Serological Methods for Detection of Polymyxa graminis, an Obligate Root Parasite and Vector of Plant Viruses

A purification procedure was developed to separate Polymyxa graminisresting spores from sorghum root materials. The spores were used as im-munogen to produce a polyclonal antiserum. In a direct antigen coating enzyme-linked immunosorbent assay (DAC ELISA), the antiserum could detect one sporosorus per well of the ELISA plate. In spiked root samples, the procedure detected one sporosorus per mg of dried sorghum roots. The majority of isolates of P. graminis from Europe, North America, and India reacted strongly with the antiserum. Interestingly, P. graminis isolates from the state of Rajasthan (northern India), from Pakistan, and an isolate from Senegal (West Africa) reacted weakly with the antiserum. The cross-reactivity of the serum with P. betae isolates from Belgium and Turkey was about 40% of that observed for the homologous isolate. There was no reaction with common fungi infecting roots or with the obligate parasite Olpidium brassicae. However, two isolates of Spongospora sub-terranea gave an absorbance similar to that observed with the homologous antigen. The DAC ELISA procedure was successfully used to detect various stages in the life cycle of P. graminis and to detect infection that occurred under natural and controlled environments. A simple procedure to conjugate antibodies to fluorescein 5-isothiocyanate (FITC) is described. Resting spores could be detected in root sections by using FITC-labeled antibodies. The potential for application of the two serological techniques for studying the epidemiology of peanut clump disease and for the characterization of Polymyxa isolates from various geographical origins is discussed.     

Evidence for two predominant viral lineages,recombination and subpopulation structure in begomoviruses associated with yellow vein mosaic disease of okra in India

Yellow vein mosaic disease (YVMD) caused by whitefly‐transmitted begomoviruses is an economically significant viral disease of okra. In this study, a survey of begomoviruses associated with YVMD was carried out in eight states and two union territories of India. A total of 92 full‐length DNA‐A components were sequenced and characterized. Sequence comparisons and population structure analysis revealed the existence of four begomovirus species. Two novel species were detected with several recombinationally derived genome fragments that probably originated from begomoviruses known to infect malvaceous and non‐malvaceous hosts. Among the four species, Bhendi yellow vein Maharastra virus (BYVMaV) and Bhendi yellow vein Madurai virus (BYVMV) were found to be predominant in okra, with BYVMV having a pan‐India distribution. There was evidence for a high degree of genetic variability and subpopulation structure within these four species. Neutrality tests suggested the occurrence of purifying selection acting upon these populations. The results of the current study have uncovered the diversity and genetic structure of okra‐infecting begomoviruses in India and generated potentially useful information for developing management strategies for YVMD.     

Resistance to blast (Magnaporthe grisea) in a mini-core collection of finger millet germplasm

Blast caused by Pyricularia grisea [teleomorph: Magnaporthe grisea] is an economically important and widespread disease of finger millet in the world. Host resistance is the most economical and effective means of combating this disease as finger millet is predominantly grown by resource-poor and marginal farmers. At the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), we evaluated a finger millet mini-core collection of 80 germplasm accessions (about 1 % of the total germplasm collection representing major trait variability) for blast resistance both in the field and greenhouse. Field evaluation was done using a refined screening technique that included new improved rating scales for leaf, neck and finger infection. Sixty six of the 80 accessions showed combined resistance to leaf, neck and finger blast in two seasons (2009 and 2010) of field screening. A highly significant and positive correlation was found between neck and finger blast ratings (r?=?0.92), whereas small but significant correlations were found between leaf blast and neck blast (r?=?0.25) and between leaf blast and finger blast (r?=?0.30). These accessions were also screened for leaf blast resistance in the greenhouse by artificial inoculation of seedlings to confirm field observations. Fifty-eight of the 80 accessions were resistant to leaf blast in the greenhouse screen as well. These resistant accessions represented one wild (africana) and four cultivated races (vulgaris, plana, elongate and compacta) of finger millet that originated from 13 countries in Asia and Africa and exhibited considerable diversity for agronomic traits, such as maturity period, plant height and panicle type. These blast resistant accessions from the mini-core collection would be useful in finger millet disease resistance breeding programs.     

Insecticidal, antifeedant and oviposition deterrent effects of the essential oil and individual compounds from leaves of Chloroxylon swietenia DC

Essential oil from the leaves of Chloroxylon swietenia DC. was obtained by hydrodistillation and cold extraction, and the chemical composition was determined by gas chromatography and gas chromatography-mass spectrometry. The major identified components were limonene, geijerene, pregeijerene, germacrene D and trans-beta-ocimene. Laboratory bioassays of the essential oil and four constituents of essential oil isolates were evaluated for insecticidal, antifeedant and oviposition deterrent effects on tobacco cutworm, Spodoptera litura (F.). Toxicity was determined by topical application of the isolates at varying concentrations. Pure oil, geijerene and pregeijerene were found to be more toxic, with LD50 values of 28.6, 35.4 and 40.7 microg per larva respectively. Maximum feeding deterrence was noted for geijerene and pregeijerene, with DC(50) values of 82.5 and 95.1 microg cm(-2) respectively. Furthermore, pure oil, geijerene and pregeijerene displayed oviposition deterrence, even at low concentrations. These results indicate that these natural products may lead to useful, biodegradable, environmentally safe insect control agents.     

Induced systemic protection against tomato late blight elicited by plant growth-promoting rhizobacteria

ABSTRACT Two strains of plant growth-promoting rhizobacteria (PGPR), Bacillus pumilus SE34 and Pseudomonas fluorescens 89B61, elicited systemic protection against late blight on tomato and reduced disease severity by a level equivalent to systemic acquired resistance induced by Phytophthora infestans or induced local resistance by chemical inducer beta-amino butyric acid (BABA) in greenhouse assays. Germination of sporangia and zoospores of P. infestans on leaf surfaces of tomato plants treated with the two PGPR strains, pathogen, and chemical BABA was significantly reduced compared with the noninduced control. Induced protection elicited by PGPR, pathogen, and BABA were examined to determine the signal transduction pathways in three tomato lines: salicylic acid (SA)-hydroxylase transgenic tomato (nahG), ethylene insensitive mutants (Nr/Nr), and jasmonic acid insensitive mutants (def1). Results suggest that induced protection elicited by both bacilli and pseudomonad PGPR strains was SA-independent but ethylene- and jasmonic acid-dependent, whereas systemic acquired resistance elicited by the pathogen and induced local resistance by BABA were SA-dependent. The lack of colonization of tomato leaves by strain 89B61 suggests that the observed induced systemic resistance (ISR) was due to systemic protection by strain 89B61 and not attributable to a direct interaction between pathogen and biological control agent. Although strain SE34 was detected on tomato leaves, ISR mainly accounted for the systemic protection with this strain.     

In vitro plant regeneration of non-toxic Jatropha curcas L.: Direct shoot organogenesis from cotyledonary petiole explants

Jatropha curcas, the energy plant has attained great attention in recent years because of its biodiesel production potential; however, oil and deoiled cakes are toxic. A non-toxic variety of J. curcas is reported from Mexico. A simple and efficient protocol has been developed for plant regeneration using cotyledonary petiole explants of non-toxic variety of J. curcas. The percentage of induction of shoot buds (59.11%), and the number of shoot buds (5.01) per explant was achieved on Murashige and Skoog’s (MS) medium supplemented with 2.27 μM thidiazuron (TDZ). These induced shoot buds multiplied when subcultured on MS medium supplemented with 10 μM kinetin (Kn), 4.5 μM 6-benzyl aminopurine (BAP), and 5.5 μM α-naphthaleneacetic acid (NAA) for 4 weeks and subsequent elongation achieved on MS medium supplemented with 2.25 μM BAP and 8.5 μM indole-3-acetic acid (IAA). Shoots more than 2 cm long were harvested and cultured on MS medium containing different concentrations and combinations of IBA, IAA, NAA, and 0.25 mg L^?1 activated charcoal, and 19.91% rooting was achieved in 15 μM IBA, 5.7 μM IAA, and 16.5 μM NAA after 4 weeks with more than 90% survival rate.     

Rhizobacteria‐mediated resistance against the blackeye cowpea mosaic strain of bean common mosaic virus in cowpea (Vigna unguiculata)

BACKGROUND: The present study investigated the effect of seven Bacillus‐species plant‐growth‐promoting rhizobacteria (PGPR) seed treatments on the induction of disease resistance in cowpea against mosaic disease caused by the blackeye cowpea mosaic strain of bean common mosaic virus (BCMV). RESULTS: Initially, although all PGPR strains recorded significant enhancement of seed germination and seedling vigour, GBO3 and T4 strains were very promising. In general, all strains gave reduced BCMV incidence compared with the non‐bacterised control, both under screen‐house and under field conditions. Cowpea seeds treated with Bacillus pumilus (T4) and Bacillus subtilis (GBO3) strains offered protection of 42 and 41% against BCMV under screen‐house conditions. Under field conditions, strain GBO3 offered 34% protection against BCMV. The protection offered by PGPR strains against BCMV was evaluated by indirect enzyme‐linked immunosorbent assay (ELISA), with lowest immunoreactive values recorded in cowpea seeds treated with strains GBO3 and T4 in comparison with the non‐bacterised control. In addition, it was observed that strain combination worked better in inducing resistance than individual strains. Cowpea seeds treated with a combination of strains GBO3 + T4 registered the highest protection against BCMV. CONCLUSION: PGPR strains were effective in protecting cowpea plants against BCMV under both screen‐house and field conditions by inducing resistance against the virus. Thus, it is proposed that PGPR strains, particularly GBO3, could be potential inducers against BCMV and growth enhancers in cowpea. Copyright © 2009 Society of Chemical Industry