The present study was conducted to develop the effective disinfection protocol for the in vitro micropropagation of strawberry (Fragaria x ananassa Duch) with the use of shoot tips, runner tips, nodal segments and leaf segments as explants. The explants used in this study were surface sterilized using antibiotics, fungicides and other sterilants for different time durations. Although using the same sterilants, the most effective and successive way of using sterilants is different upon the time duration for each sterilant. In this study, two sterilization protocols were used and each protocol included same fungicide and antibiotics concentrations for the same time durations but there were slightly different concentrations and time durations of other sterilants. The present investigation revealed that the most effective way of sterilization protocol which were observed on the nodal segments while treated with protocol II including (10ml/L) fungicide solution for 2 hours, (500mg/L) concentration of ciprofloxacin for 1 hour, (20%) chlorox solution with two drops of Tween 20 for 5 mins, (70%) ethanol solution for 5 mins and (0.1 %) mercuric chloride solution for 4mins. However the same sterilants using the same sterilization time did not give raise the survival rate for runner tip explants, because these treatments resulted in tissue necrosis and contamination and then finally the death of the explant materials. And also, the explants of shoot tips and leaf segments were not shown the effective result compared with using nodal segments. So, for the micropropagation of field grown strawberry, the sterilization protocol II was suite for the nodal segments used as explants for the culture initiation on MS basal medium.
Banana propagation is dependent on propagation by tissue culture for industrial purposes. Due to lack of native endophytes in tissue culture plantlets, reintroduction of beneficial microorganisms to tissue culture plantlets become popular as a useful foundation for improving the level of establishment, increasing plant growth parameters, protecting the plantlets against pest and diseases and overall performance for field plantation. In the present study, rhizospheric and endophytic four bacterial species (Pseudomonas fluorescens, Bacillus putida, Bacillus pumilus, Bacillus amyloliquefaciens were isolated from different parts of healthy banana plants such as root, leaf and pseudostem. Banana tissue culture plantlets were cultured for root formation stage on MS medium and used as host for artificial inoculation. Then, bacterial inoculation was carried out with tissue culture plantlets both in vitro and ex vitro conditions. All banana plantlets used for this experiment were successfully acclimatized and survived in the greenhouse. The experiment consisted of three treatments: (1) bacterial inoculum with in vitro rooted plantlets for 2 weeks (2) bacterial inoculum with ex vitro (hardening) plantlets (3) control with sterilized distilled water. Among these treatments, in vitro inoculation affected significantly increased plant height, number of leaves and plant girth for all data collection times with the highest increment indices: (24.0, 7.0, and 7.3). Ex vitro inoculation showed the second highest increment indices: (22.0, 6.9 and 6.3) for plant height, number of leaves and plant girth, respectively. The lowest number of plant growth parameters were observed in control treatment for all four data collection time (19.0, 6.2, and 6.0). According to the results, application of mixtures of bacterial inoculum was effective for enhanced plant growth parameters of tissue culture banana plantlets under greenhouse condition. In addition, two weeks artificial inoculation during in vitro plantlets stage was found the most suitable method for application of microbes with tissue culture banana plantlets which can promote the microbial rhizosphere of banana prior to field plantation.
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