The cDNA clone (pNP24) coding for a protein induced by exogenous NaCl has been isolated from a tomato root cDNA library with the use of an inosine containing synthetic oligomer. The authenticity of the clone has been established by comparing the sequence of the clone to the NH2-terminal sequence of the protein which has been purified to homogeneity by HPLC. The nucleotide sequence of pNP24 reveals a 5' signal sequence, an open reading frame of 718 nucleotides, a 3' AT rich untranslated region containing a probable polyadenylation signal sequence, and a poly A stretch. The mature polypeptide sequence as deduced from the nucleotide sequence reveals a protein with a molecular weight of 24226. This protein has been named NP24. It is slightly basic and has an unusually high number of cysteines (15). Northern blot analyses reveal that the abundance of mRNA for NP24 is at least 100-fold greater in tomato suspension cells in log phase grown in medium with NaCl than in cells grown in the control medium. The mRNA for NP24 is below the level of detection in roots of young control tomato plants until several weeks after germination but it is induced earlier and to higher levels in roots stressed by 0.171 M NaCl. Thus salt stress accelerates the accumulation of message in tomato roots. A comparison of the steady state levels of mRNA for NP24 to the accumulation of NP24 by immuno analyses indicates that the accumulation of this protein is determined by its mRNA level. The protein is not secreted and is localized within the cytoplasm or the soluble fraction of the nucleus, vacuole, or microbodies. NP24 has a high degree of homology (58%) with thaumatin, a protein which has considerable value as an artificial sweetener.

Summary. Theca and granulosa layers were isolated from the preovulatory follicles of prepubertal gilts which were untreated (Group A), killed 72 h after 1000 i.u. PMSG (Group B), killed 84 h after PMSG (Group C), or killed 84 h after PMSG + 500 i.u. hCG given at 72 h (Group D). The tissues from individual follicles were cultured for 24 h alone (C), with FSH (F) or with LH (L), and the content of immunoreactive relaxin in the culture media was measured by RIA. Concentrations of relaxin-like material were close to the limit of detection of the assay in all granulosa cell cultures, and in the thecal cultures from the untreated gilts. However, thecal cultures from all 19 treated gilts produced relaxin. The mean \m=+-\ s.e.m. concentrations (pg/follicle) in Groups AC, BC, CC and DC were 26\m=.\5\m=+-\3\m=.\04,93\m=.\1 \ m=+-\ 4\m=.\6, 138 \ m=+-\ 16\m=.\4and 285\m=.\6 \ m=+-\ 54\m=.\1 respectively. Therefore relaxin production was stimulated by PMSG (P < 0\m=.\05), with hCG treatment in vivo leading to a further significant increase (P < 0\m=.\05). In-vitro treatment with gonadotrophins had no effect in Groups A, C and D, but in Group-B gilts LH produced a significant (P < 0\m=.\05) rise in relaxin levels. These studies indicate that the theca is the principal source of relaxin in the porcine preovulatory follicle. The increased production before ovulation suggests that relaxin may be involved in follicular growth or rupture.

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Two experiments were conducted to examine the effect of the feeding of a high-energy diet during early gestation on survival of porcine conceptuses. In the first, 91 second-estrus gilts were inseminated 24 h after detection of standing estrus and randomly assigned to a recommended energy (RE: 22.2 MJ of DE/d) or high-energy diet (HE: 36.0 MJ of DE/d) obtained by addition of cornstarch to RE. Gilts were slaughtered 9, 10, or 11 d later, and numbers of corpora lutea (n = 1,268) and recovered conceptuses (n = 977) were recorded. Conceptuses were photographed, and surface areas were measured using an image analysis system. Standard deviations (SD) for mean areas from 82 litters with 899 embryos were used as a measure of size variability within and between litters, and 472 of these were cultured for measurement of estrogen (E) concentrations by enzymeimmunoassay. Conceptus survival was similar for RE and HE groups (76.2 +/- 21 vs 78.8 +/- 17.5%; P > .05). Mean surface areas and SD increased (P < .05) between d 9 and 11, but they were the same for both diets (P > .1). No difference was detected between diets in E produced by d 9 and 10 blastocysts after 24 h in vitro (P > .1). In contrast, d-11 RE embryos produced more E than did HE embryos after 3 h (.77 +/- .11 vs .37 +/- .11 ng/mL; P < .01) and 24 h (3.02 +/- .40 vs 1.29 +/- .39 ng/mL; P < .01).(ABSTRACT TRUNCATED AT 250 WORDS)