EMS处理唐菖蒲无性系变异的研究

发布时间：2018-04-02 11:35

本文选题：唐菖蒲　切入点：EMS突变体　出处：《东北农业大学》2014年硕士论文

【摘要】：唐菖蒲{Gladiolus hybridus Hort)是鸢尾科(lridaceae)唐菖蒲属(Gladiolus)多年生球茎类植物,是世界上著名的四大切花之一,唐菖蒲切花生产是我国花卉产业的重要组成部分。目前,我国唐菖蒲种球主要从国外引进,国内培育和繁殖较少。唐菖蒲的遗传基础研究几乎在零水平上,利用化学诱变育种,通过筛选,构建唐菖蒲突变体库,为选育优良形态学和生理生化特点变异的突变体,培育具有自主知识产权的新品系或品种,扩大可利用种质资源范围奠定基础。本研究以唐菖蒲主栽品种“超级玫瑰”(Rose Supreme)为试材,在EMS处理以花瓣为外植体建立的再生体系的基础上,进行组培苗的继代培养与驯化移栽,并在第三生长周期进行突变植株的筛选,在营养生长阶段与生殖生长阶段进行形态学鉴定,并对筛选出的形态变异株进行生理指标的测定,为今后进一步开展唐菖蒲离体诱变育种及突变体的筛选与鉴定提供了重要的依据,通过化学诱变剂处理愈伤组织扩大变异谱,也为发现观赏性状优良、抗能能力强的新品种提供新的新途径,研究的主要内容和结果如下： (1)用唐菖蒲“超级玫瑰”花瓣为外植体建立的再生体系中,进一步优化唐菖蒲再生体系,得出如下结果,愈伤组织诱导的最佳培养基为2,4-D3.0mg/L+6-BA0.5mg/L,诱导体细胞胚最佳培养基配方为MS+2,4-D1.0mg/L+TDZ0.5mg/L,优化后的生根培养基为MS+IBA0.5mg/L,最佳驯化基质为草炭土：蛭石：珍珠岩的比例为2：1：1。 (2)用不同浓度的EMS对唐菖蒲花瓣胚性愈伤组织进行了3w的混培法诱变处理,在数据中得出,在所有的处理中EMS浓度处于7mg/L且处理时间为1w的存活率最接近于半致死剂量,为50.22%。确定EMS的半致死浓度为7mg/L,植株驯化成活后统计变异株时发现,EMS浓度为9mg/L处理时,出现的变异率最高,为11.4%. (3)在对诱变唐菖蒲的第三生长周期进行形态学上的鉴定,在营养生长阶段,在同一条件下播种,7株出苗时间晚,最晚可达到晚于对照株2个半月；4株出现了株型的变化；5株植株叶子变宽加长,茎秆变粗；4株无子球植株；7株出现子球生长量与直径生长量大。在生殖生长阶段,22号植株颜色与对照相比颜色明显加深,变为红色,花瓣边缘处无褶皱；31号植株花瓣颜色变为淡粉色,形态变为长椭型；42号植株花瓣颜色与对照相比颜色明显变浅,变为淡粉色,花径大小对照相比明显变小。且诱变后的花粉均存在不同程度的花粉壁破裂,花粉空瘪,花粉生活力低等现象。其中共筛选出25株形态变异株。 (4)在对筛选出的25株形态变异株进行抗氧化酶、总可溶性蛋白、可溶性糖及丙二醛含量活性的测定,发现经EMS处理后各项生理指标均有显著性变化,其中有11株与对照有极显著变化,分别为15号、22号、31号、37号、42号、50号、53号、55号、59号、65号、66号。
[Abstract]:Gladiolus (Gladiolus hybridus Hort) is a perennial bulbous plant of the genus Gladiolus (Iridaceae). It is one of the four famous cut flowers in the world. The production of cut gladiolus flowers is an important part of the flower industry in China. Gladiolus species were mainly introduced from abroad in China, but few were bred and propagated in China. The genetic basis of Gladiolus was studied almost at zero level, and the mutant library of Gladiolus was constructed by chemical mutagenesis and screening. In order to select the mutants with excellent morphologic, physiological and biochemical characteristics, to cultivate new varieties or varieties with independent intellectual property rights, and to expand the range of available germplasm resources. In this study, the main cultivar of Gladiolus, Rose Supreme, was used as the test material. Based on the regeneration system established by EMS treatment with petals as explants, the plantlets of tissue culture were subcultured and domesticated and transplanted. The mutant plants were screened in the third growth cycle, morphological identification was carried out in the vegetative growth stage and reproductive growth stage, and physiological indexes of the selected morphological variant plants were measured. It provides an important basis for further screening and identification of gladiolus in vitro mutation breeding and mutants. By means of chemical mutagens treatment of callus to expand the spectrum of variation, also for the discovery of good ornamental traits. New varieties with strong resistance to energy provide new approaches. The main contents and results of the study are as follows:. 1) in the regeneration system of Gladiolus "super rose" petals established by explants, the regeneration system of Gladiolus was further optimized, and the following results were obtained. The optimum medium for callus induction was 2C4-D3.0 mg / L 6-BA0.5 mg / L, the best medium for inducing somatic embryogenesis was MS 24-D 1.0 mg / L TDZ0.5 mg / L, the optimized rooting medium was MS IBA0.5 mg / L, and the optimum domesticated medium was peat soil: vermiculite: perlite: 21: 1. (2) the embryogenic callus of Gladiolus flower petals was treated with different concentrations of EMS for 3 weeks. The results showed that the survival rate of EMS concentration was at 7mg/L and the survival rate of 1 week treatment was the closest to the half lethal dose in all treatments. The median lethal concentration of EMS was determined to be 7 mg / L, and the highest variation rate was found when the concentration of 9mg/L was treated with 9mg/L after plant domestication. 3) morphological identification of the third growth cycle of Gladiolus mutagenesis was carried out. In the vegetative growth stage, 7 seedlings were seeded under the same conditions. The changes of plant type were observed in 4 plants at the latest, which were later than that of the control plants, and the leaves of 5 plants became longer and wider than those of the control plants. In the reproductive growth stage, the color of No. 22 plant became more red than that of the control. There was no fold at the edge of the petals; the petal color of plant 31 became light pink and the shape became long ellipse; the color of the petals of plant 42 became lighter and lighter pink than that of the control. The flower diameter of the control was obviously smaller than that of the control, and there were some phenomena of pollen wall rupture, pollen hollowness and low pollen viability after mutagenesis. A total of 25 morphological variant plants were screened out. The contents of antioxidant enzyme, total soluble protein, soluble sugar and malondialdehyde were measured in 25 selected morphological variant strains. The results showed that the physiological indexes of 25 strains treated with EMS were significantly changed, and the results showed that there were significant changes in the contents of antioxidant enzymes, total soluble protein, soluble sugar and malondialdehyde (MDA). Among them, 11 strains were significantly different from the control, which were 15, 22, 31, 37, 42, 50, 53, 55, 59, 65 and 66, respectively.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：S682.24

【参考文献】