| Narcissus tazetta L. is a kind of perennial plant with high economicand orinamental value. But for its long term asexual reproduction, the reproduction of narcissus was greatly limited. Causing the virus accumilation, severe degradation and resistance weakened. Therefore, improving the resistance of narcissus, accelerating the production of high-quality virus-free seedling and broadly popularing the new varieties, is the imperative methods for the sustainable development of narcissus. In this research, we take ’Yunxiang’ narcissus and White Flower Ⅱ narcissus bulds as material, and the effects of different culture media in vitro culture on the white flower Ⅱ and ’Yunxiang’ Narcissus were studied, This study will contribute to production of virus free seedling, theoretical support and technical guidance for popularization and application of the preservation of germplasm resources, and also lay the foundation for improvement of germplasm. And the resistance gene fragment of WD40 familly have been screened from ’Yunxiang’ Narcissus transcriptome established by the narcissus research group from horticulture college, FAFU. The open reading frame was cloned by RT-PCR, and the gene expression under three abiotic stress was analyzed through qPCR technology. This research will provide useful information for the improvement of germplasm, The main results are as follow:1. During this experiment, we explored the explant disinfection method of narcissus ’Yunxiang’ and white flower Ⅱ, and the most suitable culture medium of them were selected. The bulb plates of ’Yunxiang’ and white flower Ⅱ are sterilized best when they are soaked in 25% sodium hypochlorite solution for about 30 minutes and 25 minutes respectively. MS+1.5 mg/L 6-BA+0.5 mg/L 2,4-D is the optimum medium for bulblets inducing of ’Yunxiang’, and the best medium for adventitious buds inducing of white flower Ⅱ is MS+0.5 mg/L 6-BA+0.1 mg/L 2,4-D. MS+5.0 mg/L 6-BA+0.5 mg/L NAA and MS+3.0 mg/L 6-BA+0.5 mg/L NAA offer the best condition for ’Yunxiang’ and white flower II narcissus seperately in the aspect of callus differentiation. MS+1.5 mg/L 6-BA+0.5 mg/L 2,4-D is the best subculture medium for both ’Yunxiang’ and white flower II narcissus. The optimum rooting medium for ’Yunxiang’ narcissus is 1/2MS+0.05 mg/L IBA+0.1 mg/L NAA, and it’s the same for white flower Ⅱ.The construction of vitro plant regeneration system of ’Yunxiang’ and white flower II narcissus can offer technology supports for transgenic breeding, original species conservation, germplasm conservation and new varieties promotion of narcissus in the future.2. WD40 gene from ’Yunxiang’ Narcissus was obtained by RT-PCR (GenBank login number:KU193745), and the characteristic of its structure were analyzed and the function was predicted through bioinformatics methods and software, WD40 gene contains a 420 bp open reading frame, encoded 140 amino acids, is a hydrophobic protein. Phylogenetic tree analysis showed that, the gene encoding protein have closer relation with the phoenix dactylifera, Musa acuminata subsp. Malaccensis and Zea mays.3. High temperature, salt, ABA stress were treated on ’Yunxiang’ Narcissus, and the real time PCR results showed that, WD40 gene relative expression volume increase in different degrees. The most obvious expression of the Material is the WD40 with high temperature treatment. We predicate the gene may involved in the high temperature, salt, hormone and other abiotic stress. |
PDF Full Text Request