Calf production from vitrified bovine sexed embryos following in‐straw dilution

The objective of this study was to develop an in‐straw dilution method suitable for direct transfer of vitrified bovine sexed embryos. Embryo sexing was performed by molecular diagnosis. Several sexed and vitrified‐warmed embryos were transferred after evaluation of morphologically embryonic survival at warming and in‐straw dilution (Evaluation group). The other embryos were immediately directly transferred to recipients without first being expelled from the straws after in‐straw dilution (Non‐evaluation group). The pregnancy rates of vitrified sexed embryos were 38.7% and 34.8% in the Evaluation group and Non‐evaluation group, respectively, which were not significantly different. The viability of lower quality embryos before vitrification tended to be lower (P = 0.087) than that of the higher quality embryos regardless of evaluating embryos after warming and in‐straw dilution. The abortion rates were similar, and there was no difference between the two groups (13.9% and 12.5%, respectively). These results demonstrate that vitrified bovine sexed embryos can be vitrified and diluted by the in‐straw method and that the vitrified and warmed sexed embryos can develop to term.     

Comparison of Pathogenicity and Nucleotide Sequences of 3′-terminal Regions of Bean yellow mosaic virus Isolates from Gladiolus

Sixty-four isolates of Bean yellow mosaic virus (BYMV) from cultivated and naturalized gladioli were divided into two pathogenic groups, necrotic spot (NS) and chlorotic spot (CS) groups on Chenopodium quinoa. NS-type isolates (S-22N and E-24N), CS-type isolates (S-22C and E-92C), and broad bean isolates (Sb-50C and Sb-12C) differed in their pathogenicity on Antirrhinum majus, Nicotiana benthamiana, Phaseolus vulgaris, Spinacia oleracea and Vigna unguiculata. The four gladiolus isolates were different from BYMV-B, -P, -O and C1YVV-N in their pathogenicity on these plants, while the two broad bean isolates were similar to BYMV-B, originally from broad bean. The nucleotide (nt) sequences of the 3′-terminal region of the BYMV RNA genome of the two NS-type isolates, the two CS-type isolates, the two broad bean isolates and BYMV-B, -P and -O were determined. In a phylogenetic tree based on the CP amino acid (aa) sequences, the two NS-type isolates clustered together (identity 98.4% and 98.2% at the nt and aa level, respectively). The two CS-type isolates clustered with BYMV-O (93.2 to 99.3% nt identity and 95.6 to 98.5% aa identity). The two broad bean isolates clustered with BYMV-B (99.0 to 99.5% nt identity and 98.9 to 99.6% aa identity). BYMV-P clustered with BYMV-CS (identity 97.7% and 99.3% at the nt and aa level, respectively). The obtained sequences were compared with those of the 3′-terminal regions of seven published BYMV isolates. In a phylogenetic tree based on deduced aa sequences, BYMV isolates were divided into four clusters. Received 1 July 1999/ Accepted in revised form 22 May 2000     

Effect of growth hormone on estrogen receptor binding properties in the chick oviduct in vivo

The equilibrium dissociation constant (K_d) and the maximum binding capacity (B_max) of estrogen receptor in cytosolic and nuclear fractions in oviduct of pullets following various daily injections (1–10 times) of growth hormone (GH: 50 µg/chick) were examined by Scathchard analysis of specific [³H]estradiol‐17β (E₂) binding. The K_d values of receptor in both fractions decreased after three times of GH‐injection. In the B_max values, three times of the injection caused a marked decrease in that value in the cytosolic fraction with a concomitant increase in that in the nuclear fraction, whereas the total B_max (sum of B_max in the cytosolic and nuclear fractions) did not change. A similar relationship between the K_d values in the binding of the two fractions was also observed in 4–10 times of GH‐injection. However, in 4–10 times of GH‐injection the B_max of the both fractions and total B_max was greater than that in vehicle‐injection (control). When the chicks were injected with 6–10 times of GH‐injection, the weight of oviduct was increased. No change in the plasma concentrations of E₂ was found following GH‐ and vehicle‐injections into the chicks. The results suggest that growth hormone stimulates the growth of the chick oviduct by increasing the binding affinity and capacity of estrogen receptor.     

Marker-assisted selection for forelimb-girdle muscular anomaly of Japanese Black cattle

Forelimb‐girdle muscular anomaly is a hereditary disorder of Japanese Black cattle characterized by tremors and astasia caused by hypoplasia of the forelimb‐girdle muscles. The locus responsible for this disorder has been mapped on a middle region of bovine chromosome 26. In this study, we applied marker‐assisted selection to identify the carriers of this disorder. Four microsatellite markers, DIK4440, BM4505, MOK2602 and IDVGA‐59, linked to the disorder locus were genotyped in 37 unaffected offspring of a carrier sire. Transmission of the mutant or wild‐type allele of the disorder locus of the sire to the 37 offspring was determined by examining the haplotypes of these markers. The results showed that nine and 18 of the 37 animals possessed the paternally transmitted mutant and wild‐type alleles, respectively, and therefore, the nine animals with the mutant allele were identified as carriers. We concluded that the marker‐assisted selection using these four markers can be applied for the identification of the carriers of forelimb‐girdle muscular anomaly of Japanese Black cattle.