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

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

2、体核型分析。结果如下：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

3、为 1B, 2B, 1B, 1B, 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 万寿菊论文：万寿菊属植物染色体核型分析及万寿菊 psy 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

4、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 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,

5、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 Tagetes L. accessions were studied, and he result could provide the scientific basis for 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 materia

6、ls, accord ing to tran sie nt expressi on of gus gene, the suitable kind of variety 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

7、 higher lutein content by making psy gene overexpressied. The detailed study results 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

8、+20m,2 n=2x= 24m, 2n=2x=2sm+22m,2 n=2x=24m; the len gth type was 2n=4L+6M2+10M1+4S, 2n=6L+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 of 1B, 2B, 1B, 1B, 1B, 1B, 1B. The karyotype formula of T.patula L. Golde n Gate was 2n=4x=4

9、8=4sm+44m; the len gth type was 2n=12L+8M2+16M1+12S; The KA type was of 1B. (2) Based on the marigold N-091, the N-092, N-093, the N-094 leaves and stems with axillary buds for experimental material. Results of gus gene transient expression: N-094 (86.6%) N-093 (73.3%) N-091 (43.3%) N-092 (40% . N-094 was the variety which was most easily converted. (3) For N-094, based on leaves and stems with 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 was available to be the expla nt