Endogenous and environmental factors influence 35S promoter methylation of a maize A1 gene construct in transgenic petunia and its colour phenotype

Summary 30000 transgenic petunia plants carrying a single copy of the maize A1 gene, encoding a dihydroflavonol reductase, which confers a salmon red flower colour phenotype on the petunia plant, were grown in a field test. During the growing season plants with flowers deviating from this salmon red colour, such as those showing white or variegated phenotypes and plants with flowers exhibiting only weak pigmentation were observed with varying frequencies. While four white flowering plants were shown at the molecular level to be mutants in which part of the A1 gene had been deleted, other white flowering plants, as well as 13 representative plants tested out of a total of 57 variegated individuals were not mutants but rather showed hypermethylation of the 35S promoter directing A1 gene expression. This was in contrast to the homogeneous fully red flowering plants in which no methylation of the 35S promoter was observed. While blossoms on plants flowering early in the season were predominantly red, later flowers on the same plants showed weaker coloration. Once again the reduction of the A1-specific phenotype correlated with the methylation of the 35S promoter. This variation in coloration seems to be dependent not only on exogenous but also on endogenous factors such as the age of the parental plant from which the seed was derived or the time at which crosses were made.     

Resistance to Cucumber mosaic virus in Gladiolus plants transformed with either a defective replicase or coat protein subgroup II gene from Cucumber mosaic virus

Transgenic Gladiolus plants that contain either Cucumber mosaic virus (CMV) subgroup I coat protein, CMV subgroup II coat protein, CMV replicase, a combination of the CMV subgroups I and II coat proteins, or a combination of the CMV subgroup II coat protein and replicase genes were developed. These plants were multiplied in vitro and challenged with purified CMV isolated from Gladiolus using a hand-held gene gun. Three out of 19 independently transformed plants expressing the replicase gene under control of the duplicated CaMV 35S promoter were found to be resistant to CMV subgroup I. Three out of 21 independently transformed plants with the CMV subgroup II coat protein gene under control of the Arabidopsis UBQ3 promoter were resistant to CMV subgroup II. Eighteen independently transformed plants with either the CMV subgroup I coat protein or a combination of CMV subgroups I and II coat proteins were challenged and found to be susceptible to both CMV subgroups I or II. Virus resistant plants with the CMV replicase transgene expressed much lower RNA levels than resistant plants expressing the CMV subgroup II coat protein. This work will facilitate the evaluation of virus resistance in transgenic Gladiolus plants to yield improved floral quality and productivity.     

Propyl gallate,a free radical scavenger,counteracts the benefits of exogenously applied salicylic acid and aggravates the deleterious effects of the southern bean mosaic virus in Rhizobium-nodulated Phaseolus vulgaris plants

Infection of Rhizobium-nodulated Phaseolus vulgaris by the southern bean mosaic virus (SBMV) markedly decreased the growth and nodulation of plants. Exogenous applications of salicylic acid (SA) at concentrations ≥10 μM further decreased growth and nodulation of virus-infected (V) plants. Only SA concentration of 5 μM in the solution improved the growth, nodulation, chlorophyll concentration and the catabolism of ureide in leaves of V plants. The spray of leaves with 2 mM propyl gallate (+Pg) decreased growth, nodulation as well as the chlorophyll and leaf ureide concentrations in V plants, regardless of the concentration of SA at which plants were grown. Ultrastructural damages in leaf cells caused by SBMV were also enhanced in V+Pg plants. The massive proliferation of virus particles and the presence of virus crystalline arrays within symbiosomes of nodules in V+Pg plants were attributed to proliferation of branched plasmodesmata in leaf vascular-parenchyma cell walls that facilitated virus movement. Virus particles were never observed in leaf and nodule tissues of V plants not sprayed with Pg. Exogenous applications of SA hindered while Pg increased the symbiotic performance of H plants, pointing out the complexity to be addressed in breeding for virus resistance in Rhizobium-nodulated beans.     

Detection of epigenetic variation in tissue-culture-derived plants of <Emphasis Type="Italic">Doritaenopsis</Emphasis> by methylation-sensitive amplification polymorphism (MSAP) analysis

Somaclones exhibiting variations with flower characteristics were recovered from the tissue-culture-derived plants of Doritaenopsis. Two molecular techniques, random amplified polymorphic DNA (RAPD) and methylation-sensitive amplification polymorphism (MSAP) analyses, were used to characterize the somaclones. RAPD analysis, using 100 randomly selected primers, failed to differentiate variants and normal plants, even though some primers (six out of 100 primers) exhibited 6–10 distinct banding patterns. However, MSAP analysis revealed the differences in the DNA methylation patterns in the normal and variant plants which were correlated with phenotypic variation. In all, 311, 337, 366, and 343 fragments were obtained with normal and V1, V2, and V3 variant plants, respectively; each representing recognition site cleaved by either or both of the isoshizomers were amplified using 12 combination of primers. A total of 36 (11.6%), 77 (22.9%), 73 (19.9%), and 47 (13.7%) sites were found to be methylated at cytosine in the genomes of normal and V1, V2, and V3 variant Doritaenopsis plants. This study demonstrates usefulness of MSAP to detect DNA methylation events in tissue cultured Doritaenopsis plants.     

Pollen-mediated transgene flow in the wind-pollinated grass species tall fescue (Festuca arundinacea Schreb.)

Information regarding gene flow in wind-pollinated, outcrossing forage grasses is essential for any future releases of value-added transgenic cultivars. Experiments on pollen dispersal was carried out by growing transgenic tall fescue (Festuca arundinacea) in a central plot, surrounded by exclosures containing recipient plants up to a distance of 200 m from the central source plants in eight directions. The central transgenic tall fescue plants carried a chimeric hygromycin phosphotransferase gene (hph) and a chimeric -glucuronidase gene (gusA). Seeds were collected from the recipient plants and germinated seedlings were used for high throughput DNA isolation and polymerase chain reaction (PCR) analysis. More than 21,000 seedlings were PCR analyzed for the experiments conducted in three years. Transgenes were detected in recipient plants at up to 150 m from the central transgenic plot. The highest transgene frequencies, 5% at 50 m, 4.12% at 100 m and 0.96% at 150 m, were observed north of the central plot, the prevailing wind direction. Lower transgene frequencies were detected in other directions, particularly at 100 m and 150 m distances. No transgene was detected at 200 m distance in any direction. Transgene flow was less effective or ineffective when recipient plants were further away from the central donor plants. Southern blot hybridization analysis confirmed the transgenic nature of the PCR positive plants. A supplementary experiment demonstrated that transgene flow can be controlled by placing transgenic plantings downwind and long distances from non-transgenic seed increases, thus allowing tall fescue breeding and transgene development programs to be conducted concurrently at the same research station.     

Development and practice of a glasshouse screening technique for resistance of wheat to the aphid Sitobion avenae

Differences in resistance to S. avenae were confirmed amongst eight spring wheat stocks in glasshouse tests. Taking these stocks as standards, a glasshouse screening technique was developed in which 15 plants of each entry were arranged randomly in one block and aphids were scattered over the whole at the stem-extension phase. When the infestation of individual plants was scored about 2 wk later, resistant and susceptible stocks were well differentiated. Provided that scoring was completed before or at ear emergence, separation of resistant entries was more efficient with older plants. Ranking of the standard stocks was little influenced by other variations of technique or environment. Twenty-six out of 91 previously untested spring wheat breeding lines appeared to be moderately resistant to S. avenae, suggesting that progress in the selection of resistant cultivars will depend more on the further development of suitable techniques than on the availability of heritable resistance.     

Induction of resistance to Clavibacter michiganensis subsp. michiganensis

Tomato plants pre-inoculated with the avirulent strain NCPPB 3123 of Clavibacter michiganensis subsp. michiganensis (Cmm) were protected largely against challenge infection by virulent strains of Cmm. Effectiveness of this protective effect was mainly dependent on the inoculation sites, the bacterial cell concentration used for pre- and challenge inoculations, and the time interval between both inoculations. This defence reaction was systemic and stable throughout the whole growing season. Resistance can also be induced by pre-inoculation of heat-killed bacteria or application of isolated EPS of the strain 3123. Strain 3123 spreads out in tomato plants in the same manner as virulent Cmm isolates, but its colonization of tomato fruits and seeds was substantially lower. Papillary to spherical electron dense particles were observed at the tonoplast in parenchyma cells of the vascular system of tomato plants inoculated with the strain 3123. Numerous investigations carried out to examine the ability of 3123 to induce resistance in other host/pathogen-systems showed that it was only specific for tomato/Cmm.     

Fungi associated with black pepper plants in Quang Tri province (Vietnam), and interaction between Meloidogyne incognita and Fusarium solani

During a survey of nurseries and plantations of black pepper plants in Quang Tri province in Vietnam during the rainy season of 2007, nine fungal taxa were isolated from the roots of the black pepper plants. Fusarium solani was found in about one out of four black pepper root samples examined but not in the nurseries and also not from black pepper plants younger than five years growing in plantations. Since in these nurseries about one out of two black pepper plants examined had yellow leaves, this observation suggests that another pathogen must be the initial cause of the yellowing of the leaves. A likely pathogenic candidate is M. incognita which was extracted from every single black pepper plant examined in the nurseries. During the same survey, we also observed that F. solani was not isolated from the roots of black pepper plants that did not had yellow leaves and that the percentage of black pepper plants with yellow leaves increased with increased frequency of occurrence of F. solani. This observation indicates that F. solani plays a role in the yellowing of the leaves of black pepper plants in a later stage of the development of the plants. The results of a greenhouse experiment showed the negative effects inoculation with M. incognita alone or in combination with F. solani may have on the percentage of black pepper plants with yellow leaves and on plant growth. No effect of inoculation with F. solani before, at the same time, or two weeks after inoculation with M. incognita on root galling and nematode reproduction was observed.     

Density‐dependent effects on initial growth of a branching coral under restoration

Coral reef restoration aims to help threatened coral ecosystems recover from recent severe declines. Here we address whether coral fragments should be out‐planted individually or in larger aggregations. Theory suggests alternative possible outcomes: whereas out‐plants within aggregations might suffer from heightened negative interactions with neighbors (e.g. competition for space), they may alternatively benefit from positive interactions with neighbors (e.g. buffering wave disturbances). On a degraded reef in the Caribbean (St. Croix, USVI), using out‐plants of the critically endangered staghorn coral Acropora cervicornis, we experimentally tested how aggregation density (1–20 out‐planted coral fragments spaced at approximately 5 cm) influenced initial coral growth (over 3 months). Coral growth declined as a function of aggregation size, and out‐plants within larger aggregations had fewer and shorter secondary branches on average, indicative of horizontal competition for space. Our results therefore suggest that wide spacing of individuals will maximize the initial growth of out‐planted branching corals.     

Use of atrazine sensitive leguminous plants as biological indicators to evaluate the atrazine degradation efficiency of a bacterial inoculum

Atrazine sensitive leguminous plants were grown in a soil spiked with atrazine and augmented with an atrazine-degrading bacterium, Arthrobacter sp. strain MCM B-436, to ascertain its degradative efficiency. Germination and survival of plants was correlated with atrazine removal from soil. This experiment was carried out at laboratory as well as field level, showing consistent results. This bioindicator approach serves as an efficient measure for atrazine removal and could be easily adapted to determine atrazine degradation efficiency of other microbial strains.     

Association between cacti and nurse perennials in a heterogeneous tropical dry forest in northwestern Mexico

Abstract. The Sierra de La Laguna, Baja California Sur harbors the only tropical dry forest of the Peninsula. An inventory of the perennial plants was carried out at the eastern part of this mountain range. Plant spatial distribution was analyzed considering the species composition and vertical stratification of plant clusters in a study plot of 2 500 m². A plant cluster was defined as a group of at least three individuals with a maximum distance between plants of 1 m. Soil physical-chemical characteristics and soil surface temperatures were compared between shade and sun conditions. The tropical dry forest of Baja California Sur presents a highly aggregated pattern of species distribution. Most of the perennial plants grow clumped, yet not all of the six cactus species recorded were significantly associated with them. Pattern analysis revealed that only Stenocereus thurberii and Ferocactus peninsulae were aggregated and associated to perennial plants. There is no replacement pattern between cacti and perennial plants. Nutrient content, including soil fertility, between shade and sun conditions does not seem to explain the patchy distribution of perennial plants, while the differences in solar radiation between these conditions appear to be restrictive and determinant of this spatial pattern of distribution.     

Root Rot and Wilt of Kangaroo Paw (Anigozanthos manglesii) Caused by Pythium myriotylum (Drechs.) in Israel

A root rot and wilt disease of Anigozanthos manglesii (Kangaroo Paw) grown in greenhouses in Israel, for exporting as cut flowers to Europe, was characterized. Pythium myriotylum (Drechs.) and Rhizoctonia solani (Kühn) were the prevalent pathogens in diseased plants collected from commercial greenhouses. Fusarium oxysporum, Fusarium spp. and Myrothecium sp. were also isolated, but P. myriotylum or R. solani were not detected in samples from symptomless plants in tissue cultures (Australian origin) or plants at different stages in the nursery; non‐pathogenic F. oxysporum and Fusarium spp. were detected in several samples. In pathogenicity tests carried out in pots, plant mortality occurred 7 days after inoculation with P. myriotylum. In a field experiment carried out in methyl bromide‐fumigated soil, the incidence of dead plants following inoculation with P. myriotylum alone was 22% 10 days after inoculation, increasing to 78% after an additional 25 days. The incidence of dead plants following inoculation with R. solani alone was only 5% and in plants inoculated simultaneously with both pathogens, disease incidence was 88% 35 days after inoculation. Mortality reached 90–100% in plants inoculated with P. myriotylum, either singly or combined with R. solani 60 days after inoculation, whereas in plants inoculated with R. solani it was 5%. The maximum mortality in plants inoculated with R. solani was 25%, 76 days after inoculation. These results clearly demonstrate that P. myriotylum was the dominant pathogen in the root rot and wilt of A. manglesii.     

‘Hertwig Effect’ in plants: induced parthenogenesis through the use of irradiated pollen

Summary Studies of four combinations of Nicotiana involving four species, N. langsdorffii (N. l.), N. alata (N. a.), N. glutinosa (N. g.) and N. tabacum (N. t.), have shown that parthenogenetic haploid and diploid maternal individuals may arise with the use of male gametes (pollen) treated with high doses of ionising radiation in plants, similar to that found in animals (Hertwig Effect). At lower doses (10–20 Kr) rapidly diminishing numbers of seedlings were produced and many of these died soon after germination or before reaching maturity. In the intraspecific combination N. l. × N. l., viable seeds were produced only at the lower doses of 10 and 15 Kr. In the interspecific combination N. l. × N. a., at lower doses, all plants that came to bloom showed variable hybrid morphology. There were no plants resembling the female parent. In the combination N.t. × N.a., at lower doses there were rare surviving plants which were maternal dihaploid (1 plant out of 4 at 15 Kr) or tetraploid N. tabacum (all 5 plants at 20 Kr). All surviving plants at higher doses (50 and 100 Kr) were maternal tetraploids. In the combination N.t.×N.g., plants produced at lower than 20 Kr were almost all either aneuploid or triploid hybrids. Dihaploid, maternal N. tabacum plants appeared at 20 Kr and higher doses. After 50 Kr the large proportion of plants produced were maternal dihaploid or tetraploid N. tabacum.     

Development of transgenic rice pure lines by particle bombardment‐mediated transformation and genetic analysis‐based selection

Mature seed‐derived callus from an elite Chinese japonica rice cv. Eyl 105 was transformed with a plasmid containing the selectable marker hygromycin phosphotransferase (hpt) and the reporter β‐glucuronidase (gusA) genes via particle bombardment. After two rounds of selection on hygromycin (30 mg/l)‐containing medium, resistant callus was transferred to hygromycin (30 mg/l)‐containing regeneration medium for plant regeneration. Twenty‐three independent transgenic rice plants were regenerated from 127 bombarded callus with a transformation frequency of 18.1%. All the transgenic plants contained both gusA and hpt genes, revealed by PCR/Southern blot analysis. GUS assay revealed 18 out of 23 plants (78.3%) proliferated on hygromycin‐containing medium had GUS expression at various levels. Genetic analysis confirmed Mendelian segregation of transgenes in progeny. From R2 generations with their R1 parent plants showing 3:1 Mendelian segregation, we identified three independent homozygous transgenic rice lines. The homozygous lines were phenotypically normal and fertile compared to the control plants. We demonstrate that homozygous transgenic rice lines can be obtained via particle bombardment‐mediated transformation and through genetic analysis‐based selection.     

Phyto-Extraction of Nickel by Linum usitatissimum in Association with Glomus intraradices

Plants show enhanced phytoremediation of heavy metal contaminated soils particularly in response to fungal inoculation. Present study was conducted to find out the influence of Nickel (Ni) toxicity on plant biomass, growth, chlorophyll content, proline production and metal accumulation by L. usitatissimum (flax) in the presence of Glomus intraradices. Flax seedlings of both inoculated with G. intraradices and non-inoculated were exposed to different concentrations i.e., 250, 350 and 500 ppm of Ni at different time intervals. Analysis of physiological parameters revealed that Ni depressed the growth and photosynthetic activity of plants. However, the inoculation of plants with arbuscular mycorrhizae (G. intraradices) partially helped in the alleviation of Ni toxicity as indicated by improved plant growth under Ni stress. Ni uptake of non- mycorrhizal flax plants was increased by 98% as compared to control conditions whereas inoculated plants showed 19% more uptake when compared with the non-inoculated plants. Mycorrhizal plants exhibited increasing capacity to remediate contaminated soils along with improved growth. Thus, AM assisted phytoremediation helps in the accumulation of Ni in plants to reclaim Ni toxic soils. Based on our findings, it can be concluded that the role of flax plants and mycorrhizal fungi is extremely important in phytoremediation.     

The transmissibility of a potyvirus by an aphid vector: reliability of measurements under controlled conditions

Reproducibility of measurements of the transmission rate of a given potyvirus by a given aphid vector was assessed using a standardised transmission procedure. The plant-virus-vector system studied was: Cucurbita pepo - Papaya ringspot virus - Aphis gossypii. Several elements concerning the rearing and handling of the aphids were analysed. One of these, the climatic conditions under which the aphids were reared, influenced the transmission rate of the virus. Six trials were carried out to measure the rate of transmission of the virus from a defined leaf surface area. The measurements appeared to be reliable. A two year experiment was carried out to compare transmission rates at different dates. Although strong variations were observed in some source plants, the transmission rates obtained for samples of 10 source plants were stable, with a 10% coefficient of variation. The results show that transmissibility of the virus is a stable property which is independent of the trial date. They are discussed in relation to standardised elements of the transmission process.     

INTRAPOPULATION DIFFERENTIATION IN ANNUAL PLANTS. III. THE CONTRASTING EFFECTS OF INTRA- AND INTERSPECIFIC COMPETITION

The roles of intraspecific and interspecific competition in producing differentiation within populations of Veronica peregrina were studied in two populations under controlled, greenhouse conditions. In nature, each population spans an environmental gradient across the center and sides of a temporary, vernal pool in California. Individuals at the center are subjected to intense intraspecific competition produced by high densities (to 30 seedlings/cm²) generated by quasi-simultaneous germination (90% of seeds germinate in one week). Individuals at the periphery are subjected to interspecific competition with grasses, which shade out the Veronica 4–6 weeks after the onset of winter growth. I predicted that 1) when grown under immediate intraspecific competition in the greenhouse, offspring of plants from the central subpopulation (C) would perform better (i.e., grow larger and produce more seeds) than those from the periphery (P) and that 2) when grown under delayed interspecific competition provided by Agrostis tenuis and Lollium multiflorum, offspring of plants from the periphery would perform better than those from the center. Both predictions were confirmed. The center-periphery differences were pronounced and statistically significant in an undisturbed population (V-2), while in a population disturbed by yearly plowing (V-3), the differences tended to be consistent with those in V-2 but seldom significant. Distribution of variability tended to be positively skewed and/or leptokurtic in subpopulations grown under “foreign” competition (i.e., intraspecific for P plants and interspecific for C plants) but was normally distributed following exposure to “familiar” competition. Timing of competition affected many results. There were four additional significant differences between the central and peripheral subpopulations. 1) Germination rate: the faster rate in central plants can be advantageous under immediate intraspecific competition. The slower rate in peripheral plants can be advantageous under conditions of erratic and unpredictable soil moisture. 2) Response to nutrient competition: central plants were more sensitive to N-deficiency and peripheral plants were more sensitive to P-deficiency. 3) Allocation of biomass: central plants allocated a greater proportion of biomass to seeds, while peripheral plants allocated a greater proportion of biomass to leaves under all growing conditions. 4) Root elongation: at the seedling stage, central plants have longer roots, while at the adult stage, peripheral plants have longer roots (but not more root mass). Most components of this complex pattern of differentiation are interpretable in an adaptive context. Other results defy simple explanations and underline the importance of phenotypic plasticity, which was pronounced in the competition experiments.     

Microhabitat choice and diet of rodents in Maynugus irrigation field, northern Ethiopia

An ecological study on the choice of microhabitat and diet of small mammals was carried out in Maynugus irrigation field of the Tigray region in northern Ethiopia. Abundance of rodents was high during the reproductive phase of the agricultural plants. Mastomys erythroleucus occurred more frequently in vegetable fields, whereas Arvicanthis dembeensis was more frequent in habitats with monocot plants. Scrotal males and reproductive females consumed more food by weight than abdominal males and quiescent females. Arvicanthis dembeensis consumed more food than M. erythroleucus. Both the species consumed more food during the main rainy season. All the cultivated plants were attacked by these rodents with a degree of difference in relation to season, species, sex, age and sexual condition. As rodent attack was more intense during the fruiting phase of the plants, rodent control management should be carried out during the flowering phase.     

我国白裤瑶饲料植物的民族植物学研究

白裤瑶非常喜欢饲养和食用当地黑猪，对饲料植物的采集和管理具有独特而丰富的经验和知识。该研究旨在对白裤瑶地区饲料植物及其有关的本土知识进行民族植物学编目，在2019—2021年期间，该文作者多次前往当地进行饲料植物的民族植物学调查，利用了关键人物访谈、半结构式访谈和小组讨论，同时该研究还通过打分排序和统计植物利用频度来筛选出综合利用价值高的饲料植物。结果表明：(1)白裤瑶传统饲料植物共104种，隶属于42科85属；被引用植物种数最多的科是菊科(16种，占总种数的15.38%),其次为荨麻科(13种，占总种数的12.5%)。(2)枝叶(56.73%)、全草(26.92%)和叶(8.65%)为当地饲料植物的主要利用部位，大多数饲料植物是草本植物(88.46%),多数饲料植物(占总数的64.42%)一年四季都可以采集，保障了家猪食物来源充足。(3)饲料植物的主要加工方法是将其切碎或打碎后煮熟饲喂。(4)当地被受访者引用最多的物种是玉蜀黍、构树、番薯、桑、金荞麦、稻。(5)根据综合打分排序，在当地具有较高综合评价的饲料植物分别为构树、番薯、桑、金荞麦、南瓜。白裤瑶对饲料植物用于养猪的传统知识非常...     

Transgenic tobacco plants containing <Emphasis Type="Italic">Bt</Emphasis> and <Emphasis Type="Italic">GNA</Emphasis> genes

A new plant expression vector (pBSbtCry1Ac-GNA) containing two insect resistant genes, a synthetic chimeric gene SbtCry1Ac encoding the insecticidal protein CrylAc and a gene GNA encoding snowdrop lectin (Galanthus nivalis agglutinin) was constructed. Transgenic tobacco plants containing these two genes were obtained through Agrobacterium-mediated transformation of tobacco leaf discs. Results from PCR detection and genomic DNA Southern blot analysis indicated that both SbtCrylAc gene and GNA gene were integrated into the genome of these plants. Results of Western blot analysis indicated that these two proteins were expressed in the analyzed plants. Bioassays of Myzus persicae and Helicoverpa assulta on detached leaves of transformed tobacco plants were carried out. The average aphid inhibition rate of these plants tested at 12 d post-infestation was 71.9 %. The average H. assulta mortality of these plants tested at 6 d post-infestation was up to 89.8 %. The kanamycin resistance of the T₁ progeny of these transgenic plants was analyzed and a typical 3:1 segregation was observed.