Survival analysis of germination data in response to temperature for Ornithopus species and other temperate pasture legumes.

Cool temperatures can limit productivity of temperate grazing systems as poor pasture growth rates in winter create feed shortages for livestock. Ornithopus spp. (serradella) are broadly adapted annual pasture legumes that produce high-quality forage in soil types considered marginal for other temperate legume species. However, serradella establishment is perceived to be difficult in cool-season environments. We used survival analysis to compare germination rate and seedling emergence for two serradella species (yellow serradella and French serradella) against three reference species (Medicago sativa, M. polymorpha and Trifolium subterraneum ) in four temperature treatments (10/5, 15/10, 20/15 and 25/20°C; max/min). We also compared shoot relative growth rate and photosynthetic rate at 15/10°C (cool) and 23/18°C (warm). Cool temperatures (10/5, 15/10°C) did not slow germination rates for serradella relative to the reference species, but warm temperatures (20/15, 25/20°C) delayed emergence and reduced post-emergent shoot growth rates. Once established, Ornithopus spp. had similar mean photosynthetic rates and stomatal conductance at cool temperatures to the reference species. We conclude that, contrary to common perception, cool temperatures did not adversely influence germination, emergence, or early growth of Ornithopus spp. relative to the reference species.

A chronology of subterranean clover burr detachment mechanics and implications for seed harvestability.

Seed retention has not been evaluated for subterranean clover (Trifolium subterraneum L.), because its geocarpic seed-bearing burrs are currently harvested by suction systems. Development of improved harvest methods requires knowledge of subterranean clover seed retention characteristics and their changes with plant development. This study evaluates burr attachment and peduncle tensile strength during burr maturity until plant senescence across the three subspecies: subterraneum (cv. Dalkeith), yanninicum (cv. Monti) and brachycalycinum (cv. Mawson). Peduncle tensile strength was greater than burr-to-peduncle attachment strength for each subspecies, with peak mean peduncle strength 30-130% greater than peak mean burr-to-peduncle attachment strength. Both strength measurements decreased significantly (greater than 50% for each subspecies) as the plant senesced, which was associated with reductions in burr moisture content, and burr and peduncle diameters. Microscopy indicated a ductile to brittle transition as peduncles senesced, reducing energy absorption and increasing the likelihood of failure at defects. These results are important for the commercial production of subterranean clover seed and suggest it may be possible to harvest seed before plant senescence with dig-invert machinery, similar to that used for peanut harvesting. However, this approach would require harvesting prior to maximum seed development and the implications for seed viability and yield need to be further evaluated.

Carbon trading for phosphorus gain: the balance between rhizosphere carboxylates and arbuscular mycorrhizal symbiosis in plant phosphorus acquisition.

Two key plant adaptations for phosphorus (P) acquisition are carboxylate exudation into the rhizosphere and mycorrhizal symbioses. These target different soil P resources, presumably with different plant carbon costs. We examined the effect of inoculation with arbuscular mycorrhizal fungi (AMF) on amount of rhizosphere carboxylates and plant P uptake for 10 species of low-P adapted Kennedia grown for 23 weeks in low-P sand. Inoculation decreased carboxylates in some species (up to 50%), decreased plant dry weight (21%) and increased plant P content (23%). There was a positive logarithmic relationship between plant P content and the amount of rhizosphere citric acid for inoculated and uninoculated plants. Causality was indicated by experiments using sand where little citric acid was lost from the soil solution over 2 h and citric acid at low concentrations desorbed P into the soil solution. Senesced leaf P concentration was often low and P-resorption efficiencies reached >90%. In conclusion, we propose that mycorrhizally mediated resource partitioning occurred because inoculation reduced rhizosphere carboxylates, but increased plant P uptake. Hence, presumably, the proportion of plant P acquired from strongly sorbed sources decreased with inoculation, while the proportion from labile inorganic P increased. Implications for plant fitness under field conditions now require investigation.

Putting the P in Ptilotus: a phosphorus-accumulating herb native to Australia.

BACKGROUND AND AIMS: Ptilotus polystachyus (green mulla mulla; ptilotus) is a short-lived perennial herb that occurs widely in Australia in arid and semi-arid regions with nutrient poor soils. As this species shows potential for domestication, its response to addition of phosphorus (P) and nitrogen (N) was compared to a variety of the domesticated exotic perennial pasture herb Cichorium intybus (chicory), 'Puna'. METHODS: Pots were filled with 3 kg of an extremely nutrient-deficient sterilized field soil that contained 3 mg kg(-1) mineral N and 2 mg kg(-1) bicarbonate-extractable P. The growth and P and N accumulation of ptilotus and chicory in response to seven rates of readily available phosphorus (0-300 mg P pot(-1)) and nitrogen (N) (0-270 mg N pot(-1)) was examined. KEY RESULTS: Ptilotus grew extremely well under low P conditions: shoot dry weights were 23, 6 and 1.7 times greater than for chicory at the three lowest levels of P addition, 0, 15 and 30 mg P pot(-1), respectively. Ptilotus could not downregulate P uptake. Concentrations of P in shoots approached 4% of dry weight and cryo-scanning electron microscopy and X-ray microanalysis showed 35-196 mM of P in cell vacuoles in a range of tissues from young leaves. Ptilotus had a remarkable tolerance of high P concentrations in shoots. While chicory exhibited symptoms of P toxicity at the highest rate of P addition (300 mg P pot(-1)), no symptoms were present for ptilotus. The two species responded in a similar manner to addition of N. CONCLUSIONS: In comparison to chicory, ptilotus demonstrated an impressive ability to grow well under conditions of low and high P availability. Further study of the mechanisms of P uptake and tolerance in ptilotus is warranted.

Location and quantification of phosphorus and other elements in fully hydrated, soil-grown arbuscular mycorrhizas: a cryo-analytical scanning electron microscopy study.

• Concentrations of phosphorus (P), potassium (K), magnesium (Mg) and calcium (Ca) were determined in situ in fully hydrated arbuscular mycorrhizas by cryo-analytical scanning electron microscopy. The field- and glasshouse-grown plants (subterranean and white clovers, field pea and leek) were colonized by indigenous mycorrhizal fungi. • The [P] in intraradical hyphae was generally 60-170 mM, although up to 600 mM was recorded, and formed strong linear relationships with [K], up to 350 mM, and [Mg], up to 175 mM. Little Ca was detected. The turgid branches of young arbuscules contained 30-50 mM P, up to 100 mM K and little Mg. Collapsing arbuscule branches and clumped arbuscules had greatly elevated Ca (30-250 mM), but otherwise differed little from young arbuscule branches in elemental concentration. • The [P] was low or undetectable in 86% of uncolonized cortical cell vacuoles, but was generally elevated in vacuoles surrounding an arbuscule and in the liquid surrounding hyphae in intercellular spaces. • Our results suggest that both young arbuscules and intercellular hyphae are sites for P-transfer, that Mg2+ and K+ are probably balancing cations for P anions in hyphae, and that host cells may limit arbuscule lifespan through deposition of material rich in Ca.

Regression of extensive pulmonary metastases in mice by adoptive transfer of antigen-specific CD8(+) CTL reactive against tumor cells expressing a naturally occurring rejection epitope.

In this study, we developed a mouse model of adoptive immunotherapy reflecting immune recognition of syngeneic tumor cells naturally expressing an endogenous rejection Ag. Specifically, in a pulmonary metastases model, we examined the potency and maintenance of an antitumor CD8(+) CTL response in vivo, as well as its effectiveness against an "extensive" tumor burden. The approach taken was to first generate tumor-specific CTL from mice challenged with the CMS4 sarcoma coadministered with anti-CTLA4 mAb, which has been shown to facilitate the induction of Ag-specific T cell responses in vivo. An H-2L(d)-restricted nonamer peptide, derived from an endogenous murine leukemia provirus was identified as a CMS4-reactive CTL epitope based upon the following: CTL cross-recognition of another syngeneic tumor cell line (CT26 colon carcinoma) previously characterized to express that gene product; sensitization of Ag-negative lymphoblasts or P815 targets with the peptide; and by cold target inhibition assays. In vivo, the adoptive transfer of CMS4-reactive CTL (> or =1 x 10(6)) resulted in nearly the complete regression of 3-day established lung metastases. Furthermore, mice that rejected CMS4 following a single adoptive transfer of CTL displayed antitumor activity to a rechallenge 45 days later, not only in the lung, but also at a s.c. distal site. Lastly, the adoptive transfer of CTL to mice harboring extensive pulmonary metastases (> 150 nodules) led to a substantial reduction in tumor burden. Overall, these data suggest that the adoptive transfer of tumor-specific CTL may have therapeutic potential for malignancies that proliferate in or metastasize to the lung.

Juvenile rheumatoid arthritis: linkage to HLA demonstrated by allele sharing in affected sibpairs.

OBJECTIVE: To test for linkage between the HLA region and juvenile rheumatoid arthritis (JRA), with stratification by onset and course types, in a cohort of affected sibling pairs (ASPs). METHODS: Eighty pairs of siblings with JRA who were registered with the Research Registry for JRA ASPs (sponsored by the National Institute of Arthritis and Musculoskeletal and Skin Diseases) were typed for HLA-DR. The observed ratio of sharing of none, one, or both parental DR alleles was compared against the expected ratio of 1:2:1 by goodness-of-fit chi-square tests. A group of 265 unrelated control subjects served as a comparison population for HLA-DR allele frequencies among patients, by Fisher's exact test. RESULTS: Overall, there was excess sharing of 2 DR alleles among ASPs with JRA. The observed ratio of sharing 0, 1, or 2 DR alleles was 8:40:32, instead of the expected ratio of 20:40:20 (P < 0.001). When stratified by JRA onset type, excess allele sharing was demonstrated among ASPs who were concordant for onset type (P = 0.002). This was true for both pauciarticular and polyarticular onset. When stratified by disease course, excess allele sharing was also demonstrated among ASPs who were concordant for disease course (P < 0.001). This was true for both the pauciarticular and the polyarticular course. Among the 32 ASPs who shared two DR alleles, 5 pairs had both DR8 and DR11, which was significantly more frequent (P < 0.0001) than the incidence in the control group (n = 0). CONCLUSION: This study of an independent cohort of multiplex families confirms the previously reported linkage between pauciarticular JRA and the HLA-DR region that was identified using a different analytic method in a cohort of simplex families. Additionally, this study establishes evidence for linkage between polyarticular JRA and the HLA-DR region.