| In this study, Tianbaojiao (Musa acuminata cv. Tianbaojiao, AAA group) were used as experimental materials to study the key technology for factory production of banana seedlings. Firstly, seedlings were induced in vitro from Tianbaojiao explants, then LED light sources were used to study the proliferation and rooting of Tianbaojiao seedlings and ISSR molecular marker technology was used to detect genetic stability of Tianbaojiao tube seedlings with different subculture time. Based on these, a new model of factory production for banana seedlings were constructed and analyzed.1 The establishment and optimization of in vitro regeneration system in Tianbaojiao1.1 The establishment of sterile system of TianbaojiaoIn the experiment, Tianbaojiao buds were used as explants. The buds were cut into 8 cm3 and then were treated with 75% C2H5OH for 30 s and 0.1% HgCl2 for 17 min, finally washed with water for 5-7 times, and put on MS medium with 4.0 mg/L 6-BA,0.5 mg/L NAA,30 g/L sugar and 6 g/L agar for induction culture. The induction rate was as high as 64.51% after 30 days.1.2 The studies on optimization of proliferation system of Tianbaojiao plantlets in vitroâ‘ The effects of different culture methods on the proliferation of Tianbaojiao. In this experiment, the solid and liquid culture conditions were compared for optimization of Tianbaojiao proliferation. L9 (34) orthogonal treatments were used in the solid medium, which compared the different 6-BA, NAA and MS concentrations on the proliferation. Results showed that:the MS concentrations had the biggest impact on the proliferation of in vitro plantlets, the optimal proliferation medium was:MS+5.0 mg/L 6-BA+0.4 mg/L NAA+30 g/L sugar+6 g/L agar, under which the proliferation coefficient was up to 3.57; the completely randomized method was used in the liquid culturewith comparing the liquid volume and inoculum density on the proliferation, and the results showed that when the liquid volume is 22 ml and the inoculation density was 5 per bottle, the proliferation coefficient was largest (4.30).â‘¡The effects of LED light sources on the proliferation of Tianbaojiao. All the Tianbaojiao seedlings were cultured on the MS medium with 5.0 mg/L 6-BA,0.4 mg/L NAA,30 g/L sugar and 6g/L agar under white, warm white, red, blue, green and yellow light, respectively, to compare the different effects of LED light qualities on the proliferation, using fluorescent lamp as control. The results showed that the warm white light was the most optimum LED light source for the propagation of Tianbaojiao, under which the proliferation coefficient was 4.11.â‘¢The effect of illumination intensity on the proliferation of Tianbaojiao. All the Tianbaojiao seedlings were cultured on the MS medium with 5.0 mg/L 6-BA,0.4 mg/L NAA,30 g/L sugar and 6g/L agar under LED warm white light with five different illumination intensities(1000 lx,2000 lx, 3000 1x,4000 1x and 5000 1x). The results showed that 20001x is the most suitable illumination intensity for the proliferation of Tianbaojiao tube seedlings.1.3The optimization of rooting in Tianbaojiaoâ‘ Hormone and sugar concentration affected the rooting of Tianbaojiao. The Tianbaojiao seedlings with similar vigours and sizes were used as materials to analyze the effects of different concentrations of white sugar, NAA and IBA on the rooting. The results showed that:the optimal combination for rooting in Tianbaojiao was:1/2 MS+1.5 mg/L IBA+0.5 mg/L NAA+20 g/L sugar+6 g/L agar+1 g/L AC.â‘¡Different LED light sources had impacted on Tianbaojiao rooting. The sterile Tianbaojiao seedlings with similar vigours and sizes were used as materials to analyze the effect of different LED light qualities on the rooting. Six kinds of LED light qualities were:White light, Warm white light, Red light, Blue light, Green light and Yellow light and the common fluorescent light was used as control. All seedlings were cultured on the same medium (1/2MS+1.5 mg/L IBA+0.5 mg/LNAA+20 g/L sugar+1.0 g/L AC). The results showed that the most favorable light for rooting was LED warm white light.â‘¢LED light intensity had effected on the rooting of Tianbaojiao. The sterile Tianbaojiao seedlings with similar vigours and sizes were used as materials to analyze the effect of different LED light intensities on the rooting.5 kinds of light intensity were:1000 1x,2000 1x,3000 1x, 4000 1x and 5000 1x. All seedlings were cultured on the same medium (1/2 MS+1.5 mg/L IBA+ 0.5 mg/LNAA+20g/L sugar+1.0 g/L AC). The results showed that the most suitable light intensity for Tianbaojiao seedling rooting was 4000 lx.1.4 The transplantion of rooted plants of TianbaojiaoThe rooted plantlets were used as materials to compare the effects of different soil matrix on the survival rate of transplanted plantlets. The experimental results showed that:the best time for acclimatization was 7 days and it not only had the highest survival rate, but also had a faster recovery time.The optimal soil matrix was peat soil+river sand+garden soil,whose ratio was 2:2:1, and the survival rate was as high as 96.3%.2 The ISSR genetic stability analysis of Tianbaojiao in different subculture cyclesIn the experiment, the DNA was extracted from the banana leaves of Generations 0, Generations 5, Generations 10, Generations 12, Generations 13, Generations 14 and Generations 15 for detecting the ISSR genetic stability.70 DNA samples were used as templates to PCR with 12 ISSR primers, which produced 96 bands in total. The results showed that:the variation rates of the 10th and 12th generation were both 4.8%, and that of 13th,14th and 15th generation were 6.7%, 9.7% and 16.5%, respectively. Tianbaojiao plantlets had some variations in the subculture, which were relatively stable within the 12th generation. However, the variation rate increased gradually later, and the genetic stability remained relatively stable within the 13th generation.3 The estaiblishment of the factory production model for the in-vitro plantlets and economic benefit analysis in TianbaojiaoThe main programs for industrial production of in vitro plantlets of Tianbaojiao: Establishing sterile lines from suckersâ†'buds proliferationâ†'rooting cultureâ†'acclimatizationâ†' transplantationâ†'sales. The cost of established production system was analyzed:the total cost for an annual output of 1 million strains of tube seedlings is 221173.9 yuan, equivalent to the cost of the seedlings of 0.189 yuan/plant, including the cost for seed stage (174038.21 yuan, which account for 79.90%). According to the market sale price of every plant with 30 cm height is 0.5 yuan, the total sale price of 1 million strains was 500000, and the profit for one year was 278826.1 yuan. |
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