This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.Abstract Purple passion fruit (Passiflora edulis Sims) is one of the introduced tropical plants, an increasing interest has arisen due to its distinctive taste and attractive flavor. It is expected that passion fruit production and planted area will increase gradually in the years ahead because of high profitability and consumer's demands of healthful ingredients. So we tried to investigate the effect of plant growth regulators and antioxidants on in vitro plant regeneration and callus induction from leaf explants of passion fruit for an establishment of optimal mass propagation system. Young leaf explants of purple passion fruit were cultured in Murashige and Skoog (MS) medium containing different growth regulators and antioxidant additives to induce the shoot organogenesis. After 8 weeks, the highest embryogenic callus formation rate was obtained in MS medium supplemented with 1 mg･L -1 6-benzylaminopurine (BAP) and 2 mg･L -1 2,4-dichlorophenoxyacetic acid (2,4-D), furthermore, the shoot development via organogenesis was also observed. Silver nitrate (AgNO 3 ), which was added into the medium to minimize the adverse effects of leached phenolics, was effective for reduction of medium browning and sudden explant death. In the medium supplemented with 1 mg･L -1 BAP and 1 mg･L -1 gibberellic acid (GA 3 ), shoots were most vigorously regenerated and elongated. Most shoots rooted successfully in half strength medium with 1 mg･L -1 indol-3 acetic acid (IAA), and more than 90% of plantlets survived after 4-month acclimatization period.

The genus Cypripedium, known as lady's slipper orchid, has been considered to have a considerable marketability as a potted and garden plant with beautiful flowers; however, this species is becoming endangered and, in some places extinct, due to habitat destruction and illegal collections. As such, an optimized artificial propagation system is necessary for its conservation and horticultural cultivation. For the establishment of the in vitro proliferation of the endangered Cypripedium macranthos Sw., native to Korea, the effects of medium strength and sucrose concentrations on germination, protocorm formation and seedling growth were investigated through asymbiotic seed culture. The highest germination rate and protocorm formation rate were obtained with a 1/4 MS medium; higher MS medium strengths did not generate the favorable conditions required for germination and protocorm growth. The overall growth characteristics of roots and buds developed from protocorms were best in cultures of 1/4 MS medium. On this medium containing 10 g•L-1 sucrose, both the germination rate and protocorm formation rate increased significantly. The general growth properties of seedlings developed on the medium with 10 g•L-1 sucrose were best, showing the highest bud formation rate and root number. Our results demonstrate that the 1/4 MS medium, supplemented with 10 g•L-1 sucrose, could improve in vitro germination, and facilitate the growth of seedlings developed from the protocorms of C. macranthos Sw.

To find the optimal propagation condition for endangered Cypripedium macranthos Sw., also known as lady's slipper orchid, the effect of various organic additives on in vitro germination, protocorm formation and seedling growth was investigated during asymbiotic seed culture. When 100 ml·L-1 coconut water was added to the basal medium, the highest germination rate and protocorm formation rate were achieved, with 70.8% and 74.2% respectively. Supplementation of phloem sap from birch tree or maple tree also showed a facilitating effect to improve the germination and protocorm development. With 100 ml·L-1 birch sap or maple sap, both the germination and protocorm formation rates were roughly more than 65% and 68%. The roots and buds of the seedlings grew vigorously in the medium containing 100 ml·L-1 coconut water or phloem sap, in particular, their bud formation rates increased by more than 70%. Addition of banana powder and peptone could not create a more significantly favorable culture condition, and non-addition had the worst results. Our results demonstrated that proper organic amendments such as coconut water and phloem sap might be preferred to in vitro germination and the growth of seedlings developed from the protocorm of C. macranthos Sw. during asymbiotic seed culture.

Mulberry (Morus sp.) of the family Moraceae is very economically important in Asian countries including Korea, because its leaf and fruit have been commercially used in sericulture and horticultural industries. Therefore it is necessary to develop the optimal production system for rapid and cost-effective propagation of mulberry. Our studies focused on establishing an acclimatization method for the successful plantlet production of new cultivar 'Cheongsu' which was transferred ex vitro after in vitro culture. In particular, effect of abscisic acid (ABA) addition into the last subculture medium on plantlet response to subsequent ex vitro transfer and its growth was investigated. During acclimatization, stomatal conductance and transpiration rate of ABA-pretreated plantlets were significantly lower than those of non-treated plantlets. Net photosynthetic rate of ABA-pretreated plantlets decreased after ex vitro transfer but increased after 14 days, and it was mostly higher than that of non-treated plantlets. Moreover, relative water content as well as chlorophyll contents and its ratio were also higher in ABA-pretreated plantlets. On the other hand, proline was considerably higher than in control plantlets. After 1 month of ex vitro transfer, survival rate of ABA-pretreated plantlets was 85.6%, which increased by 29.1% in comparison with control (56.5%). More vigorous growth was also observed in ABA-pretreated plantlets. From these results, it was found that application of ABA to the last subculture medium could improve acclimatization and promote survival of mulberry plantlets after ex vitro transfer, inducing water stress tolerance and alleviating abiotic stresses.Keywords Mulberry, Abscisic acid, Ex vitro transfer, Acclimatization, Water stress Introduction Mulberry (Morus sp.) is an economically important tree grown in India, China, Korea and several Asian countries where its foliage is used as food for silkworms in sericulture industry (Vijayan et al. 2012). It is also commercially valuable in the horticulture, food and cosmetic industries, in particular, its phytochemical and medicinal properties such as antioxidants (Yen et al. 1996) and hypoglycemia compounds (Kelkar et al. 1996) have been widely used as healthcare products. These days it is cultivated for fruit production, which is used for human consumption including jam, jelly, frozen desserts, pulp, juice and wine (Koyuncu 2004). Mulberry fruit is regarded as a traditional medicine for dysentery, constipation and avulsed teeth due to rich phenolic acids and flavonoids (Arfan et al. 2012; Lee and Bae 2011).Plant micropropagation has been widely applied to agriculture industry for large scale production of economically important and valuable species. But its commercial use is limited in many species due to the low plantlet survival rates during the acclimatization (Pospisilova et al. 2009a, b), which are known to be related to an abnormal morphology, anatomy and physiology under in vitro culture conditions (Dias et al. 2013a). Poor functioning of...

: These studies were conducted to discriminate Cynanchum wilfordii, C. auriculatum, Metaplexis japonica, and Polygonum multiflorum by morphological characters. Cynanchum wilfordii, C. auriculatum, and Metaplexis japonica(Asclepiadaceae) were easily distinguished from Polygonum multiflorum by twining stem to the left, producing opposite leaves, having laticiferous stems and leaves at cutting cross-sectionally, and lacking lignification of the stem. Four species had different morphological characters of flower. Comparing four species to fruit type, three species of Asclepiadaceae had follicle, while Polygonum multiflorum had achene with three wings. Follicle shape was lanceolate, narrowly lanceolate, and widely lanceolate in C. wilfordii, C. auriculatum, and Metaplexis japonica, respectively. Whereas seed color of four species was dark brown similarly, seed shape and 1,000 seed weight were useful characters to discrimate between three species of Asclepiadaceae and Polygonum multiflorum. Shape and color of root were thickened and yellowish brown in C. wilfordii and C. auriculatum. They were thin and long and white in Metaplexis japonica, and fusiform and reddish brown in Polygonum multiflorum. They would be useful characters to distinguish Cynanchum spp., Metaplexis japonica, and Polygonum multiflorum, however, impossible characters to discriminate between C. wilfordii and C. auriculatum.