ELISA identification ofRhizobium strains by use of enzymelabeled protein A

Kishinevsky, B.; Maoz, Arieh

doi:10.1007/bf01567132

Cited by 21 publications

(12 citation statements)

References 10 publications

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“…The simplified indirect ELISA procedure (Kishinevsky & Maoz 1983;Fuhrmann & Wollum 1985) is a reliable technique which could be used in variously-equipped laboratories.…”

Section: Discussionmentioning

confidence: 99%

“…The remaining nine antisera (7/20E, 31/14N, 31/9N, 22/6N, 30/4N, 34/6NE, 25/3N, 28/15N and 10/14N) were produced from Bradyrhizobium isolates obtained in Thailand during the course of this study, using the methods of Kishinevsky & Gurfel (1980). The indirect ELISA of Engwal & Perlmann (1971), with the modifications of Kishinevsky & Maoz (1983) and Fuhrmann & Wollum (1985), was employed for serological typing of Bradyrhizobium cultures.…”

Section: Serological Proceduresmentioning

confidence: 99%

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World Journal of Microbiology and Biotechnology 2008

Dover

Large

Qureshi

et al. 2008

World J Microbiol Biotechnol

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ELISA and antibody adsorption tests were applied to determine the minimal somatic antigen constitution of 243 strains of Bradyrhizobium sp. (Arachis) using 12 antisera. The 243 indigenous bradyrhizobial isolates were from 15 sites in four regions of Thailand. A total of 29 serogroups were identified. Most (80%) of the isolates tested had at least one heat-stable antigen in common with strain 280A, forming a so-called 280A serocluster. At 11 of 15 sites tested, 53 to 100% of the isolates fell into one or two predominant serogroups. The serological properties of the indigenous bradyrhizobia were not related to the cropping history of the cultivated fields from which they were isolated.Key words: Antigens, antisera, Arachis, Bradyrhizobium, ELISA, peanut, serological groups.Peanut (Arachis hypogaea L.) is naturally nodulated by the slow-growing rhizobia (Bradyrhizobium spp.) that are widespread in tropical soils (Ezedinma 1963), the so-called 'peanut rhizobia '. Boonkerd et al. (1991) reported that native peanut rhizobia isolates, collected in the main peanutgrowing areas of Thailand, range widely in their ability to fix N 2 . In some areas, most bradyrhizobia isolated from peanut nodules were less effective than the inoculum strain, THA 205, recommended for use in Thailand. Substitution of ineffective nodules, formed by indigenous peanut rhizobia, with nodules from a more effective inoculant strain should therefore increase the symbiotic performance of inoculated plants. To approach this problem, a better understanding of the nature of native peanut rhizobial populations is necessary. Although the antigenic specificity of peanut rhizobia has been demonstrated by several authors (Van Der Merve & Strijdom 1973;Dadarval of al. 1974) Materials and Methods Isolation of Indigenous RhizobiaSoil samples were collected from four regions of Thailand: North, North-east, East and Central Plain. Different cropping areas were chosen in each region: (1) continuously cropped with peanuts; (2) continuously cropped with non-legumes; and (3) non-cultivated fields. Soil samples, collected from the total of 15 sites, came from one uncultivated field, two fields cropped with non-legumes, three fallow fields and nine fields cropped with legumes. The nine fields of legumes contained peanuts (five), mungbean (one), cowpea (one), soybean (one), or a mixture of other legumes (one). Soil samples were obtained by randomly collecting 10 core samples from each field. All soil cores from one site were pooled and mixed, passed through a 6.5-mm sterile sieve. A sub-sample (3 kg) was then put immediately into an ice box (4°C) and left until tested (≤ 7 days). Two kg of each soil sample were mixed with 2 kg autoclaved sand and put in clean plastic containers. Peanut seeds (cv. Tainan 9), surface-sterilized in 2% (w/v) sodium hypochlorite and pre-germinated at 28°C on 1.5% (w/v) water agar (Difco) plates for 60 h, were then planted at six seeds per container. The seedlings were thinned to three per container 7 days after germination. All pots were i...

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“…The simplified indirect ELISA procedure (Kishinevsky & Maoz 1983;Fuhrmann & Wollum 1985) is a reliable technique which could be used in variously-equipped laboratories.…”

Section: Discussionmentioning

confidence: 99%

Section: Serological Proceduresmentioning

confidence: 99%

World Journal of Microbiology and Biotechnology 2008

Dover

Large

Qureshi

et al. 2008

World J Microbiol Biotechnol

View full text Add to dashboard Cite

show abstract

“…To confirm their occupancy by the test rhizobial strains, nodules from each Cyclopia species were pooled for each treatment and a sub-sample of 20 nodules analysed using the indirect enzyme-linked immunosorbent assay (ELISA) technique modified from Kishinevsky and Maoz (1983). This method has proved reliable in differentiating the 4 Cyclopia test strains from one other and in differentiating strains UCT40a, UCT44b and UCT61a from other rhizobia (Spriggs, 2004).…”

Section: Nursery Trialmentioning

confidence: 99%

Symbiotic performance of selectedCyclopia Vent. (honeybush) rhizobia under nursery and field conditions

Spriggs

Dakora

2009

Symbiosis

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Three newly selected strains of Cyclopia rhizobia together with an inoculant strain, which has never been tested in the field with adequate experimental design, was assessed under both nursery and field conditions for symbiotic performance. The three new test strains were initially selected for their superior Nz-fixing abilities under glasshouse conditions, and then evaluated in this study for field performance. Cyclopia subternata Vogel and Cyclopia genistoides (L.) R. Br., which have the potential for producing high quality honeybush tea, were used as host plants in both the nursery and field studies.The effect of seedling inoculation at the nursery level was also examined for the four test strains under nursery conditions. The inoculation of cuttings under nursery conditions produced.significant increases in shoot biomass, shoot %N and shoot N content. More specifically, inoculating C. subternata with strains UCT44b and UCT61a significantly increased shoot biomass and N content relative to strain PPRICI3. Strains UCT44b and UCT61a also showed better nodulation with C. subternata cuttings compared to strains UCHOa and PPRICI3. Field inoculation of Cyclopia increased all growth parameters relative to the uninoculated control, except for leaf %N. Cyclopia subternata inoculated with strains UCT44b, UCT40a and UCT61a produced significantly lower b 15 N values than the uninoculated C. subternata reference plant. Using the 15N natural abundance method, C. subternata was estimated to be gaining about half of its N from N z fixation, while C. genistoides obtained less than half of its N from symbioticnutrition.

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“…(Brady)rhizobium competition has been studied so far by examining the antibiotic resistance (Josey et al 1979;Eaglesham 1987), serological properties (Schmidt et al 1968;Dudman 1971;Schmidt 1974;Kingsley and Boh100l 1981;Kishinevsky and Maoz 1983;Kang et al 1991), and by using transposon Tn5 (Brockman et al 1991), etc. Recently, Wilson et al (1995) have developed a new technique, using f3-glucuronidase (GUS) transposon for marking (Brady)rhizobium and tracking the behavior and competitive ability for nodulation of strains in various plants.…”

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confidence: 99%