferous forests, were observed on the root systems. Mycorrhizal diversity differed between inoculated (5 taxa) and control (8 taxa) seedlings. Ectomycorrhizal fungi which occurred naturally in the nursery on control seedlings (Thelephora terrestris and Suillus sp.) did not survive after outplanting. Both inoculated and naturally occurring Laccaria species, as well as Cenococcum geophilum, survived on the old roots and colonized the newly formed roots, limiting the colonization by other naturally occurring fungi. Other fungi, such as Paxillus involutus, Scleroderma citrinum and Hebeloma sp. preferentially colonized the old roots near the seedling's collar. Russulaceae were found mainly in the middle section of the root system. Mycorrhizal colonization by Laccaria species on inoculated seedlings (54%) was significantly greater than on controls (13%) which were consequently dominated by the native fungi. Significant differences (up to 239%) were found in the growth of inoculated seedlings, especially in root and shoot weight, which developed mainly during the second year after outplanting. Seedling growth varied with the species of mycorrhizae and with the degree of root colonization. Competitiveness and effectiveness of the introduced strain on improving growth performances of seedlings are discussed.     

不同磷源对云南松幼苗生长和磷吸收量的影响

磷是控制生命过程的重要元素,植物在生长过程中需要大量的磷,低磷常导致一些植物发生适应性变化.云南松(Pinus yunnanensis Franch.)对低磷土壤环境表现出了很强的适应能力,广泛分布并正常生长于贫瘠的低磷红壤上,研究不同磷源对云南松幼苗生长和磷吸收量影响,对揭示云南松的低磷适应机理具有重要参考意义.试验所用培养云南松幼苗的种子采集自云南省通海县秀山森林公园内的健壮云南松林.试验研究了不同磷源对云南松幼苗茎高、主根长、生物量、根冠比、磷含量的影响,结果表明:(1)不同磷源处理间云南松幼苗茎高(F=2.352,P=0.067)、主根长(F=1.775,P=0.151)、茎叶生物量(F=1.359,P=0.269)、根系生物量(F=2.807,P=0.035)和总生物量(F=1.017,P=0.427)几个参数并没有表现出实质性差异,云南松幼苗生长在不同磷源处理间的差异不明显;(2)不同磷源处理下云南松幼苗根冠比的大小顺序依次为普通磷源KH2PO4>钙磷Ca3(PO4)2>铝磷AlPO4>无磷CK>铁磷FePO4 · 4H2O,幼苗体内磷含量的高低顺序依次为普通磷源KH2PO4>铁磷FePO4 · 4H2O>铝磷AlPO4>钙磷Ca3(PO4)2>无磷CK;(3)普通磷源比其它磷源更能够被云南松幼苗吸收利用;(4)云南松幼苗地下部分磷含量始终比地上部分磷含量高.     

广东松幼苗存活率和生长特征的初步研究

2006年5月至2008年7月,在南岭国家级自然保护区测定了不同年龄、不同种植方式的广东松(Pinus kwangtungensis)幼苗的存活率、株高和基径等指标,以期为国家保护植物广东松的保育提供资料。结果表明,实验期间袋栽苗存活率下降明显,从开始时的100%下降到结束时的50%;裸植苗与移植苗的存活率从100%下降到80%左右。总月均株高、基径的增量,袋栽苗、裸植苗、移植苗依次为1.01、1.39、2.76cm;0.17、0.20、0.64mm。年均株高、基径增值,移植苗、裸植苗、移植苗依次为32.91、17.22、12.77cm;7.64、2.31、2.0mm。随着年龄的增长,各组幼苗的株高和基径增长均加快,株高与基径之间存在着极显著的线性相关关系,广东松幼苗至10a生仍属快速生长期。     

土施L-蛋氨酸、L-苯基丙氨酸、L-色氨酸对玉米生长和养分吸收的影响

通过盆栽试验,研究了3种植物生长调节剂前体物质对玉米生长发育和养分吸收的影响,并确定了其适宜用量.结果表明,土施L蛋氨酸(L-MET)、L-苯基丙氨酸(L-PHE)和L-色氨酸(L-TRP)能不同程度地增加玉米株高、地上部和地下部干重,提高玉米根系活力、体内硝酸还原酶和过氧化氢酶的活性,促进玉米对氮、磷、钾、锌养分的吸收.在所有供试浓度中,以土施L-MET0.0185～0.185mg·kg^-1、L-PHE0.2mg·kg^-1和L-TRP0.03～0.3mg·kg^-1效果最佳,而且L-PHE和L-TRP对玉米生长的促进作用和提高养分的吸收能力均优于L-MET.     

Gas exchange and growth responses of ectomycorrhizal Picea mariana, Picea glauca, and Pinus banksiana seedlings to NaCl and Na2SO4

Abstract: Black spruce (Picea mariana), white spruce (Picea glauca), and jack pine (Pinus banksiana) seedlings were inoculated with Hebeloma crustuliniforme or Laccaria bicolor and subjected to NaCl and Na₂SO₄ treatments. The effects of ectomycorrhizas on salt uptake, growth, gas exchange, and needle necrosis varied depending on the tree and fungal species. In jack pine seedlings, ectomycorrhizal (ECM) fungi reduced shoot and root dry weights and in the ECM white spruce, there was a small increase in dry weights. Sodium chloride treatment reduced net photosynthesis and transpiration rates in the three studied tree species. However, NaCl-treated black spruce and jack pine colonized by H. crustuliniforme maintained relatively high photosynthetic and transpiration rates and needle necrosis of NaCl-treated black spruce seedlings was reduced by the ECM fungi. Higher concentrations of Na+ were found in shoots compared with roots of the three examined conifer species. ECM fungi reduced the concentrations of Na+ mainly in the shoots and this reduction was greater in plants treated with NaCl compared with Na₂SO₄. Shoots contained generally higher concentrations of Cl^- compared with roots. In the NaCl-treated black spruce and white spruce, both ECM species significantly reduced Cl^- concentrations. Our results point to overall greater phytotoxicity of NaCl compared with Na₂SO₄ and support our earlier findings which demonstrated beneficial effects of ECM fungi for woody plants exposed to NaCl stress.     

Effects of mutual interaction of Laccaria laccata with Trichoderma harzianum and T. virens on the morphology of microtubules and mitochondria

Summary. Organelles are known to respond to challenges caused by many stress factors. The morphology of the microtubular cytoskeleton and mitochondria during mutual interaction in coculture of Laccaria laccata with Trichoderma harzianum and T. virens were examined. Hyphae from the interaction region were sampled between 4 and 12 days of growth. Microtubules were labelled with a specific antibody and mitochondria with 3,3′-dihexyloxacarbocyanine iodide, and the organelles were examined microscopically. The morphology of microtubules and mitochondria were similar in all three fungi. Microtubules were arranged in long arrays parallel to the hyphal axis and mitochondria formed an interconnected network. In hyphae growing within the interaction zone, microtubules became wavy and eventually fragmented or depolymerised, and mitochondria also became fragmented. The effects were time-dependent. In general, the organelles of all three fungi were affected during the interaction, but L. laccata was affected the least and to the same extent by each of the saprotrophic fungi. The saprotrophic fungi were affected by L. laccata to a similar extent at 4 and 8 days of interaction. Our results suggest that the studied fungi antagonistically affect each other at the cellular level, although the mechanisms involved remain to be elucidated. Correspondence: M. Zadworny, Laboratory of Root Pathology, Institute of Dendrology, Polish Academy of Sciences, Parkowa 5, 62-035 Kórnik, Poland.     

外源调节剂对淹涝水稻幼苗株高及碳水化合物消耗的影响

为研究淹涝条件下水稻幼苗株高及碳水化合物消耗对不同外源生长调节剂的响应,本试验选用籼型常规稻IR64和导入耐淹涝基因Sub1的IR64-Sub1为试验材料,秧龄20 d时喷施1-氨基环丙烷-1-羧酸(ACC)、多效唑(PB)、赤霉素(GA)3种外源生长调节剂,以喷施清水为对照(CK).喷施处理2 d后进行0、4、8、12、16 d没顶淹涝胁迫,淹涝胁迫结束后常温恢复7 d,取样分析不同外源生长调节剂对水稻成活率、株高、叶绿素降解及恢复、地上部非结构性碳水化合物(NSC)消耗的影响.结果表明:淹涝导致水稻幼苗株高显著增长,叶片SPAD值快速下降,叶片可溶性糖迅速消耗,但耐淹涝品种IR64-Sub1淹水前茎鞘NSC含量明显高于IR64,淹涝中NSC消耗速率低于IR64,淹水结束后地上部淀粉含量高于IR64.外源PB处理显著抑制水稻幼苗株高增长、叶绿素降解及NSC消耗,提高存活率,且对IR64-Sub1效果更为显著.外源GA处理水稻幼苗叶绿素降解、株高增长和NSC消耗最快,植株恢复能力最低,耐淹涝能力最差,但与IR64相比,GA对IR64-Sub1淹涝耐性的抑制明显减弱.外源ACC促进伸长效果明显低于外源GA处理.淹水前喷施PB可有效抑制植株水下伸长,延缓叶绿素降解,减缓NSC消耗,保留更多NSC,为淹水胁迫解除后水稻快速恢复提供有利条件,这对于易涝地区减轻涝渍危害具有重要意义.     

二氧化氮对樟树幼苗生长及光合作用的影响

采用开顶式人工熏气装置,对1年生樟树幼苗进行了为期2个月不同体积分数NO₂(0.1、0.5和4.0 μl·L^-1)熏气试验,研究其对幼苗生长及光合作用的影响.结果表明:0.5和0.1 μl·L^-1 NO₂处理促进了樟树幼苗生长,而4.0 μl·L^-1 NO₂处理则抑制其生长.各处理樟树幼苗叶片净光合速率(P_n)日变化呈不对称的双峰型曲线,存在光合“午休”现象;在光合日进程中,0.5 μl·L^-1 NO₂处理使叶片P_n提高,最大值达8.542 μmol CO₂·m^-2s^-1,4.0 μl·L^-1NO₂处理的大多数时段使P_n降低,而0.1 μl·L^-1 NO₂处理对P_n的影响则依时段而不同;0.5和4.0 μl·L^-1 NO₂处理提高了叶片气孔导度(G_s)和胞间CO₂浓度(C_i)的最大值和最小值,0.1 μl·L^-1 NO₂处理提高了C_i的最大值和最小值,降低了G_s的最大值和最小值.熏气处理中、后期,0.5μl·L^-1 NO₂处理叶片的日均净光合速率显著高于其他处理.在熏气处理前期,0.5和4.0 μl·L^-1 NO₂处理使叶片最大PSⅡ的光能转换效率(F_v/F_m)显著下降;在熏气处理后期,4.0 μl·L^-1 NO₂处理的叶片F_v/F_m仍显著低于对照.     

Effects of aluminium on growth and cation uptake in seedlings of Eucalyptus mannifera and Pinus radiata

This experiment was designed to examine the effects of aluminium (Al) on the growth of Pinus radiata (D. Don) and Eucalyptus mannifera subsp. mannifera (Mudie) seedlings in culture solutions in a glasshouse to help explain the failure of radiata pine trees on some acid, low fertility soils in Australia on which the native eucalypts flourish. Aluminium (Al) in culture solution increased the growth of roots and shoots of seedlings of both species but while growth of the eucalypt continued to increase with increases in Al to 2.222 μM, growth of the pine was largest at 370 μM Al. In addition to total root length, specific root length (length per unit dry weight), a measure of fineness of the root, increased in the eucalypt seedlings as the substrate Al increased. Growth of the shoots and roots of the pine in the absence of any added Al was extremely poor suggesting that Al, in low concentrations, may be an essential element or ameliorate some other factors in solution culture at low pH. Root and shoot concentrations of K increased with increasing Al, whilst Ca and Mg Concentrations decreased and Mn concentrations were unaffected in both species. Tissue Ca and Mg concentrations were 2 to 3 times higher in the eucalypt seedlings than the pine at all levels of added Al due to greater uptake of these elements by the eucalypt. In contrast, at the highest concentration of Al in the medium, shoot Al concentrations were lower in the cucalypt than in the pine due to a greater proportion of Al being retained in the eucalypt roots. These differences between the seedlings in terms of root growth and tissue cation concentrations may help explain the ability of eucalypt species to maintain vigorous growth on acid soils high in Al and low in Ca and P, where growth of the pines failed.     

Effect of bacteria isolated from composts and macrofauna on sorghum growth and mycorrhizal colonization

Summary Beneficial plant–microbe interactions in the rhizosphere are primary determinants of plant health and soil fertility. The effect of combined inoculation of plant growth-promoting bacteria, Bacillus circulans EB 35, Serratia marcescens EB 67 and Pseudomonas sp. CDB 35 and arbascular mycorrhizal fungi, Glomus spp. on sorghum growth and mycorrhizal colonization was investigated. Plant growth observations taken at 45 days after sowing (DAS) revealed that all the three strains applied along with arbascular mycorrhizae (AM) improved plant biomass from 17 to 20% and mycorrhizal colonization from 25 to 35%. Further studies at 90 DAS also showed improvement in plant growth parameters measured. It was apparent that all the three strains stimulated plant and root growth in combination with AM and infection of sorghum roots with mycorrhizae at 45 DAS was equal to or even greater than the AM + rock phosphate (RP) inoculation at 90 DAS. This shows the possible reduction of AM culturing period to 45 days compared to its 3-month culturing in the pot cultures.     

Effects of applying stem-shortening plant growth regulators to leaves on root elongation by seedlings of wheat, oat and barley: mediation by ethylene

Several plant growth regulators (PGRs) commonly used in practicalfarming to restrict shoot height and control lodging were examined for theirimpact on root growth in naturally short or tall cultivars of barley (cvs.Kymppi and Saana), oat (cvs. Veli and Pal), and wheat (cvs. Mahti and Tjalve).The possible involvement of ethylene in the responses was also examined. Shootswere sprayed at the two-leaf stage with the gibberellin biosynthesis inhibitorsCycocel (chlormequat chloride) (CCC) or Moddus (Trinexapac-ethyl) (TE), or withthe ethylene-releasing agent Cerone (ethephon) (ETH) at 0, 0.1, 1, 10 or 50times the recommended agricultural rate (RR). Root elongation and ethyleneproduction by roots or shoots were unaffected by CCC at all application ratesorby TE at ×0.1 or ×1.0 RR. At ×10 and ×50 RR, TE wasinhibitory to root extension but did not increase ethylene biosynthesis bytheseroots or the shoots. ETH at ×0.1 or ×1.0 RR did not affect rootextension or ethylene production in roots or shoots. At all higher rates ofapplication ETH stimulated ethylene production strongly in shoots and roots ofall three species, while root elongation was retarded severely in barley,moderately in oat and only slightly in wheat. These differences in elongationresponse are attributed to differences in sensitivity to ethylene released byethephon. Accordingly, root elongation in wheat was only slightly affected whenethylene gas was supplied at concentrations up to 100 ppm for 3d. In contrast, root elongation in barley was strongly inhibitedbyethylene, with oat demonstrating an intermediate responsiveness.