تاثير التعقيم مصدر الجزء النباتي ومنظمات النمو في اكثار نبات الكاردينيا المتقزم Gardenia jasminoides Ellis صنف Radicans خرج الجسم الحي واقلمتة == Effect of Sterilization, The Source of Explants and growth regulators on propagation of dwraf Gardenia jasminoides Ellis. cv. Radicans In vitro and Acclimatization
Author name:
لمياء حسين موسى عايش المازني
Supervisor name:
هدى عبد الكريم عبد الودود الطه
General topic:
Agricultural sciences
Specific topic:
Plant - Gardening
Degree:
Master
University:
University Of Basrah - Faculty Of Agriculture
Language:
Arabic
University location:
Basrah
First pages:
31T1851 - p.pdf
Abstract:
This work was conducted in Plant Tissue Culture Laboratory, Agriculture Collage, Basrah University, Basrah Governorate, Iraq during the period 23/ October/ 2013 to 15/ March/ 2015. The aims of this study can be summarized to : (a) set up a protocol for establishment of sterilization techniques and effect the source of explants in micropropagation of dwarf gardenia (Gardenia jasminoides Ellis.) cultivar 'Radicans', (b) study the effect of different growth regulators auxins and cytokinins on multiplication and rooting of regenerants , and (c) set up a procedure for plantlet acclimatization .Results can be summarized as follow : 1. Contamination is first obstacle facing the micropropagation for dwarf gardenia. In order to overcome this problem, a sterilization procedure was developed as a base line to determine the relative effectiveness of various disinfectants which is described in this study. Observations indicate that only 26.60% healthy clean cultures was obtained when 40% of sodium hypochlorite solution used singly. But, when explants disinfected first with 40% sodium hypochlorite followed by treating with 0.1% mercuric chloride for 10 minute the percentage rate of aseptic cultures increased to 56.60%. Therefore, the successful protocol was adopted in this study.2. (a) Results revealed that High response to tissue culture , shoot numbers , length, leaf width and leaf numbers (100%, 1.30 shoots/explant, 8.4 - 1.55 cm and 2.66 leaves/explant respectively) were obtained when the shoot tips and nodal explants cultured on MS medium supplemented with 3 mg/L BA+ 0.2mg/L NAA+ 0.2mg/L GA3 each. While, maximumsuccessful response of nodal explant to tissue culture 80% and 100% was observed in MS medium enriched with BA 3 and 4 mg/L BA. However, granular callus was formed on the lower part of the nodes, but after a time this callus turned brown and hyperhydric (vitrified).(b) Maximum successful response of shoot tips in vitro 100% was noticed in MS medium enriched with different concentrations of TDZ 0.5, 1, 1.5 and 2 mg/L in the presence of NAA and GA3 0.2mg/L each, but the high rate of shoot number 1.8 and length 1.18 cm obtained on MS medium containing 1 mg/L TDZ. Where, the rate value of leaf width and numbers were increased only on MS medium fortified with low concentrations of TDZ 0.5 and 1mg/L. On the contrary, nodal explants cultured on same medium failed to responding, except the ones cultured on MS medium containing 1mg/L TDZ , showed moderate rate value of average growth responding66. 06%.3. (a) Results also revealed that the culture of shoot tips on medium supplemented with a combination of 3 mg/L BA, 0.2 mg/L NAA and 2 mg/L GA3 gave the highest value of average shoots number, leave number, and leaf width (2.50 shoots/explant, 6 leaves/plant and 1.7cm respectively). But, at this formula shoots became hyperhydic and leaves fall down. Whereas, highest shoot length 1.75 cm was recorded on MS medium enriched with 1 mg/L BA.(b) Highest shoot and leaf numbers and leaf width recorded on MS medium containing 1 mg/L TDZ in the presence of NAA and GA3 0.2mg/L each 4.33 shoots/explant, 6.00 leaves/ explant and 1.30 cm respectively. Whereas, highest shoot length 1.50 cm recorded on MS medium fortified with 0.5 mg/L TDZ. However, small amount of white brownish callus raised around the nodes grown on medium containing 0.5 mg/L TDZ, whilebig amount of the same callus was recognized around the nodes grown on MS medium containing 1.5 mg/L TDZ.4. (a) The present study has revealed that abundant primary callus formation (100%) could be achieved only from young leaf segments (basal and middle segments) within 10.3 and 15.3 days in MS medium containing 3 mg/L 2,4 - D and 0.2 mg/L BA, and under dark conditions.1) Transferring the primary callus to MS medium enriched with 3mg/L 2,4 - D for 8 weeks under light conditions led to induce somatic embryos, that can be identified by their nodular consistency and green color.2) While, shifting the primary callus to MS medium enriched with 3 or 5mg/L BA + 0.2mg/L 2,4 - D led to converting the primary callus to compact callus, and at the end of incubation period, small adventitious shoots was generating.(b) Results also showed that white brownish granular callus initiated under dark conditions on the base of the leaf petioles (basal segment) explants grown on MS medium containing 5 mg/L BA+0.2mg/L NAA, then the callus increased to covered whole explant during the end of incubation period. However, adventitious shoots was generated from primary callus when shifted to MS medium fortify with 1 or 2 mg/L BA. Results also revealed that the middle segment of the leaf explants failed to form callus on MS medium containing different concentrations of BA used in this study.(c) It was observed that in vitro shoots segments 1.5 cm length produced 100% callus on 1/2 MS medium enriched with 8 mg/L NAA within 8 - 9 days of inoculation. However, transferring this callus to full strength of MS medium supplemented with 1 or 2 mg/L BA led to generating small green adventitious shoots.5. (a)Maximum root formation rate 100% with high number of roots 7.33/plantlet, root length 7.33cm and number of secondary roots (8.00/ main root), were obtained within a short period 13.33 days Also, it was found that the length of shoots and number of leaves were increased to 12 cm and 11.33 leaves/plantlet, respectively.(b) On ½MS medium containing different concentrations of NAA 2, 4, 6 and 8 mg/L in the presence of 0.2mg/L BA, adventitious roots formed with rate 100%. These roots emerged indirectly from the edge of shoots with callus induction phase and developed into white thick and wide roots which grew and branched inside medium. However, maximum value of morphological traits for root was obtained when the shoots grown on MS medium containing 8 mg/L NAA in the presence of 0.2mg/L BA, while the less period of root formation was recorded when the NAA concentration decreased to 2mg/L.6. Two procedures of acclimatization have been attempted to obtain full healthy plantlets ex vitro. Results showed that the soaking of rooted plants for 10 - 14 days in pure water, nearly 60% of plantlets were survived. Whereas, treated the plantlet with 2% PEG 6000 before transplanting to soil increased the survival rate to 100%, Also this concentration of PEG was decreased the precentage of wather loss from leaves of gardenia plants through the acclimatization stage 12.8%.
