Response of Stylosanthes guianensis to endomycorrhizal fungi inoculation as affected by lime and phosphorus applications

The effect of 3 different species of vesicular-arbuscular mycorrhizal fungi on the growth of Stylosanthes guianensis (Aubl.) Sw. cultivated in a sterilized acid and dystrophic soil (Quartzipsament), with 4 levels of lime (0; 0.27; 0.63 and 1.10 meq Ca²⁺/100 g air-dried soil, as Ca(OH)₂) and 2 P levels (0 and 20 mg P/kg soil, as KH₂PO₄) was evaluated under greenhouse conditions. Plants were harvested 40, 60, and 80 days after planting. Stylosanthes guianensis was highly mycotrophic, especially in soil without P fertilization. Mycotrophism was highest in plants inoculated with Acaulospora scrobiculata in soil receiving no P fertilizer and with 0.63 meq Ca²⁺/100 g air-dried soil. Shoot growth increment was as high as 5129% at the third harvest. Inoculation with Glomus macrocarpum presented intermediate results, whereas inoculation with Gigaspora margarita had no significant effect on plant growth. Root per cent colonization and shoot dry weight, as well as root percent colonization and shoot to root ratio were significantly correlated. The occurrence of S. guianensis in very acid and dystropic soils, containing toxic levels of Al³⁺, requires the association with VAM fungi for the plant tolerate such conditions.     

Somaclonal variation from Sorghum bicolor (L.) Moench cell culture

Plant Cell, Tissue and Organ Culture (PCTOC) - Sorghum bicolor (L.) Moench, plants were regenerated from 4 to 5 month old callus cultures originally derived from seedling explants. Somaclonal...     

Plant regeneration from leaf-derived callus cultures of the tropical pasture legume Stylosanthes scabra Vog.

Callus cultures of 5 genotypes of S. scabra Vog. were optimally established from leaf tissue on Murashige and Skoog (MS) basal medium supplemented with 0.5–2.0 mg l^-1 2, 4-Dichlorophenoxy acetic acid (2, 4-D) and 1.0–2.0 mg l^-1 6-benzylaminopurine (BAP). On media containing 2, 4-D only, calli were soft, and rhizogenesis occurred on calli of 4 genotypes. Calli formed on media containing BAP only, but not with kinetin only. In the presence of 2, 4-D, BAP inhibited rhizogenesis and promoted better callus growth than kinetin. High frequency shoot induction was achieved for 3 genotypes on MS +2.0 mg l^-1 BAP. Roots formed on shoots when sub-cultured on half-strenght MS without growth regulators. The form of cytokinin used in the callus induction media appeared to affect subsequent shoot organogenesis. Genotypic differences were observed for shoot organogenesis. There was some morphological variation evident among regenerants.     

Response ofStylosanthes guianensis to endomycorrhizal fungi inoculation as affected by lime and phosphorus applications

The nutrient concentration in the shoots ofStylosanthes guianensis (Aubl.) Sw. cultivated in a sterilized acid and dystrophic soil (Quartzipsament) amended with 4 levels of lime (0; 0.27; 0.63 and 1.10 meq Ca²⁺/100 g soil, as Ca(OH)₂), 2 levels of P (0 and 20 mg P kg^-1 soil, as KH₂PO₄) and not-inoculated or inoculated with 3 vesicular-arbuscular (VA) mycorrhizal fungi was evaluated under greenhouse conditions. The effectiveness of the different fungal species in increasing the nutrient concentration in the shoots varied with the different edaphic conditions. In general, mycorrhiza formation was associated with increases in the concentrations of most of the nutrients analyzed. Under the experimental conditions, the increments in nutrient concentration were higher overall in plants inoculated withAcaulospora scrobiculata. Inoculation ofS. guianensis with VA mycorrhizal fungi was also associated with alterations in the ratios of nutrients in the shoots, which might be important in understanding and explaining the tolerance of mycorrhizal plants to nutritional stresses such as Al and Mn toxicity.     

The use of carbon isotope ratios to evaluate legume contribution to soil enhancement in tropical pastures

Soil carbon distribution with depth, stable carbon isotope ratios in soil organic matter and their changes as a consequence of the presence of legume were studied in three 12-year-old tropical pastures (grass alone —Brachiaria decumbens (C₄), legume alone —Pueraria phaseoloides (C₃) and grass + legume) on an Oxisol in Colombia. The objective of this study was to determine the changes that occurred in the¹³C isotope composition of soil from a grass + legume pasture that was established by cultivation of a native savanna dominated by C₄ vegetation. The¹³C natural abundance technique was used to estimate the amount of soil organic carbon originating from the legume. Up to 29% of the organic carbon in soil of the grass + legume pasture was estimated to be derived from legume residues in the top 0–2-cm soil depth, which decreased to 7% at 8–10 cm depth. Improvements in soil fertility resulting from the soil organic carbon originated from legume residues were measured as increased potential rates of nitrogen mineralization and increased yields of rice in a subsequent crop after the grass + legume pasture compared with the grass-only pasture. We conclude that the¹³C natural abundance technique may help to predict the improvements in soil quality in terms of fertility resulting from the presence of a forage legume (C₃) in a predominantly C₄ grass pasture.     

Somatic cell culture of rice cultivars with different grain types: Somaclonal variation in some grain and quality characters

Variations in some grain and quality characters in progenies of regenerated rice plants were studied. Grain length and weight decreased significantly, yet gel consistency increased. Variations in these quantitative characters of all cultivars studied were consistent, showing the tendencies of the variations. Grain protein contents of the somaclones were higher in one cultivar. Variability of most traits was increased by combining low-dosage radiation and tissue culture.     

Root nodulation and nitrogen accumulation and partitioning in legume crops as affected by soil solarization

Soil solarization is a preplanting technique used in hot climates to control weeds and soilborne pathogens consisting of mulching the soil surface with polyethylene sheets. The increase in temperature associated with solarized soil could affect nitrogen availability for grain legume crops through effects on nitrogen fixing soil microorganisms or other mechanisms. To examine the effects of solarization on natural root nodulation and nitrogen accumulation and partitioning in the plant, two solarization field experiments were carried out over two planting seasons, involving genotypes of both faba bean (Vicia faba) and chickpea (Cicer arietinum). The effect of sowing date was also studied in the first season. Solarization increased the maximum soil temperature by 9–10 °C in the first, and by 13–15 °C in the second season. At 5 cm below the solarized soil surface, a temperature of over 46 °C prevailed for 146 and 280 h over the two respective seasons, while this temperature was not attained in unmulched soil. Solarization delayed the initiation of nodulation and consistently reduced the nodule number per host plant, but generated an approximate doubling of mean nodule weight. The total nodule mass per plant was not affected by the treatment in the first season, but was reduced in the second season. Solarization significantly increased the concentrations of NO₃^– -N, Na⁺, Zn²⁺, Ca²⁺ and K⁺ in the soil extract, and the total nitrogen accumulated in the whole plant. This latter increase was due to both higher plant growth and a greater plant nitrogen concentration. The increased nitrogen level in the plant was not uniform with respect to plant component, varying from 57% in the roots to 198% in the pods and seeds. The plants grown in non-solarized soil accumulated about 31% of their total N content in the shoots of the parasitic weed Orobanche crenata. Solarization dramatically improved grain yield by 300–900% in both seasons and in all genotypes studied, due to increased N availability in soil, N accumulation in plants, improved plant growth, and complete control of the parasite weed O. crenata. On the basis of these beneficial effects, soil solarization, which avoids site contamination and is suited to organic farming, should be a good opportunity in Mediterranean areas where the level and stability of grain yields are low, and the infestation of O. crenata is high.     

Cloning of a 9-<Emphasis Type="Italic">cis</Emphasis>-epoxycarotenoid dioxygenase gene (<Emphasis Type="Italic">SgNCED1</Emphasis>) from <Emphasis Type="Italic">Stylosanthes guianensis </Emphasis>and its expression in response to abiotic stresses

Abscisic acid (ABA) regulates plant adaptive responses to various environmental stresses. Oxidative cleavage of cis-epoxycarotenoids catalyzed by 9-cis-epoxycarotenoid dioxygenase (NCED) is the main regulatory step in the biosynthesis of ABA in higher plants. A NCED gene, SgNCED1, was cloned from the dehydrated leaves of Stylosanthes guianensis. The 2,241-bp full-length SgNCED1 had a 1,809-bp ORF, which encodes a peptide of 602 amino acids. The deduced amino acid sequence of SgNCED1 protein shared high identity with other NCEDs. At the N-terminus of the SgNCED1 located a chloroplast transit peptide sequence. DNA blot analysis revealed that SgNCED1 was a single copy gene in the genome of S. guianensis. The relationship between expression of SgNCED1 and endogenous ABA level was investigated. The expression of SgNCED1 was induced in both leaves and roots of S. guianensis under drought stress. Dehydration and salt stress induced the expression of SgNCED1 strongly and rapidly. The ABA accumulation was coincidently induced with the SgNCED1 mRNA under drought, dehydration and salt stress. The expression of SgNCED1 and ABA accumulation were also induced under chilling condition.     

Somaclonal variation in celery: screening for resistance to Fusarium oxysporum f. sp. apii

Summary Two methods were used to screen putative Fusarium-resistant celery (Apium graveolens L.) plantlets from cell culture: placing plantlets on a mycelial mat for one month or planting them directly in Fusarium-infested soil. Resistant phenotypes were identified with both methods, but the plants grown on the mycelial mat died before they reached reproductive maturity. Four plants, K, T-2, T-3, and R-R₁ from the soil screen, survived and produced viable seed. Tests of self-pollinated progeny, in field and greenhouse conditions, showed that T-2, T-3, and R-R₁ were superior to the original cultivar, 5270R, with respect to disease resistance, as measured by vascular discoloration and plant height. Chi-square analysis of progeny scores for root and crown decay showed that the new variation was heritable and appeared to be conditioned by more than one locus.     

Induction of a protective antibody response to FMDV in mice following oral immunization with transgenic <Emphasis Type="Italic">Stylosanthes</Emphasis> spp. as a feedstuff additive

The expression of antigens in transgenic plants has increasingly been used as an alternative to the classical methodologies for the development of experimental vaccines, and it remains one of the real challenges in this field to use transgenic plant-based vaccines effectively as feedstuff additives. We report herein the development of a new oral immunization system for foot and mouth disease with the structural protein VP1 of the foot and mouth disease virus (FMDV) produced in transgenic Stylosanthes guianensis cv. Reyan II. The transgenic plantlets were identified by polymerase chain reaction (PCR), Southern blotting, and northern blotting; and the production of VP1 protein in transgenic plants was confirmed and quantified by western blotting and enzyme-linked immunosorbent assays (ELISA). Six transformed lines were obtained, and the level of the expressed protein was 0.1–0.5% total soluble protein (TSP). Mice that were orally immunized using studded feedstuff mixed with desiccated powder of the transgenic plants developed a virus-specific immune response to the structural VP1 and intact FMDV particles. To our knowledge, this is the first report of transgenic plants expressing the antigen protein of FMDV as feedstuff additives that has demonstrated the induction of a protective systemic antibody response in animals. These results support the feasibility of producing edible vaccines from transgenic forage plants, and provide proof of the possibility of using plant-based vaccines as feedstuff additives.     

Screening for plant growth-promoting rhizobacteria in Chamaecytisus proliferus (tagasaste), a forage tree-shrub legume endemic to the Canary Islands

Rhizobacteria with properties related to plant growth-promotion were isolated from the rhizosphere of the perennial legume Chamaecytisus proliferus ssp. proliferus var. palmensis (tagasaste) growing in field conditions. Samples were collected in two localities of the Tenerife Island: La Laguna and El Tanque, NE and NW at 600 and 1000 meters above sea level, respectively, and in two seasons, winter and summer. The strains were isolated by using culture dependent procedures, and identified by phenotypic (culturable and biochemical) and genotypic (ERIC-PCR fingerprinting) features. The rhizosphere isolates formed a diverse community of mainly Gram-negative bacteria, with members of genera Pseudomonas, Burkholderia and Sphingomonas being predominant. A high level of selectivity was found in the rhizosphere environment as compared to the non-rhizosphere soil where Gram-positives were more abundant. Species richness (number of species) and species abundance were related to the sampling season and the locality, thus, samples obtained in winter at both sites had larger counts than samples obtained in summer, and the higher species richness was found in La Laguna. The species Pseudomonas fluorescens showed the highest number of properties related to plant growth promotion (PGP): 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, phytohormone production, nitrogen fixation, fungal growth inhibition and cyanogenesis, thus it seems to be the most suitable microorganism to be tested in PGP-field experiments.     

Genome size and polyploidy variation in the tropical hardwood genusTerminalia (Combretaceae)

Chromosome complements and 2C DNA amounts of six species ofTerminalia have been studied.Terminalia oliveri, T. myriocarpa andT. arjuna are diploid (2n = 24),T. chebula andT. bellirica are tetraploid (2n = 48),T. muelleri shows a triploid number (2n = 36). Two well demarcated groups of species are recognizable on the basis of chromosome length and 2C DNA values which range from 3.60 pg (T. oliveri) to 12.80 pg (T. bellirica) showing a 3.5-fold difference. Differences of DNA per basic genome or per chromosome are greatest (1.97-fold) betweenT. oliveri andT. arjuna. Two species groups (1)T. oliveri andT. chebula, and (2)T. myriocarpa, T. arjuna, T. muelleri, T. bellirica, therefore are well differentiated by DNA per basic genome, irrespective of polyploidy. The mean values of the two groups are 1.81 pg and 3.34 pg, respectively, showing a 1.84-fold difference. Within diploids and tetraploids there is 1.97-fold and 1.76-fold variation, respectively.     

Growth and yield traits of sorghum grown on acid soil at varied aluminum saturations

Genetic manipulation of crops to tolerate mineral stresses is a practical approach to improve productivity of tropical acid soils. Both acid soil tolerant (AS-T) and susceptible (AS-S) sorghum [Sorghum bicolor (L.) Moench] genotypes were grown in the field on an acid ultisol at Quilichao, Colombia, South America at 60% (60-Al) and 40% (40-Al) Al saturation to evaluate plants for growth and yield traits. Except for days to flowering and root mass scores, AS-T genotypes showed no differences in growth (plant height, head length and width, second internode length and diameter, and acid soil toxicity rating) and yield (total and stover dry matter yields, grain yield, head yield, seeds per head, and 100-seed weight) traits when plants were grown at 60-Al or 40-Al. Plants grown at 60-Al were delayed in flowering and had lower root mass scores. The AS-S genotypes showed improvement for the growth and yield traits when grown at 40-Al compared to 60-Al. The growth and yield traits of the AS-S genotypes were usually less favorable for plants grown at 40-Al than the same traits were for the AS-T genotypes grown at 60-Al. Harvest indices (ratio of grain to total plant yield) were no different for the genotypes grown at 40-Al, and only slightly higher for the AS-T genotypes grown at 60-Al. Sorghum genotypes more tolerant to acid soil conditions showed favorable growth and yield traits when grown under relatively severe acid soil (60-Al, pH 4.1) conditions. Certain sorghum genotypes were able to adapt and effectively produce grain when grown on acid soils with few inputs to reduce acid soil toxicity problems. Published as Paper No. 6690, Journal Series, Mississippi Agricultural and Forestry Experiment Station. Funding received in part from the International Sorghum/Millet Collaborative Research Support Program (INTSORMIL CRSP) through US Agency for International Development (USAID) grant AID/DAN-1254-G-SS-5065-00 and project Nos. MS-111 (MSU) and NE-114 (UNL).     

Somaclonal variation in a maize inbred line is not associated with changes in the number or location of Ac-homologous sequences

Summary Somaclonal variation (tissue culture-induced mutations) may result, in some instances, from the activation of transposable elements. This study was conducted to determine whether somaclonal variants in the Zea maize L. inbred line FR27rhm were associated with movement of the transposable element Activator (Ac). Ten variants, seven of which from genetic analyses fit a single recessive gene model and three which did not due to a low number of mutant plants, were selected for analysis. Total DNA from these and from uncultured FR27rhm seedlings were examined by Southern blot analysis using the internal 1.6-kb Hind III fragment derived from the cloned Ac7 element as a probe. By using a restriction endonuclease which does not cut within the element, the number and distribution of the copies of Acrelated sequences in the FR27rhm genome could be determined. From the number of bands seen in the blots, we conclude that the FR27rhm inbred contains large numbers of Ac -related sequences. However, the pattern of bands seen in the ten variants and in the uncultured seedlings were identical, indicating that there had been no movement of any of the Acrelated sequences to cause the tissue culture-induced mutations.Contribution from Department of Agronomy, University of Illinois, Urbana, IL 61801, supported by funds from the Illinois Agricultural Experiment Station and Illinois Foundation Seeds     

桉树与红锥混交对土壤养分及林下植物功能群的影响

桉树人工林对生态环境的影响一直是全球性争议的热点问题,桉树与珍贵乡土树种混交的生态环境效应备受关注。为探究桉树混交营林措施对林地土壤养分及林下植物功能群的影响,该研究以桉树纯林(PE)、桉树×红锥混交林(MEC)和红锥纯林(PCH)为对象,开展了林下植物群落及环境因子的调查测定。研究结果显示：不同林分的土壤理化性质具有显著差异,混交林的土壤pH、有效氮(AN)和有效磷(AP)含量显著高于纯林;而土壤含水量(SMC)、有机碳(SOC)、总氮(TN)含量及C∶N和C∶P在混交林中没有显著优势,并呈现PEMEC>PCH的趋势。混交林显著增加林下木本植物功能群(WFG)的物种丰富度,而PCH显著增加蕨类植物功能群(FeFG)的物种丰富度。混交林的WFG和禾草植物功能群(GFG)的重要值均显著高于红锥纯林,而红锥纯林FeFG的重要值显著高于混交林。主坐标分析结果表明,MEC与PE的林下植物功能群组成差异不显著,但与PCH的差异显著;冗余分析结果揭示了AN和AP是WFG占优势的主要影响因子,SMC、TN...     

Increased root length and branching in cotton by soil application of the plant growth regulator PGR-IV

Field and growth chamber studies were used to determine the effect of in-furrow application of PGR-IV on root and shoot development, and yield of cotton. In the field study, an in-furrow application of PGR-IV @ 73 mL ha^–1 at planting increased yield by 18% compared to the untreated control, and by 11% compared to 2-foliar applications of 292 mL/ha^–1 each at pinhead square stage of flower development and at first flower appearance. Growth chamber studies revealed that the in-furrow applications of PGR-IV @ 1.131L/plant dramatically increased root length (+47%), root dry weight (+29%), number of lateral roots per plant (+75%), and nutrient uptake one week after planting. These differences were still apparent five weeks later at pinhead square but to a lesser degree. The yield enhancement from the foliar applications was associated with increases in leaf growth, nutrient uptake, and boll number, whereas the yield enhancement from the soil application was associated with enhanced root growth and nutrient uptake. The positive effect of PGR-IV on root growth and accelerated early-season growth could have very substantial benefits in cotton production.     

T-DNA rearrangements due to tissue culture: somaclonal variation in crown gall tissues

After three years of apparent stability in tissue culture, the single cell derived shooty crown gall line sNT1.013 produced a revertant shoot which had switched from non-rooting (Rod⁺) and octopine synthesizing (Ocs⁺) to Rod^- Ocs^-, indicating that in this revertant TL-DNA genes 4 (causing the Rod⁺ trait) and gene 3 (causing the Ocs⁺ trait) had been inactivated. Southern blots revealed that the inactivation of these T-DNA genes was the result of a considerable rearrangement of DNA sequences, accompanied by deletions and possibly also by DNA amplifications. This study for the first time unambiguously proves that foreign genes which have been introduced via Agrobacterium tumefaciens can, at a low frequency, be inactivated after T-DNA integration because of reorganization of T-DNA sequences during tissue culture. This can be considered as an event of somaclonal variation.     

Genetic variability in host plant adaptation of the green peach aphid, Myzus persicae

The green peach aphid, Myzus persicae (Sulz.), is polyphagous on over 400 plant species in more than 50 families. Phenotypic plasticity of individuals and genetic variability in the population presumably contribute to this polyphagy. The genetic variability in field populations of M. persicae was assessed with respect to their adaptation to sugar beets and potatoes. An analysis of more than 1 000 clones, sampled during 1980, 1981 and 1982 from different host plants in the field, revealed a wide genetic variability in host plant adaptation to sugar beets as well as to potatoes. Both traits seem to be inherited independently from each other and do not correlate with clone-specific host plant preference of apterous adults. The aphid M. persicae can be characterized as a polyphagous insect species with a wide, continuously distributed variability and a broad phenotypic plasticity. A general differentiation of herbivorous species into generalists and specialists tends to ignore the genetic component in the complex of insect-plant relationships.

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Zusammenfassung Die Grüne Pfirsichblattlaus Myzus persicae (Sulz.) lebt polyphag an über 400 Pflanzenarten in mehr als 50 Pflanzenfamilien. Ein breites Nahrungsspektrum einer Art ergibt sich jedoch aus der phänotypischen Plastizität des Individuums oder einer Klonlinie und der genetischen Variabilität der Population. Felpopulationen der Grünen Pfirsichblattlaus wurden auf ihre genetische Variabilität bezüglich der Wirtspflanzenanpassung an Rübe und Kartoffel untersucht. Eine Analyse von mehr als 1 000 Klonen, die über die Jahre 1980, 1981 und 1982 im Rheinland gesammelt wurden, lassen eine breite Variabilität in der Wirtspflanzenanpassung der Population erkennen. Beide Merkmale scheinen unabhängig voneinander vererbt zu werden und zeigen keine Beziehung zum Wirtswahlverhalten adult apterer Läuse der entsprechenden Klone. Die Art M. persicae kann daher charakterisiert werden als eine polyphage Insektenart mit einer breiten genetischen Variabilität und einer grossen phänotypischen Plastizität. Eine generelle Differenzierung von Herbivoren in Generalisten und Spezialisten vernachlässigt die genetische Komponente in der komplexen Beziehung zwischen Insekten un ihren Wirtspflanzen.

     

Arbuscular mycorrhizal fungi and plant symbiosis in a saline-sodic soil

The seasonality of arbuscular mycorrhizal (AM) fungi–plant symbiosis in Lotus glaber Mill. and Stenotaphrum secundatum (Walt.) O.K. and the association with phosphorus (P) plant nutrition were studied in a saline-sodic soil at the four seasons during a year. Plant roots of both species were densely colonized by AM fungi (90 and 73%, respectively in L. glaber and S. secundatum) at high values of soil pH (9.2) and exchangeable sodium percentage (65%). The percentage of colonized root length differed between species and showed seasonality. The morphology of root colonization had a similar pattern in both species. The arbuscular colonization fraction increased at the beginning of the growing season and was positively associated with increased P concentration in both shoot and root tissue. The vesicular colonization fraction was high in summer when plants suffer from stress imposed by high temperatures and drought periods, and negatively associated with P in plant tissue. Spore and hyphal densities in soil were not associated with AM root colonization and did not show seasonality. Our results suggest that AM fungi can survive and colonize L. glaber and S. secundatum roots adapted to extreme saline-sodic soil condition. The symbiosis responds to seasonality and P uptake by the host altering the morphology of root colonization.     

Somaclonal variation of the mitochondrial ATPase subunit 6 gene region in regenerated triticale shoots and full-grown plants

Comparative hybridization analyses of total DNA from fertile and cytoplasmic male-sterile (CMS) triticale plants which had been regenerated from embryogenic callus cultures revealed the organization and variation of the mitochondrial atp6 gene region. In order to compare different developmental phases, we analysed mitochondrial DNA (mtDNA) from both the shoots and full-grown regenerants. Somaclonal variants were identified on the basis of differences in the mtDNA from fertile and CMS triticale. Several shoots as well as all of the full-grown plants analysed showed somaclonal variation. This phenomenon could be traced back to having primarily orginated from the influence of the nuclear background, which give rise to a stoichiometric increase in a rye-specific orf25 gene copy, and a tissue culture-induced combination of fertile and CMS-specific mtDNA organization of the atp6 gene area. The latter event is probably caused by the homologous recombination of repetitive sequences that may be accompanied by selective amplifications.