conditions and additional experimental details are given in Carter et al. (2005) . Treatments were replicated three times. Chemicals. Chiro -inositol, myo -inositol, D-pinitol, and other chemicals unless mentioned were obtained from Sigma
Mehmet Nuri Nas and Paul E. Read
Microshoots of four hazelnut genotypes grown in vitro on Nas and Read medium (NRM) containing various combinations of CuSO4 • 5H2O and myo-inositol were successfully rooted and acclimatized ex vitro without any need of in vitro hardening treatments. Dipping of shoot bases in 1000 ppm indole-3-butyric acid (IBA) solution for 5 or 10 seconds followed by placement of shoots in plant growth regulator free NRM gave rise to formation of roots as early as 8 days. Shoots treated for 5 and 10 seconds rooted similarly, and depending on genotype, 88% to 98% rooting was observed within 15 days after treatment with IBA. Ex vitro survival of shoots three months after in vitro-root induction was 73% when shoots were treated with IBA for 5 seconds and 66% when shoots were treated for 10 seconds. The highest ex vitro survival rate (97%) 3 months after root induction was observed when shoots were treated with IBA solution for 10 seconds, and then cultured directly in peat pellets. Shoots developed good roots, and grew up to 70 cm in height 3 months after root induction. The potential use of rooting and acclimatization protocol for commercial micropropagation of hazelnut is presented.
Hongmei Du, Zhaolong Wang, Wenjuan Yu, and Bingru Huang
). The decrease of melibiose content was also found in arabidopsis ( Rizhsky et al., 2004 ) under drought stress. However, the relationship between lower melibiose content and drought stress was unknown. The relative content of myo-inositol significantly
Whei-Lan Teng, Yann-Jiun Liu, and Tai-Sen Soong
An efficient method for the regeneration of shoots directly from cell suspensions of three commercial cultivars of lettuce (Lactuca sativa L. cv. Great Lakes 659-700, Salad Bowl, and Prize Head) is described. Cell suspensions were prepared by osterizing cotyledon-derived callus for 60 seconds. The effects of callus quality, light intensity, carbohydrate type and concentration, auxins, and cytokinins on cell growth and differentiation in the suspension culture were examined. Among these factors, callus quality and carbohydrates were the most critical. The optimal medium for regeneration of shoots in suspension culture was SH (Schenk and Hilderbrandt) basal medium containing 1000 mg myo -inositol/liter, 1.5% glucose, 0.44 μm BA, and 0.54 μm NAA. The pH of the medium was adjusted to 5.8. Under such condition, hundreds of shoots could be produced from 50 to 55 mg (dry wt) of cell aggregates within 2 weeks. Chemical names used: α-] naphthaleneacetic acid (NAA); indole3-acetic acid (IAA); 6-benzylaminopurine (BA).
Mahmoud B. Arif and Houchang Khatamian
Surface sterilized stem nodal sections of western soapberry (Sapindus drummondii Hook. & Arn.) were cultured on Murashige and Skoog (MS) medium. The basal media consisted of one half and full strength MS medium each supplemented with the following (mg-1): Nicotinic acid 0.5, pyridoxine Hcl 0.5, Glycine 2.0, myo-inositol 100, sucrose 30,000 and agar 8000. Each medium also was supplemented with either 0, 0.01, 0.1, 1.0 and 10 mg/l Thidiazuron (TZD) or 0, 0.5, 2.0 and 5 mg/l 6-Benzyladenine (BA). The pH of all media was adjusted to 5.8 ± 0.1. The culture media were autoclaved at 120°C at 1.5 Kgcm-1 pressure for 15 min.
The highest percentage of nodal sections resulting in shoot regeneration occurred on 1/2 MS with TZD at 0.01 mg/l and MS medium containing 0.5 mg/l of BA Increasing the TZD concentration above 0.1 mg/l resulted in callus formation on cut surfaces.
P. Healey, T.J Ng, and F.A. Hammerschlag
Mesophyll cells are desirable targets for studying responses to pathogens or pathogen-induced toxins. Based on host-pathogen or host-toxin interaction studies at the cellular level it can be determined whether a toxin can be used as a selective agent. Suspension cells are suitable selection units for in vitro selection of potentially useful somaclonal variants. Protocols for the isolation of muskmelon mesophyll and suspension cells were developed in order to study the effects of roridin E, a toxin produced by Myrothecium roridum, on leaf spot tolerant and sensitive muskmelon cultivars. Viable mesophyll cells were obtained by exposing leaf tissue to 1% cellulysin and 5% macerase in B5 medium with 0.4M sucrose for one hour. Viable suspension cells were maintained a medium consisting of MS salts, 3% sucrose, 3 (μM thiamine·HCl, 555 μM myo-Inositol, 28 μM kinetin and 9 μM IAA. Fluorescein diacetate was used to determine viability over time. Membrane stability was monitored by measuring changes in the fluorescence of cells stained with Merocyanine 540 (MC 540), an optical probe for changes in transmembrane electrical potential (PD).
Gerson R. de L. Fortes, Nilvane T.G. Müller, Janine T.C. Faria, Luciana B. Andrade, and Marisa de F. Oliveira
Asparagus is a vegetable that presents an increase in yield when propagated by meristem culture. On the order hand, the rooting phase in asparagus is greatly affected by the previous phase, i.e,. multiplication. This species presents a better rooting performance when callus is formed at the shoot base. So, the aim of this work was to evaluate treatments during the multiplication phase, which also leads to callus formation at the shoot base. The initial explants came from shoots being cultivated in vitro. It was tested kinetin at: (0.0, 0.5, 1.0, 1.5, and 2.0) μM; ancymidol at (0.0 and 0.5) μM and NAA at (0.0 and 0.5) μM for both genotypes, which were cultured in a MS medium added to sucrose (30 g·L–1), agar (6.0 g·L–1) and myo-inositol (100.0 m g·L–1). Shoots bearing two buds were inoculated in 10-ml test tubes and placed in a growth room for 30 days when they were evaluated. The addition of kinetin significantly improved the number of buds and at 1.3 μM this growth substance presented the best results as number of shoots is concerned. NAA application promoted a negative effect on spear bearing. The addition of ancymidol in this phase did not improve the bud multiplication. It was shown that clone M14 performed better than the hybrid cv. Deco as multiplication is concerned.
Gerson R. de L. Fortes, Luciana B. Andrade, Marisa de F. Oliveira, Nilvane T.G. Müller, and Janine T. C. Faria
The potato cultivar Cristal has recently been released by the CPACT/EMBRAPA Breeding Program. Such cultivar was selected for having high dry matter and low sugar content, which makes it desirable for the chip industry. However, this is a recalcitrant cultivar as far as in vitro multiplication is concerned. The aim of this work was to improve the rate of multiplication for this cultivar when it was submitted to different MS salt and sucrose concentrations in the culture media. Two-bud microcuttings were inoculated in test tubes (20 × 150) mm with 10 ml MS media at 3/4-, 1/2-, and full-strength and MS vitamins added to: myo-inositol (100 mg·L–1), agar (7.0 g·L–1) and sucrose as follows: 10, 20 and 30 g·L-1. Each treatment was repeated eight times and each replicate had eight explants. After inoculation the whole material was kept in a growth room at 25 ± 2°C, 16-hr photoperiod and 2000 lux. The evaluation was done 35 days later. It was found and increase in the number of buds as the sucrose concentration in the media decreased. As far as MS salts are concerned no difference in bud number was observed. The rate of multiplication was slightly higher for MS media at full strength and sucrose at low concentration (10 g·L–1). This treatment could be recommended for this cultivar.
Gerson R. de L. Fortes, Luciana B. Andrade, Janine T.C. Faria, Marisa de F. Oliveira, and Nilvane T.G. Müller
The potato cultivar Cristal recently released by the CPACT/EMBRAPA Breeding Program has high dry matter and low reduce sugars. These are desirable characteristics as industry processing is concerned. Nevertheless, this is a recalcitrant cultivar. The meristem culture is difficult to establish along with a very low multiplication rate. The aim of this work was to improve the multiplication rate for this cultivar. Two-bud microcuttings derived from apical, mid, and basal regions were inoculated in test tubes with 10 ml MS culture media and vitamins as follows; myo-inositol (100 mg·L–1); sucrose (10 g·L–1). No growth regulator was added. All treatments were placed in a growth room in a 16-hour photoperiod; 25 ± 2°C and 2000 lux. One month later, although it was observed that the final growth was more pronounced for basal microcuttings, no difference could be detected for number of shoots and multiplication rate. It was concluded that it makes no difference whatsoever kind of microcutting is used to start the micropropagation process.
Alessandro Chiari and Mark P. Bridgen
Meristems from three different positions were excised from in vitro plants of Alstroemeria genotype A30. Explants were removed from the most-distal vegetative shoot apical meristems, rhizome tip apical meristems, and rhizome tip axillary meristems. Meristems were cultured on four different media to compare the effect of meristem position and medium on the ability to produce Alstroemeria rhizomes from meristems. The meristem culture media were Murashige & Skoog salts plus 8.39 μM pantothenic acid, 1.19 μM thiamine, and 0.55 mm myo-inositol (MSM), MSM plus 8.88 μM of 6-benzylaminopurine (BA), MSM plus 8.88 μM BA, and 0.72 μM gibberellic acid (GA3), and MSM plus 0.72 μM GA3. Meristems that were removed from the vegetative shoot apices did not develop rhizomes on any medium. Rhizome tip apical meristems developed less than 10% rhizomes when subcultured on media containing BA and GA3. However, rhizome tip axillary meristems developed rhizomes on all media with best results achieved when the medium was supplemented with BA.