加勒比松种源苗期DNA甲基化多样性分析

为了解加勒比松(Pinus caribaea)种源的遗传多样性,利用甲基化敏感扩增多态性技术对加勒比松3个变种17个种源的DNA甲基化多样性进行了研究。结果表明,56对引物组合共扩增出425条谱带,其中多态性谱带422条,多态性百分率为99.25%。加勒比松种源幼苗半甲基化比率比全甲基化比率稍高,洪都拉斯加勒比松、古巴加勒比松和巴哈马加勒比松的DNA甲基化率分别为22.39%、22.29%和22.35%,差异不显著。加勒比松的DNA序列遗传多样性(H=0.4376)高于DNA甲基化多样性(H=0.3274),Mantel检验表明,基因组遗传变异与表观遗传变异不存在相关性(r=-0.171,P=0.16)。表观聚类与遗传聚类间存在较大差异,两种聚类分析结果均未将3个加勒比松变种分开。这表明加勒比松变种间的表观遗传变异极为丰富,能为加勒比松遗传改良提供优良种质资源。     

Production and regeneration of protoplasts from the mycorrhizal fungus Suillus granulatus

Experiments were performed with the mycorrhizal fungus Suillus granulatus to define the parameters for production and regeneration of protoplasts. Protoplasts were released at frequencies between 1 and 3×10⁷/ml from mycelium 3 to 7 days old. The best osmotic stabilizer for protoplast release was MgSO₄ (0.7 m). To optimize protoplast release and regeneration an enzyme (Novozym 234) concentration 1.7 mg/ml was chosen, with a digestion time of 1 to 2 h. Regenerated colonies formed mycorrhizae within 60 days after inoculation in Pinus caribaea var. hondurensis seedlings.     

Recovery of plants from cryopreserved embryogenic cell suspensions of Pinus caribaea

Summary Embryogenic cell suspension cultures of Pinus caribaea var. hondurensis have been cryopreserved in liquid nitrogen for up to four months, using sucrose and dimethylsulfoxide as cryoprotectants. Post-thaw growth was obtained after a short lag phase. Removal of the remaining liquid around the cells using a filter disc favoured subsequent regrowth of the cells. These reestablished cultures maintained an embryogenic potential similar to non-frozen cultures. The embryos produced were able to regenerate into plants, which are now growing in a greenhouse.Abbreviations BA 6-benzyladenine - DMSO dimethylsulfoxide - FDA fluorescein diacetate - MS Murashige and Skoog - 2,4-D 2,4-dichlorophenoxyacetic acid - PAR photosynthetically active radiation     

Phylogeography of Pinus subsection Australes in the Caribbean Basin

Background and Aims

Four species of Pinus subsection Australes occur in the Caribbean Basin: P. caribaea, P. cubensis, P. maestrensis and P. occidentalis. This study analyses the phylogeography of these species to assess possible colonization events from Central America to the islands and subsequent population expansions during glacial periods driven by both drier climate and larger emerged land areas.

Methods

Allele size data were obtained for plastid microsatellites for 314 individuals from 24 populations, covering the distribution range of subsection Australes in the Caribbean Basin.

Key Results

In total, 113 plastid haplotypes were identified. The highest genetic diversity was found in populations of P. caribaea. Overall, Caribbean Basin populations fit the isolation by distance model. Significant phylogeographical structure was found (R_ST = 0·671 > permuted R_ST = 0·101; P < 0·0001). The haplotype network and a Bayesian analysis of population structure (BAPS) indicated different Central American origins for P. caribaea var. bahamensis and P. caribaea var. caribaea plastids, with Central America populations in northern and south-eastern groups. Sudden expansion times for BAPS clusters were close to three glacial maxima.

Conclusions

Central America contains ancestral plastid haplotypes. Population expansion has played a major role in the distribution of genetic diversity in P. caribaea var. hondurensis. Two colonization events gave rise to the P. caribaea var. bahamensis and P. caribaea var. caribaea lineages. Plastid variation in the eastern species (P. cubensis, P. maestrensis and P. occidentalis) evolved independently from that in P. caribaea var. caribaea. Incomplete lineage sorting between P. cubensis and P. maestrensis is apparent. Inferred expansion times for P. caribaea var. bahamensis and for the eastern lineages correspond to glacial maxima, whereas those for P. caribaea var. hondurensis correspond to the beginning of the temperature decrease that led to Marine Isotope Stage 8.     

Effect of restricted watering and its combination with root pruning on root growth capacity,water status and food reserves ofPinus caribaea var.hondurensis seedlings

Summary Six-month oldPinus caribaea var.hondurensis seedling were subjected to various regimes of restricted watering and their combinations with root pruning to determine a suitable technique of producing seedlings with optimum physiological condition for successful bare-root planting.From the range of treatments tested, plants subjected to watering once every 9 days (D9) were found to be the hardest in terms of highest root growth capacity, leaf water content, root starch content and in having a moderately low transpiration rate.P. caribaea seedlings subjected to this preconditioning treatment may be more able to survive bare-root planting.     

Influence of aluminum on in vitro formation of Pinus caribaea mycorrhizae

The ability of Pinus caribaea var. hondurensis to form mycorrhizae was determined in vitro with seven species of ectomycorrhizal fungi in the presence of six levels of Al (added as AlCl₃) in a nutrient solution. The time required for mycorrhizal formation, the number of mycorrhizal root tips and the percent mycorrhizae were measured after 15, 30, 60, 90 and 120 days. Cenococcum graniforme was susceptible to Al toxicity at all Al concentrations. Pisolithus tinctorius and Suillus sp. were depressed at lower but stimulated at higher Al concentrations. The inverse was shown for Rhizopogon reaii and Hebeloma cylindrosporum. Tolerance to Al was verified for R. nigrescens and H. crustuliniforme. Pisolithus tinctorius had the largest mycorrhizal capacity, defined as the sum of the values for time, percent and number of mycorrhizae. Ectomycorrhizal fungi appeared to ameliorate Al damage to plant roots even in treatments where no mycorrhizae formed. Inoculation of pine seedlings with Al-tolerant mycorrhizal fungi is likely to improve reforestation efforts in highly-weathered tropical soils.     

The response ofPinus caribaea mor. var. Hondurensis seedlings to nitrogen,phosphorous and potassium fertilizers

Summary A soil pot culture experiment was conducted to study the response ofPinus caribaea var. hondurensis seedlings to the N, P, K fertilizers in a 3³ factorial combination of nutrients in four replicates. For this purpose, seedlings were grown in plastic pots in soils collected from Bahau pine experimental area. These soils belong to Durian series and are generally poor in nutrients.The seedlings were supplied with 112 kg/ha, 336 kg/ha and 560 kg/ha (100 lb/ac, 300 lb/ac and 500 lb/ac) of N, P, K in solution form in all possible combinations. Periodic growth measurements were taken and dry matter production was estimated at the conclusion of the experiment.The results indicate that phosphorous is the most important nutrient for the height growth of seedlings. The absolute height increment under the best treatment combination (N₁P₃K₁) was about threefolds (25.7cm) compared to 8.8 cm in the control. It was also found that higher levels of N and K, though not beneficial to height growth, resulted in better radial growth and more dry matter production compared to control plants. However, highest levels of N and K in the present experiment produced inhibitory influence as regards height growth even in the presence of low level of phosphorous.The study indicates that application of phosphorous at least at the rate of 560 kg/ha per plant is necessary on this type of soil to boost the initial height growth ofPinus caribaea var. hondurensis. Nitrogen at the rate of 336 kg/ha and potassium at the rate of 112 kg/ha will induce better radial growth.     

Über die weite Verbreitung lichenisierter Sippen vonDictyochloropsis und die systematische Stellung vonMyrmecia reticulata (Chlorophyta)

Catinaria grossa is lichenized withDictyochloropsis splendida var.gelatinosa, var. nova. When cultured isolated from the fungus the cells of this variety are covered individually by thick gelatinous envelopes. The phycobionts ofMegalospora gompholoma subsp.gompholoma andM. atrorubicans subsp.australis belong to a new variety ofD. symbiontica, i.e. var.pauciautosporica, which preferentially reproduces by zoo- and aplanospores. The phycobiont ofPseudocyphellaria aurata does not belong toMyrmecia reticulata as formerly thought, but toDictyochloropsis symbiontica. Specimens from one locality in Tenerife and from New Zealand are lichenized with a strain ofD. symbiontica var.symbiontica, those from another locality in Tenerife with a strain ofD. symbiontica var.pauciautosporica. These strains differ in certain characters from other lichenized strains of these varieties.

     

A taxonomic study of KoreanSmilacina

Several taxonomic problems of KoreanSmilacina were examined by statistical methods, based on morphological data. Consequently,S. davurica described in Korean flora differs greatly from originalS. davurica, and closely related toS. bicolor morphologically. Therefore, the present authors treated this new taxon as a variety ofS. bicolor, S. bicolor var.flavovirens N. S. Lee et J. Y. Kim var. nov (“Yondusomdae” in Korean). The distribution in South Korea ofS. bicolor var.bicolor is known as only Mt. Chiri, but this species is collected in Mt. Dukyou, also. The bounds of morphological variations ofS. japonica var.japonica, the short plant size and no hairs on the stem, were revealed. The morphological characteristics ofS. japonica var.japonica andS. japonica var.mandshurica is obscure to distinguish. According to the present study,S. japonica var.mandshurica differs fromS. japonica var.japonica in plant size, size of adaxial leaf epidermal cell, ratio of style/ovary length and shape of stigma. The unrecorded taxon in Korean flora,S. robusta, was given a new Korean name as “Kunsomdae.” AlthoughS. japonica var.mandshurica is the variety ofS. japonica var.japonica, it is more closely related toS. robusta than toS. japonica var.japonica.     

国外松种间杂交后代生长和形态性状变异及相关性分析简

利用随机区组设计营建的试验林资料,对6种树龄5年的国外松杂种后代生长和形态性状变异及其相关进行了分析,结果显示：国外松种间杂交后代不同物种,同一物种的不同家系,同一家系内的不同单株均表现出丰富的多样性,单株材积和树干通直度的变异系数达30%~50%;而其它形态、生长性状变异系数为2%~30%。这些杂交种丰富的生长和形态变异为多目标选择奠定了基础。以湿地松（Pinus elliottii Engelman var. elliottii）为母本的杂交松针叶长度变短,同时湿地松×洪都拉斯加勒比松（P.caribaea Morelet var. hondurensis(Senecl.) Barrett et Golfari）还表现出良好的干形性状（通直度好,分枝角小等）。以火炬松（P.taeda L.）为母本的杂交松对针叶的改良效果不明显,但是火炬松×洪都拉斯加勒比松单株平均材积大、干形通直、分枝角小、侧枝细,说明这个杂交类型具有生长快、树形紧凑等优良特性。相关分析显示生长快、针叶短、分枝角度小、树干通直度好这几个性状可以同步改良。研究认为国外松种间杂交后代生长性状不低于母本,在形态性状上湿地松×洪都拉斯加勒比松与母本相比针叶较稀疏,针叶长度较短,分枝角较小,因此比母本更能抵抗雨雪冰冻等自然灾害,这对于我国南方湿地松抗逆性育种具有重要的意义。     

The taxonomy of some Swartzieae (Leguminosae, subfam. Papilionoideae) from southeastern brazil

Three new species of Swartzieae are described and illustrated:Swartzia alternifoliolata, S. capixabensis, andZollernia cowanii. Swartzia apetala var.blanchetii and var.subcordata are considered to be synonyms ofS. apetala var.apetala, andS. grazielana a synonym ofS. macrostachya var.macrostachya. Keys to southeastern Brazil members ofSwartzia andZollernia are provided.     

Effect of the nitrogen source on the biosynthesis, composition, and structure of the exopolysaccharides ofAureobasidium pullulans (de Bary) Arnaud

The effect of the nitrogen source and the C/N ratio of the growth medium on the biosynthesis, composition, and structure of the exopolysaccharides (EPSs) ofAureobasidium pullulans (de Bary) Arnaud var.aubasidani Yurlova var. nov. andA. pullulans var.pullulans was studied.A. pullulans var.pullulans andA. pullulans var.aubasidani strains synthesized the maximum amounts of EPSs in the presence of, respectively, a reduced nitrogen source ((NH₄)₂SO₄) and an oxidized nitrogen source (NaNO₃) in the medium. The data presented confirm the validity of using the chemical composition and structure of the major cetavlon-precipitated fraction ofA. pullulans EPSs for the characterization of intraspecies taxa.     

The fine structure of motile cells in the generaUlvaria andMonostroma,with special reference to the taxonomic position ofMonostroma oxyspermum (Ulvophyceae,Chlorophyta)

The zoospores and isogametes ofUlvaria obscura var.blyttii, the isogametes ofMonostroma bullosum, and the anisogametes ofM. grevillei have a flagellar apparatus with counterclockwise absolute orientation and terminal caps, and therefore belong to theUlvophyceae. On the basis of the absence or presence of body scales and the morphologies of certain flagellar apparatus components,Ulvaria obscura var.blyttii is retained in theUlvales, whileM. bullosum, M. grevillei andM. oxyspermum are referred to theUlotrichales. Differences in scale morphology, certain flagellar apparatus components, and early thallus ontogeny support the transfer ofM. oxyspermum to the genusGayralia. Mating structures and their positional relationships within the cell are described from the gametes examined. A plasmalemma-associated plaque that may be a degenerate mating structure occurs in someG. oxysperma motile cells.     

Another look at Hymenoxys subgenus Plummera (Asteraceae: Heliantheae: Gaillardiinae) from Arizona and New Mexico

The recent merger ofPlummera intoHymenoxys is followed here. The two previously recognized species ofPlummera are here treated as a single polymorphic species ofHymenoxys, H. ambigens, consisting of three varieties: var.ambigens from the Mescal, Santa Teresa, and Pinaleno Mountains of southeastern Arizona; var.floribunda from the Chiricahua, Dos Cabezas, Dragoon, Little Dragoon, and Mule Mountains of southeastern Arizona; and var.neomexicana described here from the Animas and Peloncillo Mountains of southwestern New Mexico.     

An attachment tip and pili-like structures in insect- and plant-pathogenic spiroplasmas of the class Mollicutes

Ultrastructural studies using scanning electron microscopy (SEM), negative-staining transmission electron microscopy (TEM), and thin-sectioning TEM on four species of Spiroplasma, in vitro and/or in vivo, indicated that their helices commonly possess one tapered end (tip structure) and one blunt or round end. These tip structures appeared morphologically different from the rest of the helix, exhibiting an electron-dense conical or rod-shaped core. In thin sections of the midgut of the leafhopper Dalbulus elimatus, the tip structures of Spiroplasma kunkelii in the midgut lumen were mostly aligned between microvilli, perpendicular to the apical plasma membrane of epithelial cells. These tip structures appeared frequently attached or closely apposed to the plasma membrane, in which cup-shaped invaginations close to the tips were observed. Pleomorphic forms of spiroplasma, enclosed in membranous vesicles, were found in the cytoplasm of the midgut epithelial cells. These findings suggest that the tip structure may be involved in the orientation and attachment of spiroplasma helices in relation to their host cells, and thus may be functionally comparable to the attachment organelle of mycoplasmas. Additionally, pili-like structures were observed by negative-staining TEM on the surface of Spiroplasma melliferum, and in thin sections of S. kunkelii infecting the leafhopper vector Dalbulus gelbus. Abbreviations CSS Corn stunt spiroplasma - SEM Scanning electron microscopy - TBS Tris-buffered saline - TEM Transmission electron microscopy     

Comparative study of leaf carbon gain in saplings ofThujopsis dolabrata var.hondai andQuercus mongolica var.grosseserrata in a cool-temperate deciduous forest

Seasonal courses of leaf CO₂ gas exchange in a growing season were examined in saplings ofThujopsis dolabrata var.hondai andQuercus mongolica var.grosseserrata in a cool temperate deciduous forest. Between the two tree species there were no large differences in the light compensation point of leaf photosynthesis, except for the season of new leaf expansion. However, light-saturated rates of net photosynthesis were obviously high inT. dolabrata var.hondai. EvergreenT. dolabrata var.hondai saplings had large photosynthetic production in two seasons, before the emergence of new foliage and after foliage fall of the overstory deciduous trees, because of the significantly high solar radiant energy penetrating under the forest canopy during the seasons. Saplings of deciduousQ. mongolica var.grosseserrata were heavily shaded throughout the growing season by foliage of the overstory trees, which resulted in a low daily surplus production. The annual surplus production of leaves in the growing season was estimated to be 2300 mmol CO₂ m⁻² inT. dolabrata var.hondai and −100 mmol CO₂ m⁻², slightly negative, inQ. mongolica var.grosseserrata. These results supported the high survivability ofT. dolabrata var.hondai saplings and the high mortality ofQ. mongolica var.grosseserrata in the deciduous forest.     

Karyology of theScorzonera (Compositae) species from the Iberian Peninsula

A karyological study of 15 taxa ofScorzonera L. from the Iberian Peninsula has been made. The chromosome numbers found inS. hispanica var.pinnatifida, S. baetica, S. reverchonii, S. angustifolia, S. laciniata var.calcitrapifolia and var.subulata (2n = 14) are new. Diploid cytotypes with 2n = 14 and 2n = 12 prevail, andS. hispanica var.crispatula is the only taxon which exhibits autopolyploidy (2n = 14, 28). x = 7 is considered to be the base chromosome number within the genus, with x = 6 being derived from it by translocation. This and detailed karyotype analyses allow to group the Iberian Peninsula species ofScorzonera into three groups.     

雷州半岛尾叶桉和湿加松人工林的蒸腾耗水规律

为正确认识大径材桉树及湿加松耗水规律,为地区人工林树种选择、栽培及抚育提供指导,应用TDP热扩散探针技术,对10年生尾叶桉和湿加松树干液流进行连续监测,并同步测定各气象因子,分析了雷州半岛地区尾叶桉和湿加松蒸腾耗水的日变化特征和季节变化规律,并与气象因子建立了相关模型。结果表明:尾叶桉和湿加松边材液流均表现出典型的昼高夜低的单峰型日变化特征,各月平均液流速率不同,且旱雨季差异显著;其中峰值尾叶桉雨季(0.127 cm/min)和旱季(0.096 cm/min)分别是湿加松雨季和旱季的1.30倍和1.57倍;日平均液流速率尾叶桉雨季(0.045 cm/min)和旱季(0.033 cm/min)分别是湿加松雨季和旱季的1.27倍和1.54倍;启动时间和迅速下降时间雨季两树种间差异不大,但旱季尾叶桉要提前湿加松约1—1.5 h启动,并晚0.5—1 h迅速下降。影响两树种边材液流速率的主要气象因子相同。尾叶桉人工林年平均单株日耗水量为12.79 L/d,是湿加松的1.33倍,林分蒸腾耗水量尾叶桉(582.16 mm)和湿加松(483.24 mm),分别占同期年降雨量的34.2%和28.4%,且两树种旱雨季蒸腾耗水量均雨季显著大于旱季。     

Neue Chrysophyceen aus dem nordmährischen Quellengebiet

New taxa ofChrysophyceae from small heleocrene and limnocrene springs are described and pictured. The delicate flagellates were observed and drawn in living stage immediately after collection. Some of them show peculiar morphology and striking structure of organelles. The following new taxa are described in this paper:Chromulina nasuta, Chromulina pavlikii, Chromulina taeniata, Chromulina oligochrysis, Chromulina pyrenoidosa, Monochrysis stigmatica, Ochromonas oligochrysis, Ochromonas taeniata, Pseudokephyrion hypermaculatum, Pseudokephyrion hyalinum var.tubiforme, Chrysococcus ellipsoideus, Epipyxis borealis var.monoplastida.