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Fundamental Studies on Somatic Fusion of Alfalfa and Sweet Clover

Author: MaoYaNi
Tutor: ZhangDeZuo;SunJuan
School: Gansu Agricultural University
Course: Grassland
Keywords: Alfalfa Leucanthous Melilot Callus Somatic embryos Protoplast Somatic cell fusion
CLC: S541.9
Type: Master's thesis
Year: 2010
Downloads: 60
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Abstract


Alfalfa protein content of about 17% of the total amount of dry matter to 20% of the vitamin and mineral content rich, sweet clover resistant to drought, cold, salinity, poor resistance to the test by both organizations culture study, a prerequisite for somatic cell fusion, and has a high nutritional value of alfalfa and sweet clover strong resistance of new varieties accumulated scientific technical material. Of this study Gannon the Giants, WL324 and Suntory four alfalfa varieties through somatic embryos ways to establish a process of regeneration system, observed Gannon somatic embryos cytology structure; explore the basic medium, culture time and hormones dialogue flowers and sweet-scented osmanthus callus induction; Gannon and white flowers and sweet-scented osmanthus cotyledons materials dissociation protoplast and its culture. The main findings are as follows: the test optimal induction of alfalfa callus screened by culturing explants, hormones, light intensity and other factors, the research was supported by a complete set of tissue culture regeneration system and somatic embryos Cytological observation. The results show that: (1) the optimal incubation time Gannon callus induction 14d, explants hypocotyl hormone combination of 2.0 mg / L 2,4-D 0.2 mg / L KT; different light conditions no significant effect on callus induction rate was 100%, light and dark alternating conditions induced callus growth faster, more exuberant color slightly green or green and yellow, but higher rates of browning. (2) the transfer of callus to somatic embryogenesis medium, four varieties of alfalfa callus induction under conditions of light and dark alternating easier to form somatic embryos to Gannon somatic embryos highest incidence , between 20.83% to 56.25%, and then turn to the Giants, Suntory and WL324; the best somatic embryogenesis medium for UM 0.1 mg / L NAA 0.5 mg / L KT, the occurrence rate of 56.25%; study also found the right amount of PEG6000 promote somatic embryogenesis to 6% PEG6000. (3) cytological observations suggest that somatic embryogenesis experienced globular, heart-shaped embryo, torpedo embryo and cotyledon stage 4. This test studied basic medium of Leucanthous Melilot, explants, training time, and the effects of hormones on callus induction. The results show that: UM medium callus induction rate is the best basic medium; 3 explants hypocotyl callus growth most productive, highest induction rate is the best explants; callus to reach exuberant period after induction 21d, the highest induction rate is the best time to cultivate; higher the concentration of 2,4-D and KT, the harder, the more dry the formation of callus, cotyledon and hypocotyl callus induction rate lower, while the radicle the first to promote after inhibition. The trial Gannon varia alfalfa and white flowers and sweet-scented osmanthus cotyledon material solutions from cultured protoplasts, the results show that: Gannon 1 to 2.5% R-10 cellulase 0.5% R-10 Segregation enzymatic highest the protoplast yield obtained from the 12h, 46.75 × 106 / g for the optimal enzyme solution portfolio; white flowers and sweet-scented osmanthus cellulase 2.0% 0.5% R-10 the macerozyme 0.5% pectinase for 24h the protoplasm body production was 5.00 × 106 / g. Medicago Varia and white flowers and sweet-scented osmanthus protoplast medium of KM8P 1.5 mg / L 2,4-D 0.2 mg / L KT, small callus culture to visible long after February.

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