First report of Alternaria sp. PMK1: causative agent of leaf spot disease on parthenium weed

Congress grass, Parthenium hysterophorus L. (Asteraceae, tribe Heliantheae), is an erect and much branched annual or ephemeral herb, known as a pest for environmental, medical and agricultural reasons. Survey of fungal pathogens of P. hysterophorus was conducted in Kurukshetra and its adjoining areas, Haryana, India. A leaf spot pathogen Alternaria sp. PMK1 was isolated. Koch's postulates were performed and proved that this isolate was pathogenic to this weed. This is the first report of this pathogen causing leaf spot on parthenium weed.     

Host range evaluation of the leaf-feeding beetle,Zygogramma bicolorata and the stem-boring weevil,Listronotus setosipennis demonstrates their suitability for biological control of the invasive weed,Parthenium hysterophorus in Ethiopia

The invasive weed, parthenium (Parthenium hysterophorus L.) (Asteraceae: tribe Heliantheae), damages agriculture, adversely impacts biodiversity and is hazardous to human and animal health in Ethiopia. The host range of two natural enemies, a leaf-feeding beetle, Zygogramma bicolorata (Coleoptera: Chrysomelidae) and a stem-boring weevil, Listronotus setosipennis (Coleoptera: Curculionidae) was evaluated for biological control of the weed in Ethiopia. The specificity of Z. bicolorata and L. setosipennis was assessed against 29 and 31 non-target plant species, respectively. The host range of Z. bicolorata and L. setosipennis was first assessed using no-choice tests to examine their oviposition and feeding response on non-target plants. Although oviposition by Z. bicolorata occurred on six non-target species in four Asteraceae species in no-choice tests, it was significantly lower than on parthenium and no larvae developed. Zygogramma bicolorata nibbled the leaves of one of the five niger seed (Guizotia abyssinica L. – an oil seed crop closely related to parthenium) cultivars tested, but feeding and oviposition were significantly less than on parthenium. Furthermore, choice tests indicated that Z. bicolorata did not oviposit nor feed on G. abyssinica when parthenium was present. In no-choice tests, L. setosipennis did not oviposit on any of the non-target species assessed. Mean oviposition on parthenium was 39.0?±?3.4 eggs per plant whereas no eggs were laid on any of the 31 species tested. Based on these and other host range tests, permission was obtained to field release Z. bicolorata and L. setosipennis in Ethiopia.     

Isolation of disease-tolerant cotton (Gossypium hirsutum L. cv. SVPR 2) plants by screening somatic embryos with fungal culture filtrate

We report an in vitro selection method that has led to isolation of Fusarium wilt and Alternaria leaf spot disease-tolerant plantlets in cotton (Gossypium hirsutum L. cv. SVPR2). Embryogenic callus was isolated from hypocotyl explants of cotton cultured on 5–50% Fusarium oxysporum culture filtrate-fortified callus induction medium. Somatic embryos tolerant to fungal culture filtrate (FCF) were isolated from this embryogenic callus on somatic embryo regeneration medium fortified with 40% FCF. Sixteen plantlets were selected as FCF-tolerant from 34 somatic embryos tested, which corresponds to about 47% success rate. The FCF-tolerant plants were analyzed for disease tolerance by challenging them with spores of F. oxysporum and Alternaria macrospora. Four plants were selected as F. oxysporum tolerant from a total of 24 plants tested. The selected plants showed an enhanced survival rate compared with the control when they were grown in earthen pots inoculated with 1 × 10⁵ spores/mL of F. oxysporum. From the FCF-tolerant plants, another nine randomly selected plantlets were challenged with spores of A. macrospora in order to test their tolerance to Alternaria leaf spot disease. The number of lesions per leaf significantly decreased from 8.2 to 0.9 and the lesion lengths were also reduced from 2.8 to 1.2 mm per leaf spot in these plants. Electrophoresis analysis of extracellular proteins from the FCF-tolerant plants showed enhanced secretion of proteins in the range of 24–36 kDa. Isozyme analysis by of FCF-tolerant plants by using native gels showed the presence of chitinase. Quantitative analysis showed that there was 13-fold increase in a chitinase activity in the selected FCF-tolerant plants compared to the control plants. Our results show that over-expression of chitinase enzyme leads to enhanced disease resistance against F. oxysporum and A. macrospora.     

Host range and risk assessment of Zygogramma bicolorata,a defoliating agent released in South Africa for the biological control of Parthenium hysterophorus

Parthenium hysterophorus (parthenium) is a weed of international importance and is spreading rapidly in sub-Saharan Africa. Consequently, it has been targeted for biocontrol in South Africa since 2003. Based on precedents elsewhere in the world, the defoliating beetle Zygogramma bicolorata was prioritised as a candidate agent. Although no-choice tests, involving some 48 test plant species, indicated a significant preference for parthenium, significantly reduced feeding and oviposition was recorded on some species. Multiple-choice tests resolved many of these non-target results; however, Helianthus annuus (sunflower) was still selected for oviposition and feeding. Of the 12 sunflower cultivars tested, four were selected for oviposition, while two were selected for oviposition and feeding. These six cultivars were then subjected to larval development trials, together with three native and two weed species (in the Asteraceae). These trials showed high levels of complete development on parthenium, significantly reduced development on sunflower cultivars, and partial development on only one of the weed species. Finally, a risk assessment was conducted on the six sunflower cultivars to quantify Z. bicolorata feeding and reproductive performance. Feeding risk calculations revealed these cultivars to have an extremely low risk (<0.2%) of supporting Z. bicolorata feeding and development. Similarly, reproductive risk calculations showed a very low risk (<0.16%) of supporting viable Z. bicolorata populations. These data are supported by findings from both the native (Mexico) and introduced ranges (Australia, India) of Z. bicolorata, where it has never been recorded as a pest of sunflower. These considerations were accepted by the regulatory authorities and in August 2013, Z. bicolorata became the second insect agent to be released in South Africa for the biocontrol of parthenium.     

Interference effects of Cassia uniflora Mill on Parthenium hysterophorus L.

In some parts of Karnataka and Maharashtra states (India), a leguminous undershrub, Cassia uniflora has effectively replaced the problematic weed Parthenium hysterophorus (parthenium). The studies revealed that the phenolic leachates from different parts of C. uniflora, specially from germinating seeds significantly inhibited the germination of parthenium seeds and retarded the growth of seedlings from the successfully germinated parthenium seeds. At field level, seedlings of C. uniflora outcompeted the seedlings of summer generation of parthenium causing reduction in height, dry weight and number of inflorescences of the latter which ultimately resulted in reduction of their seed output. Because of its longer life span, C. uniflora effectively prevented the establishment of parthenium plants of the winter generation in its area of growth. Results indicate that C. uniflora replaces parthenium through interference.     

Rearing the Mexican beetle Zygogramma bicolorata (Chrysomelidae: Coleoptera) on a semi-synthetic diet

A semi-synthetic diet utilising parthenium leaf powder was evaluated for rearing the Mexican beetle Zygogramma bicolorata Pallister under in vitro conditions at 26±1°C, 65% RH. Significant differences were observed in the biological parameters of the beetle viz., the grub survival (54.3%), pupation (47.2%), fecundity (54.6/day) and total developmental period (37.7 days), when reared on the semi-synthetic diet compared to those when reared on natural parthenium leaves (91.5, 85.5, 74.8 and 29.8, respectively). The feasibility of utilising the semi-synthetic diet for mass production of the beetle is discussed.     

Evaluation of Alternaria alternata ITCC4896 for use as mycoherbicide to control Parthenium hysterophorus

Mycoherbicides are specialty biotechnology products which employ the use of fungi or fungal metabolites as non-chemical alternatives, thereby reducing the input of harmful chemicals to control noxious weeds. The present communication emphasises the potential of an indigenous isolate of Alternaria alternata ITCC 4896 as a mycoherbicide for the global weed Parthenium hysterophorus. Of the various spore concentrations tested by in vitro detached leaf bioassay, 1 × 10⁶ spores/ml was the most effective, inducing 89.2% leaf area damage on the seventh day and was further tested by the whole plant bioassay. Both in vitro detached leaf assay and whole plant bioassay exhibited a similar trend in disease development, exhibiting 50% damage at 96 hours post-treatment. However, 100% mortality was observed in the whole plant bioassay on the seventh day. This is the very first report on the bioweedicidal potential of Alternaria alternata ITCC 4896 (LC#508) for use as a mycoherbicide for Parthenium hysterophorus.     

Invasion status and impacts of parthenium weed (Parthenium hysterophorus) in West-Central region of Bhutan

Parthenium weed (Parthenium hysterophorus L.) is an invasive alien plant species that is spreading rapidly all over the world. With globalization and climate change, it will continue to spread, posing threats to agriculture, biodiversity and the environment. To determine its invasion and impacts in Bhutan, an investigation was undertaken in West-Central Bhutan using a questionnaire survey, complemented by MaxEnt modelling. Most farmers (19–30%) recall having seen parthenium for the last 11–15 years, or more. Infestations of parthenium differed significantly (P?<?0.05) across three districts for dryland, orchards, fallow land and pastures, but not in rice fields and on roadsides. The most intensely invaded land types were fallow land, pastures, and roadsides, with farmers perceiving it as ‘somewhat common’(score?~?3) and ‘common’ (score?~?4) weed. Farmers’ perceptions of the invasiveness of parthenium weed were high at 77% (Wangdue), 80% (Punakha), and 88% (Tsirang), considering it as at least ‘somewhat invasive’, or higher. Only 13–25% of respondents scored the weed as ‘highly invasive’. Parthenium invasion significantly (P?<?0.01 to 0.001) escalated the cost of production, reduced fodder quality, supressed native species and caused allergies to the locals in the study districts. Further, MaxEnt modelling of parthenium distribution was undertaken, using the parthenium presence-only data and bioclimatic variables, under 2 CO₂ emission scenarios of the Intergovernmental Panel on Climate Change (IPCC), i.e. Representative Concentration Pathways (RCP2.6 and 8.5). The projections showed discernible spatial changes in climate suitability over 2070, mostly in the mid- and high-altitude zones, indicating aggressive incursion into Bhutan’s important agro-ecological landscapes. The findings, highlight the urgent need to initiate parthenium management strategies for food production and natural resources.

     

Does Zygogramma bicolorata Pallister really affects the growth,density and reproductive performance of Parthenium hysterophorus L?

Zygogramma bicolorata Pallister (Coleoptera: Chrysomelidae) is a satisfactory potential candidate for controlling Parthenium hysterophorus L. The study was conducted in order to evaluate the bio-control potential of Z. bicolorata to satisfactory reduced the density of parthenium weed in natural field conditions. Therefore, beetles were released in the twelve study sites @ 12 adults and/or larvae/m² in their respective sites on 5th, 10th, 15th and 20th day after initial field releases. Observations were made on defoliation by the beetle, perthenium plant height, plant biomass, leaf injury rating and soil seed reservoir. Present study showed that field releases of Z. bicolorata considerably lowering down the parthenium density at all treated sites. At larval release sites by the 60th day of observation a complete defoliation was noted, however at adult release sites it took 80 days for complete defoliation. Moreover, significantly higher number of developmental stages i.e. eggs, larvae and adults were noted in the adults released sites than to larval released sites and the controls. It was also noted that plant height and its density, flowers & leaves production, biomass of parthenium plant and seed reservoir in soil was negatively affected in treated sites as compared to untreated sites. Thus, we have concluded that adults can be utilized in biocontrol program because population build-up was greater in those sites where adults were released.     

Natural host range,thrips and seed transmission of distinct Tobacco streak virus strains in Queensland,Australia

Diseases caused by Tobacco streak virus (TSV) have resulted in significant crop losses in sunflower and mung bean crops in Australia. Two genetically distinct strains from central Queensland, TSV‐parthenium and TSV‐crownbeard, have been previously described. They share only 81% total‐genome nucleotide sequence identity and have distinct major alternative hosts, Parthenium hysterophorus (parthenium) and Verbesina encelioides (crownbeard). We developed and used strain‐specific multiplex Polymerase chain reactions (PCRs) for the three RNA segments of TSV‐parthenium and TSV‐crownbeard to accurately characterise the strains naturally infecting 41 hosts species. Hosts included species from 11 plant families, including 12 species endemic to Australia. Results from field surveys and inoculation tests indicate that parthenium is a poor host of TSV‐crownbeard. By contrast, crownbeard was both a natural host of, and experimentally infected by TSV‐parthenium but this infection combination resulted in non‐viable seed. These differences appear to be an effective biological barrier that largely restricts these two TSV strains to their respective major alternative hosts. TSV‐crownbeard was seed transmitted from naturally infected crownbeard at a rate of between 5% and 50% and was closely associated with the geographical distribution of crownbeard in central Queensland. TSV‐parthenium and TSV‐crownbeard were also seed transmitted in experimentally infected ageratum (Ageratum houstonianum) at rates of up to 40% and 27%, respectively. The related subgroup 1 ilarvirus, Ageratum latent virus, was also seed transmitted at a rate of 18% in ageratum which is its major alternative host. Thrips species Frankliniella schultzei and Microcephalothrips abdominalis were commonly found in flowers of TSV‐affected crops and nearby weed hosts. Both species readily transmitted TSV‐parthenium and TSV‐crownbeard. The results are discussed in terms of how two genetically and biologically distinct TSV strains have similar life cycle strategies in the same environment.     

Complementing biological control with plant suppression: Implications for improved management of parthenium weed (Parthenium hysterophorus L.)

Parthenium hysterophorus L. is a weed of global significance that has become a major weed in Australia and many other parts of the world. A combined approach for the management of parthenium weed using biological control and plant suppression, was tested under field conditions over a two-year period in southern central Queensland. The six suppressive plant species, selected for their demonstrably suppressive ability in earlier glasshouse studies, worked synergistically with the biological control agents (Epiblema strenuana Walker, Zygogramma bicolorata Pallister, Listronotus setosipennis Hustache and Puccinia abrupta var. partheniicola) present in the field to reduce the growth (above ground biomass) of parthenium weed, by between 60–86% and 47–91%, in Years 1 and 2, respectively. The biomass of the suppressive plants was between 6% and 23% greater when biological control agents were present than when the biological control agents had been excluded. This shows that parthenium weed can be more effectively managed by combining the current biological control management strategy with selected sown suppressive plant species, both in Australia and elsewhere.     

Tamarindus indica L. leaf is a source of allelopathic substance

The allelopathic potential of the Tamarindus indica L. leaf was investigated through bioassay guided studies using several weed and edible crop species. Both radicle and hypocotyl growth of all the plant species tested was strongly inhibited by the tamarind leaf using a sandwich method. The growth of weed species was reduced more than that of edible crop species. Among the weed species, barnyard grass followed by white clover, and in the edible crop species, lettuce followed by radish ranked top in terms of growth inhibition. Different concentrations of tamarind leaf crude water-soluble extract exhibited a strong inhibition in all the plant species tested and, by contrast, the magnitude of inhibition in the weed species was higher than in edible crop species and ranged from 30–75%. The 10% concentration of the tamarind leaf crude water-soluble extract was most potent against growth of seedlings. The concentrations of the nutrient components were linearly correlated with an increase in the concentration of tamarind leaf crude water-soluble extract. No significant changes in either pH or EC were found in the variations of different concentrations of tamarind leaf crude water-soluble extracts. As compared to control, growth of both radicle and hypocotyl in weed (barnyard grass and white clover) and in edible crop (lettuce and radish) species were significantly reduced when blended tamarind leaves at different concentrations were incorporated into the growth medium. The inhibitory magnitude increased with an increase in the concentration of the tamarind leaf. In terms of growth inhibition, among these tested plants, weed species particularly barnyard grass were most sensitive to the allelochemicals exuded from blended tamarind leaves. When the blended tamarind leaves were removed from the growth medium, all the seedlings grew quickly and the percentage of recovery was between 76–97% of the corresponding controls. Reduction in the fresh and dry weight of these 4 plant species was observed under the experimental conditions, and ranged between 33–42% and 40–53% in the radicle and hypocotyl, respectively. The fresh and dry weight, and total chlorophyll content declined significantly in the incorporated tamarind leaf treatments. Compared to the control, the highest drop in the chlorophyll content of 60% in barnyard grass was observed with the 10% concentration of the leaf treatment. These results clearly indicate that the tamarind leaf contains one or more strong biologically active allelochemical(s) that function as true growth regulator(s) and is involved in plant growth regulation, particularly in weed species.     

Xanthomonas campestris pv.parthenii pathovar nov. incitant of leaf blight of parthenium

A new bacterial leaf blight disease of parthenium (Parthenium hysterophorus L.) is described for the first time. The disease-causing bacterium was isolated and its morphological, physiological and biochemical characters were determined. The pathogenicity of bacterium is apparently limited only to parthenium. The pathogen was identified asXanthomonas campestris pv.parthenii pathovar nov. on the basis of morphological, physiological, biochemical and pathogenic characteristics.     

Leaf scarring by the weevils Neochetina eichhorniae and N. bruchi enhances infection by the fungus Cercospora piaropi on waterhyacinth, Eichhornia crassipes

Additive or synergistic effects among introduced and native insect and plant pathogen agents are necessary to achieve biological control of waterhyacinth (Eichhornia crassipes), a globally damaging aquatic weed. In field plots, plants were infested with waterhyacinth weevils (Neoechetina bruchi and N. eichhorniae) and leaves were scarred by weevil feeding. Subsequent infection by the fungal pathogen Cercospora piaropi caused necrotic lesions to form on leaves. Necrosis development was 7.5- and 10.5-fold greater in plots augmented with both weevils and C. piaropi and weevils alone, respectively, than in plots receiving only C. piaropi. Twenty-four days after weevil infestation, the percentage of laminar area covered by lesions on third-youngest and oldest live leaves was elevated 2.3–2.5-fold in plots augmented with weevils. Scar density and necrosis coverage on young leaf laminae were positively correlated, even though antipathogenic soluble peroxidases were elevated 3-fold in plots augmented with weevils alone or weevils and C. piaropi. Combined weevil and fungal augmentation decreased shoot densities and leaves per plant. In a no-choice bioassay, weevil feeding on oldest but not young leaves was reduced 44 two weeks after C. piaropi inoculation. Protein content and peroxidase activities were elevated 2–6-fold in oldest leaves three weeks after inoculation. Augmentation with both waterhyacinth weevils and C. piaropi led to the development of an additive biological control impact, mediated by one or more direct interactions between these agents, and not plant quality effects.     

Comparative Efficacy of Weed Control Practices for Parthenium Weed and Sunflower Crop under Varying Tillage Systems

Parthenium poses serious threat to modern crop production system and necessitate evaluating control practices for its effective management. Efficacy of different weed control practices for controlling parthenium was explored in conventional and deep tillage systems in the field conditions. Hand hoeing (20 and 35 days after emergence), S-Metolachlor (pre-emergence herbicide), sorghum straw mulch @ 5 tons ha^-1 and combination of hand hoeing and sorghum straw mulch (hand hoeing at 20 and straw mulch at 35 days after emergence) were used as weed control practice. Weedy check where no weed control measure was applied was also included in this experiment for comparison. Results concluded that the all weed management treatments significantly reduced parthenium density, its fresh and dry biomass during both the years of study as compared to weedy check. Maximum sunflower achene yield was recorded in hand hoeing (20 and 35 days after emergence) in combination with deep tillage. So, mold bold plough used for the purpose of deep tillage should be encouraged for better control of parthenium and higher achene yield of sunflower crop (3293.3 kg ha^-1 in 2017 and 3221.3 kg ha^-1 in 2018). Moreover, is also inferred that total dose of herbicide might be reduced by using hoeing and mulching in an integrated way.     

Transfer of Alternaria macrospora from Cotton Seed to Seedling: Light and Scanning Electron Microscopy of Colonization

Alternaria macrospora is transferred from an infected seed, through the plant stem to the cotyledon and sporulates on its surface. However, at the beginning of the process, there is no difference between resistant and susceptible plants. In later stages, lower levels of fungal mycelium and sporulation were detected on the seedlings of a resistant cultivar. The fungus grew inside the plant from the underground to the upper parts. The mycelia which originated from surface spores could penetrate, through stomata and result in leaf colonization. A. macrospora sporulated ultimately on plant surfaces whereas the internal leaf tissue was colonized by mycelium only. This study suggests that cotyledon infection is a result of seed infection.     

Geographical variation and genetic analysis of leaf shape inCrepis tectorum (Asteraceae)

Canonical variate analysis of plants raised in a uniform environment was used to study the pattern of geographical variation in leaf shape ofCrepis tectorum (Asteraceae). The diversity in leaf shape was much greater among populations confined to areas with exposed bedrock in the Baltic region than among weed populations scattered throughout Europe and Canada. A Ward's clustering linked outcrop populations from the archipelago of SW. Finland and the islands of Öland (Sweden) and Saaremaa (Estonia) due to the deeply lobed leaves characterizing these populations, while outcrop populations along the coast of E. Sweden were grouped due to their weakly lobed, narrow and dentate leaves. Most of the weed populations were grouped together but there was no tendency for the variation in this group to be related to habitat or geographical location. A mosaic of variation reflected in sharp (random) differentiation among local populations was superimposed on the large-scale ecogeographical pattern.—Crossing data indicated that most of the variation in degree of leaf dissection is governed by one major gene with deeply lobed leaves dominant over weakly lobed leaves. I suggest that the simple pattern of inheritance may have favoured rapid evolutionary changes in leaf shape, particularly in the Baltic area which emerged relatively late from the sea. Genetic correlations may have constrained the pattern of variation at higher taxonomic levels, since some of the trait associations detected in a segregating F₂ generation were also found at the among-population level.     

Distributions of invasive weed Parthenium (Parthenium hysterophorus L.) in the University campus Peshawar KPK

A very interesting survey was performed to measure the distribution of invasive Parthenium (Parthenium hysterophorus L.) weed in the University Campus, Peshawar, the capital city of Khyber Pakhtunkhwa province. The University Campus comprised on; The University of Peshawar, Islamia College University, The University of Agriculture Peshawar, Pakistan Forest Institute and the New Developmental Research Farm. In these selected areas Parthenium weed distribution was measured in particular the academic, residential and research farm. Moreover, the survey also investigated the impact of parthenium weed upon native weeds flora inside the University campus. A 1m² quadrat was used to collect the data. During the survey, a total of 32 weeds species were recorded belonging to 18 different families and 32 genera, the life cycle of 23 weeds were annual while 9 weeds have a perennial. The data were recorded on absolute density (%), relative density (%), absolute frequency (%), relative frequency (%) and importance values (%) of parthenium weed and others flora. The collected data show that parthenium weed was the most dominant species having 76.8 m^?2 plants density in the NDF and (32.0 m^?2) at Islamia College University. However, the highest frequency (100%) of Parthenium was observed for The University of Agriculture, while minimum (80%) occurred in the Agronomy field. A More, the relative frequency of Parthenium was maximum (25%) in Islamia College; whereas, it's minimum (12.9%) at Agronomy field. Finally, the Importance Value of Parthenium indicates the highest (45.98%) at Dairy Farm followed by (40.33%) at the Horticulture field's area. In short, the Parthenium weed becomes more problematic and replaces all the native flora in the non-crop area specially Dicanthium annulatum L. a native grass species on the ridges of the field being replaced by parthenium. Due to no proper management and high growth rate, nowadays it becomes like a superweed in the KPK and especially in Pakistan. So, the management of Parthenium weed requires the call attention from the Govt. policymaker and proper quarantine inspection to conserve the native flora and field crops to get maximum crop yield.     

Investigating the mechanisms of glyphosate resistance in <Emphasis Type="Italic">Lolium multiflorum</Emphasis>

Evolved resistance to the herbicide glyphosate has been reported in eleven weed species, including Lolium multiflorum. Two glyphosate-resistant L. multiflorum populations were collected, one from Chile (SF) and one from Oregon, USA (OR), and the mechanisms conferring glyphosate resistance were studied. Based on a Petri dish dose–response bioassay, the OR and the SF populations were two and fivefold more resistant to glyphosate when compared to the susceptible (S) population, respectively; however, based on a whole-plant dose–response bioassay, both OR and SF populations were fivefold more resistant to glyphosate than the S population, implying that different resistance mechanisms might be involved. The S population accumulated two and three times more shikimic acid in leaf tissue 96 h after glyphosate application than the resistant OR and SF populations, respectively. There were no differences between the S and the glyphosate-resistant OR and SF populations in ¹⁴C-glyphosate leaf uptake; however, the patterns of ¹⁴C-glyphosate translocation were significantly different. In the OR population, a greater percentage of ¹⁴C-glyphosate absorbed by the plant moved distal to the treated section and accumulated in the tip of the treated leaf. In contrast, in the S and in the SF populations, a greater percentage of ¹⁴C-glyphosate moved to non-treated leaves and the stem. cDNA sequence analysis of the EPSP synthase gene indicated that the glyphosate-resistant SF population has a proline 106 to serine amino acid substitution. Here, we report that glyphosate resistance in L. multiflorum is conferred by two different mechanisms, limited translocation (nontarget site-based) and mutation of the EPSP synthase gene (target site-based).     

The physiological significance of the leaf nodules of psychotria

Summary In 6-month growth experiments it was found that leaf-nodulatedPsychotria mucronata seedlings grown in N-poor soil showed a restricted growth and developed severe nitrogen-deficiency symptoms in the leaves. Plants in the same soil supplied with NO₃-N showed healthy growth and dark green leaves. Detached Psychotria leaves bearing leaf nodules exposed to an atmosphere containing N¹⁵-labelled nitrogen gas or acetylene gas gave no evidence of nitrogen fixation, either in the light or in the dark or in both in succession. Therefore nitrogen fixation is probably not associated with the leaf nodules. Chlorophyll retention was observed around the leaf nodules in senescent Psychotria leaves. Psychotria leaf-nodule discs placed on oat leaves cause chlorophyll retention in the oat leaves below the discs. As chlorophyll retention is a common bioassay for cytokinins, these results indicate that a cytokinin-like substance is involved. With the aid of autoradiography and C¹⁴-labelled α-amino-isobutyric acid it was shown that this amino acid accumulates in the leaf nodules. Such directed transport is also a property of cytokinin.