Fingerprinting the Australian rhizobial inoculant mother cultures using refined PCR protocols yields beneficial inoculant management applications

2005 ◽  
Vol 45 (3) ◽  
pp. 141 ◽  
Author(s):  
A-M. Vachot-Griffin ◽  
J. E. Thies

Monitoring the success of rhizobial inoculation requires reliable identification of the introduced strains in nodules and when recovered from field soil. The polymerase chain reaction (PCR) coupled with the use of either random or directed primers has increasingly become the molecular method of choice for characterising bacteria at the strain level. We have investigated the use of 5 markers (REP, ERIC, BOXA1R, RPO1 and IGS) commonly used for PCR fingerprinting to characterise rhizobia bacteria used in the manufacture of rhizobial inoculants in Australia. PCR with random primers often yields inconsistent results because most protocols do not specify stringent cycling and non-cycling parameters. We have increased the stringency and improved the specificity of reaction conditions for 4 of the 5 markers tested. Optimised protocols were then used to fingerprint the 39 strains of rhizobia bacteria held in the 1998 mother culture collection of the Australian Legume Inoculant Research Unit (ALIRU). Results for 34 strains using at least one marker are presented. Although the mother cultures of these inoculant strains undergo numerous quality assurance tests annually, it was not until PCR fingerprinting was applied that 2 strains, believed to be unique, were found to be identical. In the subsequent investigation, we determined that the 2 strains were originally unique but that a mix-up in the cultures had occurred at least 3 years before our analysis. Use of serology, plant infection tests and field tests were not sufficient to detect this problem. The use of PCR fingerprinting with optimised protocols has now been incorporated into the annual quality assurance regime used by the ALIRU who monitor strain quality for the Australian rhizobial inoculant industry. Higher quality rhizobial inoculant for use by Australian farmers is a beneficial outcome of this work.

2018 ◽  
Vol 205 ◽  
pp. 00010 ◽  
Author(s):  
Haryono ◽  
Heri Triluqman Budisantoso ◽  
Edi Subkhan ◽  
Yuli Utanto

This research aims to analyze the implementation of learning quality assurance at school and develop its model based on the applied educational technology. The research unit of analysis encompassing several junior high school, senior high school and vocational school in Semarang, Kendal, and Kudus district. The research results (1) school had implemented the learning quality assurance including the planning, implementation, monitoring, and evaluating process, although the documentation is still desultory, (2) the learning quality assurance based on the applied educational technology had been developed as a reference to define (a) the scope of quality assurance at school, (b) the quality policy and quality assurance organization at school, (c) the standard of learning quality and its achievement strategy, and (d) the manual for learning quality and its instrument, and (3) the learning quality assurance based on the applied educational technology is worth implemented on the learning quality assurance process at school.


2014 ◽  
Vol 8 (2) ◽  
pp. 255-271 ◽  
Author(s):  
D. De Medici ◽  
T. Kuchta ◽  
R. Knutsson ◽  
A. Angelov ◽  
B. Auricchio ◽  
...  

1997 ◽  
Vol 2 (1) ◽  
pp. 44-46 ◽  
Author(s):  
Biljana Čuljković ◽  
Sabera Ruždijić ◽  
Ljubisav Rakić ◽  
Stanka Romac

2011 ◽  
Vol 29 (32) ◽  
pp. 4279-4285 ◽  
Author(s):  
David J. Dabbs ◽  
Molly E. Klein ◽  
Syed K. Mohsin ◽  
Raymond R. Tubbs ◽  
Yongli Shuai ◽  
...  

Purpose HER2 (ERBB2) status is an important prognostic and predictive marker in breast carcinoma. In recent years, Genomic Health (GHI), purveyors of the Oncotype DX test, has been separately reporting HER2 by reverse transcription polymerase chain reaction (RT-PCR) to oncologists. Because of the lack of independent evaluation, this quality assurance study was undertaken to define the concordance rate between immunohistochemistry (IHC)/fluorescent in situ hybridization (FISH) and GHI RT-PCR HER2 assay. Methods All patients at three participating laboratories (Magee-Womens Hospital [Pittsburgh, PA], Cleveland Clinic [Cleveland, OH], and Riverside Methodist Hospital [Columbus, OH]) with available HER2 RT-PCR results from GHI were included in this study. All IHC-positive and equivocal patient cases were further evaluated and classified by FISH at respective laboratories. Results Of the total 843 patient cases, 784 (93%) were classified as negative, 36 (4%) as positive, and 23 (3%) as equivocal at the three institutions using IHC/FISH. Of the 784 negative patient cases, 779 (99%) were also classified as negative by GHI RT-PCR assay. However, all 23 equivocal patient cases were reported as negative by GHI. Of the 36 positive cases, only 10 (28%; 95% CI, 14% to 45%) were reported as positive, 12 (33%) as equivocal, and 14 (39%) as negative. Conclusion There was an unacceptable false-negative rate for HER2 status with GHI HER2 assay in this independent study. This could create confusion in the decision-making process for targeted treatment and potentially lead to mismanagement of patients with breast cancer if only GHI HER2 information is used.


2006 ◽  
Vol 52 (6) ◽  
pp. 570-574
Author(s):  
Ching-Fu Lee ◽  
Daniel Yuen Teh Liu ◽  
Ming Tsong Lai ◽  
Tzong-Hsiung Hseu

Strain W-10, originally identified as Trichoderma koningii, and its supposed mutant G-39, published for production and gene expression of cellulase and xylanase, demonstrated morphological characteristics distinct from those of T. koningii, respectively. To clarify the identification derived from morphological characteristics, several methods were used, including electrophoretic karyotyping, internal transcribed spacer (ITS) analysis of rDNA, and polymerase chain reaction (PCR) fingerprinting using the universal primer L45. All the molecular characteristics showed that strains G-39 and W-10 were identical to T. reesei and T. longibrachiatum, respectively. The results strongly supported that T. koningii G-39 and W-10 should be reassigned as T. reesei and T. longibrachiatum, respectively. Strain G-39 should be considered a mutant from T. reesei QM9414 whose spores were contaminated with those of strain W-10 during a laboratory operation. According to this, we declare that T. koningii G-39 and W-10 must be renamed as T. reesei and T. longibrachiatum, respectively.Key words: PCR fingerprinting, electrophoretic karyotypes, ITS, Trichoderma.


2012 ◽  
Vol 25 (9) ◽  
pp. 1230-1237 ◽  
Author(s):  
Katja Zuther ◽  
Jörg Kahnt ◽  
Jan Utermark ◽  
Julia Imkampe ◽  
Simon Uhse ◽  
...  

The smut fungus Sporisorium reilianum occurs in two varieties (S. reilianum f. sp. reilianum and S. reilianum f. sp. zeae) that cause head smut disease on sorghum and maize, respectively. Prior to plant infection, compatible haploid sporidia of S. reilianum fuse to form infectious dikaryotic hyphae that penetrate the leaf surface, spread throughout the plant, and reach the inflorescences, in which spore formation occurs. To elucidate the basis of host specificity of the two S. reilianum varieties, we compared disease etiology of S. reilianum f. sp. reilianum and S. reilianum f. sp. zeae on sorghum and maize. Both varieties could penetrate and multiply in both hosts. However, red spots appeared on inoculated leaves after sorghum infection with S. reilianum f. sp. zeae. Using matrix-assisted laser desorption-ionization time of flight analysis of leaf extracts, we show that sorghum reacts with the production of the red and orange phytoalexins luteolinidin and apigeninidin upon colonization by S. reilianum f. sp. zeae but not by S. reilianum f. sp. reilianum. Using in vitro growth assays, we demonstrate that luteolinidin but not apigeninidin slows vegetative growth of both S. reilianum f. sp. zeae and S. reilianum f. sp. reilianum. However, the phytoalexin biosynthesis gene SbDFR3 is only induced in sorghum after infection with S. reilianum f. sp. zeae, as shown by quantitative real-time polymerase chain reaction. This suggests that regulation of luteolinidin biosynthesis determines infection success of S. reilianum on sorghum.


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