Flora Palinologica Italiana,Sezione Aeropalinologica: S 205 -Pinus pinea L. (Pinaceae)

Summary According to the program Palynological Italian Flora, Aeropalynological section, the pollen morphological card ofPinus pinea L. is presented. The study is carried out on pollen coming from three Italian localities and regards fresh and acetolyzed pollen. For each sample, measurements are carried out on 30 fresh pollen grains in glicerol jelly with fuchsin and on 30 acetolyzed pollen grains in water/glicerol (1/1); general observations regard 1000 fresh and 1000 acetolyzed pollen grains/sample. Some observations on the main differences between fresh and acetolyzed pollen are mentioned.

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Riassunto Nell'ambito della Flora Palinologica Italiana, Sezione Aeropalinologica, è presentata la scheda morfopalinologica diPinus pinea L. nella versione su polline fresco e polline acetolizzato, su tre campioni di diversa provenienza. Vengono notate le principali differenze tra polline fresco e polline acetolizzato.

     

Variability in the 11S globulin fraction of seed storage proteins ofHelianthus (Asteraceae)

The seed storage globulins from sixHelianthus and four hybrids were studied using mono and bidimensional gel SDS electrophoresis (+ 2 mercaptoethanol). The polypeptide composition of each subunit was determined. Different pairs are specifically expressed according to the species studied. Three typical patterns were discriminated. All the studied species exhibit five subunits: two of them are expressed in all the species (₁₁ and ₂₂). The subunit corresponding to the ₁₁ pair is present inH. petiolaris and in the three populations ofH. annuus studied. The _2b2 pair is common toH. annuus andH. argophyllus. H. petiolaris presents two specific _2a2 and ₄₄ pairs andH. annuus a specific ₃₃ pair. InH. argophyllus _{11 33} or ₄₄ are never observed but are replaced by ₁₃ and ₃₁ pairs. Some globulins, poorly represented, are of forms but present chains of higher molecular weights (in the range 54–56 kDa). Expressing variations in the banding patterns between these species by the use of a similarity index reveals complete identity between the three populations ofH. annuus. Identity between the twoH. petiolaris studied is also observed.H. annuus andH. argophyllus appear to be closer to each other thanH. petiolaris concerning the seed storage globulins.     

The glycolipid specificity ofErythrina cristagalli agglutinin

The interaction of¹²⁵I-labeledErythrina cristagalli agglutinin (ECA) with neutral glycosphingolipids on thin layer chromatograms was examined by the overlay technique followed by radioautography. The lectin bound topara-globoside with a sensitivity about 10 times higher than to lactosylceramide or globoside, in agreement with the specificity of the lectin forN-acetyllactosamine. The lower limit of detection ofpara-globoside was about 0.66 nmol. The specific binding of ECA to this glycolipid was confirmed by a highly sensitive enzyme-linked lectin assay (ELLA), utilizing the horseradish peroxidase-avidin-biotin system for detection of bound lectin. Overlays of neutral glycosphingolipid extracts from human erythrocyte membranes and from human granulocytes with ECA demonstrated that the lectin can be employed for the detection of small amounts ofpara-globoside in biological materials also in the presence of excess globoside. No staining was obtained when thin layer chromatograms of neutral glycosphingolipid extracts from rabbit erythrocyte membranes were overlayed with¹²⁵I-ECA. Afterin situ treatment of the chromatograms with -galactosidase, the lectin bound to several components, one of which had a mobility corresponding to that of the pentahexosylceramide Gal3Gal4GlcNAc3Gal4Glc1Cer, the major neutral glycosphingolipid of rabbit erythrocytes, thus providing further evidence for the specificity of ECA forpara-globoside.Abbreviations GSL glycosphingolipid(s) - CDH lactosylceramide, Gal4Glc1Cer - CTH trihexosylceramide, Gal4Gal4Glc1Cer - GLOB globoside, GalNac3Gal4Gal4Glc1Cer - PG para-globoside, Gal4GlcNAc3Gal4Glc1Cer - AsG_M1 asialo-G_M1, Gal3GalNAc4Gal4Glc1Cer - FORS Forsmann antigen, GalNAc3GalNAc3Gal4Gal4Glc1Cer - CPH pentahexosylceramide, Gal3Gal4GlcNAc3Gal4Glc1Cer - ECA Erythrina cristagalli agglutinin - SBA soybean agglutinin - PBS phosphate-buffered saline - PVP-40 polyvinylpyrrolidone M.W. 40000 - BSA bovine serum albumin - HRP-avidin horseradish peroxidase conjugated to avidin - ELLA enzyme-linked lectin assay - ELISA enzyme-linked immunosorbent assay - PMNL polymorphonuclear leukocytes - HPTLC high performance thin layer chromatography     

Variation in Ambrosia pollen concentration in Southern and Central Poland in 1982–1999

The aim of the study was to investigate theAmbrosia pollen concentrations inselected Polish cities and for Kraków torelate it to some meteorological factors.Sampling was carried out in Kraków in1982–1997 and in Rabka in 1992–1996 with theuse of the gravimetric method. In Zakopane,Kraków, Ostrowiec witokrzyski,Warszawa and Pozna in 1995–1996 both thegravimetric and volumetric methods (Burkardtrap) were employed. In Kraków themonitoring has been performed since 1994 usingthe volumetric method. The results show theragweed pollen presence in August and Septemberwith the tendency to appear more frequently inAugust in some years. In Kraków (1994–1999)Ambrosia pollen was found either in thelast two weeks of August or in the first twoweeks of September which seems to be a regularand repeatable pattern every two years.Seasonal fluctuations of Ambrosia pollenconcentration do not show a clear increasingtendency except at Warszawa and Ostrowiecwitokrzyski in 1996 and at Krakówin 1999. Percentage of Ambrosia pollen inannual sums of total pollen is very low anddoes not exceed 1% except at Ostrowiecwitokrzyski in 1996 (1.2%) and atKraków in 1999 (2%). For Kraków theanalysis of some meteorological factors (Tmax,Tmin, precipitation, wind direction) wasperformed. High temperature and lack of rain orlow precipitation correlate well with ragweedpollen concentrations. During the Ambrosia pollen seasons ESE, E, S, SE, WSW, SWwind directions prevailed which could suggest along-distance transport from Ukraine, the CzechRepublic, Slovakia and also from Hungary, one ofthe most ragweed-polluted countries.     

Regional Differentiation in the Dynamics of the Pollen Seasons of Alnus, Corylus and Fraxinus in Poland (Preliminary Results)

In 1995–1996 a study of pollen concentrations of Corylus, Alnus and Fraxinus was performed at seven sites in regions of different climatic and natural conditions. The aim of the study was to determine whether regional differences in the course and duration of pollen seasons occur in Poland. The study was performed using a volumetric method. Several phases during the pollen season were defined for each taxon (1, 2.5, 5, 25, 50, 75, 95, 97.5, 99% of annual total) and duration of the pollen seasons was calculated using 98 and 90% methods. Dynamics and duration of the pollen seasons and a start of particular phases of the season were compared among sites. On the basis of the preliminary analysis it could be supposed that regional differences were most evident in the case of Corylus. The pollen season of this taxon started the earliest in Pozna where thermal conditions were most favourable and the latest in Gdask, a place at the furthest to the north ( ², 0.05). In montane regions (Zakopane, Rabka) last phases of the season were significant extended ( ², 0.05). Probably it results from secondary pollen deposition and a long-distance transport by montane wind. In case of Fraxinus the significant regional differences in the start of the pollen season were not found. The study supported that weather conditions have the substantial influence on the start of consecutive phases of the pollen season.     

Development of high frequency multiple shoot formation in Persian clover (<Emphasis Type="Italic">Trifolium resupinatum</Emphasis> L.)

An efficient and reliable micropropagation system for Persian clover (Trifolium resupinatum L.) was developed using different explants and media. Node, hypocotyl and cotyledonary node explants were cultured on Murashige and Skoog (MS) medium supplemented with combinations of either 6-benzyladenine (BA) and indole-3-butyric acid (IBA) or BA, Kinetin (KIN) and IBA. Direct multiple shoots developed within 6weeks in all explants in most media tested. The best shoot multiplication capacity was obtained from cotyledonary node explants on MS medium containing 7.1M BA and 1M IBA or 14.1M BA and 1M IBA. Elongated shoots were rooted on either MS medium alone or combination with different concentrations of indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) and -naphthaleneacetic acid (NAA). High rooting was achieved in half strength MS medium containing 8M IBA.     

Entwicklungsgeschichte und Ultrastruktur von Pollenkitt und Exine bei nahe verwandten entomophilen und anemophilen Angiospermensippen derAlismataceae,Liliaceae, Juncaceae,Cyperaceae, Poaceae undAraceae

Some closely related members of the monocotyledonous familiesAlismataceae, Liliaceae, Juncaceae, Cyperaceae, Poaceae andAraceae with variable modes of pollination (insect- and wind-pollination) were studied in relation to the ultrastructure of pollenkitt and exine (amount, consistency and distribution of pollenkitt on the surface of pollen grains). The character syndromes of pollen cementing in entomophilous, anemophilous and intermediate (ambophilous or amphiphilous) monocotyledons are the same in principal as in dicotyledons. Comparing present with former results one can summarize: 1) The pollenkitt is always produced in the same manner by the anther tapetum in all angiosperm sub-classes. 2) The variable stickiness of entomophilous and anemophilous pollen always depends on the particular distribution and consistency of the pollenkitt, but not its amount on the pollen surface. 3) The mostly dry and powdery pollen of anemophilous plants always contains a variable amount of inactive pollenkitt in its exine cavities. 4) A step-by step change of the pollen cementing syndrome can be observed from entomophily towards anemophily. 5) From the omnipresence of pollenkitt in all wind-pollinated angiosperms studied one can conclude that the ancestors of anemophilous angiosperms probably have been zoophilous (i.e. entomophilous) throughout.

     

Cowdria Ruminantium Antibodies in Acaricide-Treated and Untreated Cattle Exposed to Amblyomma Variegatum Ticks in The Gambia

An indirect enzyme-linked immunosorbent assay (ELISA), based on the major antigenic protein 1 fragment B (MAP1-B) of Cowdria ruminantium, was used to assess seroprevalence in cattle in The Gambia. Two groups of 20 N'Dama and 20 Gobra zebu cattle were monitored for 12 months with flumethrin treatment and for another 10 months without acaricidal treatment. Two groups of 20 N'Dama and 20 Gobra cattle served as untreated controls. During the period of acaricidal treatment, the cumulative proportions of positive serum samples were 25.6±5.6% (±confidence interval) and 34.7±6.8% in treated N'Dama and Gobra cattle respectively; the proportion of positive sera in untreated cattle was 52.2±6.9% in N'Damas and 61.4±7.3% in Gobras. Within breed, difference in antibody prevalence between treated and untreated cattle was significant (P<0.001) but between breed differences were not significant. In the 10 months following suspension of acaricide application, there was an increase of proportion of positive serum samples in previously treated N'Dama and Gobra cattle. In both previously treated and untreated animals the peak of positive seroreactions occurred during and subsequent to the period of activity of Amblyomma variegatum adults. Cumulative seroprevalences in previously treated N'Dama and Gobra cattle were 32.6±6.9% and 44.7±8.5%, respectively; in untreated animals seroprevalence was 38.6±7.2% in N'Dama and 65.3±8.4% in Gobra cattle. Throughout the study period, within the N'Dama breed, the seropositive rate in previously treated cattle did not differ from that in untreated animals. Conversely, within the Gobra breed, the number of positive seroreactions was higher (P<0.002) in untreated animals than in previously treated cattle. These results provide a support for designing A. variegatum and heartwater control strategies, if necessary, in The Gambia in relation to cattle breeds.     

Ultrastructural investigations on Lycopersicum peruvianum pollen activation and pollen tube organization after self-and cross-pollination

No differences have been observed in vivo between Lycopersicum peruvianum compatible and incompatible pollen during activation and pollen tube emission and organization, that is until 4 h and 30 min after pollination. During pollen activation the main events are the setting free of rough endoplasmic reticulum (RER) cisterns which were stacked in the mature pollen, the increase in the number of polysomes, and a great activity of the dictyosomes. Immediately after germination of the vegetative nucleus and the generative cell move into the tube, the generative cell diviting to form the male gametes; the tube then becomes organized in four zones. This series of changes is similar to what has already been observed in vitro except that in vitro the generative cell remains undivided and the whole process from seeding to tube organization takes 3 h instead of 4 h and 30 min after pollination, as it does in vivo. Our findings are compatible with the main models of the tube inhibition mechanism proposed till now.Abbreviations RER rough endoplasmic reticulum - GC generative cell - VN vegetative nucleus - GP germinative pore Research performed under C.N.R. (Italian National Research Council) program Biology of Reproduction     

Memorization and delayed expression of regulatory messages in plants

Plantlets of Bidens pilosus L., considered to be basically symmetrical, can be lateralized (A/B) by being administered a symmetry-breaking signal such as puncturing one of the plant cotyledons. The induced asymmetry remains latent as long as the plants have not been made permissive, i.e. as long as the plant apex is left functioning. When the apex has been removed (plant decapitation), the latent asymmetry is expressed by one of the cotyledonary buds (a/b) statistically beginning to elongate before the other. The interval of time between delivering the symmetry-breaking signal and making the plant permissive is the memorization-time, t. Memorization can be quantified by using a precedence index, q, the values of which range from 0 (no detectable asymmetry with regard to bud growth) to ±1 (bud growth perfectly asymmetric in favour of either bud b or a). Even for memorization times, t, up to 14 d, q-values up to 0.4 (or even larger) are observed. Various experimental characteristics (e.g. light, temperature, presence or absence of the root system) but not the plant age can affect the q-values, at the moment when the treatments are performed, at least in the range of 6 to 25 d. Combining several puncturing treatments either increases or decreases the q-values, depending on the nature of these treatments and the time-intervals, t, between them. Symmetrically removing both cotyledons in the minutes following the puncturing of one of them does not significantly alter the results, which means that the symmetry-breaking message is rapidly transported and memorized within the plant. Non-traumatic asymmetrical treatments (droplets of saline solutions, light-gradients) can also act as symmetry-breaking signals and be memorized. Plants other than Bidens are likely to possess similar memorization ability, although the q-values observed up to now have not been very large.     

Structure of a new nonasaccharide isolated from human milk: VI2Fuc,V4Fuc,III3Fuc-p-lacto-N-hexaose

The structure of a new nonasaccharide isolated from human milk has been investigated. By using methylation analysis, FAB-MS and¹H-and¹³C-NMR spectroscopy as basic methods of structural investigation, this oligosaccharide was identified as VI²--Fuc,V⁴-Fuc,III³--Fuc-p-lacto-n-hexaose: Fuc1-2Gal1-3[Fuc1-4]GlcNAc1-3Gal1-4[Fuc1-3]GlcNAc1-3Gal1-4Glc.Abbreviations COSY correlation spectroscope - DP degree of polymerisation - FAB-MS fast atom bombardment-mass spectrometry - HPLC high performance liquid chromatography - NMR nuclear magnetic resonance - GLC gas-liquid chromatography     

Natural abundance of 15N in tropical plants with emphasis on tree legumes

Natural abundance of ¹⁵N ( ¹⁵N) of leaves harvested from tropical plants in Brazil and Thailand was analyzed. The ¹⁵N values of non-N₂-fixing trees in Brazil were +4.5±1.9, which is lower than those of soil nitrogen (+8.0±2.2). In contrast, mimosa and kudzu had very low ¹⁵N values (–1.4+0.5). The ¹⁵N values of Panicum maximum and leguminous trees, except Leucaena leucocephala, were similar to those of non-N₂-fixing trees, suggesting that the contribution of fixed N in these plants is negligible. The ¹⁵N values of non-N₂-fixing trees in Thailand were +4.9±2.0. Leucaena leucocephala, Sesbania grandiflora, Casuarina spp. and Cycas spp. had low ¹⁵N values, close to the value of atmospheric N₂ (0), pointing to a major contribution of N₂ fixation in these plants. Cassia spp. and Tamarindus indica had high ¹⁵N values, which confirms that these species are non-nodulating legumes. The ¹⁵N values of Acacia spp. and Gliricidia sepium and other potentially nodulating tree legumes were, on average, slightly lower than those of non-N₂-fixing trees, indicating a small contribution of N₂ fixation in these legumes.     

Inoculation of sugar mill by-products compost with N2-fixing bacteria

Inoculation of sugar mill by-products compost with N₂-fixing bacteria may improve its quality by increasing total N and available P. Compost was inoculated with Azotobacter vinelandii(ATCC 478), Beijerinckia derxii (ATCC 49361), and Azospirillumsp. TS8, each alone and all three together. Numbers of all N₂-fixing bacteria in compost declined from an initial population of 5×10⁵cellsg^–1 during incubation. The population of Azotobacter declined to approximately 2×10²cellsg^–1 and the population of Beijerinckia and Azospirillum declined to approximately 9×10³ and 3.5×10⁴cellsg^–1 respectively, at day 50. Inoculation with N₂-fixing bacteria increased acetylene reduction, total N by 6–16 and available P by 25–30% in comparison to the uninoculated control. Increasing the N content and P availability of compost increases its value and there may be additional benefit from providing N₂ fixing bacteria.     

An improved micropropagation system for Chrysanthemum based on Pluronic F-68-supplemented media

Supplementation of MS-based medium containing 4.4 M 6-benzyladenine and 2.7 M -naphthaleneacetic acid with 0.001–0.1% (w/v) of the non-ionic, co-polymer surfactant, Pluronic F-68, significantly (p<0.05) increased the mean fresh weight gain of cultured leaf explants of chrysanthemum (Dendranthema grandiflora) Tone Maid and Early Charm by a maximum of 74% and 34%, respectively. The percentage of individual explants giving adventive shoots was also stimulated by Pluronic F-68; for cv. Early Charm, 0.001% (w/v) Pluronic F-68 induced the maximum response, whereas for cv. Tone Maid, the maximum response occurred with 0.01% surfactant. Shoot regeneration from explants was also enhanced at these concentrations of surfactant, compared to explants cultured in the absence of Pluronic F-68.Abbreviations BA 6-benzyladenine - MS Murashige & Skoog 1962 - NAA -naphthaleneacetic acid     

Glycosylation with thioglycosides activated by dimethyl(methylthio)sulfonium tetrafluoroborate: synthesis of two trisaccharide glycosides corresponding to the blood group A and B determinants

Two trisaccharide glycosides,p-trifluoroacetamidophenylethyl 3-O-(2-acetamido-2-deoxy--d-galactopyranosyl)-2-O-(-l-fucopyranosyl)--d-galactopyranoside andp-trifluoroa-cetamidophenylethyl 2-O-(-l-fucopyranosyl)-3-O-(-d-galactopyranosyl)--d-galactopyranoside, corresponding to the human blood group A and B determinants, were synthesized. A key fucosylgalactosyl disaccharide derivative was glycosylated with galactosaminyl or galactosyl donors, respectively. Dimethyl (thiomethyl)sulfonium tetrafluoroborate was used for thioglycoside activation in coupling reactions.     

Abbau der 4-Chlorbenzoesäure durch eineArthrobacter-Species

Zusammenfassung EineArthrobacter-Species, die 4-Chlorobenzoesäure als einzige Kohlenstoffquelle verwerten kann, gibt beim Wachstum auf dieser Verbindung 4-Hydroxybenzoesäure und Protocatechusäure ins Medium ab. Der weiter Abbau des aromatischen Ringes erfolgt durch meta-Spaltung. Beim Wachstum derArthrobacter-Species auf Benzoesäure trit im Medium cis,cis-Muconsäure auf. In diesem Fallewird also der ortho-Weg eingeschlagen. Die Enzyme für beide Abbauwege sind induzierbar.

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Degradation of 4-Chlorobenzoic acid by anArthrobacter species

AnArthrobacter sp. growing on 4-Chlorobenzoic acid as its sole source of carbon excretes 4-hydroxybenzoic acid and protocatechuic acid into the culture medium. Protocatechuic acid is further attacked by meta-cleavage. During growth of theArthrobacter sp. on benzoic acid cis-cis muconic acid can be isolated from the medium, suggesting the involvement of the ortho-cleavage pathway. The enzymes both for the meta- and the ortho-cleavage pathway are inducible.

Dem Andenken an Professor Bernhauer gewidmet     

N-andO-alkylation of glycoconjugates and polysaccharides by solid base in dimethyl sulphoxide/alkyl lodide

O-Methylation of simple neutral oligosaccharides is readily accomplished in dimethyl sulphoxide containing solid sodium hydroxide and methyl iodide [Cincanu I, Kerek F (1984) Carbohydr Res 131209-17]. This procedure has been extended to 2-acetamido-2-deoxy sugars and sialic acid-containing oligosaccharides. CompleteO-andN-methylation was in most cases achieved in 15 min. Esterification of carboxylic groups in uronic acids was fast and resulted in concomitant -elimination. The method is also suitable for methylation of glycoproteins and glycosphingolipids. Polysaccharides can also be methylated by this technique. Analysis of the products by gas-liquid chromatography and mass spectrometry showed no degradation products.Abbreviations lacto-N-tetraose LcOse₄, Gal3GlcNAc3Gal4Glc - lacto-N-fucopentaose III III³Fuc-nLcOse₄, Gal4[Fuc3]GlcNAc3Gal4Glc - trihexosylceramide GbOse₃Cer, Gal4Gal4Glc1-1Cer - globoside GbOse₄Cer, GalNAc3Gal4Glc1-1Cer - FAB-MS fas atom bombardment mass spectrometry     

The addition of Dasypyrum villosum (L.) Candargy chromosomes to durum wheat (Triticum durum Desf.)

Summary Six monosomic addition lines were produced in which different Dasypyrum villosum (L.) Candargy chromosomes were added to the chromosome complement of Triticum durum Desf. cv. Creso. Each added alien chromosome was found to have a specific effect on plant morphology and fertility. Transmission rate varied widely (from 7.5 to 27.7%) among the six univalent chromosomes. Different monotelosomic addition plants derived by a relatively high frequency of chromosome misdivision were isolated. The addition lines should be useful for studying Dasypyrum chromosome homoeology and the introduction of alien variation into durum and common wheats.Research supported by a grant from the Italian Research Council for Finalized Project IPRA. Sub-project Plant Breeding, Paper No. 1095     

Cryptopolyploidy inBunias (Brassicaceae) revisited — A flow-cytometric and densitometric study

The concern of the present analysis is the hypothetical cryptopolyploidy, a concept basically of historical interest only, but discussed again by Battaglia (1996) in a recent treatment of the term and its historical background. Melinossi (1935), while reanalyzing erratic observations on the crucifersBunias erucago andB. orientalis by Jaretzky (1928a), found 2n = 14 in both species but twice the chromosome volume inB. erucago compared withB. orientalis. Melinossi considered cryptopolyploidy inB. erucago, i.e., she discussed pairwise fused chromosomes on a tetraploid basis or endoreduplicated (and thus binemic) chromosomes in this species. Cryptopolyploidy has also been claimed by Pannocchia-Laj (1938) inVinca difformis (Vincaceae). Battaglia (1996) criticized the term cryptopolyploidy because, in his opinion, the genuinely polyploid status of these plants is not hidden (crypto) but phenotypically (from herbarium specimens) recognizable. He coins the term phenopolyploidy, i.e., phenotypic polyploidy disagreeing with the karyotype numerically evalated. We measured genome size ofB. orientalis andB. erucago (both 2n = 14) by Feulgen densitometry and propidium iodide flow cytometry. Surprisingly,B. erucago (the annual species with 2.13 pg, 1 C) turned out to have only 0.81-fold the DNA amount ofB. orientalis (the perennial species with 2.64 pg, 1 C). Therefore, any kind of genetically polyploid status inB. erucago is out of the question. Only speculative significance can be ascribed to the terms cryptopolyploidy and phenopolyploidy.     

A simple method for maintaining high multiplication of Narcissus shoot cultures in vitro

A simple method for stimulating and maintaining high in vitro multiplication of Narcissus shoot clump cultures was developed. Shoot clumps were subjected either to normal cutting where leaves were trimmed to 20 mm in length at the beginning of each culture passage or to severe cutting where shoot clumps were cut down to the basal plate region removing all green tissue. Severe cutting at the beginning of each culture passage initially doubled the leaf multiplication, compared to normal cutting, but the difference between cutting treatments declined in successive passages. The improvement in leaf multiplication was maintained when shoot clumps were subjected to severe cutting only at every other culture passage, with no cutting in the alternate recovery passages. In vitro multiplication was increased by severe cutting in all seven Narcissus cultivars which were tested.Abbreviations NAA-1 naphthylacetic acid - BAP benzylaminopurine