The Effect of Flower Maturity and Harvest Timing on Floral Extract fromBoronia megastigma(Nees)

Development of floral organs during maturation of flower budsinto fully open boronia flowers is described. The petals andfunctional anthers attain their maximum size prior to the non-functionalanthers and the stigma. Organoleptic properties of the floralextract change with successive stages of bud development. Theconcentrations of extract and volatiles in the extract (% byf. wt) increase as buds mature, the extract concentration beinghighest in large buds and open flowers and the concentrationof volatile compounds being highest in open flowers. The rateof flower and extract development was measured. Yields of flowermaterial and floral extract per plant, and the concentrationof total volatiles including ß-ionone reach maximumlevels when 70% of flowers have reached anthesis. All measuredfactors decline after this point, except extract concentration(% of f. and d. wt) which is maintained up to 83% open flowers. Boronia megastigma(Nees); brown boronia; Rutaceae; flower development; floral extract; solvent extraction; ß-ionone; essential oils     

Emission of Volatiles From Brown Boronia Flowers: Some Comparative Observations

Extensive research has focused on the concentration of aglyconeswithin brown boronia (Boronia megastigma) flowers, however emissionof volatiles into the headspace above these flowers is not welldocumented. Using solid-phase microextraction (SPME) to trapvolatiles and GCMS analysis, we observed 23 volatiles in theheadspace above buds and flowers throughout flower maturation,above dissected floral organs and above whole plants held for36 h under either continuous light, continuous dark or 12 hlight:12 h dark:12 h light treatments. Fully-opened flowersemitted the most complex mixture of volatiles and in the greatestquantity, with a rapid decline in senescent flowers. Caryophyllene,humulene and bicyclogermacrene declined as flower buds matured;ß-ionone increased. From the individual floral organs,emission from the petaline anthers comprised 38% of total emissionsfrom the (calculated) ‘whole flower’, with 27% contributedby the petals and 10.5% by the stigma. Monoterpenes dominatedthe headspace from the calyx; dodecyl acetate, methyl jasmonateand (Z)-n-heptadec-8-ene were relatively predominant in emissionsfrom the androecium. ß-Ionone, the major floral volatilein brown boronia, dominated volatiles emitted from the stigma(87%). However, the relatively tiny petaline anthers, activein pollen production and high in carotenoids, contributed thegreatest overall amount of ß-ionone to emission fromthe whole flower. There were three different patterns in emissionof volatiles from plants in response to different light conditions:(1) emission patterns identical irrespective of light environment,with maximum emission in the ‘endogenous’ dark period,i.e. when the plant would normally have been in the dark (-pinene);(2) similar emission in all treatments, with an increase anddecline over a period of 26 h (5-acetoxy linalool, cyclic ß-ionone,dodecyl acetate and (Z)-n-heptadec-8-ene); and (3) emissionin all treatments but enhanced in the dark, with a 27.5 h periodin some cases (cyclic ß-ionone endoperoxide, dihydroß-ionone, ß-ionone, and ‘total volatiles’).Preliminary evidence is presented for endogenous control ofemission of a number of volatiles such as -pinene, with perhapsdiurnal control of others such as ß-ionone. Copyright2000 Annals of Botany Company Boronia megastigma, brown boronia, SPME, headspace, floral volatiles, ß-ionone     

Biosynthesis of Volatiles in Brown Boronia Flowers after Harvest: Effect of Harvest Time and Incubation Conditions

Brown boronia flowers (Boronia megastigma)were incubated at25 °C after harvest, increasing the concentration of a solventextracted product by up to 25% (d. wt basis) and floral volatilesby up to 300% (d. wt). Open flowers produced more volatilesduring post-harvest incubation than flower buds, whole flowersproduced more than flowers treated to simulate harvester damage,and fresh flowers produced more than frozen and thawed flowers.Flowers incubated in bags purged with air produced more extractand volatiles than those purged with nitrogen gas; ß-iononewas particularly depleted in nitrogen-purged flowers. Flowersand buds from three successive harvests (68, 82 and 90% openflowers) were incubated at several temperatures for up to 24h. The greatest increase in the concentration of floral extractoccurred in flowers harvested when 90% were open and incubatedfor 4 h at 12 °C, or 14 h at 23 °C. The largest increasein total volatiles occurred in material harvested at 82% openflowers and incubated for 25 h at 12 °C, or 13 h at 23 °C.The concentration of extract and volatiles declined after prolongedpost-harvest incubations. Brown boronia flowers produce volatilesand other components of the floral extract after harvest whenflowers with mature, intact enzyme systems are well suppliedwith oxygen, at temperatures at or below 25 °C. As flowersbecome visibly senescent, their ability to produce volatilesafter harvest declines.Copyright 1998 Annals of Botany Company Boronia megastigma(Nees); brown boronia; essential oils; floral volatiles; ß-ionone; phenology; post-harvest storage.     

Flower and Volatile Oil Ontogeny in Boronia megastigma

The volatile oils of flowers of Boronia megastigma Nees. areimportant commercially but little has been published regardingtheir localization or the dynamics of their synthesis. In thisstudy we examined the histochemistry, anatomy and volatile oilcomposition of the floral organs of plants from a native populationby SEM. Lysigenous glands occurred in all organs except the stigma andandroecium and were associated with the presence of - and ß-pineneand limonene in tissue extracts. These compounds increased inconcentration throughout fruit development. Petals, stamens,staminodes and stigmas were glandular and contained phenolicdeposits, a dense cytoplasm and prominent intercellular spaces.Extracts of these tissues contained dodecanol, ß-iononeand dodecyl acetate, the concentrations of which were greatestat the time of stigma receptivity. The location and timing oftheir synthesis together with their biological activity suggeststhat these volatile oils may have a role in pollination.Copyright1995, 1999 Academic Press Boronia megastigma Nees., Rutaceae, essential oils, volatile oils, reproductive biology, -pinene, ß-pinene, limonene, ß-ionone, dodecyl acetate, heptadecene     

Intraspecific Variation in Oil Components ofBoronia megastigmaNees. (Rutaceae) Flowers

Intraspecific variation in the oil composition ofBoronia megastigmaNees.(Rutaceae) was examined. Boronia absolute is extracted fromblossom primarily for use as a food additive. A major componentis ß-ionone andB. megastigmais one of the commercial,natural sources of this compound. Genotypes superior in productionof ß-ionone and low in monoterpene hydrocarbons weresought from natural populations in the south west of WesternAustralia as part of a breeding programme. Flowers were collectedfrom 25 plants in each of 29 different populations. Blossomwas extracted with ethanol and analysed using a gas liquid chromatographfitted with ionisation detectors. The contents of ß-ionone,dodecyl acetate, -pinene, ß-pinene and limonene inthe oil extract were compared. Intra-population variation wasas great as inter-population variation and no distinct chemotypeswere found. Considerable variation existed in the content ofcomponents. The highest ß-ionone content was 1787mg g^-1f. wt. Some genotypes contained all five components analysed,others lacked one or more of the monoterpenes: -pinene, ß-pineneor limonene. Principle components analysis indicated that contentsof ß-ionone and dodecyl acetate were associated andthat they were distinct from the content of the monoterpenes,which were associated with each other. Natural shading was associatedwith lower levels of monoterpenes but other oils were unaffected.Young plants contained less pinenes than older plants and oldplants contained the most dodecyl acetate. Vigorous plants producedmore pinenes. Red flowers contained the least ß-iononeand dodecyl acetate.Copyright 1999 Annals of Botany Company Boronia megastigma,boronia, Rutaceae, oil, -pinene, ß-pinene, limonene, ß-ionone, dodecyl acetate, monoterpenes, chemotypes.     

Floral volatiles play a key role in specialized ant pollination

Chemical signals emitted by plants are crucial to understand the ecology and evolution of plant–animal interactions. Scent is an important component of floral phenotype and represents a decisive communication channel between plants and floral visitors. Floral volatiles promote attraction of mutualistic pollinators and, in some cases, serve to prevent flower visitation by antagonists such as ants. Despite ant visits to flowers have been suggested to be detrimental to plant fitness, in recent years there has been a growing recognition of the positive role of ants in pollination. Nevertheless, the question of whether floral volatiles mediate mutualisms between ants and ant-pollinated plants still remains largely unexplored. Here we review the documented cases of ant pollination and investigate the chemical composition of the floral scent in the ant-pollinated plant Cytinus hypocistis. By using chemical-electrophysiological analyses and field behavioural assays, we examine the importance of olfactory cues for ants, identify compounds that stimulate antennal responses, and evaluate whether these compounds elicit behavioural responses. Our findings reveal that floral scent plays a crucial role in this mutualistic ant–flower interaction, and that only ant species that provide pollination services and not others occurring in the habitat are efficiently attracted by floral volatiles. 4-oxoisophorone, (E)-cinnamaldehyde, and (E)-cinnamyl alcohol were the most abundant compounds in Cytinus flowers, and ant antennae responded to all of them. Four ant pollinator species were significantly attracted to volatiles emitted by Cytinus inflorescences as well as to synthetic mixtures and single antennal-active compounds. The small amount of available data so far suggest that there is broad interspecific variation in floral scent composition among ant-pollinated plants, which could reflect differential responses and olfactory preferences among different ant species. Many exciting discoveries will be made as we enter into further research on chemical communication between ants and plants.     

Regulation of Floral Terpenoid Emission and Biosynthesis in Sweet Basil (<Emphasis Type="Italic">Ocimum basilicum</Emphasis>)

Past studies have focused on the composition of essential oil of Ocimum basilicum leaves, but data on composition and regulation of its aerial emissions, especially floral volatile emissions, are scarce. We studied the chemical profile, within-flower spatial distribution (sepals, petals, pistils with stamina, and pedicels), diurnal emission kinetics and effects of exogenous methyl jasmonate (MeJA) application on the emission of floral volatiles by dynamic headspace collection, and identification using gas chromatography-mass spectrometry (GC–MS) and proton-transfer reaction mass spectrometry. We observed more abundant floral emissions from flowers compared with leaves. Sepals were the main emitters of floral volatiles among the flower parts studied. The emissions of lipoxygenase compounds and monoterpenoids, but not sesquiterpene emissions, displayed a diurnal variation driven by light. Response to exogenous MeJA treatment of flowers consisted of a rapid stress response and a longer-term acclimation response. The initial response was associated with enhanced emissions of fatty acid derivatives, monoterpenoids, and sesquiterpenoids without variation of the composition of individual compounds. The longer-term response was associated with enhanced monoterpenoid and sesquiterpenoid emissions with profound changes in the emission spectrum. According to correlated patterns of terpenoid emission changes upon stress, highlighted by a hierarchical cluster analysis, candidate terpenoid synthases responsible for observed diversity and complexity of released terpenoid blends were postulated. We conclude that flower volatile emissions differ quantitatively and qualitatively from leaf emissions, and overall contribute importantly to O. basilicum flavor, especially under stress conditions.     

Homeotic, Meristic and Cytological Floral Mutants of Thryptomene calycina (family Myrtaceae)

Twenty plants with various phenotypic abnormalities to the flowerswere selected from very large populations of Thryptomene calycinain the Grampian and Black Ranges. Most of these had impairedreproductive function. Normal flowers were epigynous with fivesepals, five petals, five anthers, a single style and two anatropousovules. The mutants were two partially male sterile, tetraploidplants with large flowers, one of which occasionally producedadditional flowers from the leaf axils with peduncles as wellas pedicels; one plant which produced a proportion of hexapetaloidflowers with six stamens; three gross mutants with fleshy, bracteoidpointed petals and sepals, no stamens, vestigial styles andstigmas, exposed ovules and no inferior ovary; one plant withfleshly, bracteoid pointed sepals, vestigial style and stigmabut with exposed ovular structures replaced by four to fivesterile ovules generally inside an abnormal ovary; two plantswith reduced ovary diameter and sterile ovules, shortened style,five reduced sepals and petals and five to eight anthers; threeanthocyanin-free plants; three plants with pink sepals; twoplants with half-sized flowers which produced a proportion offasciated stems; one plant which occasionally produced flowerswithout pedicels which virtually resulted in organs which wereleaf-flower composites; two plants which produced sepals andpetals which contained chlorophyll and prematurely senesced,and had partial substitution of petals by anthers.Copyright1993, 1999 Academic Press Thryptomene calycina, Myrtaceae, Victorian lace flower, floral mutations, mutants, homeotic, meristic, tetraploid, fasciation, male sterility, cut flowers     

Effects of Daylength on Floral Characteristics of Six White Clover (Trifolium repens) Varieties

The effect of daylength upon floral initiation, flower position,and associated features was measured in six white clover (Trifoliumrepens) varieties Most plants initiated flowers in the longest days (16 h). Thevarieties of more southerly origin and with large leaves hada greater spread of response to daylength. Increasing daylength increased the number of buds becoming inflorescencesin all varieties except Milkanova in which the reverse was true.Peduncle length and nectar production were both greatest inthe longest days. Trifolium repens, white clover, daylength, flower initiation, flower position     

异质生境对二型花柱植物雪地报春花性状分化的影响

该研究以生长在新疆北部的二型花柱植物雪地报春(Primula nivalis)为研究材料,对7个低海拔(1 657~2 013m)森林居群和7个高海拔(2 400~2 700m)草原居群的花型比例进行调查,对花部特征进行检测,并对长、短花柱植株在不同海拔居群的花性状分化进行了比较研究,以揭示该类群植物的演化与环境的关系。结果表明:(1)在高海拔草原居群中长花柱植株比例高于短花柱植株。(2)比较草原和森林生态系统中的雪地报春居群,长花柱花形态特征间存在显著差异,但短花柱花形态特征间的差异不显著。(3)高海拔草原居群中长花柱花的花冠筒长度、雄蕊长度、雌雄蕊距离、雌雄异位程度以及雌雄蕊空间位置是导致花性状分化的主要因素。(4)在草原居群中,长花柱花的雄蕊、雌雄异位程度及花冠口至雄蕊距离的变化减少了长短花柱花间的互补式对应性。研究认为,在异质生境中长花柱花性状的变化是导致该物种长短花柱花性状分化的主要因素。     

Floral Biology, Pollination, Pistil Receptivity, and Pollen Tube Growth of Teak (Tectona grandis Linn f.)

Teak flowers are weakly protandrous and pollen is shed withina few hours of flower opening. Pollen is tricolpate and 29 µmin diameter. The papillate stigma is of the wet type and isreceptive from 1100–1300h. The style is hollow throughoutits length. Nectar and pollen are the major floral rewards forpollinators. The major pollinators areCeratina sp. which carryteak pollen on most parts of their bodies, especially the specializedhair structures (scopal brushes) on the tibia. The most effectivepollination period in terms of flowers pollinated and pollenper flower is between 0900 and 1300h. At 1300h the number ofpollen per flower is the highest, ranging from 1–36 (average7). Pollen tubes grow very fast. Within 2 h after pollination8% of the pollen tubes have reached the micropylar end of theovule and pollen tubes first enter the embryo sac at 8 h. Onlyone to two pollen tubes enter the micropyles of a flower. Although78% of flowers were pollinated in open-pollination, the lowfruit set (3.5%) suggests that there are factors other thanpollination limiting fruit set. The main factor appears to bea high amount of selfing, and self-incompatibility occurs whenpollen tubes are arrested at the lower portion of the ovary. Tectona grandis ; floral biology; pollen tube growth; pollination; receptivity; pollinators     

Patterns and Polarity in Floral Meristem and Floral Organ Initiation

The initial event in plant floral organogenesis is bract specification, followed by floral meristem (FM) initiation in bract axils, but initiation signals and the interplay between both lateral organs remain unelucidated. Floral organs are initiated on the flanks of the outgrowing FM and the enormous diversity in floral morphology throughout the plant kingdom reflects variations in organ position, meristy and ontogeny. Classical models of floral development have focused on Arabidopsis, which has mostly actinomorphic flowers, and Antirrhinum, which exhibits zygomorphy, although neither species is typical or representative of angiosperm flower diversity. Although the ABCE model defines a centripetal model of organ identity establishment in different whorls, the characterization of floral organ initiation in many species has relied on their morphological appearance, due to a lack of founder cell-specific markers. Recent progress in early Arabidopsis floral development using histology, molecular markers and mutants has led to refinements of existing floral organ initiation paradigms. In Arabidopsis, sepals initiate unidirectionally, in a temporal window characterized by the absence of CLAVATA3 and WUSCHEL stem cell markers and are partly dependent on PRESSED FLOWER function, whereas initiation of inner-whorl organs occurs centripetally. Arabidopsis mutants reveal that the FM is highly polarized along an ab-/adaxial axis and a comparison of floral development in Arabidopsis and Antirrhinum suggests that heterochrony of conserved gene functions has been evolutionarily adaptive.

This review discusses current views on FM and organ specification signals, the gene regulatory networks that underlie floral meristem polarity, and analogies between the development of floral and leaf primordia as lateral organs. Alternative stem-cell proliferation mechanisms and the bifurcation of founder cell populations can help to explain the diversity in floral diversity throughout the plant kingdom and underpin comparative evolutionary biology and macroevolution. An analysis of plants with divergent body plans at the level of organ specification is urgently needed.     

APETALA1启动子驱动AtIPT4在转基因拟南芥中表达导致花和花器官发育异常

细胞分裂素对拟南芥（Arabidopsis thaliana）花分生组织细胞的分裂和分化具有重要作用。本研究利用APETALA1（AP1）特异启动子在花分生组织和第1、2轮花器官中表达细胞分裂素合成酶（isopentyl transferase,IPT）基因IPT4,研究细胞分裂素对花和花器官发育的影响。在pAP1∷IPT4转基因植株中出现了花密集和花器官数目增多等现象。原位杂交和GUS组织染色结果发现,在pAP1∷IPT4转基因植株中,花分生组织特征决定基因LEAFY（LFY）与花器官特征决定基因AP1、PISTILLATA（PI）和AGAMOUS（AG）的表达量均有不同程度的提高。研究结果表明在拟南芥中表达pAP1∷IPT4影响其花和花器官的正常发育。     

Effects of Gibberellic Acid on Floral Development in vivo and in vitro in the Cleistogamous Species, Lamium amplexicaule L.

Using established developmental markers in the corolla and androeciumof Lamium amplexicaule L., we investigated the apparent inductionof open (chasmogamous — CH) from closed (cleistogamous— CL) flowers after application of GA₃ (0.1 mM). In vivotreatment of potentially CL flower primordia caused cell expansionbut not the increased cell division in anthers and corolla necessaryto convert a CL into a CH flower. Floral primordia that appearedto be of undetermined floral type were grown in vitro on a basalmedium supplemented with kinetin (0.1 p.p.m.) and grew to maturityas CL flowers. On media additionally supplemented with GA₃,flowers underwent anthesis, but no true CH flower was produced.Gibberellins appear to be directly responsible for anthesisin the CH flower; but additional, as yet unknown growth factorsare involved in the switch from CL to CH floral form in a developinginflorescence. floral morphogenesis, Lamium amplexicaule L, henbit, cleistogamy, gibberellin, kinetin, anthesis     

Characterization of the Rice Floral Organ Number Mutant fon3

A spontaneous rice mutant named floral organ number 3 (fon3) had major mutations in floral organ numbers. Genetic analysis indicated thatfon3 acted as a single recessive gene. Microscopic observation showed that the number of floral organs infon3 increased centripetally. For example, the number of pistils was the more frequently increased than organs in the outer whorls. Homeotic conversion of lodicules and glumes into palea/lemma-like organs was observed in some flowers. Scanning electron microscopy observation showed that the size of flower meristems was maintained the same or similar until the lemma primordium started to differentiate, at which time the floral meristem became enlarged, suggesting abnormal development of the inner whorls of rice florets. The relationship offon3 with other similar rice mutants is discussed.     

APETALA1 启动子驱动AtIPT4 在转基因拟南芥中表达导致花和花器官发育异常

细胞分裂素对拟南芥(Arab idopsis thal iana)花分生组织细胞的分裂和分化具有重要作用。本研究利用APETALA1(AP1)特异启动子在花分生组织和第1、2轮花器官中表达细胞分裂素合成酶(isopentyl trans ferase, IPT)基因IPT4, 研究细胞分裂素对花和花器官发育的影响。在pAP1::IPT4转基因植株中出现了花密集和花器官数目增多等现象。原位杂交和GUS组织染色结果发现, 在pAP1::IPT4转基因植株中, 花分生组织特征决定基因LEAFY (LFY)与花器官特征决定基因AP1、PISTILLATA (PI )和AGAMOUS (AG)的表达量均有不同程度的提高。研究结果表明在拟南芥中表达pAP1::IPT4影响其花和花器官的正常发育。     

Temporal relationship between emitted and endogenous floral scent volatiles in summer‐ and winter‐blooming Jasminum species

Jasminum spp. is cultivated for their fragrant flowers used in essential oil production and cosmetic uses. An attempt was made to study the temporal variations in floral scent volatiles composition including emitted, free endogenous and glycosyl‐linked volatile compounds from two summer‐blooming species namely, Jasminum auriculatum and Jasminum grandiflorum as well as from two winter‐blooming species namely, Jasminum multiflorum and Jasminum malabaricum. The overall emitted volatile organic compounds (VOCs) were found to be highest when the matrix Porapak Q 80/100 was used with dichloromethane (DCM) as elution solvent. The floral volatile emission from bud to senescence exhibited nocturnal maxima pattern for both the summer‐blooming species. Both the winter‐blooming species emitted its highest concentration at noon. The free endogenous concentrations of all VOCs were low when corresponding emitted concentrations were high. Enzymatic treatment of petal extract revealed that several aromatic volatiles including aromatic alcohols and monoterpenols are synthesized and stored in the flowers as water‐soluble glycosides; these compounds were shown to accumulate in higher amounts in flowers at late bud stage. These findings indicate the utilization of the precursors, i.e. the volatile‐conjugates, through hydrolysis followed by their release as free‐volatiles at flower opening stage. The outcome as a whole suggests a linkage among the temporal pattern of emitted volatiles, free‐endogenous volatiles and glycoside‐bound volatile compounds in all above studied Jasminum spp. and provided an overview of their floral volatilome.     

Characterization of the Rice Floral Organ Number Mutant fon3

A spontaneous rice mutant named floral organ number 3 (fon3) had major mutations in floral organ numbers. Genetic analysis indicated that fort3 acted as a single recessive gene. Microscopic observation showed that the number of floral organs infon3 increased centripetally. For example, the number of pistils was the more frequently increased than organs in the outer whorls. Homeotic conversion of lodicules and glumes into palea/lemma-like organs was observed in some flowers. Scanning electron microscopy observation showed that the size of flower meristems was maintained the same or similar until the lemma primordium started to differentiate, at which time the floral meristem became enlarged, suggesting abnormal development of the inner whorls of rice florets. The relationship of fort3 with other similar rice mutants is discussed.     

Floral volatile profile in Pleurothallidinae,an orchid subtribe pollinated by flies: ecological and phylogenetic considerations

The Pleurothallidinae are recognized as the largest orchid group pollinated by Diptera. This subtribe has been focus of ongoing phylogenetic studies, in which the floral biology and mating systems of their representatives have been recently assessed and some synapomorphies pointed out, all of them regarding features or processes occurring in flowers. Data about the flower volatiles are still scarce, although it may offer valuable information about the floral biology of this subtribe. Here, we collected and analyzed the volatiles emitted from flowers at anthesis. We selected seven Brazilian species representing both late and early-divergent clades of Pleurothallidinae aiming to describe the chemical profile of flower volatiles and relate the identified compounds to the reproductive biology of the selected species. The outstanding feature regarding the floral scent profile is the ubiquitous occurrence in remarkable concentrations of alkanes, a situation not found in Laellinae, a closed subtribe of Pleurothallidinae. We also present a discussion concerning the ecological aspects of the flower volatiles and their pollinators.     

新疆胡颓子属植物花部性状的变异及其分类学价值

新疆胡颓子属植物栽培历史久、变异丰富、种下变异类型多、分类较为困难,因此有必要进一步发掘该属落叶组植物的分类学特征,尤其是对相对保守的繁殖器官的性状进行筛选。该研究以形态较为稳定的繁殖器官为切入点,分析花部性状的变异特点,筛选稳定性状,以期为解决该属的分类难题提供依据。先采用定株采集和同花标记的方法,对比分析了柱头形态、花盘先端有毛无毛、花柱与雄蕊的长短比、花被裂片程度四个性状的稳定性。结果表明:花盘先端有毛无毛、花柱与雄蕊的长短比、花被裂片程度三个性状较稳定,可以作为分类依据;柱头形态在单花中表现稳定,但在单株中出现极大的变异,不具有分类价值;然后采取新疆南、北疆地区的胡颓子属落叶组的尖果沙枣、大果沙枣和沙枣这三种来验证花部性状的稳定性,证明了上述结论的正确性,同时也是对该属分类研究实践的应用。该结果可为胡颓子属的经典分类学和形态学研究以及植物的开发和利用等提供参考依据。