Gene delivery is one of the keen interests in animal industry as well as research on gene functions. Some of the in vivo gene delivery techniques have been successively used in various tissues for the gene therapy and transgenesis. Despite intensive efforts, it still remains to overcome problems of limited local and regional administration and low transgene expression. To improve the efficiency of gene delivery, a new procedure was tested. We injected exogenous DNA containing LacZ into the female or male gonads and then pulsed electric field. Electroporated gonads showed positive X-gal staining in many seminiferous tubules of the porcine fetal gonads. Exogenously introduced LacZ genes were also expressed in female porcine gonad. In addition, we demonstrated efficient gene delivery in gonad of adult mouse. Furthermore, we succeed to generate genetically modified germline cells showing GFP and positive X-gal signals. The results suggest that the newly developed gene delivery is an effective way of in vivo transfection in mammals. The developed gene delivery procedure should be useful in producing transgenic animals when combined with primary cell culture and nuclear transplantation.

In mammals, male and female germline stem cells are derived from primodial germ cells. Despite many efforts to identify stem cells from gonads, there has been little successe to identify germline stem cells yet. In this study, we isolate and characterized porcine germline stem cells using only stem cell markers that are prevalently expressed in various tissues. Gonadal cells derived from both male and female formed colonies and showed AP activities and different lectin binding properties. Pluripotency of germline stem cells was also identified by positive signals against putative stem cells markers such as SSEA-1 and SSEA-3. In addition, nestin was also found in primary gonad cells that have a similar morphology to the AP-positive cells. The nestin expression suggests that the germline stem cells may have similar expression of the prevalent stem cell markers found in other tissues. The demonstration of nestin expression together with pluripotent cell markers calls further investigation of the possible differentiation of nestin-positive cells into neurons.

An organic apple (Malus x domestica Borkh.) orchard was established to study the interaction effects of ground cover management systems (GMS) and nutrient sources (NS) on soil and tree nutrient status and tree growth. Trees received one of four GMS: 1) green compost (GC), 2) wood chips (WC), 3) shredded paper (SP), and 4) mowand- blow (MB). Across all GMS, one of three NS was applied: A) a commercial organic ferlilizer (CF), B) poultry Iitter (PL) ， and C) control (NF). Overall, GMS had greater effects on the variables than did NS. GC mulch supplied greater nutrients, followed by WC, SP, and MB mulches. SP trees had lower foliar [N] in the first two years than the GC and WC trees. GC- and WC-treated trees had larger trunk cross sectional area than the SP and MB trees.