Evaluation of phosphate-solubilizing bacteria on the growth and grain yield of rice (Oryza sativa L.) cropped in northern Iran.

AIMS: This study aimed to evaluate the efficiency of four phosphate-solubilizing bacteria (PSB) on the growth and yield of rice under different soil conditions. METHODS: Bacterial strains were Rahnella aquatillis (KM977991), Enterobacter sp. (KM977992), Pseudomonas fluorescens and Pseudomonas putida. These studies were conducted on different rice cultivars ('Shiroodi', 'Tarom' and 'Tarom Hashemi') in both pot and field experiments. Measurements started from transplanting and continued throughout the growing season in field experiments. RESULTS: Single PSB inoculations in field trials increased grain yield, biological yield, total number of stems hill(-1) , number of panicles hill(-1) and plant height by 8·50-26·9%, 12·4-30·9%, 20·3-38·7%, 22·1-36·1% and 0·85-3·35% in experiment 1, by 7·74-14·7%, 4·22-12·6%, 6·67-16·7%, 4·0-15·4% and 3·15-4·20% in experiment 2 and by 23·4-37%, 16·1-36·4%, 30·2-39·1%, 28·8-34% and 2·11-4·55% in experiment 3, respectively, compared to the control. Our results indicate that the application of triple super phosphate together with PSB inoculations resulted in reducing the use of chemical fertilizers (about 67%) and increasing fertilizer use efficiency. CONCLUSIONS: This study clearly indicates that these PSBs can be used as biofertilizers in ecological rice agricultural systems. SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this is first report on the association of Rahnella aquatilis with rice and also the application of a mathematical model to evaluate the effect of PSBs on rice growth.

First Report of Hyphodermella rosae Causing Dry Fruit Rot Disease on Plum in Iran.

Plum (Prunus domestica) and peach (P. persica) are widely grown, often in alternate rows with citrus, in the Mazandaran Province of Iran. In June 2011, a dry fruit rot of plum was observed in several production regions in Mazandaran Province (35°47'N, 50°34'E). Initial symptoms at pit-hardening stage appeared as dark brown, circular, necrotic spots from 2 to 5 cm in diameter. They later developed into a dry fruit rot. Severe symptoms occurred during June and July when warm weather (temperature around 28°C) and high relative humidity (RH) (>85%) were present. Marketable yield losses reached 50% to almost 100% in many orchards. To isolate the causal organism, symptomatic fruits were surface disinfested for 1 min in 0.5% active chlorine, washed thoroughly with sterile distilled water, and segments were plated on potato dextrose agar (PDA) amended with 50 mg/liter of streptomycin sulfate and incubated at 25°C for 3 days. The fungus Hyphodermella rosae (Bresadola) Nakasone was consistently isolated (37 isolates from 79 samples) and identified on the basis of morphological characteristics on PDA. Basidiomata were effuse, resupinate, 15 × 10 mm, crustaceous, tubercules small with apical bristles, and light orange to grayish orange. Subhymenium was up to 30 µm thick, composed of vertically arranged, short-celled, nonagglutinated hyphae; subhymenial hyphae were 3 to 4 µm in diameter. Basidiospores were ellipsoid, 7.5 to 8.5 × 4.5 to 5.5 µm (100 determination), and their cell walls were thin, hyaline, and smooth (1). Genomic DNA was extracted from mycelium with a DNA extraction kit (Qiagen, Hilden, Germany) according to the manufacturer's directions and grown on potato dextrose broth for 4 days at 28°C. The rDNA region was amplified with the primers ITS4 (5'-TCCTCCGCTTATTGATATGC-3') and ITS5 (5'- GGAAGTAAAAGTCGTAACAA-3') (4) and the PCR product was sequenced. Nucleotide BLAST analysis of the amplified 627-bp fragment confirmed a 99% similarity with the sequence of H. rosae (GenBank Accession No. JN593086). A pathogenicity test was conducted with isolate MA4099 by placing 5-day-old mycelial plugs grown on PDA at the surface of healthy fruit (n = 6) incubated under >85% RH at 25°C for at least 4 days until the appearance of symptoms, which were similar to those displayed under orchard conditions. Control fruits, inoculated with blocks of PDA plugs, remained intact and symptomless. Reisolation from inoculated fruit samples consistently yielded the inoculated fungus, completing Koch's postulates. The genus Hyphodermella has been reported to be causing wood rot on apricot (2) and sweet and sour cherry (3). To our knowledge, this is the first report of H. rosae causing dry fruit rot on a stone fruit species in the world. References: (1) K. K. Nakasone. Mycologie, 29:231, 2008. (2) J. M. Ogawa et al. Diseases of Apricot (Prunus armeniaca L.). The American Phytopathological Society, St. Paul, MN, 2003. (3) J. K. Uyemoto et al. Diseases of Sweet Cherry (Prunus avium L.) and Sour Cherry (P. cerasus L.). IS-MPMInet, http://www.ismpminet.org/resources/common/comment/cherry.asp , accessed June 2012. (4) T. J. White et al. Page: 315 in: PCR Protocols: A Guide to Methods and Application. M.A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

Life cycle characteristics of six Artemia populations from Iran.

The present study was conducted to achieve the life cycle characteristics of six Artemia populations (one bisexual and five parthenogenetic) from Iran. The cysts of parthenogenetic strains were collected from Maharlu, Incheh, Varmal and Qom salt lakes and Lagoons at the periphery of Lake Urmia. Cysts of the bisexual Artemia urmiana were collected from the Lake Urmia. All cysts strains were hatched using the standard procedures and the nauplii from all populations were reared at 80 g L(-1) at laboratory conditions. Survival and total length of the Artemia were measured on days 8, 11, 14, 17, 20 and 23 of culture. Randomly selected adult animals were studied for eight reproductive and four life span characteristics. The findings showed that parthenogenetic Artemia from Maharlu, Incheh lakes and from Lagoons at the vicinity of the Lake Urmia had significantly highest (p < 0.05) values of survival rate (73 and 62.8%, respectively) compared to bisexual A. urmiana and parthenogenetic strains from Qom and Varmal lakes (49.6, 29.2 and 23.2%). No significant differences were observed in all growth strain populations when cultured under similar laboratory conditions. Artemia populations from Maharlu, Qom lakes and from Lagoons in many occasions had significantly highest (p < 0.05) reproductive values compared to other three populations including the bisexual A. urmiana. The results showed the highest heterogeneity and intrapopulation variations among parthenogenetic population strains.

Analysis of the biological and molecular variability of Watermelon mosaic virus isolates from Iran.

Watermelon mosaic virus (WMV) is one of the most important viruses that causes different symptoms in Cucurbitaceae. WMV is a potyvirus with a worldwide distribution, but occurs most commonly in temperate and Mediterranean regions. Cucurbit species grown in Yazd, Esfahan, West Azerbaijan, Hormozgan, and Kerman provinces were surveyed for the relative incidence of WMV in 2004-2005. A total of 757 symptomatic cucurbit and 31 weed species were collected and assayed for infection with WMV. Of 788 leaf samples from cucurbit and weed plants, 190 samples were positive by double antibody sandwich ELISA (DAS-ELISA) using specific polyclonal antibody. Among the weed species tested only colocynth (Citrullus colocynthis) was found to be infected with WMV. The coat protein (CP) gene from 18 representative isolates was PCR amplified, cloned, sequenced, and compared with the sequences available in GeneBank. Phylogenetic analysis using 778 nucleotide long sequences of the coat protein gene showed that these isolates fell into two; groups I and II. Only one isolates (KER.JI.1) was classified in the group II. This isolate had a wider host range and infected Nicotiana debneyii and Datura metel. None of the other 17 isolates could infect these two species. Members of group I were divided into three subgroups; A, B, and C. The subgroup I(B) appears to be a new subgroup comprising only of the Iranian isolates. Phylogenetic analysis based on 200 nucleotides coding for the N-terminal segment of the CP showed that all Iranian isolates except KER.JI.1 clustered with the previously reported WMV strains. All Iranian isolates had a DAG amino acid triplet which is involved in aphid transmissibility. This is the first report on sequence analysis of the nearly full-length CP cDNA clones of WMV isolates from Iran.

Human dietary exposure to fumonisin B1 from Iranian maize harvested during 1998-2000.

Fumonisin B(1) (FB(1)) is the most abundant of the fumonisin mycotoxins, mainly produced in maize by F. verticillioides and F. proliferatum. A previous study on the FB1 contamination of maize harvested in Mazandaran and Isfahan Provinces of Iran in 1998 and 1999 demonstrated contamination in both provinces. This present study was undertaken to further investigate the variation in levels of contamination and to estimate possible levels of human exposure to fumonisins in Iran. The mean level of FB(1) in 49 visually healthy maize samples collected from Mazandaran Province during 2000 was 6.14 mg/kg, which is higher than that found during 1998 and 1999 (2.27 and 3.18 mg/kg, respectively). Although these levels are higher than the Iranian legislative limits for fumonisins in maize intended for humans, the relatively low estimated consumption of maize in Iran (3.3 g/person/day) implies that average exposures (0.011 and 0.215 microg/kg body weight/day in Isfahan and Mazandaran, respectively) are within the provisional maximum tolerable daily intake of 2 microg/kg body weight/day set by the Joint FAO/WHO Expert Committee on Food Additives. Nevertheless, certain sections of the population who may consume higher amounts of maize or who may replace all or some of their consumption of other cereals with maize, could well exceed this limit.

Witches'-broom of Bakraee (Citrus reticulata hybrid) in Iran.

Symptoms, resembling those of witches'-broom disease of lime (WBDL), consisting of proliferation and growth of spindly shoots, rosetting, development of stunted, pale green, cupped leaves, and long thin thorns were observed on some bakraee (Citrus reticulata hybrid) trees, the major rootstock species in Ghazi-Ghale (Hormozgan Province) and Kahnouj (Kerman Province) in southern Iran. Vein clearing, mottling, and mild to severe yellowing of leaves was evident on some rosetted flushes. The disease occurred sporadically, and only some branches of the tree displayed the symptoms. Sour lime (Citrus aurantifolia) trees with symptoms of WBDL were also encountered in adjacent citrus groves in the area where an association of "Candidatus Phytoplasma aurantifolia" with diseased trees was previously established. Total nucleic acids were extracted from symptomatic as well as apparently healthy bakraee (1). The nucleic acids were subjected to nested polymerase chain reaction (PCR) amplification with the universal primers R16mF2/R16mR1, followed by R16F2n/R16R2 (2). The PCR program consisted of an initial denaturation at 94°C for 2 min, followed by 40 cycles of 1 min at 94°C, 2 min at 55°C, and 3 min at 72°C. After a final extension of 10 min at 72°C, the samples were electrophoresed on agarose gels, and the DNA fragments were stained with ethidium bromide and visualized under ultraviolet light. An expected 1.2-kb product was amplified in DNA preparations from broomed bakraee and WBDL-affected sour lime trees. No such amplicon was detected in similar DNA extracts of asymptomatic trees. These results demonstrated an association of a phytoplasma with witches'-broom disease of bakraee. However, its relationship with "Candidatus Phytoplasma aurantifolia" remains to be determined. References: (1) S. L. Dellaporta et al. Plant Mol. Biol. Rep.1:19, 1983. (2) I. M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998.

Pseudobacciger harengulae from the Atlantic herring Clupea harengus: a new host and locality record.

During the summer and autumn of 1994, 1995 and 1996, 406 juvenile herring caught off the Swedish west coast were examined for parasites. Amongst those found was the digenean Pseudobacciger harengulae, which represents new host and locality records for this parasite. Pseudobacciger harengulae has been reported from several species of clupeiformes, mostly from tropical and temperate regions of the Atlantic, Pacific and Indian Oceans. The morphology of P. harengulae is described and compared with earlier descriptions of P. harengulae and P. manteri. The possible relationships between P. harengulae and P. manteri are discussed and the validity of the P. manteri is questioned. Most of the specimens (75%) of P. harengulae were found in the pyloric caeca and the remainder (25%) in the intestine.

Fumonisin B(1) in maize harvested in Iran during 1999.

The fumonisin B(1) (FB(1)) contamination of maize collected in two areas of Iran during 1999 was determined. The 20 maize samples from Mazandaran Province, situated on the Caspian littoral of Iran, consisted of random samples of farmers' lots and were all contaminated with FB(1) at a mean level of 3.18 mg kg(-1) (range 0.68-7.66 mg kg(-1)). The 10 samples (of the same maize cultivar) from Isfahan Province in central Iran were purchased as maize cobs in local retail markets and had mean FB levels of 0.22 mg kg(-1) (mean of all samples, 6/10 samples positive, range <0.01-0.88 mg kg(-1)). The FB levels in Mazandaran, an area of high oesophageal cancer, were significantly (p < 0.0001) higher than the FB levels found in maize from Isfahan, an area of low oesophageal cancer in Iran.

Natural occurrence of fumonisins in corn from Iran.

Corn collected in the Mazandaran and Isfahan Provinces of Iran was analyzed for fumonisin B(1) (FB(1)), fumonisin B(2) (FB(2)), and fumonisin B(3) (FB(3)). The samples from Mazandaran Province, situated on the Caspian littoral of Iran, were random samples from farmers' corn lots collected in September 1998, whereas those from Isfahan Province, situated further south in the center of Iran, were bought as corn cobs in the local retail market during October 1998. All 11 samples from Mazandaran showed high levels of fumonisin contamination with FB(1) levels between 1.270 and 3.980 microg/g, FB(2) levels between 0.190 and 1.175 microg/g, and FB(3) levels between 0.155 and 0.960 microg/g. Samples from Isfahan showed lower levels of contamination with eight of eight samples having detectable FB(1) (0.010-0.590 microg/g), two of eight samples having detectable FB(2) (0.050-0.075 microg/g), and two of eight samples having detectable FB(3) (0.050-0.075 microg/g). This is the first report of fumonisin contamination of corn from Iran, in which samples from the area of high esophageal cancer on the Caspian littoral have been shown to contain high levels of fumonisins.

Pseudanthocotyloides heterocotyle (van Beneden, 1871) Euzet & Prost, 1969 (Monogenea: Polyopisthocotylea: Mazocraeidae), a parasite of herring Clupea harengus L. and sprat Sprattus sprattus L. (Teleostei: Clupeidae).

A total of 690 herring Clupea harengus L. and 88 sprat Sprattus sprattus L. caught off the west coast of Sweden, in the North Sea and off the west and south coasts of the United Kingdom, were examined for gill parasites. The monogenean Pseudanthocotyloides heterocotyle (van Beneden, 1871) Euzet & Prost, 1969 was found in 38 (5.5%) herring and one (1.1%) sprat. The parasite was significantly (P<0.05) more common off the west coast of Sweden than elsewhere and most specimens (62.5%) were found on the pseudobranchs. Only the smaller herring were infected. P. heterocotyle is redescribed and its taxonomy discussed, together with the possibility of host and parasite misidentification in previous reports.

Pathology and morphology of Ichthyophonus hoferi in naturally infected fishes off the Swedish west coast.

The pathology and morphology of Ichthyophonus hoferi was studied in naturally infected Atlantic herring Clupea harengus, in sprat Sprattus sprattus, and in flounder Pleuronectes flesus from the west coast of Sweden. The pathogen was found in all organs examined, with the intensity of infection varying in different organs of the different fish species. Two main phases in the life of infecting parasites were identified, 'active' and 'passive', the latter being able to switch to active. The active phase of the infection in herring was usually accompanied by a lean and slender appearance of the body, a drastic decrease in intestinal fat, emaciation of the somatic muscles, swelling of the visceral organs, poor quality of flesh texture and a distinctive off-odour. The most characteristic macroscopic sign of ichthyophonosis in herring and flounder was the occurrence of creamy white nodules on the heart. The infection causes a chronic systemic granulomatous inflammation. The nature of the granulomatous inflammation was host- and tissue-dependent. The pathogenicity of the parasite in its active form and the side effects of host defence cells were also reflected in dramatic tissue damage and loss of structure and function of the infected organs. Three kinds of spores were identified: 'un-developing spore', 'developing spore' and 'plasmodio-spore'. The formation and spread of 'plasmodia', from plasmodio-spores, as a secondary infection agent is documented. Transmission electron microscopy revealed I. hoferi to be multinucleated, containing different organelles and structures. These included a cell wall, an undulating cell membrane, a thin paramural endoplasm, an endoplasmic reticulum, polymorphic but usually spherical mitochondria with short tubulo-vesicular cristae, dictyosomes with plate-like cristernae, large electron-dense lipid droplets and electron-lucid vacuoles, probably containing glycogen.