Acid-adaption by a medic microsymbiont: new insights from the genome of Sinorhizobium medicae WSM419

The poor availability of nitrogen is one of the principal factors limiting global biomass. Legumes are vital components of agricultural systems because of their ability to associate symbiotically with root nodule bacteria (RNB) and subsequently fix atmospheric nitrogen to a form that can be utilised by the plant partner. Furthermore, this symbiotic relationship provides available soil nitrogen for subsequent non-leguminous crops. This RNB-legume interaction is affected by a number of environmental factors. Progressive acidification of agricultural soils is one of the big challenges in agriculture as soil acidity negatively impacts legume productivity. One genus of RNB, Sinorhizobium, is particularly acid-sensitive causing a major reduction in Medicago productivity in acidic soils. Due to the importance of Medic pasture production, alternative strains have been captured, and are still being captured, from the genetic pool that display superior acid tolerance characteristics. This presentation will focus on the acid-tolerant species S. medicae (previously known as S. meliloti) and in particular on the previously used commercial inoculant WSM419

Technical Issues Relating to Agricultural Microbial Genetic Resources (AMiGRs), Including their Characteristics, Utilization, Preservation and Distribution : draft information paper

Draft information paper prepared for the Genetic Resources Policy Committee (GRPC) of the CGIAR for submission to the 11th Regular Session of the Commission on Genetic Resources for Food and Agriculture (CGRFA), 11–15 June 2007. At it's 16th Session in September 2004, the Genetic Resources Policy Committee (GRPC) of the CGIAR recommended that the CGIAR’s System-wide Genetic Resources Programme (SGRP) and the Centres work together to determine the scope of microorganisms relevant to food and agriculture, as a first step in analyzing policy on access, use and conservation of culture collections within the CGIAR system and outside. The GRPC subsequently commissioned a background study on technical issues related to developing harmonized management policies, guidelines and practices concerning acquisition, use and distribution of Agricultural Microbial Genetic Resources (AMiGRs). This study was carried out by Prof. John Howieson of Murdoch University, Australia. The resulting paper was considered by the GRPC in subsequent meetings and circulated to FAO, the CGIAR and other interested organizations for comments. It was finally approved, with some changes, by the GRPC at it's 21th Session in April 2007, for submission as a draft information paper to the 11th Regular Session of Commission on Genetic Resources for Food and Agriculture (CGRFA), 11–15 June 2007. The GRPC added four recommendations that it developed based on inputs from the paper and other sources. The recommendations are included in the "conclusions" of the paper. These same recommendations were provided to the 11th CGRFA by way of an oral intervention

Rapid in situ evolution of nodulating strains for Biserrula pelecinus L. through lateral transfer of a symbiosis island from the original mesorhizobial inoculant

Diverse rhizobia able to nodulate Biserrula pelecinus evolved following in situ transfer of nodA and nifH from an inoculant to soil bacteria. Transfer of these chromosomal genes and the presence of an identical integrase gene adjacent to a Phe tRNA gene in both the inoculant and recipients indicate that there was lateral transfer of a symbiosis island

Application of next-generation sequencing for rapid marker development in molecular plant breeding: a case study on anthracnose disease resistance in Lupinus angustifolius L.

Background In the last 30years, a number of DNA fingerprinting methods such as RFLP, RAPD, AFLP, SSR, DArT, have been extensively used in marker development for molecular plant breeding. However, it remains a daunting task to identify highly polymorphic and closely linked molecular markers for a target trait for molecular marker-assisted selection. The next-generation sequencing (NGS) technology is far more powerful than any existing generic DNA fingerprinting methods in generating DNA markers. In this study, we employed a grain legume crop Lupinus angustifolius (lupin) as a test case, and examined the utility of an NGS-based method of RAD (restriction-site associated DNA) sequencing as DNA fingerprinting for rapid, cost-effective marker development tagging a disease resistance gene for molecular breeding. Results Twenty informative plants from a cross of RxS (disease resistant x susceptible) in lupin were subjected to RAD single-end sequencing by multiplex identifiers. The entire RAD sequencing products were resolved in two lanes of the 16-lanes per run sequencing platform Solexa HiSeq2000. A total of 185 million raw reads, approximately 17 Gb of sequencing data, were collected. Sequence comparison among the 20 test plants discovered 8207 SNP markers. Filtration of DNA sequencing data with marker identification parameters resulted in the discovery of 38 molecular markers linked to the disease resistance gene Lanr1. Five randomly selected markers were converted into cost-effective, simple PCR-based markers. Linkage analysis using marker genotyping data and disease resistance phenotyping data on a F8 population consisting of 186 individual plants confirmed that all these five markers were linked to the R gene. Two of these newly developed sequence-specific PCR markers, AnSeq3 and AnSeq4, flanked the target R gene at a genetic distance of 0.9 centiMorgan (cM), and are now replacing the markers previously developed by a traditional DNA fingerprinting method for marker-assisted selection in the Australian national lupin breeding program. Conclusions We demonstrated that more than 30 molecular markers linked to a target gene of agronomic trait of interest can be identified from a small portion (1/8) of one sequencing run on HiSeq2000 by applying NGS based RAD sequencing in marker development. The markers developed by the strategy described in this study are all co-dominant SNP markers, which can readily be converted into high throughput multiplex format or low-cost, simple PCR-based markers desirable for large scale marker implementation in plant breeding programs. The high density and closely linked molecular markers associated with a target trait help to overcome a major bottleneck for implementation of molecular markers on a wide range of germplasm in breeding programs. We conclude that application of NGS based RAD sequencing as DNA fingerprinting is a very rapid and cost-effective strategy for marker development in molecular plant breeding. The strategy does not require any prior genome knowledge or molecular information for the species under investigation, and it is applicable to other plant species

Burkholderia dipogonis sp. nov., isolated from root nodules of Dipogon lignosus in New Zealand and Western Australia

Seven strains, ICMP 19430T, ICMP 19429, ICMP 19431, WSM4637, WSM4638, WSM4639 and WSM4640, were isolated from nitrogen-fixing nodules on roots of the invasive South African legume Dipogon lignosus (subfamily Papilionoideae, tribe Phaseoleae) in New Zealand and Western Australia, and their taxonomic positions were investigated by using a polyphasic approach. All seven strains grew at 10–37 °C (optimum, 25–30 °C), at pH 4.0–9.0 (optimum, pH 6.0–7.0) and with 0–2 % (w/v) NaCl (optimum growth in the absence of NaCl). On the basis of 16S rRNA gene sequence analysis, the strains showed 99.0–99.5 % sequence similarity to the closest type strain, Burkholderia phytofirmans PsJNT, and 98.4–99.7 % sequence similarity to Burkholderia caledonica LMG 19076T. The predominant fatty acids were C18 : 1ω7c (21.0 % of the total fatty acids in strain ICMP 19430T), C16 : 0 (19.1 %), C17 : 0 cyclo (18.9 %), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c; 10.7 %) and C19 : 0 cyclo ω8c (7.5 %). The polar lipid profile consisted of a mixture of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and several uncharacterized aminophospholipids and phospholipids. The major isoprenoid quinone was Q-8 and the DNA G+C content of strain ICMP 19430T was 63.2 mol%. The DNA–DNA relatedness of the novel strains with respect to the closest neighbouring members of the genus Burkholderia was 55 % or less. On the basis of 16S rRNA and recA gene sequence similarities and chemotaxonomic and phenotypic data, these strains represent a novel symbiotic species in the genus Burkholderia, for which the name Burkholderia dipogonis sp. Nov. is proposed, with the type strain ICMP 19430T (=LMG 2841T =HAMBI 3637T)

Complete genome sequence of the Medicago microsymbiont Ensifer (Sinorhizobium) medicae strain WSM419

Ensifer (Sinorhizobium) medicae is an effective nitrogen fixing microsymbiont of a diverse range of annual Medicago (medic) species. Strain WSM419 is an aerobic, motile, non-spore forming, Gram-negative rod isolated from a M. murex root nodule collected in Sardinia, Italy in 1981. WSM419 was manufactured commercially in Australia as an inoculant for annual medics during 1985 to 1993 due to its nitrogen fixation, saprophytic competence and acid tolerance properties. Here we describe the basic features of this organism, together with the complete genome sequence, and annotation. This is the first report of a complete genome se-quence for a microsymbiont of the group of annual medic species adapted to acid soils. We reveal that its genome size is 6,817,576 bp encoding 6,518 protein-coding genes and 81 RNA only encoding genes. The genome contains a chromosome of size 3,781,904 bp and 3 plasmids of size 1,570,951 bp, 1,245,408 bp and 219,313 bp. The smallest plasmid is a fea-ture unique to this medic microsymbiont