Genetic variability in a temperate intertidal Phoronid,Phoronopsis viridis

The Phoronida are a coelomate phylum consisting of only two genera and about 12–15 described species. Phoronids probably represent the common ancestral stock of all lophophorates, and may be the most primitive living deuterostomes. Using the techniques of starch gel electrophoresis, we have studied genetic variation at 39 loci in 120 individuals of Phoronopsis viridis collected in Bodega Harbor, Bodega Bay, California. Allelic variation was found at 27 (69.2%) loci. If a locus is considered polymorphic when the frequency of the most common allele is no greater than 0.99, the proportion of polymorphic loci in the total sample is 48.7%. The average number of alleles per locus is 2.23. The expected frequency of heterozygous loci per individual on the assumption of Hardy-Weinberg equilibrium is 9.4%. There is evidence of inbreeding; the mean value of F, Wright's fixation index, is 0.21±0.02. Genetic variability in P. viridis is intermediate among marine invertebrates. The tropical clam, Tridacna maxima, has on the average 20.2% heterozygous loci per individual. At the other extreme, a brachiopod from Antarctica, Liothyrella notorcadensis, has an average of 3.9% heterozygous loci per individual. Among marine invertebrates, there seems to be a gradient of decreasing genetic variability from low to high latitudes, which may reflect their different adaptive strategies.     

Maternal serum alpha-fetoprotein screening: report of a Canadian pilot project.

A pilot project of maternal serum alpha-fetoprotein (MSAFP) screening was carried out in Ontario from 1982 to 1985 to examine the feasibility and acceptability of screening a prenatal population for open fetal neural tube defects. A total of 8140 patients at low genetic risk were screened. Patient acceptance was excellent. Blood samples were taken at 16 to 18 weeks'' gestation. If the MSAFP level was elevated, the assay was repeated and an ultrasound examination performed. Amniocentesis was offered to 67 women with unexplained persistently elevated levels. The outcome of pregnancy was known in 7473 patients (91.8%). Seven of nine known open fetal neural tube defects were detected. All were confirmed, and no unaffected fetuses were aborted on the basis of the screening results. The rates of perinatal death (6.7%), intrauterine growth retardation (11.7%) and prematurity (23.3%) were significantly higher among the patients with unexplained elevated MSAFP levels than among those with normal levels (p less than 0.001). Of 20 patients with unexplained low levels, 10 subsequently had spontaneous abortions and 10 gave birth to term appropriate-for-gestational-age infants. Seven of nine patients who gave birth to infants with autosomal trisomy had MSAFP values below the median. The findings indicate that MSAFP screening is feasible, accurate and acceptable in a low-risk area.     

The use of bifunctional biotinyl compounds to determine the arrangement of subunits in avidin

A series of bisbiotinyl diamines was synthesized with between 9 and 25 bonds between the carboxyl groups of the two biotin residues. It was found that only one of the two biotin residues could combine with avidin when there were fewer than 12 bonds between the biotin residues. Compounds with longer chains behaved in a bifunctional manner and gave rise to linear polymers of avidin, which were characterized by electron microscopy and by gel filtration. The polymers formed with the shorter-chain reagents (12, 13 or 14 bonds) were relatively unstable and could be depolymerized by weakly bound analogues of biotin. The polymers of longer-chain reagents were not depolymerized under these conditions and were only slowly affected by added biotin. When the chain length of the reagent reached 23 bonds the polymers became much shorter, suggesting that the reagent was now able to link two subunits within the same avidin molecule. From the morphology of the polymers it could be concluded that the four subunits of the avidin molecules were arranged with 222 symmetry and that they were grouped in two pairs at opposite ends of the short axis of the molecule whose dimensions were 55Ax55Ax41A.     

PHO1 family members transport phosphate from infected nodule cells to bacteroids in Medicago truncatula

Legumes play an important role in the soil nitrogen availability via symbiotic nitrogen fixation (SNF). Phosphate (Pi) deficiency severely impacts SNF because of the high Pi requirement of symbiosis. Whereas PHT1 transporters are involved in Pi uptake into nodules, it is unknown how Pi is transferred from the plant infected cells to nitrogen-fixing bacteroids. We hypothesized that Medicago truncatula genes homologous to Arabidopsis PHO1, encoding a vascular apoplastic Pi exporter, are involved in Pi transfer to bacteroids. Among the seven MtPHO1 genes present in M. truncatula, we found that two genes, namely MtPHO1.1 and MtPHO1.2, were broadly expressed across the various nodule zones in addition to the root vascular system. Expressions of MtPHO1.1 and MtPHO1.2 in Nicotiana benthamiana mediated specific Pi export. Plants with nodule-specific downregulation of both MtPHO1.1 and MtPHO1.2 were generated by RNA interference (RNAi) to examine their roles in nodule Pi homeostasis. Nodules of RNAi plants had lower Pi content and a three-fold reduction in SNF, resulting in reduced shoot growth. Whereas the rate of ³³Pi uptake into nodules of RNAi plants was similar to control, transfer of ³³Pi from nodule cells into bacteroids was reduced and bacteroids activated their Pi-deficiency response. Our results implicate plant MtPHO1 genes in bacteroid Pi homeostasis and SNF via the transfer of Pi from nodule infected cells to bacteroids.

Two members of the PHO1 family in Medicago truncatula are involved in the transport of phosphate from the infected nodule cells to the Sinorhizobium meliloti bacteroids.     

Nitrogen and carbon costs of soybean and lupin root systems during phosphate starvation

Phosphate (P) starvation is one of the most limiting nutrients to N₂ fixation in legumes. Soybeans and lupins present different climatic origins, nodule morphologies and metabolic complexities, which may have various adaptive responses to short-term P starvation. Lupins and soybeans were cultivated hydroponically for 3 weeks. Short-term P starvation was induced for 14 days by switching the P-supply to 2 μM P. During P starvation, the lupins showed a lower decline in nodular P concentrations and maintained their biological N₂ fixation (BNF), in contrast to the soybeans. The lupins also maintained their photosynthetic rates and the nodular construction and growth respiration costs under P starvation, whilst soybeans showed a decrease in photosynthetic rates and an increase in nodular construction and growth respiration costs under P starvation. There was a also a shift towards more organic acid synthesis, relative to amino acid synthesis in lupin nodules than soybean nodules under P starvation. The lupins had higher amino acid concentrations in their nodules, whilst the soybean nodules maintained their ureide levels at the expense of a decline in amino acids. These results indicate that lupins may to be better adapted to maintaining BNF during short-term P starvation than the soybeans.     

Pathway analysis of radiation-sensitive meiotic mutants ofCoprinus cinereus

We have isolated 37 radiation-sensitive mutants of the basidiomyceteCoprinus cinereus. Each mutation is recessive, and the collection defines at least ten complementation groups for survival of gamma irradiation. Four complementation groups define the genesrad3, rad9, rad11 andrad12, which are required both for survival of gamma irradiation and for meiosis. Mutants in each of these four groups fail to complete meiosis and produce mushrooms with greatly reduced numbers of viable spores. Propidium iodide staining of meiotic nuclei showed a characteristic terminal appearance for each mutant: few cells of any of the meiotic mutants progress beyond prophase I, and both condensation and fragmentation or dispersal of meiotic chromatin are frequently observed. Scanning electron micrographs showed that the meiotic mutants make varying numbers (0–6) of basidiospore initials and that few of these initials develop into mature spores. When initials are present they are always symmetrically arrayed on the basidium, regardless of initial number. In quantitative measurements of gamma ray sensitivity, double mutants of every tested combination ofrad3, rad9, rad11 andrad12 consistently showed the same gamma ray sensitivity as the more sensitive single mutant parent of the cross. Therefore, these four genes are in the same pathway for the repair of gamma radiation damage, and this pathway also represents one or more functions essential for meiosis.