Establishment of a new cell line susceptible to Cyprinid herpesvirus 3 (CyHV‐3) and possible latency of CyHV‐3 by temperature shift in the cells

A new cell line named CCF‐K104 predominantly consisting of fibroblastic cells showed optimal growth at temperatures from 25 °C to 30 °C. Serial morphological changes in the cells induced by Cyprinid herpesvirus 3 (CyHV‐3) included cytoplasmic vacuolar formation, cell rounding and detachment. Mature virions were purified from CyHV‐3‐infected CCF‐K104 cells by sucrose gradient ultracentrifugation and had a typical herpesvirus structure on electron microscopy. Infectious CyHV‐3 was produced stably in CCF‐K104 cells over 30 viral passages. Our findings showed that CCF‐K104 is a useful cell line for isolation and productive replication of CyHV‐3. A temperature shift from 25 °C to 15 °C or 35 °C did not allow serial morphological changes as observed at 25 °C for 14 days. Under the same conditions, real‐time PCR showed that CyHV‐3 was present with low viral DNA loads, suggesting that CyHV‐3 may establish latent infection in CCF‐K104 cells. Amplification of the left and right terminal repeat sequences of the CyHV‐3 genome arranged in a head‐to‐tail manner was detected by nested PCR following an upshift in temperature from 25 °C to 35 °C. The PCR results suggested that the circular genome may represent a latent form of CyHV‐3.     

锦鲤成熟系数及个体生殖力的研究

对锦鲤(Cyprinus carpiod)的成熟系数和个体生殖力进行了研究。结果显示:雌、雄锦鲤成熟系数变化规律基本一致,成熟系数在5月下旬达到最高值。2+~5+龄锦鲤个体绝对生殖力F与其年龄A、体长L、体重W呈直线相关,回归方程分别为:F=67054A+61567(R2=0.8419,P<0.01),F=8588.4L+24969(R2=0.9217,P<0.01),F=315.33W-9688.8(R2=0.9631,P<0.01)。     

温度对鲫血液生化指标和消化酶的影响

采用室内模拟方法,研究了鲫（Carassius auratus）在9 ℃、16 ℃、25 ℃、30 ℃四个温度条件下的血液学指标和消化酶活力的变化。结果显示,鲫血清无机成分中的镁、磷以显著差异在16 ℃时存在最大值,而其他成分变化不显著。在9~25 ℃水温范围内,甘油三脂、总胆固醇、总蛋白、白蛋白、球蛋白等均随水温上升而上升,并在25 ℃时达到最大值,而超过25 ℃后,甘油三脂、总胆固醇、总蛋白、白蛋白、球蛋白等含量均随水温升高而下降。在9~30 ℃水温范围内,血清中谷丙转氨酶和谷草转氨酶活性随水温上升先上升后有下降,在25 ℃时达到最大值。鲫肠道中,蛋白酶和脂肪酶活性在9~25 ℃阶段升高,25~30 ℃时下降,蛋白酶和脂肪酶的活性均在25 ℃时最高;而肝胰脏中蛋白酶和脂肪酶活性随温度变化不明显。     

cDNA cloning and characterization of the warm-temperature-acclimation-associated protein Wap65 from carp, Cyprinus carpio

We determined full-length cDNA of carp warm-temperature-acclimation-associated 65-kDa protein (Wap65). It encoded 439 amino acid residues with a signal peptide of 22 residues and showed an amino acid sequence identity of 88% to that of goldfish reported before (J. Biol. Chem. 1995. 270: 17087–17092). The number of potential N-linked glycosylation sites of carp Wap65 was two in contrast to three for goldfish. In addition, molecular mass determined by SDS-PAGE was apparently different from that of goldfish. These results suggest that the amount of oligosaccharide is different between the carp and goldfish protein. As in goldfish, carp Wap65 mRNA showed marked accumulation in hepatopancreas of the 30 °C- acclimated fish, which was 8-fold higher than that of the 10 °C-acclimated fish. Carp Wap65 showed 30% amino acid identity to mammalian hemopexins, which appeared to be considerably low in comparison with those among mammalian hemopexins (72 to 80%), or among carp Wap65 and rainbow trout hemopexin-like protein (70%). However, although mammalian hemopexins contain residues comprising the heme binding pocket, carp Wap65 lacked one of the two histidine residues to serve as heme axial ligands in hemopexins. Our data on carp protein substantiates the previous observation for goldfish and indicates that Wap65 might have some important functions in warm-temperature-acclimation of fish.     

Transmission of resistance to common scab from the diploid to the tetraploid level via 4x-2x crosses in potatoes

Summary Diploid potato clones selected for their reaction to common scab and their ability to produce 2n male gametes were used in a series of crosses to a susceptible tetraploid female parent (cv. Shepody). In addition, two tetraploid clones were also selected for their reaction to common scab and crossed with Shepody as a female parent. Results indicated that resistance to common scab can be effectively transmitted from the diploid to the tetraploid level via 4x-2x crosses. Diploid parents producing 2n pollen via either first division or second division restitution can be used to transmit scab resistance. A relatively small proportion of resistant individuals could be recovered from susceptible x susceptible crosses in both 4x-2x and 4x-4x combinations. The data support a previously developed hypothesis that scab resistance is relatively simply inherited.     

Geographical distribution of the DL1 and DL2 genes causing hybrid dwarfism in Phaseolus vulgaris L., their association with seed size,and their significance to breeding

Summary Dwarlism in F₁ hybrids has been observed in over 100 crosses of dry beans (Phaseolus vulgaris L.) at the Centro Internacional de Agricultura Tropical (CIAT), Cali, Colombia. In each cross, one parent always had small seeds and the other parent either medium or la ge ones. This apparent incompatibility between the two groups of germplasm was controlled by two complementary, dominant genes: DL₁ and DL₂. Smallseeded bean lines carried gene DL₁ and originated in Brazil, Colombia, Guatemala, and Mexico; medium for large-seeded bean lines carried gene DL₂ and were from Bolivia, Brazil, Chile, Colombia, Turkey, The United States, and West Germany. Thes two genes have probably played an important role in the evolution of dry bean forms of different seed sizes by serving as a genetic barrier or isolating mechanism, thus limiting free genetic recombination between the two germplasm groups.Apparent differences in the adaptiveness and yielding ability of the two groups of bean germplasm, smallys, medium- and large-seeded, and some breeding implications for manipulation of the genes causing F₁ hybrid dwarfism were also discovered.     

A procedure for combining a quantitative inheritance study with the first cycle of a breeding program

Summary The number of effective factors controlling the inheritance of a quantitative character can be estimated by analysis of F₃ lines where the parental lines are homozygous. The procedure is illustrated with data from a study of pod length inheritance in common bean (Phaseolus vulgaris L.). The mean pod length of Sprite, the parent with longer pods, was compared statistically with the means of F₃ progenies derived from F₂ plants obtained from the appropriate tail of the F₂ frequency distribution for pod length. The data indicate that analysis of F₃ progenies avoids much of the underestimation of the effective factor number which is characteristic of other procedures. The number of effective factors was estimated to be a minimum of 4 and the correlation between random F₂ parents and F₃ progeny means was r=0.82.Florida Agricultural Experiment Station Journal Series No. 406.     

QTL analysis of photoperiod sensitivity in common buckwheat by using markers for expressed sequence tags and photoperiod-sensitivity candidate genes

Photoperiod sensitivity is an important trait related to crop adaptation and ecological breeding in common buckwheat (Fagopyrum esculentum Moench). Although photoperiod sensitivity in this species is thought to be controlled by quantitative trait loci (QTLs), no genes or regions related to photoperiod sensitivity had been identified until now. Here, we identified QTLs controlling photoperiod sensitivity by QTL analysis in a segregating F₄ population (n = 100) derived from a cross of two autogamous lines, 02AL113(Kyukei SC2)LH.self and C0408-0 RP. The F₄ progenies were genotyped with three markers for photoperiod-sensitivity candidate genes, which were identified based on homology to photoperiod-sensitivity genes in Arabidopsis and 76 expressed sequence tag markers. Among the three photoperiod-sensitivity candidate genes (FeCCA1, FeELF3 and FeCOL3) identified in common buckwheat, FeELF3 was associated with photoperiod sensitivity. Two EST regions, Fest_L0606_4 and Fest_L0337_6, were associated with photoperiod sensitivity and explained 20.0% and 14.2% of the phenotypic variation, respectively. For both EST regions, the allele from 02AL113(Kyukei SC2)LH.self led to early flowering. An epistatic interaction was also confirmed between Fest_L0606_4 and Fest_L0337_6. These results demonstrate that photoperiod sensitivity in common buckwheat is controlled by a pathway consisting of photoperiod-sensitivity candidate genes as well as multiple gene action.     

Development of an appropriate breeding scheme for tolerance to Empoasca kraemeri in common bean

Summary Significant advances in increasing tolerance to the leafhopper Empoasca kraemeri Ross & Moore in common bean have been obtained using a new breeding scheme where yield under leafhopper attack is the principal selection criterion in the evaluation of progenies. However, to further refine this breeding scheme, a study was conducted to determine whether selection for nonprotected yield would be more effective in early versus late generations. Two selection strategies were compared. In Strategy I, early generation selections in the F₂ and F₃ were compared to Strategy II where late generation selections were made in the F₄ and F₅, with the F₂ and F₃ generations advanced using single pod descent and bulk practices, respectively. Yield trials of the F₆ lines from both selection strategies were conducted under nonprotected and insecticide protected treatments. No significant differences were detected between the two selection strategies. However, Strategy II did produce advanced lines with greater nonprotected yields than did Strategy I, with the best F₆ line, in three out of four crosses, coming out of Strategy II. Late generation selection is recommended over early generation selection. Results of the new leafhopper breeding scheme, based on yield, are compared to the old breeding scheme where selections were made using visual selection practices in early generations.     

Expected levels of heterozygosity in autotetraploid progeny populations

Summary A breeding program aimed at improving the seed yield in bean for processing was started. Six parents divergent in a range of agronomical characters were crossed according to diallel mating design. Cluster analysis performed on the composite cross population evidenced divergence between F1 hybrids and to maximize genetic recombination, the most different F1 hybrids were intercrossed. A Factor analysis was performed on 7 morphological and yield-determining traits of 15 hybrids F1 and parents. Three Factors, representing patterns of variables interpreted as productivity per plant, growth factor, and branching-productivity, were extracted. These results evidenced the inverse relationship between seed yield components (seeds/pod, pods/plant, and hundred seed weight): i.e. the selection for either variable is detrimental to the other. It may be possible to break this association if the plant construct assures high sink to satisfy all plant requirements: tall and large plants bearing numerous nodes, leaves and reproductive structures. Moreover, the selection practiced simultaneously on the yield components positively affected the seed yield trait. An index of selection and response to selection were estimated on the segregant populations. The realized gain in seed/yield reached after one cycle of selection (C1) was 6.5 g, about 20% over the base population (C0). The expected gain from selection based on the superior 5% plants in C2 families was 25.5% and approached the realized gain. The result of this paper gives evidence to support that the seed yield can be improved by an adequate selection intensity and selection index can be very useful when it is faced with the improvement of several traits at the same time.