Hybrid origin of some ornamentals of Allium subgenus Melanocrommyum verified with GISH and RAPD

 Random amplified polymorphic DNA (RAPD) and genomic in situ hybridization (GISH) methods have been used to verify the hybridogenic origin and to identify the parental species of some ornamental cultivars in the subgenus Melanocrommyum of the genus Allium. The cultivars had been selected from seed obtained after uncontrolled pollination in breeders’ fields. The combination of GISH analysis with RAPD markers is very suitable for testing the hybridogenic origin of plants and to ascertain the parental species of the hybrids in such cases. As suspected, A. macleanii and A. cristophii are the parental species of ‘Globemaster’. The parental species of cultivar ‘Globus’ are A. karataviense and A. stipitatum, and not A. cristophii and A. giganteum as has been assumed on morphological grounds. Cultivars ‘Lucy Ball’ and ‘Gladiator’ are of hybrid origin, though only one of the parental species, A. hollandicum, could be confirmed. The cultivars ‘Purple Sensation’, ‘Mount Everest’, ‘White Giant’, ‘Michael H. Hoog’ and ‘Mars’ are not hybrids since neither GISH nor RAPD suggest the presence of a second genome. ‘Purple Sensation’ belongs to A. hollandicum, ‘Mount Everest’, ‘White Giant’ and ‘Mars’ to A. stipitatum,‘Michael H. Hoog’ to A. rosenorum. Received: 3 July 1997 / Accepted: 9 October 1997     

Parasitism ofLeptinotarsa decemlineata [Coleoptera: Chrysomelidae] byEdovum puttleri [Hymenoptera: Eulophidae] in different cultivars of eggplant

Experiments were conducted to quantify parasitism of Colorado potato beetle,Leptinotarsa decemlineata (Say), by the egg parasitoid,Edovum puttleri Grissell, on 3 different cultivars of eggplant,Solanum melongena L. Levels of parasitism were higher (P<0.05) on ‘Black Pride’ than on other cultivars. The percentage of egg masses that were parasitized was 1.2-fold higher (P<0.05) on ‘Black Pride’ than on ‘Harris Special’ and ‘White’. The number of eggs per mass that were parasitized was 1.3- and 1.4- fold greater (P<0.05) on ‘Black Pride’ than on ‘Harris Special’ and ‘White’, respectively. The percentage of eggs that were parasitized per mass and percentage of emerged adult parasitoids did not differ (P>0.05) among cultivars; between 2.1- to 2.6- fold more females than males emerged from eggs on all cultivars during the growing season.Edovum puttleri suppressed the 2nd generation ofL. decemlineata on ‘Black Pride’ and ‘Harris Special’, but did not suppress populations on ‘White’.      

Late-acting self-incompatibility in tea plant (<Emphasis Type="Italic">Camellia sinensis</Emphasis>)

The self-incompatibility of tea plant (Camellia sinensis (L.) O. Kuntze) was studied with the methods of aniline blue fluorescence assay and paraffin sections. The characteristics of pollen tube elongation after hand pollination was analyzed in 4 tea cultivars, including ‘Keemenzhong’, ‘Longjing-changye’, ‘Fuding-dabaicha’ and ‘Yabukita’, under self-pollination and cross-pollination, respectively. Although there were some difference among cultivars, pollen tubes elongated through the style and reach the ovary successfully at 48 h after pollination for both cross- and self-pollen tubes in all the four cultivars of tea. Pollen tubes entered into the ovule micropyles, however, only for cross-pollination, but not for self-pollination. Pollen tubes of selfing plants, failed in fertilizing, seemed have some difficulties to enter the ovule. All of which indicated that the self-incompatibility of tea plant is a late-acting self-incompatibility system (LSI) or an ovarian sterility (OS), in which the self incompatibility was due to none self pollen tube penetrating into the ovule and no fertilization.     

Molecular mapping of leaf rust resistance gene <Emphasis Type="Italic">Rph14</Emphasis> in <Emphasis Type="Italic">Hordeum vulgare</Emphasis>

An incompletely dominant gene conferring resistance to Puccinia hordei, Rph14, identified previously in an accession of Hordeum vulgare, confers resistance to all known pathotypes of P. hordei in Australia. Knowledge of the chromosomal location of Rph14 and the identification of DNA markers closely linked to it will facilitate combining it with other important leaf rust resistance genes to achieve long lasting resistance. The inheritance of Rph14 was confirmed using 146 and 106 F₃ lines derived from the crosses ‘Baudin’/‘PI 584760’ (Rph14) and ‘Ricardo’/‘PI 584760’ (Rph14), respectively. Bulk segregant analysis on DNA from the parental genotypes and resistant and susceptible DNA bulks using DArT markers located Rph14 to the short arm of chromosome 2H. DArT marker bPb-1664 was identified as having the closest genetic association with Rph14. PCR based marker analysis identified a single SSR marker, Bmag692, linked closely to Rph14 at a map distance of 2.1 and 3.8 cm in the ‘Baudin’/‘PI 584760’and ‘Ricardo’/‘PI 584760’ populations, respectively.     

ABA and proline accumulation in leaves and roots of wild (<Emphasis Type="Italic">Hordeum spontaneum</Emphasis>) and cultivated (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> ‘Maresi’) barley genotypes under water deficit conditions

The purpose of the study was to examine water stress-induced changes in the ABA and proline contents in roots and leaves of a potentially more resistant wild accession of Hordeum spontaneum and the modern cultivar Maresi (Hordeum vulgare). Leaves of H. spontaneum had higher contents of constitutive ABA and proline in comparison to those of ‘Maresi’. A moderate water deficit resulted only in root dehydration, which was higher in ‘Maresi’. Increases of water deficit in roots coincided with an increase of ABA content in roots, followed by that in leaves. The level of proline increased only in leaves and only in the case of H. spontaneum. Under conditions of severe water stress, the root dehydration levels were similar in the both genotypes, whereas leaf dehydration was higher in ‘Maresi’. H. spontaneum, as compared to ‘Maresi’ showed an earlier increase of ABA content in the roots and accumulated more ABA in the leaves. Free proline levels in the roots increased in both genotypes but H. spontaneum exhibited a 2-fold higher proline accumulation than ‘Maresi’. In H. spontaneum the accumulation of proline in the leaves occurred noticeably earlier and to a higher extent than in ‘Maresi’. A possible connection of these modifications with water stress resistance of the investigated genotypes is discussed in this paper.     

Taxonomic status of the hominid mandible KNM-ER TI 13150 from the middle pliocene of tabarin,in Kenya

A partial mandible with two molars intact was recovered between 1981 and 1984 from deposits of the Middle Pliocene at Tabarin, in Kenya. It has been described and assigned toAustralopithecus cf.afarensis Johanson, White, andCoppens, 1978, with the condition that if ‘A. afarensis’ is revised, then the attribution may change. The taxon ‘A. afarensis’ was found to be invalid and was revised. The smaller specimens of ‘A. afarensis,’ to which the Tabarin mandible was said to be similar, were redescribed asHomo antiquus Ferguson, 1984. Since the Tabarin mandible andH. antiquus are successive transients of the same gens and are allopatric, the Tabarin hominid population is described as an earlier chronosubspecies,Homo antiquus praegens ssp. n.     

Resistance to Phytophthora fragariae var. fragariae in strawberry: the Rpf2 gene

  Phytophthora fragariae var. fragariae is the causal agent of red stele (red core) root rot in strawberry (Fragaria spp.). The inheritance of resistance to one isolate of this fungus was studied in 12 segregating populations of F.×ananassa derived from crosses between four resistant cultivars (‘Climax’, ‘Redgauntlet’, ‘Siletz’, and ‘Sparkle’) and three susceptible cultivars (‘Blakemore’, ‘Glasa’, and ‘Senga’ Sengana’). The analysis clearly supports the hypothesis of a single segregating dominant resistance gene. It is proposed that this gene be designated Rpf2. Received 12 November 1996 / Accepted: 22 November 1996     

Estimation of mortality by entomophages on exotic Harmonia axyridis versus native Adalia bipunctata in semi-field conditions in northern Italy

A semi-field experiment was carried out in two peach orchards in northern Italy to assess mortality due to predators and parasitoids on the exotic coccinellid Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) in comparison with the native coccinellid Adalia bipunctata L. (Coleoptera: Coccinellidae). The experiments were conducted in cages to avoid the possible escape of the exotic ladybird (not yet established in Italy). Two kinds of cage experiments were included: ‘exclusion cages’ (access by walking predators impeded) and ‘free cages’ (walking predators free to enter). The cages, containing all the stages of the two ladybird species, were placed in two localities and left for 24 h. All ladybird stages used for the semi-field experiments came from a laboratory rearing. The eggs of H. axyridis experienced less mortality than those of A. bipunctata. The ant workers were the most frequent predators in ‘free cages’ but A. bipunctata cannibalism on eggs was also detected. Larvae of both coccinellid species were predated equally but larval predation of L1 and L2 was higher in comparison to predation of L3 and L4. Pupae and adults of both exotic and native ladybirds were never attacked by predators. Predation on younger larval stages was higher in the ‘free cages’ in comparison with ‘exclusion cages’. No ladybird parasitisation was observed. The ‘free cage’ technique seems to provide a standardised and realistic estimation of predation impact but more studies are needed to evaluate ladybird parasitisation in semi-field conditions.     

Genetic Discrimination of <Emphasis Type="Italic">Catharanthus roseus</Emphasis> Cultivars by Pyrolysis Mass Spectrometry

Pyrolysis mass spectrometry (PyMS) is a rapid, simple, high-resolution analytical method based on thermal degradation of complex materials in a vacuum. It is widely applied to the discrimination of closely related microbial strains. Leaf samples from eight cultivars (‘Apricot Delight’, ‘Cooler Grape’, ‘Cooler Peppermint’, ‘Equator Grape’, ‘Equator Rose’, ‘Equator White’, ‘Equator White Eye’, and ‘Little Bright Eye’) of Catharanthus roseus were subjected to PyMS for spectral fingerprinting. Discriminant analysis (DA) of PyMS data enabled us to assign these cultivars to discrete clusters. A hierarchical dendrogram based on DA provided a possible relationship among them that was in general agreement with a previously reported classification of the cultivars based on DNA fingerprints. Furthermore, those belonging to the same ‘series’ were grouped into a single cluster, which previously could not be achieved through similar approaches based on Fourier transform infrared spectroscopy or ¹H NMR data. Overall results suggest that chemical differences (i.e., in pyrolysate composition) among cultivars, as detected by mass spectrometry, reflect their genetic variation.     

An <Emphasis Type="Italic">in vitro</Emphasis> method to evaluate grapevine cultivars for <Emphasis Type="Italic">Erysiphe necator</Emphasis> susceptibility

Powdery mildew, caused by the obligate biotrophic ascomycete Erysiphe necator, is one of the most destructive grapevine diseases worldwide. Cultivars of Vitis vinifera L, for wine and table grape production, are all susceptible to E. necator, whose attacks result in severe epidemics under the warm and dry conditions of the Mediterranean basin. The aim of the present study was to compare the susceptibility of different grapevine cultivars to E. necator by an in vitro assay for assessing the potentiality of this method in breeding programs for resistance to the pathogen. Leaves of 12 grapevine cultivars were spot-inoculated in vitro with about 10 conidia from five different isolates of E. necator, using colony growth and conidiation 3 wk post-inoculation as indicators of susceptibility to the disease. A remarkable difference was observed between highly susceptible cultivars like ‘Baresana’, ‘Malvasia’, ‘Bianca’, and ‘Italia’, and the less susceptible ‘Alphonse Lavallée’ and ‘Ohanez’, in accordance with their behavior in the field. No statistically significant differences were found in the virulence of E. necator isolates.     

Localization of the rice stripe disease resistance gene, Stv-bi, by graphical genotyping and linkage analyses with molecular markers

 We used graphical genotyping and linkage analyses with molecular markers to determine the chromosomal location of the rice stripe disease resistance gene, Stv-b ⁱ . The stripe resistance gene from the indica rice (Oryza sativa) cv ‘Modan’ was introgressed into several Japanese rice varieties. We found 4 RFLP markers in ‘Modan’, five susceptible parental rice varieties (‘Norin No. 8’, ‘Sachihikari’, ‘Kanto No. 98’, ‘Hokuriku No.103’ and ‘Koganebare’) and four resistant progeny varieties (‘St. No. 1’, ‘Aichi No. 6’, ‘Aoisora’ and ‘Asanohikari’). Graphical genotyping of the resistant progeny revealed a chromosomal segment ascribable to ‘Modan’ and associated with stripe resistance. The chromosomal segment from ‘Modan’ was located at 35.85 cM on chromosome 11. Linkage analysis using 120 F₂ individuals from a cross between ‘Koshihikari’ (susceptible) and ‘Asanohikari’ (resistant) revealed another 8 RFLP markers in the same chromosome. We performed a bioassay for rice stripe resistance in F₃ lines of the F₂ individuals using infective small brown planthoppers and identified an 1.8-cM segment harboring the rice stripe disease resistance gene, Stv-b ⁱ , between XNpb220 and XNpb257/ XNpb254. Furthermore, Stv-b ⁱ was linked by 0.0 cM to a RFLP marker, ST10, which was developed on the basis of the results of RAPD analysis. These DNA markers near the Stv-b ⁱ locus may be useful in marker-assisted selection and map-based cloning of the Stv-b ⁱ gene. Received: 26 September 1997 / Accepted: 4 November 1997     

Association of barley photoperiod and vernalization genes with QTLs for flowering time and agronomic traits in a BC2DH population and a set of wild barley introgression lines

The control of flowering time has important impacts on crop yield. The variation in response to day length (photoperiod) and low temperature (vernalization) has been selected in barley to provide adaptation to different environments and farming practices. As a further step towards unraveling the genetic mechanisms underlying flowering time control in barley, we investigated the allelic variation of ten known or putative photoperiod and vernalization pathway genes between two genotypes, the spring barley elite cultivar ‘Scarlett’ (Hordeum vulgare ssp. vulgare) and the wild barley accession ‘ISR42-8’ (Hordeum vulgare ssp. spontaneum). The genes studied are Ppd-H1, VRN-H1, VRN-H2, VRN-H3, HvCO1, HvCO2, HvGI, HvFT2, HvFT3 and HvFT4. ‘Scarlett’ and ‘ISR42-8’ are the parents of the BC₂DH advanced backcross population S42 and a set of wild barley introgression lines (S42ILs). The latter are derived from S42 after backcrossing and marker-assisted selection. The genotypes and phenotypes in S42 and S42ILs were utilized to determine the genetic map location of the candidate genes and to test if these genes may exert quantitative trait locus (QTL) effects on flowering time, yield and yield-related traits in the two populations studied. By sequencing the characteristic regions of the genes and genotyping with diagnostic markers, the contrasting allelic constitutions of four known flowering regulation genes were identified as ppd-H1, Vrn-H1, vrn-H2 and vrn-H3 in ‘Scarlett’ and as Ppd-H1, vrn-H1, Vrn-H2 and a novel allele of VRN-H3 in ‘ISR42-8’. All candidate genes could be placed on a barley simple sequence repeat (SSR) map. Seven candidate genes (Ppd-H1, VRN-H2, VRN-H3, HvGI, HvFT2, HvFT3 and HvFT4) were associated with flowering time QTLs in population S42. Four exotic alleles (Ppd-H1, Vrn-H2, vrn-H3 and HvCO1) possibly exhibited significant effects on flowering time in S42ILs. In both populations, the QTL showing the strongest effect corresponded to Ppd-H1. Here, the exotic allele was associated with a reduction of number of days until flowering by 8.0 and 12.7%, respectively. Our data suggest that Ppd-H1, Vrn-H2 and Vrn-H3 may also exert pleiotropic effects on yield and yield-related traits.     

Plasmid profiles of bacteriocin-producingLactobacillus isolates from African fermented foods

Fourteen of 200Lactobacillus isolates from African fermented foods,viz. ‘wara’, ‘kenkey’, ‘ugba’, ‘ogi’, ‘kunuzarki’, ‘fufu’ and ‘iru’ were found to produce bacteriocins againstL. plantarum and only three bacteriocinogenic isolates inhibited some of the food pathogens. Plasmid analysis of the 14 bacteriocin-producing lactobacilli showed that only 5 isolates harbored plasmids ranging in size from 3.1 to 55.5 kb.     

Accumulation of additive effects generates a strong photoperiod sensitivity in the extremely late-heading rice cultivar ‘Nona Bokra’

Many rice cultivars that originated from lower-latitude regions exhibit a strong photoperiod sensitivity (PS) and show extremely late heading under long-day conditions. Under natural day-length conditions during the cropping season in Japan, the indica rice cultivar ‘Nona Bokra’ from India showed extremely late heading (202 days to heading) compared to the japonica cultivar ‘Koshihikari’ (105 days), from Japan. To elucidate the genetic factors associated with such extremely late heading, we performed quantitative trait locus (QTL) analyses of heading date using an F₂ population and seven advanced backcross progeny (one BC₁F₂ and six BC₂F₂) derived from a cross between ‘Nona Bokra’ and ‘Koshihikari’. The analyses revealed 12 QTLs on seven chromosomes. The ‘Nona Bokra’ alleles of all QTLs contributed to an increase in heading date. Digenic interactions were rarely observed between QTLs. Based on the genetic parameters of the QTLs, such as additive effects and percentage of phenotypic variance explained, these 12 QTLs are likely generate a large proportion of the phenotypic variation observed in the heading dates between ‘Nona Bokra’ and ‘Koshihikari’. Comparison of chromosomal locations between heading date QTLs detected in this study and QTLs previously identified in ‘Nipponbare’ × ‘Kasalath’ populations revealed that eight of the heading date QTLs were recognized nearby the Hd1, Hd2, Hd3a, Hd4, Hd5, Hd6, Hd9, and Hd13. These results suggest that the strong PS in ‘Nona Bokra’ was generated mainly by the accumulation of additive effects of particular alleles at previously identified QTLs.     

Exogenous ethylene influences flower opening of cut roses (<Emphasis Type="Italic">Rosa hybrida</Emphasis>) by regulating the genes encoding ethylene biosynthesis enzymes

The purpose of this paper is to investigate the differential responses of flower opening to ethylene in two cut rose cultivars, ‘Samantha’, whose opening process is promoted, and ‘Kardinal’, whose opening process is inhibited by ethylene. Ethylene production and 1-aminocyclopropane-1-carboxylate (ACC) synthase and oxidase activities were determined first. After ethylene treatment, ethylene production, ACC synthase (ACS) and ACC oxidase (ACO) activities in petals increased and peaked at the earlier stage (stage 3) in ‘Samantha’, and they were much more dramatically enhanced and peaked at the later stage (stage 4) in ‘Kardinal’ than control during vasing. cDNA fragments of three Rh-ACSs and one Rh-ACO genes were cloned and designated as Rh-ACS1, Rh-ACS2, Rh-ACS3 and Rh-ACO1 respectively. Northern blotting analysis revealed that, among three genes of ACS, ethylene-induced expression patterns of Rh-ACS3 gene corresponded to ACS activity and ethylene production in both cultivars. A more dramatic accumulation of Rh-ACS3 mRNA was induced by ethylene in ‘Kardinal’ than that of ‘Samantha’. As an ethylene action inhibitor, STS at concentration of 0.2 mmol/L generally inhibited the expression of Rh-ACSs and Rh-ACO in both cultivars, although it induced the expression of Rh-ACS3 transiently in ‘Kardinal’. Our results suggests that ‘Kardinal’ is more sensitive to ethylene than ‘Samantha’; and the changes of Rh-ACS3 expression caused by ethylene might be related to the acceleration of flower opening in ‘Samantha’ and the inhibition in ‘Kardinal’. Additional results indicated that three Rh-ACSs genes were differentially associated with flower opening and senescence as well as wounding.     

Strict paternal inheritance of chloroplast DNA and maternal inheritance of mitochondrial DNA in intraspecific crosses of kiwifruit

 Previous studies have established that chloroplasts are inherited paternally in Actinidia interspecific crosses. However, fertilisation problems in interspecific crosses may affect the transmission of organelles. Six female clones, i.e. ‘Abbott’, ‘Bruno’, ‘Greensill’, ‘Hayward’, ‘Jones’, ‘Monty’, and four male clones were used to identify cpDNA polymorphisms within the cultivated kiwifruit species A. deliciosa. The restriction patterns by HpaII of a chloroplast fragment amplified by PCR with a pair of universal primers revealed a polymorphism at the intraspecific level. The inheritance of cpDNA in 143 seedlings from three intraspecific crosses in kiwifruit (Actinidia deliciosa) was studied. All offspring displayed the restriction pattern of the paternal parent, indicating that maternal inheritance of cpDNA in kiwifruit is rare at best. Strict maternal inheritance of mtDNA was confirmed in the same crosses used to investigate cpDNA transmission. Studies of cytoplasmic inheritance in the Actinidia genus represent to date the best documented report of differential organelle inheritance of cpDNA and mtDNA in angiosperms. Received: 10 November 1998 / Accepted: 14 December 1998     

Self-sterility in the mutant ‘Zigui shatian’ pummelo (<Emphasis Type="Italic">Citrus grandis</Emphasis> Osbeck) is due to abnormal post-zygotic embryo development and not self-incompatibility

‘Shatian’ pummelo (Citrus grandis Osbeck), one of the main citrus cultivars in China, is self-incompatible, and its pollen tubes are believed to be arrested in style after self-pollination.We have characterized one ‘Shatian’ pummelo mutant, named ‘Zigui shatian’ pummelo. The mutant pummelo had identical DNA ploidy level, morphology (leaf shape, stoma size and density, pollen shape and size) and developmental progress of pistil and male organs to that of the common ‘Shatian’ pummelo. However, unlike the common ‘Shatian’ pummelo, ‘Zigui shatian’ is self-compatible since its pollen tubes can self-pollinate allowing for successful fertilization. Histological analyses of ‘Shatian’ pummelo further verified abnormal post-zygotic development which led to seed abortion. Simple sequence repeats (SSR) analysis revealed polymorphism in 1 of the 120 primers screened showing that ‘Zigui shatian’ and ‘Shatian’ pummelo are different at the DNA level. Taken together, these data suggested mutant ‘Zigui shatian’ pummelo might be derived from ‘Shatian’ pummelo with self-sterility by self-incongruity after self-fertilization.     

Species of <Emphasis Type="Italic">Neohaliotrema</Emphasis> Yamaguti, 1965 (Monogenea: Ancyrocephalidae) from the pomacentrid <Emphasis Type="Italic">Abudefduf vaigensis</Emphasis> (Quoy &; Gaimard) off Pulau Langkawi,Malaysia, with a revised diagnosis of the genus and a key to its species

Four new and one unidentified species of Neohaliotrema Yamaguti, 1965 were obtained from the gills of the Indo-Pacific sergeant Abudefduf vaigensis (Quoy & Gaimard) off Pulau Langkawi, Malaysia. The five species, N. malayense n. sp., N. bombini n. sp., N. andamanense n. sp., N. parvum n. sp. and an unidentified Neohaliotrema sp. (similar to N. macracanthum Zhukov, 1976), are described and distinguished based mainly on features of the haptor. Species of this genus are divisible into two groups, the ‘maomao group’, with two pairs of morphometrically modified ‘marginal’ hooks and a fenestrated haptor, and the ‘gracile group’, with morphologically similar marginal hooks and an entire haptor. With the exception of N. bombini n. sp., the species described fit within the ‘maomao group’. It is suggested that the more complex Neohaliotrema species of the ‘maomao group’ have modified hooks 1 and 2 on a haptoral ‘isthmus’ between two large apertures, i.e. ‘windows’, whereas the less complex species lacking these features are those of the ‘gracile group’. Neohaliotrema spp. have only a single pair of pigmented eye-spots. A fenestrated haptor is unique to the Neohaliotrema spp. of the ‘maomao group’. The generic diagnosis of Neohaliotrema is amended to include new data and a key to its known species is presented.     

Infraspecific variability ofJuniperus L. (Cupressaceae) based on monoterpenoid compositions

The compositions of leaf monoterpenoids from 11 species in the Juniperus section (Juniperus chinensis var. chinensis,J. virginiana, J. communis var. montana,J. rigida, J. chinensis var. globosa,J. chinensis var. sargentii,J. chinensis ‘Kaizuka Variegata’,J. squamata ‘Wilsonii’,J. x media ‘Shimpake’,J. x media ‘Plamosa Aurea’, andJ. squamata ‘Aloderi’) were comparatively analyzed by GC-MS. Of the 24 compounds identified, α-pinene, myrcene, limonene, terpinolene, and bornyl acetate were common to all, but particular combinations differed remarkably among taxa. The simplest composition (eight compounds) was found in.J, chinensis var. chinensis; the most complex (19 compounds), inJ. x media ‘Shimpake’. Cluster analysis generated four distinctive clades within the Juniperus section. The minimum spanning network revealed thatJ. squamata ‘Wilsonii’ andJ. x media ‘Shimpake’ were most similar in their chemical makeup.