《万寿菊论文：万寿菊属植物染色体核型分析及万寿菊psy基因遗传转化体系影响因素的研究》由会员分享，可在线阅读，更多相关《万寿菊论文：万寿菊属植物染色体核型分析及万寿菊psy基因遗传转化体系影响因素的研究（5页珍藏版）》请在金锄头文库上搜索。
1、万寿菊论文：万寿菊属植物染色体核型分析及万寿菊psy基因遗传转化体系影响因素的研究【中文摘要】本研究以八个万寿菊属（Tagetes L.）品种为材料，进行染色体核型分析，从细胞学水平为万寿菊遗传育种提供科学依据。以万寿菊（Tagetes erecta L.）N-091、N-092、N-093 和 N-094为材料，根据gus基因瞬时表达情况，筛选易转化基因型及外植体种 类,在此基础上对根癌农杆菌介导的万寿菊psy基因遗传转化体系的影响因素进行研究，为实现psy基因过量表达，获得高叶黄素含量的 万寿菊转基因品种奠定基础。本研究具体结果如下：（1）对八个万寿 菊属品种进行染色体核型分析。结果如下：
2、观赏万寿菊丰收、泰山、 奇迹、完美黄、完美橙黄、印卡橙黄和印卡黄的核型公式分别为2n=2x=24=6sm+18m, 2n=2x=24=14sm+10m, 2n=2x=24=24m ,2n=2x=4sm+20m, 2n=2x=24m,2 n=2x=2sm+22m,2 n=2x=24 染色体长度组成分别为2n=4L+6M2+10M1+4S,2 n二 6L+4M2+10M1+4S,2 n=4L+8M2+8M1+4S,2 n=6L+ 6M2+6M1+6S,2 n=2x=4sm+20m,2 n=4L+8M2+8M1+4S,2 n=4L+4M2+12M1+2S。核型类型分别为1B, 2B, 1B, 1B,
3、 1B, 1B, 1B。孔雀草金门的核型公式为2n=4x=48=4sm+44n染色体长度组成为2n=12L+8M2+16M1 + 12核型类型为 1B（2）以万寿菊 N-091、N-092、N-093和N-094的叶片及带腋芽茎段为实验材料，各品种gus基因瞬时表达阳性率结果为:N-094（86.6%） N-093（ 73.3%） N-091(43.3%) N-092 (40% ,易转化的基因型为 N-094。(3)以 N-094 的叶片和带腋芽茎段为外植体，再生诱导率结果为带腋芽茎段(93.3%)叶片(50% ,表明带腋芽茎段可作万寿菊再生体系的外植 体。其诱导培养基为：MS + 6-BA
4、0.1 mg/L+ NAA 0.1 mg/L+植物凝胶 6 g/L。(4)以N-094带腋芽茎段为外植体，研究psy基因遗传转化体 系各影响因素，结果为：预培养1 d,农杆菌浓度OD600=0.8,1/2 MS悬 浮，共培养时培养基添加AS浓度为100卩mol/L,侵染采用80 W10 min 的超声波针刺法处理，黑暗共培养2 d,选择培养基中抗生素Kan浓度 为80 mg/L,Cef400 mg/L。利用上述体系，本研究侵染外植体485个, 获得4个抗性株系，经过PCR佥测,有两株呈阳性，阳性植株获得率为 0.41%。【英文摘要】Karyotypes of eight genus Taget
5、es L. accessions were studied, and he result could provide the scientificbasisfor gen etic breedi ng in the cytology level. Based on the marigold (Tagetes erecta L. ) N-091, the N-092, N-093, the N-094 for the materials, accord ing to tran sie nt expressi on of gus gene, the suitable kind of variety
6、 and explant types for conv ersi on have bee n chose n, and the factors in the Agrobacterium-mediated gen etic tran sformati on system have been studied, This will provide basic information for new marigold which with higher lutein content by making psy gene overexpressied. The detailed study result
7、s are as follows.(1) The karyotype formula of T. erecta L. Harvest , Taishan , Marval , Perfection Yellow , Perfection Orange , Inca Orange , Inca Y ellow was like:2n=2x=6sm+18m,2 n=2x=14sm+10m,2 n=2x=24m,2 n=2x=4sm+20m,2 n=2x=24m, 2n=2x=2sm+22m,2 n=2x=24m; the len gth type was2n=4L+6M2+10M1+4S,2n=6
8、L+4M2+10M1+4S,2 n=4L+8M2+8M1+4S,2 n二 6L+6M2+6M1+6S,2 n=2x=4 sm+20m,2 n=4L+8M2+8M1+4S,2 n=4L+4M2+12M1+2S;The KA type was of1B, 2B, 1B, 1B, 1B, 1B, 1B. The karyotype formula of T.patulaL. Golde n Gate was 2n=4x=48=4sm+44m; the len gth type was2n=12L+8M2+16M1+12S; The KA type was of 1B. (2) Based on th
9、emarigold N-091, the N-092, N-093, the N-094 leaves and stemswith axillary buds for experimental material. Results of gusgene transient expression: N-094 (86.6%) N-093 (73.3%) N-091 (43.3%) N-092(40% . N-094 was the variety whichwas most easily converted. (3) For N-094, based on leaves and stems wit
10、h axillary buds as the expla nts, the results of rege nerati on in ducti on rate: stems with axillary buds(93.3%) leaves(50% . Stems with axillary buds wasavailable to be the expla nt in the rege nerati on system ofmarigold.The inductive mediumfor stems with axillary buds: MS + 6-BA 0.1 mg/L + NAA0.1 mg/L + 6 g/L plant gel. (4) According
