Morphological and molecular characterization of Leptomelanconium allescheri associated with necrotic lesions on Pinus mugo needles in the Polish Tatra Mountains

Leptomelanconium allescheri was identified for the first time in Poland in 2016 in the Tatra Mts. on Pinus mugo needles. It produced numerous conidiomata on necrotic lesions on live needles. In our study, we provide detailed characteristics of disease symptoms and describe morphology of the structures of the fungus. The shape and size of macroconidia are proven to be more variable in vitro than in situ. The malt extract agar colonies of the fungus obtained from germinating macroconidia are described for the first time. Our observations prove also that, apart from macroconidia, L. allescheri produces in vitro unicellular, hyaline microconidia, 3.5–10 × 1.8–5.0 μm in size, the previously unknown aspect in the development cycle of this species. The BLAST search with ITS and LSU sequences of L. allescheri resulted in the closest match with Cenangium acuum accessions (94%–95% and 99%, respectively). Two gene based, LSU plus ITS, phylogenetic positioning of L. allescheri proves that it belongs to Pezizomycotina, Helotiales. Further, on family level, placement of L. allescheri, as well as its two closest species C. acuum and Piceomphale bulgarioides, remains undefined. The L. allescheri barcode sequences (accession numbers: ITS— MF573935 , LSU— MF573936 , IGS— MF573937 ) and cultures were submitted to NCBI GenBank and CBS Utrecht, The Netherlands culture collection.     

华山松叶栖内生真菌的分离鉴定及筛选

用常规内生真菌分离方法从健康华山松Pinus armandii Franch.针叶上分离到17种内生真菌,其中子囊菌1种、酵母菌1种、半知菌15种。通过对峙培养初步筛选出抗真菌活性的内生真菌优势种:绿木霉Trichoderma viridePers.:Fr.、拟康宁木霉T.pseudok ningiiRifai,并对这2种木霉培养滤液的抑菌作用进行初步研究。结果表明,这2种木霉培养滤液对来自于针叶的拟盘多毛孢PestalotiopsisStey.和针叶树散斑壳Lophodermium conigenum(Brunaud)Hilitz菌落生长只有较弱抑制作用,对来自于枝干的拟茎点霉Phomopsis(Sacc.)Bubak有很强的抑制作用,对拟盘多毛孢孢子萌发有不同程度抑制作用。     

灰背杜鹃菌根真菌的分离与鉴定及DGGE的应用

应用传统的培养方法分离菌根真菌,应用ITS rDNA分子生物学手段,进行了菌株的分类鉴定,并结合DGGE(变性梯度凝胶电泳)技术对菌根真菌进行检测。传统培养得到了2种与杜鹃花菌根真菌亲缘关系十分密切的真菌,分别为Phialocephala fortinii和Epacris pulchella root associated fungus。DGGE(变性梯度凝胶电泳)技术检测到1种菌根真菌为Rhizoscyphus ericae。     

辽东地区赤松落针病病原真菌的分类鉴定

对辽东主要地区发生落针病的赤松针叶上采集病害标本,室内进行分类鉴定,确定了在赤松针叶上有5种真菌,均是活体寄生。按照Minter（1981）分类标准进行了形态学种的分类鉴定,最后确定了5种真菌的分类地位,5个种分别为针叶树散斑壳菌、松散斑壳菌、光亮散斑壳菌、雪松散斑壳、小环绵盘菌。其中有3个种为我国赤松寄主上新记录种。     

北京地区柳树上光肩星天牛刻槽中真菌的分离和鉴定

文章首次利用加入链霉素的PDA培养基，从光肩星天牛在旱柳上不同时期的30个刻槽中分离到89株14个属的真菌，经鉴定，70株隶属于半和菌类的5目5科的11个属；19株隶属于子囊菌纲的3目3科的3个属，晚期刻槽内的真菌种类最丰富，共13种；早期最少，只有6种；中期居中，共10种。     

樟子松疱锈病重寄生菌的分离鉴定

樟子松疱锈病是樟子松枝干上的主要病害,为利用重寄生菌对樟子松疱锈病进行生物防治,对其病原菌松芍柱锈菌Cronartium flaccidum(Alb.et Schw.)Wint.的重寄生菌进行了分离和鉴定,得到重寄生菌拟枝孢镰刀菌Fusarium sporotrichioides Sherb.、深绿木霉Trichoderma atroviride,并测定ITS序列,获得基因登录号。经显微观察发现接种重寄生菌后锈孢子受到不同程度的破坏,且存在两种不同的作用机理。     

Temporal variation in nutrient concentrations of Pinus sylvestris needles

Temporal variation in nutrient concentrations of Scots pine (Pinus sylvestris L.) needles was studied during a three‐year period in three stands of differing stages of development. Concentrations of N, P and K varied significantly between years; this variation was related to differences in needle dry weight. Concentrations of all measured nutrients (N, P, K, Ca, Mg, Mn, Cu, Zn, Fe, B) and Al varied between seasons; this variation was related to nutrient mobility and the annual physiological cycle. Concentrations of the mobile nutrients N, P and K decreased in spring and early summer during shoot and needle elongation and increased in late summer and autumn during needle senescence and litterfall. Concentrations of Mg, Cu, Zn and B followed somewhat similar patterns. The poorly mobile nutrients Ca, Mn and Fe accumulated gradually in needles during each growing season. Needle nutrient concentrations were stable during the nonactive period.     

枣树越冬期内生真菌的分离鉴定及抑菌研究

为了明确越冬期枣树组织中内生真菌的种类和分布情况,并从中筛选出对枣缩果病病原有拮抗作用的内生菌株,从越冬期健康枣树的树皮、枣股、枝条中分离获得内生真菌225株,共鉴定出12个真菌属,其优势属为茎点霉属、链格孢属、毛壳菌属和镰刀菌属。研究结果表明:越冬期健康枣树组织中内生真菌分布普遍,种类丰富;不同品种、不同部位中内生真菌的种类和数量都有差异。室内平板对峙实验结果表明:在225株内生真菌中,有25株内生真菌对枣缩果病病原茎点霉菌1-2有抑制作用,最高抑菌率为85.45%;有14株内生真菌对病原壳梭孢菌1-5有抑制效果,最高抑菌率为76.00%;内生真菌F7和F8对两种病原菌均有抑菌效果,而且均有明显的抑菌带,这说明上述两株菌均可产生抗菌代谢物质。     

油松针叶枯黄原因调查研究

调查结果表明，承德避署山庄及其周边地区的油松针叶大量枯黄，侧枝枯死或整树枯死是由于松大蚜的严重危害所致。     

Qualitative and quantitative PCR methods using species-specific primer for detection and identification of wood rot fungi

Species-specific oligonucleotide primers for detecting wood rot fungi, Gloeophyllum trabeum, Trametes versicolor, Coniophora puteana, and Serpula lacrymans, and the primer detecting a group of related fungi to G. sepiarium were developed. These primer sequences were picked up from the internal transcribed spacer region between small-subunit rDNA and large-subunit rDNA. The species selectivities of the developed primers were checked. Real-time polymerase chain reaction (PCR) was carried out using these highly specific primers to quantitatively detect at least of 0.01 ng genome DNA of the target species. This quantitative PCR was also used to differentiate the target species DNA from mixed species DNA. A PCR-based technique using the species-specific primers would be applicable to multiple-sample assay in diagnosis of wood decay and to investigation of environmental fungal populations. Part of this article was presented at the International Symposium on Wood Science and Technology (IAWPC 2005), Yokohama, November 2005     

Changes in polyamine concentration associated with aging in Pinus radiata and Prunus persica

Endogenous polyamine concentrations in apical buds of peach (Prunus persica (L.) Batsch cv. 'Hakuho') and monterey pine (Pinus radiata D. Don) seedlings, saplings and mature trees were analyzed by high performance liquid chromatography coupled with fluorescence detection to establish physiological correspondences and disparities between the species. Although there were many differences in polyamine metabolism between peach and monterey pine, several similarities were found. Specifically, in both species, the ratio between concentrations of free polyamines and low molecular-weight polyamine conjugates was greater than and less than unity in juvenile and adult plants, respectively. Free putrescine concentration decreased with age in both species.     

Response of Pinus koraiensis seedling growth to different light conditions based on the assessment of photosynthesis in current and one-year-old needles

As one of the three major five-leaved pines in the northern hemisphere, Pinus koraiensis is the most important dominant tree species in the natural mixed-broadleaved Korean pine forests. However, the regeneration of P koraiensis under the canopy of secondary forest stands is poor because of the light limitation. This study was conducted to understand how P koraiensis seedlings adapt to different light intensities and what would be the optimum light level for their establishment and growth. Three repetition plots with four light intensities （15%, 30%, 60% and 100% of the natural incident irradiances, achieved by suspending layers of black nylon net above and surrounding the plots） were set up under natural climate conditions in a montane region in eastern Liaoning Province, Northeast China. A total of 80 P koraiensis seedlings with similar height and root collar diameter were transplanted into four plots. After one year of acclimation to the specific light conditions, the seasonal variations of the photosynthetic variables and needle traits of the current and one-year-old needles, and the growth parameters were observed under four light intensities. The results indicated that： （1） The seedling at 60% treatment exhibited the greatest growth, which agreed with the response of the light-saturated photosynthetic rates （Amax） and the dark respiration rate （Rd） in the current and one-year-old needles, i.e., Rd at 60% treatment was significantly lower than that at 100% treatment, but Amax did not differ between the seedlings at 100% and 60% treatments. （2） The P. koraiensis seedlings have a certain photosynthetic plasticity to adapt the light conditions by adjusting their needle traits and regulating the physiological processes, because Amax, Rd, light saturation point and compensation point, the needle mass area, needle nitrogen and chlorophyll contents were significantly （p〈0.05） correlated with the light intensities. Especially, Am,x at 100% and 60% treatments was significantly higher （p〈0.05） than that at 30% and 15% treatments for both current and one-year-old needles. （3） The needles of different ages played a commutative role during the growing season, i.e., the one-year-old needles played a major role for the photosynthesis in the early growing season; the current year needles did in the later growing season. This ensured the effective photosynthesis throughout the growing season. These findings suggest that P. koraiensis is the in-between heliophilous and shade-tolerant tree species at least for the seedlings up to 8 years.     

赤松幼苗接种菌根真菌生长效应的初步研究

以赤松幼苗接种松茸菌进行盆栽和圃地栽培试验,研究菌根菌感染数量和菌根菌对苗木生长效应的影响。结果表明:在盆栽条件下,三种处理根系菌根真菌的感染指数分别为80、50、3,截根接种和不截根接种分别为对照的26.7倍和16.7倍,且截根处理是不截根的1.6倍;幼苗苗高、地径生长量显著提高,干重比对照分别增加33.3%和18.5%。在圃地条件下,截根接种与不截根接种的幼苗苗高、地径生长量比对照分别提高5.4%、4.2%和14.9%、6.9%。     

呼伦贝尔退化草场土壤AM真菌多样性及与土壤因子的相关性分析

指出了草原是我国面积最大的陆地生态系统,约占陆地总面积的20%,在国家生态安全中有着重要的作用。但是由于近年来过量的放牧和毁草开荒等人为因素,使草原生态环境受到严重破坏,这严重影响了我国的社会经济发展。以内蒙古呼伦贝尔退化草原为研究对象,调查研究了不同退化程度草场的牧草根段AM真菌侵染率及多样性的变化,根据土壤因子分析引起这种变化的原因。研究结果显示:土壤含水量、土壤团聚体结构数量、总碳含量、土壤脲酶活性随草地退化程度的加深而显著下降,土壤pH值和EC值随草地退化程度的增加而显著升高,而土壤有效磷含量、土壤蔗糖酶活性和碱性磷酸酶活性则呈现出轻度退化>未退化>中度退化>重度退化的趋势。牧草根段AM真菌侵染率随退化程度加深而显著下降,并与土壤含水量、脲酶活性呈显著正相关,与土壤pH值、EC值呈显著负相关。从土壤样品DNA中扩增AM真菌特异性片段并采用DGGE技术对AM真菌多样性进行分析,发现AM真菌多样性指数、丰富度、均匀度变化均呈现轻度退化>未退化>中度退化>重度退化的趋势,且都与土壤有效磷、碱性磷酸酶活性呈显著正相关,与蔗糖酶活性呈极显著正相关。研究表明:人为干扰对草原土壤根际AM真菌群落有不同程度的影响,且于土壤因子之间存在一定的相关性。这些研究可为更好地理解草原退化原因及更有效地进行草原植被开发和修复提供可靠地理论依据。     

Effects of some seed-borne fungi on Picea glauca and Pinus strobus seeds

Effects of inoculations of nine fungi to prechilled and nonchilled seeds of Picea glauca and Pinus strobus were studied in unsterilized and sterilized nursery soils, and a peat-vermiculite-perlite mixture. All the fungi demonstrated weak pathogenicity to seeds.     

松突圆蚧危害对马尾松针叶主要营养物质的影响

通过对被松突圆蚧不同程度危害的马尾松针叶内主要营养物质的测定,表明受害马尾松针叶内总糖和可溶性糖含量随着危害程度的加重先降低后增加;氨基酸含量随着危害程度的加重而下降。研究表明,马尾松针叶内主要营养物质的含量与松突圆蚧的不同危害程度有显著的相关性。     

Pine needle consumption and bark stripping by sheep grazing annual pastures in young stands of widely spaced Pinus radiata and P. pinaster

The consumption of the pine needles and the stripping of bark by sheep grazing annual pastures in three open stands of P. radiata and one of P. pinaster were measured. The P. radiata were four years old and the densities were 250, 500 and 750 trees ha^-1. The P. pinaster included two, three and four year-old trees and the density was 440 trees ha^–1.Needles were eaten immediately after the sheep were admitted and this continued throughout the year. Within ten weeks of the start of winter grazing, sheep stocked at 7–10 ha^–1 had eaten about half of the accessible needles on both species where densities were less than500 trees ha^–1. The estimated consumption was about 260 g sheep^–1 day^–1 for the P. radiata. Bark stripping then commenced and continued throughout spring and summer on most plots. In some plots at low tree density, bark was stripped from more than 35% of the trees. In denser stands, needle consumption per tree was less and bark was stripped from less than 5% of the trees.Bark damage was most common on the smaller trees, and severity of stripping was negatively correlated with tree size. Of all the trees with bark damage, 22% of the P. pinaster and 14% of the P. radiata had more than half of the stem circumference stripped. A number of these will probably suffer both stem distortion and growth retardation, and some may die.Methods of reducing the incidence of bark damage are discussed.     

Variations in dark respiration and mitochondrial numbers within needles of Pinus radiata grown in ambient or elevated CO2 partial pressure

Within-leaf variations in cell size, mitochondrial numbers and dark respiration rates were compared in the most recently expanded tip, the mid-section and base of needles of Pinus radiata D. Don trees grown for 4 years in open-top chambers at ambient (36 Pa) or elevated (65 Pa) carbon dioxide partial pressure (p(CO2)a). Mitochondrial numbers and respiratory activity varied along the length of the needle, with the highest number of mitochondria per unit cytoplasm and the highest rate of respiration per unit leaf area at the base of the needle. Regardless of the location of the cells (tip, middle or basal sections), needles collected from trees grown in elevated p(CO2)a had nearly twice the number of mitochondria per unit cytoplasm as those grown in ambient p(CO2)a. This stimulation of mitochondrial density by growth at elevated p(CO2)a was greater at the tip of the needle (2.7 times more mitochondria than in needles grown in ambient CO2) than at the base of the needle (1.7 times). The mean size of individual mitochondria was unaffected either by growth at elevated p(CO2)a or by position along the needle. Tree growth at elevated p(CO2)a had a variable effect on respiration per unit leaf area, significantly increasing respiration in the tip of the needles (+25%) and decreasing respiration at the mid-section and base of the needles (-14% and -25%, respectively). Although a simple relationship between respiration per unit leaf area and mitochondrial number per unit cytoplasm was found within each CO2 treatment, the variable effect of growth at elevated p(CO2)a on respiration along the length of the needles indicates that a more complex relationship must determine the association between structure and function in these needles.     

Diversity and succession of adventive and indigenous vascular understorey plants in Pinus radiata plantation forests in New Zealand

The vegetation of Pinus radiata plantation forests in New Zealand was studied to examine how the indigenous flora has responded to this novel habitat. A chronosequence of stands about 5, 16 and 27 years was assessed in each of four different biogeographic regions to test the effects of several stand and site factors on the succession of vascular understorey plant communities. A total of 202 indigenous and 70 adventive vascular plant species were found across all study areas, with considerable geographic variation among forests in species composition, species richness (range 48–135 species), and the percentage of indigenous species (50–86%). Both richness and cover of adventive species decreased significantly over time, whereas richness and cover of indigenous species was highest in the oldest stands, and overall species richness was lowest at mid-rotation. The guild composition changed from dominance of grasses and forbs in young stands to dominance of ferns and understorey trees in mature stands. These temporal changes were accompanied by a decrease in light-demanding pioneer species and an increase in shade tolerant, later seral species adapted to a forest environment. Measurements of the degree of canopy closure in stands with low or high stocking and modelling of temporal changes of canopy closure indicated that these understorey plant dynamics are influenced by changes in light availability as stands age. Despite the successional changes within forests, geographic variation more strongly influenced understorey communities because stands within a forest area were grouped together in DCA and TWINSPAN analyses, along rainfall and temperature gradients. Although the canopy species of such intensively managed plantation forests is an alien element in the New Zealand flora, the sheltered forest environment of older stands allows the establishment of a mostly indigenous forest understorey community with considerable similarities to indigenous forests located nearby.     

Histopathology of primary needles and mortality associated with white pine blister rust in resistant and susceptible Pinus strobus

White pine blister rust caused by Cronartium ribicola is a damaging non‐native disease of five‐needled pines in North America. Efforts to control the disease and mitigate damage to date have been only somewhat effective. Recent efforts to improve the health of eastern white pine and reestablish the tree as a dominant species in the North Central United States have focused on identification and propagation of disease‐free eastern white pine (Pinus strobus) growing in areas with a high incidence of blister rust. Many of these selections have been shown to resist infection following artificial inoculation with C. ribicola. In this study, 13 eastern white pine families derived from controlled pollination of selections previously determined to possess putative resistance as well as susceptible selections were inoculated with C. ribicola. Mortality data from inoculation studies show superior survivability in three families with over 60% of seedlings able to survive the 52 week post‐inoculation monitoring period compared to 0–10% survival of the most susceptible families. Primary needles were collected for histological analysis from all inoculated families 4 weeks after inoculation and from selected families 6.5 weeks and 38 weeks after inoculation. Histological observations of infection sites show distinct resistance reactions in the families more likely to survive infection based on mortality data. Analysis of the reactions in susceptible families revealed extensive hyphal colonization of the vascular bundle and adjacent mesophyll cells that appear uninhibited by tree responses. In resistant families, collapsed cells adjacent to infection sites, heavy deposition of phenolic compounds and abnormal cell growth were documented more frequently and appear to play an integral role in the ability of these eastern white pine families to impede growth of C. ribicola in primary needle tissue.