REPRODUCTIVE BIOLOGY OF THE CERRO CUADRADO (JURASSIC) FOSSIL CONIFERS: PARARAUCARIA PATAGONICA

Seventy-two silicified cone specimens of Pararaucaria patagonica Wieland from the Cerro Cuadrado (Jurassic) petrified forest in Patagonia were studied by use of thin sections and reflected light. Traces from the cone axis stele to the bract and ovuliferous scale are separate at their origins. In cross section the abaxially concave ovuliferous scale trace and massive circular bract trace branch half-way out into the cone-scale complex. Two triangular-shaped bundles of fibers follow the scale trace into the laminar part of the ovuliferous scale. Both surfaces of the ovuliferous scale possess hairs which may have assisted in cone closure. Cones typically contain one flattened, winged seed per ovuliferous scale. Multi-layered seed integuments are represented by an inner fleshy layer of large cells filled with a black amorphous material, a sclerotesta consisting of wedges or fan-shaped groups of sclereids oriented at right angles to the seed surface, and a thin, often crushed fleshy layer. Seed wings consist of anastomosing, branched cells resembling glandular hairs and containing many intercellular spaces. Well-preserved polycotyledonous embryos containing up to eight cotyledons are present in several cones. Tissues of the embryo including the shoot apex, calyptroperiblem, hypocotyl, and cotyledons are described. On the basis of anatomical and morphological features, specimens of P. patagonica are compared to members of both extant and extinct families of conifers including the Araucariaceae, Taxodiaceae, Pinaceae, Cupressaceae, and Cheirolepidaceae.     

ONTOGENY OF THE SHOOT APEX OF SEEDLINGS OF PINUS PONDEROSA

Dormant seeds of ponderosa pine (Pinus ponderosa) were stratified and then planted in the greenhouse. Changes occurring in the shoot apex during germination and growth of the seedlings were observed and are described. Cell divisions in germinating embryos are first noted on the flanks of the apex near the cotyledons. From these loci, mitotic reactivation proceeds up the flanks of the apex and inward toward cells of the rib meristem. Cells are dividing actively in all regions of the shoot apex of seedlings 12 days after planting, but no cytohistological zonation is evident due to differences in staining intensity and in cell and nuclear size. In 64-day-old seedlings the mitotic rate is reduced and the characteristic zonation of pine apices begins to appear. Zonation is even more evident in older seedlings. These observations are discussed in relation to the concept of initials and to theories concerning factors which regulate growth in the shoot apex.     

广西裸子植物

广西本土生长的裸子植物共43种,包含苏铁科3种,三尖杉科4种,红豆杉科4种,罗汉松科6种,松科17种,杉科1种,柏科3种,和买麻藤科5种。虽然它们在本区分布较广,但其种类多集中于北部和中部属酸性土且气候较凉的山脉上。 油杉有数种生长在石灰岩石山上,它们比低地的松树之外的裸子植物较为耐旱。少数种类稀少且分布局限,少数为特有。 经作者研究,广西裸子植物的学名有些需要修改:Cephalotaxus hainanensis和C.griffithii应归并于C.mannii;Podocarpus imbricatus需改为DacrycarPus imbricatus,Podocarpusfteuryi或Deeusocarpus fleuryi改为Nageia fteuryi,Podocarpus nagi或Decuso-corpus nagi改为Nageia nagi,过去误定为Podocarpus brevifolius或P.wangii应改为Podocarpus pilgeri;Kereleeria pubescens应归并于K.davidiana,K.hainanensis归并于K.evelyniana,K.eyelolepis和K.oblonga归并于K.fortunei;Tsuga longibra-cteata改为Nothotsuga longibracteata;Cupressus funebris改为Chamaecyparis fune-bris,不同意把Pinus Kwangtungensis改为p.wangii var.kwangtungensis等。     

松科4属植物茎初生结构比较研究

研究比较观察了松科云杉属的云杉（Ｐｉｃｅａ ａｓｐｅｒａｔａ）、油杉属的油杉（Ｋｅｔｅｌｅｅｒｉａ ｆｏｒｔｕｎｅｉ）、雪松属的雪松（Ｃｅｄｒｕｓ ｄｅｏｄｅｒａ）、松属的海南五针松（Ｐｉｎｕｓ ｆｅｎｚｅｌｉａｎａ）和大明松（Ｐ．ｔａｉｗａｎｅｎｓｉｓ Ｈａｙａｔａ ｖａｒ．ｄａｍｉｎｇｓｈａｎｅｎｓｉｓ）５个种的幼茎初生结构。结果表明,皮下层的细胞层数、皮层细胞的组成,树脂道的分布,鞘细胞的排     

光学显微镜下松柏类和菊科花粉的分类、鉴定要点及生态意义

本文对地层中广泛分布的松柏类和菊科花粉进行初步分类,讨论各类型(属)的鉴定特征及生态意义.以气囊的有无、外壁结构和纹饰、气囊的形态和纹饰、本体与气囊过渡点的特征、帽与帽缘纹理等为鉴定要点,对杉型、柏科型、刺柏型、红豆杉型、落叶松属、铁杉属、罗汉松属、云杉属、雪松属、冷杉属、油杉属和松属(双束松型和单束松型)13个类型的松柏类植物花粉逐步分类.通过总结前人研究和统计分析,讨论落叶松属、铁杉属、罗汉松属、云杉属、雪松属、冷杉属、油杉属和松属,特别是双束松型和单束松型花粉的生态环境指代意义.依据花粉的大小、外壁表面纹饰、外壁光切面中基柱层的结构和发育程度,初步拟将菊科花粉划分为6个类型,即菊苣-蒲公英型、紫菀型、春黄菊型、风毛菊型、蓝刺头型和蒿属,并通过对6个花粉类型植物母体生态环境的归纳总结和统计分析,讨论它们的生态环境指代意义.     

Distribution of diamine oxidase activity and polyamine pattern in bean and soybean seedlings at different stages of germination

Diamine oxidase (DAO, EC 1.4.3.6.) activity and polyamine content were measured in the shoot apex, leaves, epicotyl, cotyledons, hypocotyl and roots of light-grown bean ( Phaseolus vulgaris L. cv. Lingot) and soybean ( Glycine max L. cv. Sakai) seedlings at 3 different stages of germination (5, 8 and 14 days) as well as in embryos and cotyledons from soaked seeds. No DAO activity was detected in embryos and cotyledons of either plants. In bean seedlings DAO activity was only detectable in the shoot apex, primary leaves and cotyledons, while in soybean the activity was only detectable in the hypocotyl and roots. During seedling growth, in both plants, a different pattern of DAO activity was observed. In both species spermidine and spermine were the most abundant polyamines in embryos and cotyledons. Cadaverine, absent in bean, was only detected in soybean embryos. In the seedlings of both plants, increasing gradients of putrescine, spermidine and spermine from base to shoot apex were found. A high concentration of cadaverine was present in soybean hypocotyls and roots. A possible correlation between DAO activity and the endogenous content of the preferential substrate is discussed in relation to the possible involvement of the enzyme in regulating the cellular level of polyamines.     

Distribution and hosts of Callidiellum rufipenne (Coleoptera: Cerambycidae), an asian cedar borer established in the eastern United States

The distribution and hosts of the exotic cedar-boring beetle, Callidiellum rufipenne (Motschulsky) (Coleoptera: Cerambycidae), were determined in five northeastern U.S. states by capturing adults on cedar trap logs and by rearing adults from various conifers. This beetle was detected in the coastal states of Massachusetts, Rhode Island, Connecticut, New York, and New Jersey. In these states, adults emerged from the live or dead wood of four genera and eight species of Cupressaceae; species of Pinaceae were not hosts. Through its entire range, C. rufipenne is reported to infest at least 14 species of Cupressaceae, four species of Pinaceae, and one species of Taxaceae; but, records of Pinaceae and possibly Taxaceae are suspect. Based on the number of adults that emerged from coniferous poles in a five-way choice test in the field, the infestation level was significantly greater in Chamaecyparis thyoides (L.) Britton, Sterns, and Poggenburg and Juniperus virginiana L. than in Pinus rigida Miller, Pinus strobus L., and Tsuga canadensis (L.) Carribre (last three species uninfested). In a second test of host preference in the wild, beetles infested four cupressaceous species, but not Abies balsamea (L.) Miller, Picea rubens Sargent, Pinus rigida, P. strobus, and Ts. canadensis in the Pinaceae. Infestation level was highest in Ch. thyoides, followed in decreasing order by Juniperus communis L., Thuja occidentalis L., and J. virginiana. In a comparison of live and dead J. virginiana, beetles developed to adults only in dead trees (36 beetles per tree). When trunk sections of Th. occidentalis with and without bark were offered to females in cages, beetles of the next generation emerged exclusively from wood with bark. In the Northeast, only species of Cupressaceae apparently are suitable hosts for C. rufipenne. Infestation of these species may be prevented or reduced by proper care of live plants and by debarking trees after harvesting.     

Nutrient concentrations in litterfall from some western conifers with special reference to calcium

Foliar litterfall nutrient concentrations were analysed for selected members of Taxodiaceae and Cupressaceae families andPseudotsuga menziesii for two arboreta in western Oregon and Washington. Nutrient results between arboreta show similar concentrations with the exception of magnesium, which may be the result of historical land use. Nutrient concentrations between species vary considerably.Pseudotsuga menziesii is particularly distinctive from the Cupressaceae and Taxodiaceae by retaining large amounts of phosphorus and potassium. Taxodiaceae is distinctive by high concentration of Mg while Cupressaceae retains calcium, especiallyChamaecyparis nootkatensis. Results suggest that all members of Taxodiaceae and Cupressaceae retain considerably more Ca than Pinaceae in foliar litter.     

云南松天然林区植物群落结构的空间动态研究

于2005年4月采用中点四分法对云南省禄丰县一平浪林场云南松天然林区植物群落结构及空间动态进行了实地调查研究.结果表明:从山坡上部到下部,物种的组成越来越丰富,其中的优势种群云南松属于衰退型、滇油杉属于增长型、黄毛青冈属于稳定型.根据更新层的重要值来判断,山坡上部将演替成为以滇油杉为主的单优群落;山坡中部将演替成为滇油杉和黄毛青冈为主的针叶、阔叶混交林;山坡下部将演替成为以黄毛青冈为主的常绿阔叶林.研究还表明,在林冠层中,除了滇油杉在山坡上部为聚集分布外,其他树种的分布格局均呈随机分布;在更新层中,优势种类均为聚集分布.     

MORPHOGENETIC ASPECTS OF A LEAFLESS MUTANT IN TOMATO I. GENERAL PATTERNS IN DEVELOPMENT

The embryo of the reduced form of the lanceolate mutant in tomato fails to undergo the heart-shaped stage of development. Cells in the shoot apical region of this leafless mutant lose their meristematic character and develop into mature parenchyma during embryogenesis. This early loss of meristem tissue leads to the determinate growth which is evident in the seedling. In contrast to normal, starch grains are visible with the light microscope in cells of the shoot tip of the mutant hypocotyl from early embryogeny up to and including the seedling stage, and protein bodies are abundant in the same tissue of fully developed mutant embryos. The shoot apical region in homozygous mutant embryos with cotyledons or cotyledon-like structures exhibits some cytochemical and morphological similarity with the normal shoot apex. Morphological variation in these forms appears to be in a continuous pattern. The extent of their development and consequent longevity is related to possible differences in rates of cell expansion and variation in environmental factors during the early stages of embryogenesis.     

Study of aberrant morphologies and lack of conversion of somatic embryos of chickpea (Cicer arietinum L.)

Summary Somatic embryos which originated from mature embryo axes of the chickpea (Cicer arietinum L.) showed varied morphologies. Embryos were classified based on shape of the embryo and number of cotyledons. “Normal” (zygotic-like) embryos were bipolar structures with two cotyledons and a well-developed shoot and root apical meristem, whereas “aberrant” embryos were horn-shaped, had single and multiple cotyledons, and were fasciated. Histological examination revealed the absence of a shoot apical meristem in horn-shaped embryos. Fasciated embryos showed diaxial fusion of two embryos. Secondary embryogenesis was also observed, in which the embryos emerged from the hypocotyl and cotyledonary region of the primary somatic embryo. This report documents the absence of an apical meristem as a vital factor in the lack of conversion of aberrant somatic embryos.     

The occurrence of vertical resin canals in Keteleeria, with reference to its systematic position in Pinaceae

The variation in the number of resin canals in the secondary xylem of Keteleeria fortunei (A. Murray bis) Carrière is determined. At all levels from base to top of a tree, resin canals are absent rearer the pith, but they (sporadically) appear after a certain cambial age. In addition, the occurrence of resin canals in seven named species of Keteleeria was surveyed. It was confirmed that normal vertical resin canals do occur in the mature parts of secondary xylem, but that they may be absent in the juvenile parts of secondary xylem of stems, as well as in branches and in seedlings or saplings. Earlier investigations reporting the absence of resin canals altogether in this genus may have been misled by the use of younger (immature) specimens instead of mature stems of trees. The distribution pattern of resin canals found in Keteleeria is systematically significant because it supports a new system of classification of Pinaceae, in which Keteleeria is believed to have a close relationship with Nothotsuga.     

Molecular phylogeny and evolution of host-plant use in conifer seed chalcids in the genus Megastigmus (Hymenoptera: Torymidae)

Abstract. Phylogenetic relationships amongst Megastigmus species (Chalcidoidea: Torymidae) associated with conifer seeds were inferred from DNA sequence data. Twenty‐nine species of seed chalcids were analysed using two different genes, cytochrome b (mitochondrial DNA) and the D2 domain of the 28S ribosomal DNA. Maximum‐parsimony and maximum‐likelihood analyses showed that taxa formed two monophyletic groups, one clade comprising all species associated with Cupressaceae and Taxodiaceae hosts with the exception of Chamaecyparis, and the other clade composed of species associated with Pinaceae. Species infesting Cupressaceae and Taxodiaceae seemed to be specialized to particular host genera or even to be species specific, which was consistent with a taxonomic radiation following initial host adaptation. By contrast, Megastigmus species associated with Pinaceae appeared capable of shifting onto different congeneric species or even onto a new host genus, with their evolution apparently less constrained by plant association. We hypothesized that the Megastigmus group associated with Pinaceae may have a much higher invasive potential than that related to Cupressaceae. The study also confirmed the presence of invasive Nearctic species in the Palaearctic, and demonstrated the existence of a cryptic species complex.     

AGAMOUS subfamily MADS-box genes and the evolution of seed cone morphology in Cupressaceae and Taxodiaceae

In this comparative developmental genetics study, we test hypotheses based on fossil and morphological data on reproductive organ morphology and evolution in conifers--specifically, the ovule-bearing organ in Cupressaceae and Taxodiaceae. Genes homologous to the Arabidopsis gene AGAMOUS are expressed in ovuliferous scales of spruces (Picea) throughout development. Previous studies have shown that the AGAMOUS subfamily of MADS-box genes predates the split between angiosperms and gymnosperms, and that these genes have in part conserved functions in reproductive development among seed plants, especially in the specification of identity of the ovule-bearing organs. These data indicate that their expression in conifer families other than Pinaceae might be used as markers for organs homologous to the Pinaceae ovuliferous scale. Here we have isolated putative AGAMOUS orthologs from Cupressaceae and Taxodiaceae and analyzed their expression pattern in seed cones to test for the presence of morphological homologs of ovuliferous scales. Our results were not congruent with the hypothesis that the tooth of the Cryptomeria seed cone is homologous to the Picea ovuliferous scale. Likewise, the hypothesis that the bracts of Thujopsis and Juniperus contain fused ovuliferous scales was not supported. However, we found expression of AGAMOUS homologs in the sterile bracts of Cupressaceae seed cones at late developmental stages. This expression probably represents a novel gene function in these conifer families, since no corresponding expression has been identified in Pinaceae. Our study suggests that the evolutionary history of modern conifer cones is more diverse than previously thought.     

Molecular phytogeny of conifers using RFLP analysis of PCR-amplified specific chloroplast genes

We investigated the molecular phylogeny of conifers using restriction endonuclease fragment length polymorphism of six polymerase chain reaction-amplified chloroplast genes — frxC, rbcL, psbA, psbD, trnK, and 16S. We detected 227 total site changes among species, representing 23, 26, 38, 48, 67, and 25 site changes in frxC, psbA, psbD, rbcL, trnK and 16S, respectively. The mean nucleotide substitution was 10.75% (SD 0.573) among species in five families. Forty maximally parsimonious trees were obtained using the Wagner parsimony method, and a 50% majority-rule consensus tree was obtained from them. Data analysis produced similar basic patterns when both the Wagner parsimony and the neighbor-joining methods were applied, and the main lineages were clearly separated. Taxaceae and Cephalotaxaceae species were used as the out-groups when applying Wagner parsimony methods. With the Wagner method, the consistency index was 0.510, the retention index was 0.879, and tree length was 435 steps. Our results indicated that Cupressaceae and Taxodiaceae are closely related families and that Sciadopitys verticillata is the basal lineage of Cupressaceae and Taxodiaceae. The neighbor-joining tree is similar to the 50% majority-rule consensus of the 40 Wagner parsimony trees except for the position of Keteleeria daversifolia, the Picea and Cedrus group, and the divergence within Cupressaceae.     

Fundamental features of the distribution of Coniferae in Sichuan

The abundance of Coniferae in Szechuan Province is a well-known fact, especially of the Order Pinales. There are so far 19 genera 68 species and varieties belonging to the Pinaceae, Taxodiaceae and Cupressaceae. This paper dealing with the fundamental features in the distribution of these plant-groups may be not only of scientific interest but also of great importance to forestry. According to an analysis of the influence of historic, geographic and ecological conditions on the distribution of plants, it is shown that the differantiation of geographical distribution of genera and species in these 3 familias between the eastern and the western areas is very evident. There are (Tab. 1) 18 genera and 27 species in the eastern area, including 6 monotypic genera, they are Cathaya, Pseudolarix, Metosequoia, Glyptostrobus, Fokienia and Platycladus. As the great concentration of the monotypic and paleo-endemic genera is not found elsewhere, this area therefore distinctly indicates the relic nature. It seems most probably that they came originally into existence in subtropical and warm-temperate climates of middle lalitudes in the northern hemisphere, and most of them are in the lands bordering the Pacific Ocean. On the other hand, there are 13 genera and 56 species in the western area with only one monotypic genus (Platycladus) present, and most genera are polytypic with wide ranges. The number of species in these genera here is larger than in any other areas, for example, Abies and Picea are represented 12 species each (Fig. 1-4). There are 10 species in Sabina (Fig. 5). It might be considered that each genus has developed into a great centre, In addition, it has been discussed in detail regarding the vicarious distributions in geography, verticality, and ecology of ralated species in Abies, Picea, Tsuga, Keteleeria, Larix, Pinus and Cupressus (Fig. 3-5), as well as the re lationship of these species. As shown above, these plants are also, without doubt, of very old origin. Moreover, they are most likely very much closely related with Tethys in the past, but the majority species apparently have arisen in relalively recent times. They are due to the effect of mountain-making and glaciation, principally in consequence of the cooling process of the climatic vicissitude.     

Precociously germinating rapeseed embryos retain characteristics of embryogeny

We compared the germination of Brassica napus L. embryos at three stages of development-mid-cotyledon, maturation and mature dry-to determine at which stage they acquired the capacity for normal germination and seedling development. Embryos were removed from the seed and cultured on hormone-free medium, allowing them to germinate. The transition from embryogeny to germination was monitored both morphologically and biochemically, using synthesis of 12 S storage protein as a marker of embryogeny. The mature embryos (dry seeds) set the standard for normal seedling development: radicle emergence, hypocotyl extension and cotyledon expansion occurred within 2 d and true leaves were formed within a week of germination. Rocket immunoelectrophoresis indicated that the storage proteins in seedlings from mature dry embryos were completely degraded within a week. In contrast, the midcotyledon-stage embryos appeared to germinate abnormally, retaining many embryonic characteristics. Although the roots emerged, the hypocotyls did not elongate and secondary cotyledons instead of leaves were formed at the shoot apex. Also, the seedlings continued to synthesize and accumulate storage proteins. The maturation-stage embryos did develop into normal-looking seedlings, but complete degradation of storage proteins required several weeks, presumably reflecting continued synthesis and turnover. We conclude that embryogenic and germination-specific processes can occur concurrently and that the capacity to develop as normal seedlings is acquired gradually during the maturation process.Abbreviations dpa days post anthesis - EDTA ethylenedi-aminetetraacetic acid - FW fresh weight     

The gene ENHANCER OF PINOID controls cotyledon development in the Arabidopsis embryo

During Arabidopsis embryo development, cotyledon primordia are generated at transition stage from precursor cells that are not derived from the embryonic shoot apical meristem (SAM). To date, it is not known which genes specifically instruct these precursor cells to elaborate cotyledons, nor is the role of auxin in cotyledon development clear. In laterne mutants, the cotyledons are precisely deleted, yet the hypocotyl and root are unaffected. The laterne phenotype is caused by a combination of two mutations: one in the PINOID (PID) gene and another mutation in a novel locus designated ENHANCER OF PINOID (ENP). The expression domains of shoot apex organising genes such as SHOOT MERISTEMLESS (STM) extend along the entire apical region of laterne embryos. However, analysis of pid enp stm triple mutants shows that ectopic activity of STM does not appear to cause cotyledon obliteration. This is exclusively caused by enp in concert with pid. In pinoid embryos, reversal of polarity of the PIN1 auxin transport facilitator in the apex is only occasional, explaining irregular auxin maxima in the cotyledon tips. By contrast, polarity of PIN1:GFP is completely reversed to basal position in the epidermal layer of the laterne embryo. Consequently auxin, which is believed to be essential for organ formation, fails to accumulate in the apex. This strongly suggests that ENP specifically regulates cotyledon development through control of PIN1 polarity in concert with PID.     

Role of 2,4-dichlorophenoxyacetic acid (2,4-D) in somatic embryogenesis on cultured zygotic embryos of Arabidopsis: cell expansion, cell cycling, and morphogenesis during continuous exposure of embryos to 2,4-D

The relationship between cell expansion and cell cycling during somatic embryogenesis was studied in cultured bent-cotyledon-stage zygotic embryos of a transgenic stock of Arabidopsis thaliana harboring a cyclin 1 At:β-glucuronidase (GUS) reporter gene construct. In embryos cultured in a medium containing 2,4-dichlorophenoxyacetic acid (2,4-D), following a brief period of growth by cell expansion, divisions were initiated in the procambial cells facing the adaxial side at the base of the cotyledons. Cell division activity later spread to almost the entire length of the cotyledons to form a callus on which globular and heart-shaped embryos appeared in about 10 d after culture. Anatomical and morphogenetic changes observed in cultured embryos were correlated with patterns of cell cycling by histochemical detection of GUS-expressing cells. Although early-stage somatic embryos did not develop further during their continued growth in the auxin-containing medium, maturation of embryos ensued upon their transfer to an auxin-free medium. In a small number of cultured zygotic embryos the shoot apical meristem was found to differentiate a leaf, a green tubular structure, or a somatic embryo. Contrary to the results from previous investigations, which have assigned a major role for the shoot apical meristem and cells in the axils of cotyledons in the development of somatic embryos on cultured zygotic embryos of A. thaliana, the present work shows that somatic embryos originate almost exclusively on the callus formed on the cotyledons. Other observations such as the induction of somatic embryos on cultured cotyledons and the inability of the embryo axis (consisting of the root, hypocotyl, and shoot apical meristem without the cotyledons) to form somatic embryos, reaffirm the important role of the cotyledons in somatic embryogenesis in this plant.     

Mapped DNA probes from loblolly pine can be used for restriction fragment length polymorphism mapping in other conifers

A high-density genetic map based on restriction fragment length polymorphisms (RFLPs) is being constructed for loblolly pine (Pinus taeda L.). Consequently, a large number of DNA probes from loblolly pine are potentially available for use in other species. We have used some of these DNA probes to detect RFLPs in 12 conifers and an angiosperm. Thirty complementary DNA and two genomic DNA probes from loblolly pine were hybridized to Southern blots containing DNA from five species of Pinus (P. elliottii, P. lambertiana, P. radiata, P. sylvestris, and P. taeda), one species from each of four other genera of Pinaceae (Abies concolor, Larix laricina, Picea abies, and Pseudotsuga menziesii), one species from each of three other families of Coniferales [Sequoia sempervirens (Taxodiaceae), Torreya californica (Taxaceae) and Calocedrus decurrens (Cupressaceae)], and to one angiosperm species (Populus nigra). Results showed that mapped DNA probes from lobolly pine will cross-hybridize to genomic DNA of other species of Pinus and some other genera of the Pinaceae. Only a small proportion of the probes hybridized to genomic DNA from three other families of the Coniferales and the one angiosperm examined. This study demonstrates that mapped DNA probes from loblolly pine can be used to construct RFLP maps for related species, thus enabling the opportunity for comparative genome mapping in conifers.