| M.cordata (Macleaya cordata (Willd) R.Br.) belongs to the poppy, a very important resource plant, which is rich in a large number of isoquinoline alkaloids. In this thesis,8M.cordata flower buds of different origins were applied to the study, which came from Kai hua in Zhejiang Province (MCK), Gaofang in Hunan Province (MCG), Qimen in Anhui Province (MCQ), Guangze in Fujian Province (MCZ), Kaili in Guizhou Province (MCL), Songtao in Guizhou Province (MCS), Xiushan in Chongqing municipality (MCX), and Yanling in Hunan Province (MCY).respectively for anther culture in the process of a comparative study of the relevant factors. We established a relatively perfect system for anther culture and plant regeneration, providing a precondition for the establishment of DH lines. Using the anther in vitro culture to produce haploid plants provides the material for the genome research.The results are as follows:1. An reliable and simple method was confirmed for the judgment of anther development. The mononuclear stage of anther as the material, the difference between flower bud length and diameter was the relative value judgments of anther developmental stages. The difference of bud length and diameter was approximately5mm, which represented the mononuclear middle and advanced stage. If the difference was greater than the distance of5mm, most of the anthers were in a dual-core or mature.2. The induction of different Macleaya anthers origins at different seeding densities was essentially normal distribution. The highest induction was D3-D5(240-400anthers per vessel).We took D4treatment in this study,320anthers per vessel.3. The callus induction rate was different of different origins.The highest induction rate was MCG, reaching12.6%. On callus proliferation effect, different regions also showed great diversity, the proliferation of MCG was the highest conversion rate,20.1%. However the anther proliferation conversion rates of MCL and MCS were only8.9%and5.8%less than half of the MCG. 4.The pretreatment temperature on callus induction rate was crucial. The pretreatment with35℃of2-4d could improve callus induction rate. Appropriate low temperature pretreatment was benefit for the improvement of anther induction rate. After a few days more than4d induction treatment was significantly lower than the control. Different origins of Macleaya had different sensitivity to cold pretreatment, In addition, we found that the anther callus induction rate was lower with more than5days treatments.5. Low concentrations of2,4-D and6-BA were good for Macleaya anther callus. For MCK-3and M-8, in Y1treatment, the highest rates of the two anther Macleaya origin were22.1%and26.8%. The optimum hormone combination for Macleaya callus induction was: MS+0.5mg/L2.4-D+0.5mg/L6BA.6. It was not obvious of hormone treatment for rooting. Both for MCK-3and MCG-8, MS medium without hormones was better for rooting. The day of rooting was15, and root length and total root also reached the middle level.7. The hardening and transplanting of seedlings were also studied.V nutritional soil:V sand was1:1, which were more suitable for the growth of regeneration seedlings. The survival rate was transplanting seedling was100%after28d.8.216Macleaya regenerated plants were tested by flow cytometry. Only six Macleaya haploid plants were regenerated, accounting for3%of the total sample.88%were two ploidy, and6%ploidy unusual plants, as well as a small part of the regenerated seedlings were unknown. |
PDF Full Text Request