Study on the Explant Disinfection of Root Culture of Hevea brasiliensis

[Objective] The aim was to look for an effective way of decreasing explants contamination in the root tissue culture of Hevea brasiliensis.[Method] In order to study root tissue culture of H.brasiliensis,new roots from forest section after treatment for one month were as explants,and 1 g/L carbendazim,75% ethanol,0.1% mercuric chloride and Yipeilong with different concentrations were used as disinfectants for roots disinfection.[Result] Before conventional disinfection,root explants were treated by 1 g/L carbendazim for 2.5 h,and disinfected by 75% ethanol for 30 s and 0.1% HgCl2 for 6 min,then cultured on the callus induction medium supplemented with 0.1% Yipeilong.The results showed that the contamination rate of explants decreased to 44.59%,and 25.60% explants survived after cultured for 30 d.[Conclusion] The study could provide theoretical foundation for the decrease of explant contamination in the root tissue culture of H.brasiliensis.

Influence of cytokinins, basal media and pH on adventitious shoot regeneration from excised root cultures of Albizia lebbeck

A highly reproducible and efficient in vitro shoot regeneration system was developed in a potential medicinal plant, Albizia lebbeck using root explants. Root explants from 15 day-old-aseptic seedlings were cultured on Murashige and Skoog （MS） medium supplemented with different concentrations （0.5, 2.5, 5.0, 7.5 and 10.0 μM） of 6-Benzyladenine （BA）, Kinetin （Kn）, 2-Isopentenyl adenine （2-iP） singly as well as in combination with α-Naphthalene acetic acid （NAA） （0.1, 0.5, 1.0, 1.5 and 2.0 μM）. The highest rate of shoot multiplication （16.0 ± 1.87 for the average shoot number and 5.16 ± 0.38 cm for shoot length） was achieved on MS medium supplemented with 7.5 μM BA and 0.5 μM NAA. The effects of medium type, medium strength, pH and subculture on shoot induction and proliferation were also tested. An average of 21.6±2.87 shoots per explants could be obtained following this protocol. Rooting was achieved on microshoots using half strength MS medium with 2.0 μM Indole-3-butyric acid （IBA） after four weeks of culture. The in vitro raised healthy plantlets were successfully established in earthen pots containing garden soil and grown in greenhouse with 〉80% survival rate.

Role of neurotrophic factors in enhancing linear axonal growth of ganglionic sensory neurons in vitro

Neurotrophins play a major role in the regulation of neuronal growth such as neurite sprouting or regeneration in response to nerve injuries. The role of nerve growth factor, neurotrophin-3, and brain-derived neurotrophic factor in maintaining the survival of peripheral neurons remains poorly understood. In regenerative medicine, different modalities have been investigated for the delivery of growth factors to the injured neurons, in search of a suitable system for clinical applications. This study was to investigate the influence of nerve growth factor, neurotrophin-3 and brain-derived neurotrophic factor on the growth of neurites using two in vitro models of dorsal root ganglia explants and dorsal root ganglia-derived primary cell dissociated cultures. Quantitative data showed that the total neurite length and tortuosity were differently influenced by trophic factors. Nerve growth factor and, indirectly, brain-derived neurotrophic factor stimulate the tortuous growth of sensory fibers and the formation of cell clusters. Neurotrophin-3, however, enhances neurite growth in terms of length and linearity allowing for a more organized and directed axonal elongation towards a peripheral target compared to the other growth factors. These findings could be of considerable importance for any clinical application of neurotrophic factors in peripheral nerve regeneration. Ethical approval was obtained from the Regione Piemonte Animal Ethics Committee ASLTO1(file # 864/2016-PR) on September 14, 2016.