A high efficient in vitro regeneration protocol was developed from leaf explants of the female 'Red Sun' kiwifruit (Actinidia chinensis) and the multiplication coefficient and rooting rate of adventitious buds were also optimized. This method does not require formation of callus tissues which leads to somaclonal variations. The results show that the adventitious buds developing directly from explants tissue were noticed after 30 d of culture. The maximum regeneration frequency of adventitious buds is 100% and 18.67 shoots was observed in each leaf explants when MS medium was supplemented with 3.0 mg/L BA+1.0 mg/L NAA. The optimal culture medium for bud multiplication is MS+2.0 mg/L BA+1.0 mg/L NAA+0.1 mg/L GA3 and the multiplication coefficient reached 8.63. On the rooting medium with 1/2 MS+0.8 mg/L IBA for 15 d, the adventitious plantlets were transferred into matrix perlite supplied with 1/2 MS liquid medium for 15 d and the rooting rate reached 100%. 95 out of 98 plantlets (96.94%) survived acclimatization, producing healthy plants in the greenhouse. Taken together, a highly efficient regeneration method via leaf explants of 'Red Sun' kiwifruit was successfully established. This protocol may be useful for micropropagation and genetic transformation studies of 'Red Sun' kiwifruit.
Actinidia ; growth & development ; Plant Growth Regulators ; pharmacology ; Plant Leaves ; growth & development ; Regeneration ; physiology ; Tissue Culture Techniques ; methods

Objective To explore the effect of invigorating splcen and kidney methods on the pregnancy outcome of the intrauterine insemination (IUI) in infertile men with asthenospermia.Methods a total of 78 male infertility patients with spleen and kidney deficiency type were collected,all cases were randomly allocated into three groups,group A using methods of invigorating spleen and kidney,group B using Antioxidant therapy,and group C as the blank group.There are 26 cases in each group.One course of treatment lasts for 12 weeks.Before and after treatment,the changes of sperm quality index were analyzed by computer assisted sperm analysis (CASA) and sperm morphology.The pregnaucy outcome after IUI was followed up.Results After the treatment,sperm motility and sperm forward movement (PR) difference before and after treatment in group A with was statistically significant (P ＜ 0.05).The differences between patients with semen volume,sperm concentration,sperm morphology and sperm motility parameters before and after treatment in three groups were not statistically significant (P ＞ 0.05).Clinical pregnancy rate and cumulative pregnancy rate and the live birth rate in group A were higher than that of group B and group C,but the difference was not statistically significant (P ＞ 0.05).Conclusion Invigorating spleen and kidney method can improve the sperm motility and forward movement speed of the type of patients for the treatment of male infertility patients with asthenospermia,and it is helpful to improve the pregnancy outcome of the IUI.

Bacterial canker caused by Pseudomonas syringae pv. Actinidiae is one of the most important diseases of kiwifruit (Actinidia chinensis) and leads to considerable yield losses. In order to obtain transgenic plants with resistance for 'Red Sun' kiwifruit to canker disease, a non-specific lipid transfer protein-like antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) was introduced into 'Red Sun' kiwifruit through Agrobacterium-mediated transformation. After two days of co-cultivation with A. tumefaciens strain LBA4404 harboring 35S:LJAMP2, the transformed explants were transferred to the selection medium containing 25 mg/L kanamycin+3.0 mg/L BA+1.0 mg/L NAA. The regeneration efficiency of kanamycin-resistant shoots reached to 85%. All (100%) of kanamycin-resistant shoots rooted on half-strength MS medium supplemented with 0.8 mg/L IBA and a total of 40 regenerated plantlets were obtained. PCR and histochemical GUS activity analysis show that 23 of 40 lines (57.50%) were positive, suggesting that the LJAMP2 gene was integrated into the genome of 'Red Sun' kiwifruit. Taken together, we established an efficient genetic transformation method for 'Red Sun' kiwifruit using A. tumefaciens and the transformation frequency reached 5.11%. This protocol will be useful for the genetic breeding of 'Red Sun' kiwifruit for improvement of disease resistance.
Actinidia ; genetics ; Agrobacterium ; Antigens, Plant ; genetics ; Carrier Proteins ; genetics ; Leonurus ; Plant Proteins ; genetics ; Plants, Genetically Modified ; genetics ; Transformation, Genetic