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小麦×玉米远缘杂交的初步研究
刘辉
学位类型硕士
导师陈集贤
1993
学位授予单位中国科学院西北高原生物研究所
学位授予地点中国科学院西北高原生物研究所
学位专业植物学
关键词小麦 玉米 远缘杂交 胚胎发育 单倍体 电泳分析
摘要小麦×玉米的远缘杂交是近年发展起来的一种获得小麦单倍体的方法。本论文从三方面对小麦×玉米的远缘杂交作了初步的研究工作:1.改良小麦×玉米杂交方法获得了高频率的小麦单倍体;2.从细胞胚胎学的角度观察了小麦×玉米远缘杂交的受精作用和胚胎发育;3.通过对小麦×玉米杂交后代的蛋白质及同工酶电泳,分析杂交后代中的变异。利用小麦×玉米的高成胚率和杂合了中玉米染色体自发消除的特点,可以获得小麦单倍体。由于胚乳的缺乏或早期败育,致使幼胚在发育早期即夭折,使得通过小麦×玉米获得单倍体植株很困难。于授粉后4小时用100 ppm的2,4-D 溶液处理麦穗,可以有效地促进幼胚在植株上的发育,延迟杂交胚在植株上的败育时间,从而获得大量可供培养的幼胚。用2,4-D 处理穗子后10天,从382个子房中解剖出64个(16.8%)幼胚,经胚培养得到47棵绿色植株。而未经2,4-D 处理的对照,104个子房中仅解剖出1个胚(0.96)和得到1棵植株。在西宁及张家口两地,以栽培小麦“康选9号”、模式小麦品种“中国春”为母本,超甜玉米“SS7700”为父本进行了杂交,对受精作用及胚胎发育作了细胞胎学的观察,在西宁,以“中国春”为母本的杂交,发现在被检测的76个了房中,仅5个(6.6%)发生了卵细胞的单受精,形成了胚而无胚乳,没有发现极核的单受精及双受精。在张家口,以“康选9号”为母本的杂交, 发现在63个被检测的子房中,有43(26.4%)个发生了卵细胞的单受精,形成了胚而无胚乳,7(4.3%)个发生了极核的单受精,形成了胚乳而无胚,28个(17.2%)发生了双倍受精,具有胚及胚乳;以“中国春”为母本的对照中,观察了151个子房,相应的数字为28(18.5%)、3(2.0%)、12(7.9%)。表明小麦×玉米杂交受地域之限,推测是由于气候对异种花粉管在柱头萌发和伸长的影响;栽培小麦“康选9号”和玉米杂交的受精率(47.9%)和成胚率(43.6%)均比以模式杂交母本材料“中国春”进行的杂交要高,后者的受精率和成胚率分别28.4%、26.4 观察小麦×玉米杂交中极核的受精行为,认为杂合胚乳是以三核合并的形式形成的。小麦和玉米杂交后形成的合了及胚乳核型极不稳定,来自玉米的染色体在合子及胚乳的最初几次分裂中会很快地排除,最后产生单倍体小麦胚。对以8个普通小麦为母本,2个栽培玉米为父本进行杂交所获得的F_2籽粒进行蛋白质电泳分析,发现杂交后代有五种类型的谱带结构:1.母本型,占全部测试籽粒(84粒)的22.6%;2.附加型,占14.3%;3.互补型,占15.5;4.杂种型,占30.9%;5.逸失型,占16.7%。后代的蛋白质变异主要活跃地集中在高分子量谷蛋白(HMW-Glu)区域。由于HMW-Glu各亚基已有较为明确的基因定位,因此可以推测相应基因位点是否发生了变异;同时HMW-Glu区域变异的活跃性也为筛选具优良的面粉及面包烘烤品质的杂交后代提供了可能性。对以栽培小麦“矮杆早”为母本,玉米“紫粘”为父本杂交所获得的F_2代成熟籽粒的酯酶同工酶分析,结果表明后代也有附加型及杂种型两种变异,同工酶变异出现在EST2和EST3区域。三方面的研究结果表明,小麦×玉米杂交的方法,不仅可以成功、高频率地获得小麦单倍体,同时可以达到将玉米基因向小麦染色体组转移、诱发小麦染色体发生变异的目的,在育种上具有一定的应用前景。
其他摘要The distant hybridization between wheat and maize is a method of producing haploid wheat plant which has great advancements in recent years. This thesis has reported the primary results of our research on distant hybridization between wheat and maize in three respects: 1. The production of haploid wheat plants via an improved method of wheat x maize hybridization; 2. The fertilization and embryo development in wheat x maize crosses; 3. Electrophoretic survey of proteins and esterase isoenzymes in wheat * maize hybrids. Wheat haploid plants are supposed to be produced in the bybridization between wheat and maize while the maize chromosomes eliminate rapidly. The absence and abortion of endosperms result in the abortion of hybrid embryos. So it is very difficult to produce haploid wheat plant in vivo. If spikes containing cross-pollinated florets were treated with 100 ppm 2,4-D solution 4 hours after pollination, the development of hybrid embryos will be ameliorated, the duration of embryos in vivo lengthened and it will be possible to obtain viable hybrid embryos which can be used in embryo culture. Of the 382 florets treated, 64(16.8%) embryos were obtained 10 days after treatment, and 47 plants recovered on the culture medium. In control (without 2,4-D treatment) only 1 (0.96%) embryo and 1 plant were obtained from 104 florets. In Xining and Zhangjiakou, hybridization has been done between wheat and maize, the cultiver wheat "kangxuan 9" and "Chines Spring" as the female parent, and maize "SS7700", the male parent. The result of observation on fertilization and embryo development in the crosses in Xining has shown that, only 5(6.6%) embryos were detected from 76 florets and neither polar nuclear fertilization nor double fertilization has occured. In Zhangjiacou, when cultivar "Kangxuan 9" was used as female parent, 43 embryos (26.4%) were obtained in 163 florets. 7(4.3%) had only an endosperm, 28(17.2%) had embryo and endosperm; when "Chinese Spring" was used as female parent, 28 embryos (18.5%) were obtained from 151 florets, 3(2.0%) had only an endosperm, 12(7.9%) had embryo and endosperm. Frequencies of fertilization and hybrid embryo formation in "Kangxuan 9" * maize were much higher than those of "Chinese Spring" x maize crosses. The formation of endosperm was in the way of "Three-Nucli-Fusion". The hybrid embryos and endosperms obtained were karyotypically unstable and characterized by rapid elimanation of the maize chromosomes to produce haploid wheat embryos. The potentials of wheat haploid plants production and introgression of maize genes into wheat were discussed. The analysis to soluble proteins and esterase isoenzymes in F_2 hybrid grains from wheat * maize crosses indicated that the electrophoretic pattern of protein and esterase isoenzymes were extremely different from those of their parents. The mutations of proteins actively abounded in the HMW-Glu zone. There were 5 kinds of protein patterns in the analysed hybrid grains: 1. Female parent type, 22.6% of the 84 hybrid grains were characterized by this type of protein pattern; 2. Additional type, 14.3% of the total; 3. Complementary type, 15.5% of the total; 4. Hybrid type, 30.9% of the total; 5. Omission type, 16.7% of the total. In the analysis of esterase patterns, some new bands in hybrid were also found to be those characterize male parent. The mutations of electrophoretic patterns of protein and esterase isoenzymes indicated that genetic materials of wheat have changed in the distant hybridization. Detailed and direct evidence could be obtained from RFLP analysis. The results of our research show that, the method of wide crosses between wheat and maize could not only supply a new method of producing haploid wheat plants but also make it accessible to transfer maize genes to wheat. Wheat * maize crosses could be applied in Agriculture.
页数35
语种中文
文献类型学位论文
条目标识符http://210.75.249.4/handle/363003/3324
专题中国科学院西北高原生物研究所
推荐引用方式
GB/T 7714
刘辉. 小麦×玉米远缘杂交的初步研究[D]. 中国科学院西北高原生物研究所. 中国科学院西北高原生物研究所,1993.
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