3种苜蓿花粉组织培养与单倍体鉴定

发布时间：2018-02-20 20:17

本文关键词： 花粉组织培养苜蓿花药倍性鉴定单倍体培养基　出处：《内蒙古农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：通过苜蓿花药或花粉培养获得单倍体,进一步利用纯系材料杂交育种,可充分发挥杂种优势,缩短育种年限并提高育种效率。本研究以花药为材料对草原1号杂花苜蓿、呼伦贝尔黄花苜蓿和新疆大叶紫花苜蓿进行花粉组织培养,通过固体与液体悬浮培养基、不同激素配比进行正交试验诱导花药愈伤组织,分化和生根培养后获得再生植株,建立适宜的花药组培再生体系;再生植株经气孔数初选、根尖染色体和流式细胞仪鉴定,确定染色体倍性。主要研究结果如下:(1)花药愈伤组织的诱导,草原1号杂花苜蓿固体培养基优于液体悬浮培养基,而呼伦贝尔黄花苜蓿与新疆大叶紫花苜蓿液体悬浮培养基优于固体培养基,出愈率最高值分别为83.33%、70.00%、83.33%。根据正交试验出愈率极差的分析,草原1号杂花苜蓿适宜的固体培养基为B5 +0.4 mg/L NAA+ 0.75 mg/L 2,4-D + 1.0 mg/L KT + 0.75 mg/L 6-BA;呼伦贝尔黄花苜蓿适宜的液体悬浮培养基为B5 + 0.4 mg/LNAA + 0.50 mg/L 2,4-D +3.0 mg/L KT +0.25 mg/L 6-BA;新疆大叶紫花苜蓿适宜的液体悬浮培养基为 MS + 0.2 mg/L NAA+ 2.0 mg/L 2,4-D + 3.0 mg/LKT+ 0.5mg/L 6-BA。(2)影响3种苜蓿花药愈伤组织形成的主要激素种类不同。在固体培养过程中花药愈伤形成主要因素,草原1号杂花苜蓿为KT、呼伦贝尔黄花苜蓿及新疆大叶紫花苜蓿为2,4-D;在液体悬浮培养过程中花药愈伤形成主要因素,草原1号杂花苜蓿雄性不育株为KT、雄性可育植株为6-BA、呼伦贝尔黄花苜蓿及新疆大叶紫花苜蓿为2,4-D。(3)3种苜蓿花药愈伤分化培养的适宜培养基为MS + 1.0 mg/L KT +0.2 mg/L NAA,分化率 58.33%-73.33%。(4)草原1号杂花苜蓿雄性不育植株、呼伦贝尔黄花苜蓿及新疆大叶紫花苜蓿花药培育再生植株中单倍体植株分别为4株、1株、7株,分别占各自总株数的8.51%、20%、14%。
[Abstract]:Haploid was obtained through anther or pollen culture of alfalfa, and further utilization of hybrid breeding of pure lines could give full play to heterosis, shorten breeding period and improve breeding efficiency. Pollen tissue culture was carried out on Hulunbeir alfalfa and alfalfa with large leaves in Xinjiang. The callus of anther was induced by orthogonal experiment in solid and liquid suspension medium with different hormone ratios. After differentiation and rooting culture, regenerated plants were obtained. To establish a suitable regeneration system of anther tissue culture, to determine chromosome ploidy by primary selection of stomatal number, root tip chromosome and flow cytometry. The main results are as follows: 1) Callus induction of anther. The solid medium of Medicago sativa No. 1 was superior to the liquid suspension medium, while the liquid suspension medium of Hulunbeir alfalfa and Xinjiang alfalfa was superior to the solid medium. The highest rate of callus was 83.33, respectively. According to the orthogonal test, the analysis of the difference of callus rate was made. The suitable solid medium for Medicago sativa No. 1 is B50.4 mg/LNAA 0.75 mg/L 24-D 1.0 mg/L KT 0.75 mg/L 6-BA.The suitable liquid suspension medium for Hulunbeir alfalfa is B50.4 mg/LNAA 0.50 mg/L 2n4-D 3.0 mg/L KT 0.25 mg/L 6-BAand Xinjiang large leaf alfalfa. The suitable liquid suspension medium was MS 0.2 mg/L NAA 2.0 mg/L 2G 4-D 3.0 mg/LKT 0.5 mg / L 6-BA.2) the main hormone types affecting callus formation of three alfalfa anthers were different, and the main factors of anther callus formation were found in solid culture. The main factors of anther callus formation during liquid suspension culture were KT, Hulunbeier alfalfa and Xinjiang large leaf alfalfa. The suitable medium for anther callus differentiation of alfalfa was MS 1.0 mg/L KT 0.2 mg/L NAA, the male sterile plant was KT, the male fertile plant was 6-BAand Hulunbeir alfalfa and Xinjiang alfalfa were 24-D. The male sterile plant of Medicago sativa No. 1 in the grassland was 58.33% -73.33% and 58.33% -73.33%. The haploid plants of Huulunbeir alfalfa and Xinjiang large leaf alfalfa anther culture were 4 plants and 1 plant respectively, accounting for 8.51% of the total number of plants.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S541

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