Differential Postalightment Oviposition Behavior of Monarch Butterflies on Asclepias Species

The monarch butterfly, Danaus plexippus L., oviposits mainly on plants in the Asclepiadaceae, particularly within the genus Asclepias. We studied postalightment oviposition behavior of monarch females on three host species—Asclepias curassavica, A. incarnata , and A. tuberosa. After landing on the host, they used their forelegs, midlegs, and antennae to assess plant suitability. When these appendages were examined by scanning electron microscopy, contact chemoreceptor sensilla were found. In choice tests, A. incarnata was most preferred, while A. tuberosa was least preferred. However, the use of appendages varied for the different host species. Antennae were most frequently used during post-alightment behavior on A. curassavica, whereas forelegs were used more often on A. incarnata, and all three appendages were used extensively on A. tuberosa. Use of the midlegs was generally followed by use of the antennae. Tasting with either forelegs or antennae apparently may lead to egg laying on some host species. Rupture of the plant surface by midleg spines was also observed. The behavior and host preference of individual females varied significantly and may reflect differences in receptor sensitivity.     

Hybrid bridges to gene flow: a case study in milkweeds (Asclepias)

Natural hybridization occurs throughout areas of sympatry for the North American milkweeds Asclepias exaltata and A. syriaca (Asclepiadaceae), even though the formation of F1 hybrid seed is a rare event. For introgressive hybridization to proceed, F1 and advanced hybrids must be released from reproductive barriers and successfully mate with one or both parental species. I investigated the mating system of natural hybrids between A. exaltata and A. syriaca in three populations in Shenandoah National Park, Virginia. Allozyme data and a maximum-likelihood procedure were used to estimate the frequency of six genotypic classes (parentals, F1, F2, and backcrosses) of the hybridizing populations, the pollinia received by hybrid plants, and the paternal parents of seeds produced by hybrids. F1 hybrids, backcross A. syriaca, and parental A. syriaca individuals were common in three hybrid populations. Even though self-pollinations and interhybrid pollinations were common, F2 seed production and the occurrence of F2 individuals were rare in hybrid populations. Hybrid plants received more pollen from A. syriaca than A. exaltata, which resulted in the production of more backcross-A. syriaca seed than backcross-A. exaltata seed. Asclepias exaltata was rare in the hybrid populations, but A. exaltata pollinia were received by hybrids and this species sired between 15% and 36% of the seeds produced on hybrids. The potential for introgression with A. exaltata populations is lower because this species is unsuccessful as the maternal parent in interspecific and backcross hand-pollinations. The asymetry of hybridization with A. syriaca as the maternal parent is further supported by the incorporation of maternally inherited chloroplast DNA markers in hybrids. Hybrid milkweeds frequently backcross with both parental species and may be released from the reproductive barriers that limit the formation of F1 hybrids in natural populations. The direction of interspecific gene flow and introgression in milkweeds is influenced by the reproductive biology of hybrids, the constituency of the surrounding population, and failure of some crosses to produce seeds. Finally, introgressive hybridization remains an important evolutionary force even when the initial formation of F1 hybrids in natural populations is rare.     

Changes in Structure of Phytomonas elmassiani in Experimental Infections of Apocynaceae, a Presumably Foreign Plant Host

SYNOPSIS. Allamanda nereifolia and Plumeria rubra, plants belonging to the family Apocynaceae were infected experimentally with the trypanosomatid flagellate, Phytomonas elmassiani. In these presumably exotic hosts, P. elmassiani, ordinarily infecting plants belonging to the family Asclepiadaceae, underwent marked structural changes. The presence of numerous choanomastigote like organisms along with a certain proportion of promastigotes and amastigotes, would, under ordinary circumstances, result in classification of the infecting organisms as Crithidia. Obviously, the change in environment from Asclepiadaceae to the closely related Apocynaceae, exerts a profound influence on structure of the organism.     

The Relation of Migration of Oncopeltus fasciatus to Distribution of Phytomonas elmassiani in the Eastern United States*

SYNOPSIS. A total of 4,866 plants belonging to the family Asclepiadaceae was examined in the eastern part of the United States; 291 (6%) of 11 species naturally infected with Phytomonas elmassiani; 9 of these were new host records. Eight of the 9 infected species belonged to the genus Asclepias, one to Cynanchum. Infections were recorded between 25.5° and 41.0° N latitude, and from 74.0° to 90.5° west longitude. At the southernmost latitude Asclepias curassavica, an exotic annual, perennates, harbors phytomonads and is infested the year round with Oncopeltus. Usually but not always infection is correlated with the presence and abundance of insect vectors. The annual die-back of the usually infected perennial plant hosts, the uninfected emerging vernal growth, the reservoir nidus of infected plants in the southern latitudes, and the known proclivity of O. fasciatus to migrate suggest the spread of flagellosis of Asclepiadaceae thru migration of the vector hemipteron.     

不同寄主植物对幻紫斑蝶产卵选择及种群适合度的影响

通过研究幻紫斑蝶(Euploea core Cramer)成虫对6种寄主植物和2种非寄主植物的产卵选择,以及用5种寄主植物饲养下幻紫斑蝶的发育、存活和繁殖情况,为幻紫斑蝶人工规模化养殖提供理论依据。幻紫斑蝶成虫优先选择聚果榕,其次选择古钩藤和马利筋进行产卵;用黄花夹竹桃饲养的幻紫斑蝶发育历期最长,用马利筋饲养的幻紫斑蝶发育历期最短;用古钩藤、爬森藤和马利筋饲养的幻紫斑蝶蛹的质量、体长和头宽均大于其它两种寄主植物;用古钩藤、爬森藤和聚果榕饲养的幻紫斑蝶存活率较高,而黄花夹竹桃最低;用古钩藤饲养的幻紫斑蝶成虫产卵量、交配频率和卵孵化率均最高,其次为爬森藤。因此,聚果榕、古钩藤和马利筋可作为幻紫斑蝶成虫产卵的优势寄主,古钩藤和爬森藤可作为饲养幻紫斑蝶的优势寄主。     

Oviposition by Danaus plexippus in relation to cardenolide content of three Asclepias species in the southeastern U.S.A.

Abstract. 1. Female monarchs were observed in the field ovipositing on a native North American milkweek host, Asclepias humistrata L. As in a comparable Australian study on an introduced novel host ( Asclepias fruticosa L.) we found post-alighting rejection of plants with low and high cardiac glycoside concentration (CG).
2. Most oviposition took place on plants with CG in the range 200–500 μg/0.1 g dry weight. Thin-layer chromatography showed no obvious qualitative difference in cardenolide types between accepted and rejected plants, excepting an indication that rejected plants may have a higher level of more polar cardenolides.
3. In a controlled laboratory experiment comparing oviposition on a low ( A.incarnata L.) vs high ( A.curassavica L.) CG host plant species we found no relationship between CG and oviposition on the low CG species, but a negative relationship in the high CG species. This corroborates our findings on oviposition on single host species in the field.
4. We also record the first indication of a physiological cost of monarch larvae feeding on plants high in CG. There was a significant negative relationship between survival of first instar larvae and CG levels in plants. This study opens the way for further work on the association of monarch butterflies and their toxic milkweed hosts.     

The mitochondrial coxII intron has been lost in two different lineages of dicots and altered in others

The central part of the mitochondrial coxII gene was amplified from 38 different dicots and two monocots using polymerase chain reaction. In 30 of the 40 plants studied, the amplified coxll gene-fragment contains an intron, ranging from 930 bp in Capsicum (pepper) in Solanaceae to 1,635 bp in Ampelamus albidans (climbing milkweed) in Asclepiadaceae. The composition of this intron varies as revealed by Southern hybridizations using oligonucleotide probes specific to the coxII intron-regions in maize, wheat, and rice. In the Apocynaceae, Calharanthus roseus (Madagascar periwinkle) and Vinca minor (common periwinkle) lack the coxII intron, while other members of the same family (various Mandevilla species, Nerium oleander and Apocynum cannabinum) and members of the closely related Asclepiadaceae (Asclepias incarnata, Ampelamus albidans and Asclepias tuberosa) retain the intron. Analysis of these data suggest a selective loss of the coxII intron from a plant, ancestral to both Catharanthus and Vinca, after the divergence of the Asclepiadaceae and Apocynaceae. The remaining eight plants from the Brassicaceae, Cucurbitaceae, Fabaceae, and Onagraceae lacking the intron fall into a single group or clade using the phylogenetic tree proposed by Chase et al. (Annals of the Missouri Botanical Garden: 80: 528–580, 1993) based on sequence of the chloroplast rbcL gene.     

Effects of photoperiod, temperature, and host plant age on induction of reproductive diapause and development time in Danaus plexippus

Abstract 1. Diapause induction in monarch butterflies Danaus plexippus was studied using adults captured from the wild in Minnesota and Wisconsin, and individuals reared under outdoor and controlled conditions.
2. Oocyte presence in females and ejaculatory duct mass in males were used to indicate reproductive status.
3. Some wild individuals were in diapause in mid-August, and all males and females were in diapause by late August and early September respectively.
4. Individuals reared under decreasing day lengths and fluctuating temperatures were more likely to be in diapause than were individuals reared under long or short day lengths or constant temperatures.
5. Individuals fed potted old Asclepias curassavica plants were more likely to be in diapause than were those fed potted young host plants; when cuttings of Asclepias syriaca plants from the field or greenhouse pots were used, there was no effect of host plant age.
6. Extremely high temperatures increased the number of day-degrees required for development from egg to adult, while decreasing day lengths and older host plants tended to decrease the number of day-degrees required for development.
7. There appears to be a continuum of reproductive development in monarchs, with gradual declines in mean ejaculatory duct mass and oocyte production during the late summer.
8. None of the experimental treatments led to 100% diapause, and diapause was more likely to occur in monarchs subjected to more than one diapause-inducing cue.     

Differential pollinator effectiveness and importance in a milkweed (Asclepias, Apocynaceae) hybrid zone

? Premise of the study: Exceptions to the ideal of complete reproductive isolation between species are commonly encountered in diverse plant, animal, and fungal groups, but often the causative ecological processes are poorly understood. In flowering plants, the outcome of hybridization depends in part on the effectiveness of pollinators in interspecific pollen transport. In the Asclepias exaltata and A. syriaca (Apocynaceae) hybrid zone in Shenandoah National Park, Virginia, extensive introgression has been documented. The objectives of this study were to (1) determine the extent of pollinator overlap among A. exaltata, A. syriaca, and their hybrids and (2) identify the insect taxa responsible for hybridization and introgression. ? Methods: We observed focal plants of parental species and hybrids to measure visitation rate, visit duration, and per-visit pollinia removal and deposition, and we calculated pollinator effectiveness and importance. ? Key results: Visitation rates varied significantly between the 2 yr of the study. Overall, Apis mellifera, Bombus sp., and Epargyreus clarus were the most important pollinators. However, Bombus sp. was the only visitor that was observed to both remove and insert pollinia for both parent species as well as hybrids. ? Conclusions: We conclude that Bombus may be a key agent of hybridization and introgression in these sympatric milkweed populations, and hybrids are neither preferred nor selected against by pollinators. Thus, we have identified a potential mechanism for how hybrids act as bridges to gene flow between A. exaltata and A. syriaca. These results provide insights into the breakdown of prezygotic isolating mechanisms.     

Cloning of casbene and neocembrene synthases from Euphorbiaceae plants and expression in Saccharomycescerevisiae

A large number of diterpenes have been isolated from Euphorbiaceae plants, many of which are of interest due to toxicity or potential therapeutic activity. Specific Euphorbiaceae diterpenes of medical interest include the latent HIV-1 activator prostratin (and related 12-deoxyphorbol esters), the analgesic resiniferatoxin, and the anticancer drug candidate ingenol 3-angelate. In spite of the large number of diterpenes isolated from these plants and the similarity of their core structures, there is little known about their biosynthetic pathways. Other than the enzymes involved in gibberellin biosynthesis, the only diterpene synthase isolated to date from the Euphorbiaceae has been casbene synthase, responsible for biosynthesis of a macrocyclic diterpene in the castor bean (Ricinus communis). Here, we have selected five Euphorbiaceae species in which to investigate terpene biosynthesis and report on the distribution of diterpene synthases within this family. We have discovered genes encoding putative casbene synthases in all of our selected Euphorbiaceae species and have demonstrated high-level casbene production through expression of four of these genes in a metabolically engineered strain of Saccharomyces cerevisiae. The only other diterpene synthase found among the five plants was a neocembrene synthase from R. communis (this being the first report of a neocembrene synthase gene). Based on the prevalence of casbene synthases, the lack of other candidates, and the structure of the casbene skeleton, we consider it likely that casbene is the precursor to a large number of Euphorbiaceae diterpenes. Casbene production levels of 31 mg/L were achieved in S. cerevisiae and we discuss strategies to further increase production by maximizing flux through the mevalonate pathway.     

Notes on the esterase variation in Swedish Dactylorhiza incarnata s. l. (Orchidaceae)

Swedish material of Dactylorhiza incarnara s. l. shows little variation at commonly investigated allozyme loci. However, interpretable variation was found at one esterase locus. All plants investigated of D. incarnata var. cruenta from southern Sweden (with spotted leaves) were homozygous for allele a at this locus, whereas all plants investigated of D. incarnata var. ochroleuca were fixed for allele b. In D. incarnata var. incarnata , both alleles were found, although the b allele dominated. In northern Swedish material of D. incarnata s. l., only allele b was found, regardless whether the material had spotted leaves (sometimes referred to as var. cruenta ), or had unspotted leaves.
These results indicate that there is restricted gene flow between var. cruenta and the other varieties in southern Sweden, although they often grow in mixed populations. The northern Swedish material with spotted leaves appears not to be related to the southern var. cruenta .     

Molecular identification of phytoplasmas in fasciated cacti and succulent species and associated hormonal perturbation

Fasciation, a frequent phenomenon in Cactaceae, has been attributed to various causes. The present study reports on phytoplasma-induced fasciation in Euphorbia coerulescens (Euphorbiaceae), Orbea gigantea (Asclepiadaceae), Opuntia cylindrica (Cactaceae), and Senecio stapeliiformis (Asteraceae). DNA was extracted from symptomless and fasciated tissues and amplified by nested PCR using universal primers P1/P7 followed by R16F2n/R16R2 produced amplicons of 1.2 Kb. The nucleotide sequence analyses of the amplicons indicated that fasciated plants were infected by phytoplasma. Phylogenetic analysis placed the cacti fasciation phytoplasmas in 16SrII group. The hormonal content of symptomless and fasciated tissues including indole-3-acetic acid (IAA), kinetin (Kin), N⁶-benzyladenine (BA), abscisic acid (ABA), and gibberellic acid (GA₃) was quantified by high performance liquid chromatography (HPLC). The results indicated that fasciation in O. gigantea was correlated with the accumulation of Kin and IAA increasing five and two times, respectively, as compared to symptomless tissue. However, there was no consistent pattern of hormones in other fasciated species (E. coerulescens, O. cylindrica, and S. stapeliiformis), suggesting that different plant species might have different mechanism to develop fasciation associated with phytoplasma infection.     

施肥干扰对陕北黄土丘陵区三个典型群落结构组成的影响

在陕北黄土丘陵区,为了加快当地植被的恢复速度,结合当地条件,通过小区施肥实验与野外调查,研究了施肥对不同演替阶段的3个典型群落:猪毛蒿(Artemisia scoparia)、长芒草(Stipa bungeana)和铁杆蒿(Artemisia sacrorum)结构组成的影响。结果表明:(1)与对照相比,施肥后3个群落中物种的重要值发生改变。猪毛蒿群落中,1年生植物狗尾草(Setaria viridis)重要值减小,而多年生植物阿尔泰狗哇花(Heteropappus altaicus)、冰草(Agropyron cristatum)等重要值增大,成为群落中的主要物种;长芒草群落,与对照相比,施肥降低了优势种长芒草的重要性,而达乌里胡枝子(Lespedeza dahurica)的重要值明显增大;铁杆蒿群落则随着肥量增加,铁杆蒿的重要值表现为逐渐上升,与对照相比,群落中的伴生种二裂萎陵菜(Potentilla bifurca)、茭蒿(Artemisia giraldii)在处理区的平均重要值有所下降,而演替后期物种达乌里胡枝子的重要值增大。(2)与对照相比,通过施肥,除铁杆蒿群落高肥区密度外,3个群落的高度、生物量和密度均有显著提高,表现为高度和生物量在高肥区达最大,密度在中肥区达最高。3个群落相比,演替前期的猪毛蒿群落提高幅度最大,长芒草次之,铁杆蒿最小。(3)施肥改变了3个群落的物种多样性。群落类型不同,其变化趋势亦不同。猪毛蒿和铁杆蒿群落Patrick丰富度指数在中肥区达最高,随着施肥量的增加,Margalef(Ima)、Shannon-Wiener(Isw)和Pielou(J)指数产生不同程度的下降,而Berger-Parker优势度指数(I)逐渐增大;长芒草群落则随着施肥量的增加,Patrick指数、Ima、Isw和J均有不同程度的上升,而优势度指数I呈下降趋势。     

Use of drift substrates to characterize marine fungal communities from the west coast of Portugal

This survey reports the occurrence, diversity and similarity of marine fungi associated with five categories of drift substrates (Arundo donax, Phragmites australis, Spartina maritima, "other stems" and driftwood) collected on four sandy beaches of the western coast of Portugal. "Other stems" and driftwood are composite samples with a variety of identified and unidentified pieces of non-woody and woody substrates respectively. Fifty-six taxa were identified, including 38 Ascomycota and 18 anamorphic fungi. Twenty-six taxa were generalists; however several cases of "substrate recurrence" were identified. The very frequent fungi differed among the categories of studied substrates, with the exception of Corollospora maritima, very frequent on four categories. Except for S. maritima, P. australis and driftwood, cases of multiple fungal colonization were rare. S. maritima was the single substrate with five different marine fungi on one sample, as well as with the highest number of very frequent fungi, highest percentage of colonization and average number of fungi per sample. Driftwood presented the highest value of fungal richness (37 taxa) and A. donax the lowest (22 taxa). ANOSIM analysis of similarity showed that all substrates supported different fungal communities with the exception of the pair P. australis/"other stems". The effect of sample size on estimated fungal richness was tested, and the results let us conclude that, although most of the sporadic fungi (<1% occurrence) will be detected only in a very large number of samples, 60 samples of A. donax and "other stems" and 70 samples of all the other substrates may suffice to assess their respective representative marine mycota.     

Cloning and expression of cytochrome P450 enzymes catalyzing the conversion of tyrosine to p-hydroxyphenylacetaldoxime in the biosynthesis of cyanogenic glucosides in Triglochin maritima

Two cDNA clones encoding cytochrome P450 enzymes belonging to the CYP79 family have been isolated from Triglochin maritima. The two proteins show 94% sequence identity and have been designated CYP79E1 and CYP79E2. Heterologous expression of the native and the truncated forms of the two clones in Escherichia coli demonstrated that both encode multifunctional N-hydroxylases catalyzing the conversion of tyrosine to p-hydroxyphenylacetaldoxime in the biosynthesis of the two cyanogenic glucosides taxiphyllin and triglochinin in T. maritima. This renders CYP79E functionally identical to CYP79A1 from Sorghum bicolor, and unambiguously demonstrates that cyanogenic glucoside biosynthesis in T. maritima and S. bicolor is catalyzed by analogous enzyme systems with p-hydroxyphenylacetaldoxime as a free intermediate. This is in contrast to earlier reports stipulating p-hydroxyphenylacetonitrile as the only free intermediate in T. maritima. L-3,4-Dihydroxyphenyl[3-(14)C]Ala (DOPA) was not metabolized by CYP79E1, indicating that hydroxylation of the phenol ring at the meta position, as required for triglochinin formation, takes place at a later stage. In S. bicolor, CYP71E1 catalyzes the subsequent conversion of p-hydroxyphenylacetaldoxime to p-hydroxymandelonitrile. When CYP79E1 from T. maritima was reconstituted with CYP71E1 and NADPH-cytochrome P450 oxidoreductase from S. bicolor, efficient conversion of tyrosine to p-hydroxymandelonitrile was observed.     

Antimicrobial activity of fatty acid methyl esters of some members of Chenopodiaceae

Fatty acid methyl ester (FAME) extracts of four halophytic plants, viz. Arthrocnemum indicum, Salicornia brachiata, Suaeda maritima and Suaeda monoica belonging to the family Chenopodiaceae, were prepared and their composition was analyzed by GC-MS. The FAME extracts were also screened for antibacterial and antifungal activities. The GC-MS analysis revealed the presence of more saturated fatty acids than unsaturated fatty acids. Among the fatty acids analyzed, the relative percentage of lauric acid was high in S. brachiata (61.85%). The FAME extract of S. brachiata showed the highest antibacterial and antifungal activities among the extracts tested. The other three extracts showed potent antibacterial and moderate anticandidal activities.     

Monarch butterfly oviposition on swallow-worts (Vincetoxicum spp.)

As black swallow-wort, Vincetoxicum nigrum L. Moench, and pale swallow-wort, V. rossicum (Kleopow) Barbar., spread throughout the northeastern United States and southern Canada, there is concern about the impact of these invasive plants on populations of the native North American monarch butterfly Danaus plexippus L. Recent laboratory studies in New York and Ontario, Canada, have found little or no oviposition by monarch butterflies on the two Vincetoxicum species. In Rhode Island, we found 10.5-21.7% oviposition on Vincetoxicum species relative to common milkweed Asclepias syriaca L. in choice tests and 11.9-20.3% in no-choice tests in 2 yr of laboratory testing. These results were supported by field cage trials where monarchs given a choice between V. nigrum and A. syriaca laid 24.5% of their eggs on V. nigrum. In surveys of three pasture fields in Rhode Island where relative coverage of A. syriaca exceeded that of V. nigrum by a 0.77:0.23 ratio, 15.4% of monarch eggs were found on V. nigrum plants. In V. nigrum stands with very little A. syriaca (6.25 stems/ha), monarch egg density on V. nigrum was found to be over five times greater than in the three mixed pasture fields. In none of our laboratory or field evaluations was there any survival of monarch larvae on Vincetoxicum species. It seems that in Rhode Island, Vincetoxicum species serve as an oviposition sink for monarch butterflies. These findings suggest that East Coast butterflies may differ in host selection from those in central New York and southern Ontario, Canada.     

The ecological significance of the combtoothed blenny in a coral reef ecosystem

At Heron Island reef, Great Barrier Reef Australia, biomass densities and mean wet mass of Ward's damselfish Pomacentrus wardi and the jewelled blenny Salarias fasciatus were not significantly different at 2·37 v . 2·95 g m⁻² and 8·7 v . 7·9 g, respectively. Whereas S. fasciatus significantly exceeded P. wardi in (1) total number of bites per day (3427 v . 1155), (2) the mass of epilithic algal community consumed per bite (2·19 v . 0·14 mg) and (3) total organic carbon consumed per day (487·31 v . 35·46 mg C m⁻² day⁻¹). Territorial behaviour differed also between the two species. Pomacentrus wardi chased from their territories a smaller proportion of blennies than roving grazers ( i.e . scarids, acanthurids, siganids and pomacentrids) relative to S. fasciatus . Salarias fasciatus chased c . 90% of other blennies from their territories, while chasing only c . 20% of all damsels that entered. Both P. wardi and S. fasciatus rarely chased non‐grazers. The chasing behaviour of S. fasciatus was size dependent, with resident fish chasing only individuals of its own family ( i.e . Blenniidae) that were the same or smaller size. Pomacentrus wardi may have tolerated S. fasciatus grazing within its territory, as it contributes to territory defence from other blennies. The possibility that the interaction between the two species is facilitative, rather than competitive, is discussed. It was concluded that salariine blennies play an important, and previously underestimated role in coral reef trophodynamics.     

拉杰沙希大学校园药用植物菌根研究

Forty different medicinal plants were investigated for arbuscular mycorrhizal association in the Rajshahi University Campus in Bangladesh. The results indicated that 35 different plants were infected by AM (arbuscular mycorrhizal) fungi as found by trypan blue staining procedure. The percentage of root colonization by AM fungi varied from 13.3% to 100%. Mangifera indica and Morus indica have maximum percentage of colonization (100%). The intensity of root colonization were abundant in the plants belonging to the families Anacardiaceae, Asclepiadaceae, Moraceae, Leguminosae and Apocynaceae whereas the intensity of colonization of crop roots were moderate and poor belonging to Gramineae and Leguminosae. The presence of greater number of spore in soil was always associated with the incidence of abundant mycelia. In plant roots the formation of spore and mycelia was restricted by low pH. Number of mycorrhizal fungus spores ranged between 35 to100 per 100g air dried soil in different family respective soils. The frequency of mycorrhizal fungus infection showed positive correlation with soil pH, moisture, water holding capacity, texture, total nitrogen, organic carbon, phosphorus, calcium, potassium, and magnesium. Especially phosphorus and nitrogen in the soil greatly influenced the plant root infection by AM fungi.     

Relationships among 3 Kochia species based on PCR-generated molecular sequences and molecular cytogenetics.

Forage kochia (Kochia prostrata ssp. virescens 'Immigrant' is native to the arid and semiarid regions of central Eurasia. It was introduced into the United States in 1966 as PI 314929 and released as a perennial forage shrub in 1984. Kochia americana is a perennial native to the United States, whereas Kochia scorparia is an introduced annual species that became a weed. To assess both the breeding potential and the possibility of genetic contamination, relationships among the 3 Kochia species were analyzed using random amplified polymorphic DNA (RAPD) markers, sequence tagged site (STS) marker sequences of the chloroplast NADH dehydrogenase gene (ndhF), genomic in situ hybridization (GISH), and multicolor fluorescence in situ hybridization (MC-FISH). Seventy decamer random primers yielded 458 polymorphic bands from 9 plants of K. americana, 20 plants of K. prostrata, and 7 plants of K. scoparia. Fifty-four and 55 species-specific RAPD markers were identified for K. americana and K. prostrata, whereas 80 RAPD markers were specific to K. scoparia. Based on the presence or absence of informative RAPD markers, the 3 species always grouped into 3 distinct clusters in a NTSYSpc2.01b-generated dendrogram. The same relationships were found among the 3 Kochia species based on ndhF DNA sequence divergence. Using a set of 7 STS markers that can identify each Kochia species, we did not find a single interspecific hybrid from artificial hybridizations among the 3 Kochia species. In GISH studies, chromosomes of 1 species fluoresced in green only when they were probed by genomic DNA of the same species. Cross-hybridization by genomic DNA of another species was not observed. In FISH studies using pTa71 (for 18S-5.8S-26S rDNAs) and pScT7 (for 5S rDNA) as probes, there were 1, 1 and 3 pTa71 sites and 2, 1, and 1 pScT7 sites in each haplome of K. prostrata, K. americana, and K. scoparia, respectively. It is concluded that these 3 Kochia species are so genomically distinct that gene introgression among them would be extremely rare.