Isolation, Properties and a Possible Function of a Water-Soluble Chlorophyll a/b-Protein from Brussels Sprouts

A water-soluble Chl a/b-protein (CP673) was isolated and purifiedfrom Brussels sprouts (Brassica oleracea L. var. gemmifera DC).The protein had a molecular mass of 78 kDa and an isoelectricpoint of 4.7, consisted of three or four subunits of 22 kDaand was extremely heat-stable. Although CP673 contained aboutone Chl a per protein, the blue and red absorption bands ofChl a that consisted of three or four Chl a forms with differentabsorption maxima suggested that there are several differentmodes or sites of binding for Chl a. Chl a/b ratio of largerthan 10 also indicated that Chl b is present only in a smallfraction of CP673. The heterogeneity of CP673 in terms of compositionand binding of Chl suggests that Chl is not an intrinsic componentof the Chl-protein. Homology search showed that the N-terminalamino acid sequence of CP673 is highly homologous with thatof a 22 kDa protein that accumulates in water-stressed leavesof two Brassicaceae plants, rapeseed and radish, but not withthose of the light-harvesting Chl a/b-proteins of photosynthesis.A possible function of the water-soluble Chl-protein was discussed. (Received September 17, 1996; Accepted November 18, 1996)     

Efficiency as a foraging currency in animals attaining a gain below the energetic ceiling

Previous research has found that efficiency, or, more precisely,the foraging gain ratio (FGR), is a valid currency in foragingtheory when (1) there is a limit to the energy that can beassimilated by the forager and (2) a forager is trying to meetan energy requirement. The FGR is b/ (c — c_r), whereb is the rate of metabolizable energy intake, and c and c_rare the rates of energy expenditure while foraging and resting,respectively. Here I show that, when energy expenditure hasa cost besides energy, animals should also choose the optionwith the highest FGR when they are aiming at a given positivedaily gain. The next question is which gain they should aimfor? Researchers have shown that observed intake levels ofgrowing ruminants are close to the levels predicted by maximizationof the efficiency of oxygen utilization. This currency can be approximated by (B — C + C_r) / C, where B is the daily metabolizable energy intake, and C and C_r are the total andbasal daily energy expenditures, respectively. By simulatinggrowth at different intake levels, I found that mass-specificoxygen consumption rate is indeed minimal at the observed intakelevels. This is the first study in which these efficiency measures(FGR and the efficiency of oxygen utilization) are combined.     

Estimating the capacity for improvement in risk prediction with a marker

Consider a set of baseline predictors X to predict a binaryoutcome D and let Y be a novel marker or predictor. This paperis concerned with evaluating the performance of the augmentedrisk model P(D = 1|Y,X) compared with the baseline model P(D= 1|X). The diagnostic likelihood ratio, DLR_X(y), quantifiesthe change in risk obtained with knowledge of Y = y for a subjectwith baseline risk factors X. The notion is commonly used inclinical medicine to quantify the increment in risk predictiondue to Y. It is contrasted here with the notion of covariate-adjustedeffect of Y in the augmented risk model. We also propose methodsfor making inference about DLR_X(y). Case–control studydesigns are accommodated. The methods provide a mechanism toinvestigate if the predictive information in Y varies with baselinecovariates. In addition, we show that when combined with a baselinerisk model and information about the population distributionof Y given X, covariate-specific predictiveness curves can beestimated. These curves are useful to an individual in decidingif ascertainment of Y is likely to be informative or not forhim. We illustrate with data from 2 studies: one is a studyof the performance of hearing screening tests for infants, andthe other concerns the value of serum creatinine in diagnosingrenal artery stenosis.     

Transpiration Stream of Desert Species: Resistances and Capacitances for a C3, a C4, and a CAM Plant

Transpiration rates and water potentials of three sympatricdesert perennials, a C3 subshrub (Encelia farinosa), a C₄ bunchgrass(Hilaria rigida), and a CAM succulent (Agave deserti), wereanalysed using an electrical circuit analogue that includedresistances and capacitances for the leaves, stems, and roots.The water storage capability of the organs differed considerably,capacitance ranging over 1000-fold from the thin leaves of H,rigida to the massive leaves of A. deserti, although the capacitanceper unit volume varied only 1.9-fold. The diurnal changes inwater storage could support maximum transpiration rates of H.rigida for 4 min, E. farinosa for 7 min, and A. deserti for16 h. The time constant for equilibration of water from storageto the xylem ranged from 29 s for roots of H. rigida to 52 minfor leaves of A. deserti. Resistances for such movement wererelatively low for the succulent leaves of A. deserti and wereup to about 50-fold higher for the three organs of E. farinosa.Xylem resistances calculated using the Hagen-Poiseuille lawand measured xylem dimensions were 2.1- to 2.1-fold lower thanresistances estimated from observed water potential drops, adiscrepancy which is in agreement with other published data.Contrary to data on other plants, the xylem resistances in theroots and leaves of E. farinosa and H. rigida averaged only15% of the stem xylem resistance. Key words: Capacitance, Xylem resistance, Transpiration stream, Desert     

Enzymatic Conversion of Pheophorbide a to the Precursor of Pyropheophorbide a in Leaves of Chenopodium album

Soluble proteins extracted from leaves of Chenopodium albumcatalyzed the conversion of pheophorbide a to a precursor ofpyropheophorbide a, putatively identified as C-13²-carboxyl-pyropheophorbidea. The precursor was then decarboxylated non-enzymatically toyield pyropheophorbide a. Soluble proteins and pheophorbidea, as the substrate, were required for the formation of theprecursor, and boiled proteins were enzymatically inactive.The maximum rate of conversion of pheophorbide a to the precursoroccurred at pH 7.5. The K_m for pheophorbide a was 12.5 µMat pH 7.0. Both pheophorbide b and bacteriopheophorbide a couldserve as substrates, but protopheophorbide a could not. Formationof methanol was detected during the enzymatic reaction, an indicationthat the enzyme is an esterase. Among seven alcohol analogstested, only methanol inhibited the enzymatic activity uncompetitively,with a K₁ of 71.6 mM. Mass-spectrometric (MS) analysis of theprecursor yield a peak at m/z 579 that indicated the releaseof a methyl group from pheophorbide a. It appears thereforethat the enzyme catalyzes the demethylation of the carbomethoxygroup at C-13² of pheophorbide a by hydrolysis to yield methanoland the precursor, C-13²-carboxyl-pyropheophorbide a, whichis converted to pyropheophorbide a by spontaneous decarboxylation.We have tentatively designated the enzyme "pheophorbidase".The presence of the enzyme was dependent on plant species andit was expressed constitutively. ¹Present address: Faculty of Science, Shizuoka University, Ohya,Shizuoka, 422 Japan     

Isolation of a Tobacco cDNA Encoding Sar1 GTPase and Analysis of Its Dominant Mutations in Vesicular Traffic Using a Yeast Complementation System

The cDNA clone of NtSARl, a gene encoding the small GTPase Sar1pwhich is essential for vesicle formation from the endoplasmicreticulum (ER) membrane in yeast, has been isolated from Nicotianatabacum BY-2 cells. NtSAR1 as well as AtSAR1 cDNA isolated fromArabidopsis thaliana [d'Enfert et al. (1992) EMBO J. 11: 4205]could complement the lethality of the disruption of SARI inyeast cells in a temperature-sensitive fashion. They also suppressedyeast sec12 and sec16 temperature-sensitive mutations as yeastSARI does. Using this complementation system, we analyzed thephenotypes of several mutations in plant SAR1 cDNAs in yeastcells. The expression of NtSAR1 H74L and AtSAR1 N129I showeddominant negative effect in growth over the wild-type SARI,which was accompanied by the arrest of ER-to-Golgi transport.Such dominant mutations will be useful to analyze the role ofmembrane trafficking in plant cells, if their expression canbe regulated conditionally. (Received October 29, 1997; Accepted March 17, 1998)     

Energy-dependent redox state of heme a+a3 and copper of cytochrome oxidase in perfused rat brain in situ

Using theblood-free perfused rat brain, we examined the redox behavior ofcytochrome oxidase of two chromophores, hemea + a₃ and copper.When perfusate inflow was stopped to induce global ischemia,the reduction of heme a + a₃ was triphasic,with a rapid phase, a slow phase, and a second rapid phase. Incontrast, the reduction of copper was monophasic after the rapid phaseof heme a + a₃. The triphasicreduction of heme a + a₃ was diminished by energy-depleting treatments, such as addition of an uncoupler. Thetime course of the reduction of copper was not affected by the energydepletion. During global ischemia the decrease in creatine phosphate nearly paralleled the reduction of hemea + a₃, whereas ATPremained at the control level until ~60% of hemea + a₃ was reduced inthe rapid phase. In the slow phase, ATP started to decrease with thereduction of copper. The redox behavior of copper was similar to theslow phase of the reduction of heme a + a₃ because ofthe higher oxygen affinity of copper than of hemea + a₃. Therefore,the rapid phase of the reduction of hemea + a₃ can be used asan alarm before a decrease in ATP, whereas the reduction of copperindicates a decrease in ATP under severe hypoxia. Thus the coppersignal in noninvasive near-infrared spectroscopy is a useful parameterfor the clinical setting.

     

Reproductive Isolation in a Native Population of Petunia sensu Jussieu (Solanaceae)

Reproductive isolation was studied in four syntopic speciesof Petunia sensu Jussieu (Solanaceae) at a site in Rio Grandedo Sul State, Brazil. Reciprocal artificial crossing experimentsconfirmed that a genetic barrier exists between Petunia(P. axillarisand P. integrifolia) andCalibrachoa (C. parviflora and C. heterophylla),and also between C. parviflora andC. heterophylla . Petuniaaxillaris(white, nocturnally scented flower) is geneticallycompatible with the syntopic and allotopic P. integrifolia(coloured,unscented flower). Reproductive isolation appears to be maintainedby the two species having different pollinators: nocturnallyactive hawkmoths (Manduca contracta andM. diffissa subsp. petuniae)pollinate P. axillaris while a diurnally active bee (Hexanthedasp.) pollinates P. integrifolia. Flowers of P. integrifoliaexhibit diurnal opening and closing movements synchronous withthe activity period of the bee. Other than a probable nectarrobber (a carpenter bee, Xylocopa augusti), no insect visitedflowers of P. axillaris in the day. Amounts of floral nectarin P. axillaris and P. integrifolia were within the range ofhawkmoth- and bee-pollinated flowers, respectively. Reproductiveisolating mechanisms in the genus Petunia sensu Jussieu arediscussed. Copyright 2001 Annals of Botany Company Calibrachoa, hawkmoth, Petunia, Petunia axillaris, Petunia integrifolia, pollinator, reproductive isolation, Solanaceae     

Predation by a photosynthetic dinoflagellate Gyrodinium instriatum on loricated ciliates

Feeding of a naked photosynthetic dinoflagellate, Gyrodiniuminstriatum, on loricated ciliates was investigated. Gyrodiniuminstriatum preyed on Favella azorica and Eutintinnus tubulosusby engulfment through the posterior end of the sulcus. In thecase of E.tubulosus, G.instriatum preyed on this small ciliatekeeping the original gymnodinioid cell shape. On the other hand,G.instriatum preyed on Favella taraikaensis by absorbing thecell contents of this large ciliate, which resulted in a balloonlike inflation of its body size. It seemed that G.instriatumcan change its feeding style according to the size of prey.Thus, the present study shows, by using direct observationson the feeding of G.instriatum on loricated ciliates, a reversalof energy flow processes in the food chain in which photosyntheticorganisms eat primary consumers. The growth of G.instriatumafter feeding on F.taraikaensis and E.tubulosus is also describedbriefly.     

RMA1 an Arabidopsis thaliana Gene Whose cDNA Suppresses the Yeast secl5 Mutation, Encodes a Novel Protein with a RING Finger Motif and a Membrane Anchor

To identify molecules that function in the plant secretory pathway,we screened for Arabidopsis thaliana cDNA clones that complementthe temperature-sensitive (ts), secretion-deficient sec15 mutationof yeast Saccha-romyces cerevisiae. RMA1, one of the genes obtainedin this screening, suppressed not only the ts growth of sec15but also its secretory defect. RMA1 is not a structural homologueof SEC15 but encodes a novel 28 kDa protein with a RING fingermotif and a C-terminal membrane-anchoring domain. Mutationalanalysis indicates that the RING finger motif of RMA1 is importantfor its suppression activity. In Arabidopsis plant, RMA1 isubiquitously expressed. A search for homologous proteins inthe database revealed that Arabidopsis, nematode, mouse andhuman possess close homologues of RMA1. (Received January 5, 1998; Accepted March 9, 1998)     

Comparison of Plant Telomere Locations using a PCR-generated Synthetic Probe

We have generated a telomere-specific probe by the polymerasechain reaction and used it to localize chromosome telomeresof ten unrelated angiosperm species in in situ. Concatenationof the simple monomers, 5'-(TTTAGGG)-3', derived from the sequenceof Arabidopsis thaliana telomeres, yielded a stable, versatileand reliable probe that gave a signal of high intensity followingfluorescence in situ hybridization. Most species, includingthose with known karyotype rearrangements, showed telomere labelonly at chromosome termini. These findings are discussed inthe context of the chromosomal events responsible for generatingand stabilizing karyotype change in plants.Copyright 1993, 1999Academic Press PCR, telomere repeat sequence (TRS), in situ hybridization, comparative physical mapping, karyotype evolution, genome organization, Lathyrus sativus L., Vicia sativa L. subsp. nigra (L.) Ehrh., Vigna radiata (L.) R. Wilczek cv. Berken, Nicotiana sylvestris Speg et Comes, Haplopappus gracilis (Nutt.) A. Gray, Gibasis pulchella (Kunth) Rafin., Tradescantia commelinoides Schultes. fil., Hordeum vulgare L. cv. Sultan., Hordeum vulgare L. cv. Tuleen 346, Milium vernale Bieb., Paphiopedilum insigne (Wall.) Pfitz     

Changes in the structure of a zooplankton community during a Ceratium (dinoflagellate) bloom in a eutrophic fishless pond

The community structure of zooplankton was studied in a eutrophic,fishless Japanese pond. The ecosystem was dominated by a dinoflagellate,Ceratium hirundinella, two filter-feeding clado-cerans, Daphniarosea and Ceriodaphnia reticulata, and an invertebrate predator,the dipteran Chaoborus flavicans. The midsummer zooplanktoncommunity showed a large change in species composition (theDaphnia population crashed) when a heavy Ceratium bloom occurred.It is shown that (i) the rapid density decline of D.rosea inmid-May was mainly caused by a shortage of edible phytoplankton,which was facilitated by the rapid increase in Chirundinellaabundance; (ii) the low density of D.rosea in June-July wasconsidered to be mainly caused by the blooming of Ceratium hirundinella(which may inhibit the feeding process of D.rosea), while predationby Cflavicans larvae, the changing temperature, the interspecificcompetition and the scarcity of edible algae were not judgedto be important; (iii) the high summer biomass of the planktonicCflavicans larvae was maintained by the bloom of C.hirundinella,because >90% of the crop contents of C.flavicans larvae wereC.hirundinella during this period. The present study indicatesthat the large-sized cells or colonies of phytoplankton arenot only inedible by most cladocerans, but the selective effectof the blooming of these algae can also influence the compositionand dominance of the zooplankton community, especially for thefilter-feeding Cladocera, in a similar way as the selectivepredation by planktivorous fish. The large-sized phytoplanktoncan also be an important alternative food for ominivorous invertebratepredators such as Chaoborus larvae, and thus may affect theinteractions between these predators and their zooplanktonicprey. In this way, such phytoplankton may play a very importantrole in regulating the dynamics of the aquatic food web, andbecome a driving force in shaping the community structure ofzooplankton.     

Involvement of a Calcium-Dependent Protein Kinase in Hypoosmotic Turgor Regulation in a Brackish Water Characeae Lamprothamnium succinctum

Calcium ion is a key messenger in turgor regulation of internodalcells of Lamprothamnium succinctum in response to hypoosmotictreatment. An increase in the concentration of cytosolic freecalcium ion ([Ca²⁺]_c) is prerequisite for the turgor regulation[Okazaki and Tazawa (1990) J. Membr. Biol. 114: 189], We examinedwhether or not a calcium-dependent protein kinase (CDPK) isinvolved in the Ca²⁺-mediated turgor regulation of Lamprothamniumcells. A 53-kDa CDPK which phosphorylated preferentially histoneH1 but poorly myelin basic protein or casein, was detected inthe cell extract of Lamprothamnium by an in-gel protein kinaseassay. This protein kinase was detected by Western blottingand was immunoprecipitated using an anti-Dunaliella tertiolectaCDPK antibody which can neutralize the Dunaliella CDPK activity[Yuasa et al. (1995) Plant Cell Physiol. 36: 699]. The 53-kDaCDPK was partially purified from Lamprothamnium and its activitywas shown to be inhibited by the antibody and K-252a, a proteinkinase inhibitor. Microinjection of the antibody into the cytosblof Lamprothamnium cells inhibited the decrease in turgor pressurein response to hypoosmotic treatment. However, a transient increasein [Ca²⁺]_c, which was suggested by a transient reduction ofthe velocity of cytoplasmic streaming, was induced in antibody-injectedcells after hypoosmotic treatment. Turgor regulation upon hypoosmotictreatment was inhibited when the cells were treated with K-252a.These results imply that CDPK of Lamprothamnium functions ata down-stream position of Ca²⁺-mobilization in processing turgorregulation in response to hypoosmotic treatment. ² These authors contributed equally to the work.     

Portulal: a rooting promoting substance in Portulaca leaves

A new bioassay which employs disbudded epicotyl cuttings takenfrom light grown Azukia seedlings (A. angularis) was devisedfor testing rooting promotion activity. By use of this method,the rooting promoting principle in leaves of Portulaca grandiflorawas isolated and identified with portulal which had been previouslyobtained. Portulal was reported as an inhibitor of rooting inetiolated Raphanus cuttings and has recently been determinedto be a bicyclic diterpene containing a perhydroazulene nucleus.Portulal promoted the adventitious root formation in severalkinds of plants, i.e. Azukia angularis, Vigna Catiang var. sinensis,Phaseolus Mungo and Raphanus sativus var. acanthiformis ‘Risodaikon’. The rooting process in Azukia and Raphanus cuttings seems toinclude at least two phases; a "preparatory phase" or a portulal-and gibberellin-sensitive phase and a "main phase" or an auxin-sensitiveand portulal-insensitive phase. ¹Contribution No. 16 from the Botanical Gardens, Faculty ofScience, University of Tokyo, Tokyo, Japan. (Received April 3, 1969; )     

Photosynthesis and Formation of Fats in a Diatom

A study has been made, using C¹⁴ as a tracer, of the variationswhich occur according to the physiological condition of thecells in the distribution of carbon fixed, both in the lightand in the dark, by the diatom Navicula pelliculosa (Bréb.)Hilse during periods of 30 to 300 seconds. Fixation into thefollowing cell fractions was determined: (A) material solublein 80 per cent. ethanol but insoluble in benzene, (B) materialsoluble both in 80 per cent. ethanol and in benzene, and (C)material insoluble in 80 per cent. ethanol. Carbon fixed inphotosynthesis was incorporated, rapidly and in amounts representingup to 70 per cent. of the total fixation, into fractions B andC, as well as into fraction A. Considerable variation was foundin the proportions of carbon entering the three fractions inthe light; in actively growing cells the proportion enteringfraction C preponderated over that in B, corresponding to thesynthesis of protein, whereas in nitrogen-deficient cells fixationin B was the greater, corresponding to the synthesis of fat.These patterns changed only slowly, over periods of days, followingthe transfer of cells to altered conditions of nitrate supply.However, when ammonium nitrogen was supplied to nitrogen-deficientcells a marked change in distribution of carbon fixed occurredwithin 5 minutes, fixation in fraction B falling to a low valueand that in A rising correspondingly. In cells subjected toprolonged nitrogen-deficiency, fixation in fraction B fell toa relatively low value but the proportion which this fractionformed of the total dry matter in the cells rose as a resultof an increased rate of loss from the cells of constituentsother than lipides. The distribution of carbon fixed was alsodependent on light intensity. Fixation in fractions B and Crose relatively to that in A as light intensity was increasedup to 100 foot-candles but fell again at the highest intensityused, 2,000 foot-candles. These results are discussed with particularreference to the relationship between fat accumulation and photosynthesisin algae.     

Midsummer succession of rotifer plankton in a shallow eutrophic pond

Temporal changes in the density of rotifer plankton were examinedin a shallow eutrophic pond during July to September 1990, whenabiotic environmental variables were relatively stable. In earlyJuly when Daphnia similis was abundant, rotifer populationsremained at a low density. This is probably due to interferenceby the large cladoceran, because possible food was abundantand the rotifers showed high egg ratio. After late July, whenthe large cladoceran disappeared, a striking succession wasfound in the rotifer plankton. In late July, when Filinia longisetaand Conochilus dossuarius dominated, and in mid- to late Septemberwhen F.opoliensis and Brachionus falcatus dominated, there wasa causal relationship between the density and egg ratio, suggestingthat temporal changes in rotifer density were regulated mainlyby food abundance. However, in August, changes in the densityof most rotifers were not necessarily related with those inthe egg ratio. Brachionus angularis, B.forficula and Keratellaspp. decreased or remained at a low density regardless of theegg ratio, when the population of Asplanchna brightwelli wasdeveloped. However, Polyarthra vulgaris, B.calyciflorus andH.intermedia increased or maintained a high density at thistime. Stomach contents of A.brightwelli revealed that B.angularis,B.forficula and Keratella spp. were the preferred prey, whereasP.vulgaris, B.calyciflorus and H.intermedia were not. Theseresults provide strong evidence that the structure of rotiferplankton can change strikingly within a season due to species-specificdifferences not only in diet, but also in the ability to escapepredation, even if abiotic environmental variables are stable.     

Touch-Inducible Genes for Calmodulin and a Calmodulin-Related Protein Are Located in Tandem on a Chromosome of Arabidopsis thaliana

Genes for calmodulin and calmodulin-related proteins in Arabidopsisare up-regulated by a variety of physical stimuli, which includerain, wind and touch [Braam and Davis (1990) Cell 60: 357].We have isolated five genes for calmodulin (AtCALl, 2, 3, 5,6) and one gene for a calmodulin-related protein (AtCAL4) froman Arabidopsis genomic library. Touch stimulus of Arabidopsisplants induces the accumulation of mRNA transcribed from AtCAL4andAtCAL5, but not from the other isolated genes. The two touch-induciblegenes are arrayed in tandem with a short intergenic region of700 bp but they show different organ-specific patterns of expression. (Received April 27, 1995; Accepted July 20, 1995)     

Trans-Kingdom Conjugation Between Agrobacterium tumfaciens and Saccharomyces cerevisiae, a Bacterium and a Yeast

For conjugation between prokaryotic Agrobacterium tumefaciensand eukaryotic Saccharomyces cerevisiae, we constructed twonovel conjugative plasmids. A. tumefaciens transmitted the plasmidsto S. cerevisiae with the aid of tra genes on a helper plasmid.The transmitted plasmids retained their original structure andfunction in transconjugant yeasts. The presence of Ti plasmidbarely affected the trans-kingdom conjugation. ¹A preliminary report of this work was presented in Japaneseby Yoshida et al. (1993).     

Relationships Between Climate and Flowering of Eight Herbs in a Swedish Deciduous Forest

Effects of annual variation in rainfall, temperature and humidityon flowering abundance of eight temperate woodland plants (Anemonenemorosa, Cardamine bulbifera, Lamiastrum galeobdolon,Oxalisacetosella , Ranunculus ficaria, Stellaria holostea, Viola reichenbachianaand Viola riviniana) were studied during 12 consecutive years(1989–2000) in a hornbeam (Carpinus betulus) forest insoutheast Sweden. Above-average rainfall/humidity in late summerto early autumn of the preceding year increased flowering abundancein L. galeobdolon, O. acetosella, V. reichenbachiana, V. rivinianaand, especially, in R. ficaria, but not in S. holostea and A.nemorosa. Moreover, flowering of R. ficaria and O. acetosellawas positively related to rainfall/humidity during several partsof, or the entire, preceding year. On the contrary, floweringof S. holostea and A. nemorosa was closely related to low valuesof rainfall/humidity in autumn and/or winter of the precedingyear and also to low humidity in the current year in A. nemorosa.Two long periods (3–4 years) of increasing rainfall deficitcoincided with decreasing flowering abundance in most of thespecies, but not with decreasing vegetative development. Temperaturevariability was less consistently related to flowering. A coolperiod during the preceding summer or autumn seemed importantfor flowering in L. galeobdolon, O. acetosella and the Violaspecies, although these relations were, at least partly, causedby interactions with rainfall/humidity. No significant (P <0.05) correlations were found between flowering and the conditionsprevailing in April to May—the main flowering season—ofthe current year. Copyright 2001 Annals of Botany Company Climate, flowering, rainfall, temperature, Anemone nemorosa, Cardamine bulbifera, Lamiastrum galeobdolon, Oxalis acetosella, Ranunculus ficaria, Stellaria holostea, Viola reichenbachiana, Viola riviniana