Study On Agrobacterium Mediated Transformation Of Leafy Gene Overexpression Vector Into Lily

Lily is a world famous bulb flower, is also one of the main fresh cut flowers, have an important place in the flower market. Asian lily have a large share in the market because of its gorgeous color and elegant pattern. In recent years, as the development of tissue culture in lily production breeding and molecular breeding, optimization of varieties of lily species tissue culture system is particularly important.The LEAFY genes of Arabidopsis play a key role in plant transition of flowering. From the1980s, along with research on the Arabidopsis take control way of flowering and the definition of the role of LEAFY genes as floral meristem identify genes and flower induction signal integrator in flower modulate path, LEAFY gene is widely used in higher plants flowering regulation through the transgenic approach. Different kinds of lily varies a lot in flowering time, and the long vegetative growth stage of lily has already caused obstacles for commercial production, the traditional control way is time-consuming and the effect is not significant.In this study,we use the field bulb scales as primary culture materials, and through the Sterilization Treatment, differentiation and bulb inductive medium was MS+6-BA2.0mg I-1+NAA0.2mg I-1, bulb expansion medium was2MS+NAA0.2mg I-1. In order to optimize the formulation of Asian lily rooting medium and lay a foundation for rooting and transplanting of Asian lily transgenic plants, Lilium Asiatic hybrid cv.’Pollyanna’ and Lilium Asiatic hybrid cv.’Prato’ test-tube seedlings were used to study the effect of combinations of five different hormone in different concentrations on rooting rate, root number, longest root length.we have screened relative suitable root medium through the measurement of relative index and data statistics analysis.In order to study the role of LEAFY gene in lily growth and development, constructing the Arabidopsis LEAFY gene overexpression vector, and transform LEAFY gene overexpression vector into Asian Liliy ’Elite’(lilium Asiatic CV ’Elite’)through the Agrobacterium-mediated technique, through the pre-culture3days, co-culture in dark for3days, inhibit the growth of Agrobacterium treatment and screening culture, after hygromycin B resistant gene screening,,we have gotten hygromycin B resistant plants, through PCR detection and Phenotype observation and statistics analysis. The results are as follows: 1.The results showed that: a combination of different types and concentrations of plant hormones significantly affected Asiatic lily rooting, and the rooting status of the two species had some differences.(1) The most appropriate rooting medium of Lilium cv.’Pollyanna’ was:MS+2,4-D1.0mg l-1+KT0.1mg l-1+6-BA1.0mg l-1+sucrose30g l-1+agar7g l-1; the average rooting rate of Lilium Asiatic hybrid cv.’Pollyanna’ could reach98.73%, the root number of them could reach6.53cm and the longest root length of them was4cm;(2) the most suitable rooting medium for Lilium cv.’Prato’was:MS+NAA1.0mg I-1+TDZ0.lmg l-1+sucrose30g I-1+agar7g l-1;the average rooting rate of Lilium Asiatic hybrid cv.’Prato’could reach92.73%, the root number of them could reach6.63cm and the longest root length of them was3.77cm.2.Research on Agrobacterium mediated transformation of Arabidopsis LEAFY gene overexpression vector into lily(1)Construct the LEAFY gene overexpression vector promoted by CaMV35S(2) Screening concentration of hygromycin B from explants of Asian Lily’Elite’was20mg l-1;(3) we have gotten11transformation plants;(4) Observation and statistics results show that: compared with the control plants, transgenic plants all show bulb diameter increase and the dwarf phenomenon.