【作者】 冯锐；

【导师】 张慧英；

【作者基本信息】 广西大学 ， 作物遗传育种， 2007， 硕士

【摘要】 本文利用花粉管通道法将外源DNA（大豆）导入玉米自交系，后代产生了多种多样的变异类型，变异性状十分丰富，这些变异性状有的具有供体性状，有的产生新的性状，通过对其后代农艺性状观察、叶绿素含量测定、过氧化物同工酶酶谱、营养指标测定等分析。具体结果如下：1)提取供体（高蛋白大豆富锦J05-2-075）的总DNA，配成500μg／ml浓度，分别在受体（普通玉米自交系7313）自交授粉后22h、24h、26h导入，从变异的效果来看：26h＞24h＞22h（D₁代、D₂代表现相同），其中D₁代26h处理的变异率最高，1.34％；D₂代26h处理的变异率为1.09％。2)调查田间农艺性状（如株高、穗位高、叶片数、叶片的长度与宽度、光合叶面积、雄穗分枝数、百粒重、生育期、结苞数、果穗长度和籽粒形状等），对不同于CK的植株的农艺性状进行观察、记录、统计与分析，结果表明这些变异有的表现为供体性状（如生育期延长、百粒重减轻、黄色籽粒形状椭圆似大豆等。例见图3-5中的26-D₁-6和图3-9中的26-D₂-5），有的表现为新的性状（如植株高度和叶片长宽度有变化、植株有分蘖、雄穗分枝数和雌穗数增加、籽粒形状不规则，粒小，有稃，百粒重明显减少等，例见图3-5中的26-D₁-11、26-D₁-13和图3-9中的26-D₂-3、26-D₂-19、26-D₂-21等）。3)对获得的变异株进行过氧化物酶同工酶酶谱分析，结果表明：变异株中同工酶酶谱变异大，变异后代与受体间有明显的差异(如变异株26h—D₂-19在迁移率Rf₁₃=0.87、Rf₁₄=0.89，出现了两条供体特有而受体没有的酶谱带，酶带颜色深，活性强；26h—D₂-3在迁移率Rf₁₀=0.57、Rf₁₂=0.69、Rf₁₄=0.82出现了供体和受体都没有的酶谱带。)。4)对D₂代变异株系籽粒的油分、蛋白质含量、醇溶蛋白含量和谷蛋白含量进行测定分析，按照选育高蛋白玉米的方向要求：油分、蛋白质含量和谷蛋白含量有所提高，而醇溶蛋白含量降低，从中选出符合育种目标的9个株系，分别为26h-D₂—1、26h-D₂—3、26h-D₂—6、26h-D₂—7、26h-D₂—13、26h-D₂—14、26h-D₂—15、26h-D₂—18和26h-D₂—19，这些株系与CK相比，叶绿素含量增加，增加幅度在7.12％～28.2％；油分含量增加7.65％～44.6％；粗蛋白含量增大12.46％～80.79％；醇溶蛋白从CK的55.7437％最低降至36.2187％（26h-D₂—3）；谷蛋白从CK的31.8295％最高升至51.1583％（26h-D₂—14），所选的9个株系符合育种目标的要求。综上所述：通过花粉管通道法将外源DNA导入玉米自交系，已经引起了变异，可从中获得新的优良变异品系，为创造玉米新种质开辟一条新途径。更多还原

【Abstract】 With the methods of pollen-tube-pathway, the exogenous DNA extracted from different plants （soybean） was introduced into different maize inbred lines. The progenies appear various mutant types and the mutant characters are very abundant, some of the varied characters were similar to donors, while others, resemble neither that of donors nor that of acceptor, were newly produced .After surveying the quality of field growth ,chlorophyll contains ,the peroxidase isozyme （POD）and the index of nutrition .The results are as follows:1) The DNA extracted from soybean of high protein content Fujin J05-2-075 , with the density of 500/tg/ml, was introduced common 7313 maize inbred lines, 22h,24h and 26h after pollination, the effect of variation is 26h>24h>22h（the generation of D₁ and D₂ show alike） .In the D₁ generation ,the mutant rate of 26h treatment is highest, 1.34%; the mutant rate of 26h treatment in D₂ generation is 1.09%.2) To investigate the quality of field growth such as the height of plant, the height of fruit, the number of leaf, the length and width of leaf, the number of ear-ramify, number of tassel branch , the weight of 100 grain ,the period of growth ,number of fruit ,panicle length and type of corn grain etc.), observed , recorded ,counted and analyzed the quality of field growth of mutant plants difference from CK, the results showed some of the varied characters were similar to donor such as extending the growth period , decreasing 100-grain weigh and ovum type of yellow corn grain similar with soybean etc. example: 26-D-1-6 in fig . 3-5 and 26-D2-5 in picture 3-9 , some of characters were of neither the donor nor the acceptor such as the height of plant and the length and width of leaf have change , plant own root- ramify ,the number of ear-ramify and fruit increased, irregular type of corn grain ,small ,decreasing 100-grain weigh etc. ,such 26-D₁-11,26-D₁-13 in picture 3-5 and 26-D₂-3, 26-D₂-19,26-D₂-21 in picture 3-9) .3)Analyzed the zymograms of peroxidase isozyme of the mutant plant, the results also indicated that the zymograms of peroxidase isozyme of transferred progenies were different from the recipients (the mutant plant 26h—D₂-19 exhibited two specific band, there was an increase of peroxidasic color and activity in Rf₁₃=0.87 and Rf₁₄=0.89, that only belonged to the donor plant; the mutant plant 26h—D₂-3 exhibited three specific band ,in Rf₁₀=0.57、Rf₁₂=0.69、Rf₁₄=0.82,that neither belonged donor ,nor receptor.).4) Analyzing the mutant plants of D₂ generation oil, protein, gliadin protein and alkali protein content , according to the request of high protein content maize for select and bear: such as increasing oil, protein and alkali protein content, but decreasing gliadin protein content ,select nine plant-line at present from the mutant plants of D₂ generation: 26h-D₂-1、26h-D₂-3、26h-D₂-6、26h-D₂-7、26h-D₂-13、26h-D₂-14、26h-D₂-15、26h-D₂-18和26h-D₂-19.Comparison of these mutant plant and CK, the total chlorophyll contains increased extent at 7.12%～28.2%; oil contains raised at 7.65%～44.6%; protein content added at 12.46%～80.79%; the gliadin protein content decreased from 55.7437 % of CK to 36.2187% （26h-D₂-3）; the alkali protein content most added 31.8295% of CK to 51.1583% （26h-D₂-14）, all of the nine plant-line fit for the request of the breeding.In conclusion , the exogenous DNA was introduced into the maize inbred lines through pollen-tube-pathway methods, it caused mutation and created a new way to develop new maize, which offer middle material for breeding.更多还原