多花黑麦草品种（系）农艺性状综合评价及SSR指纹图谱构建

发布时间：2018-01-29 22:34

本文关键词： 多花黑麦草新品系品种 SSR标记指纹图谱　出处：《四川农业大学》2013年硕士论文　论文类型：学位论文

【摘要】：本研究选用多花黑麦草国审品种“长江2号”、“特高”、“阿伯德”、“杰威”及杂交亲本为对照,于2010-2012年对13个多花黑麦草品种(系)同时在雅安和洪雅开展品种比较试验,通过对其农艺性状进行综合分析评价,以期筛选出优质高产、适合西南地区栽培种植的多花黑麦草新品种,为进一步区域试验和生产推广提供依据。另外,以两个多花黑麦草国家审定品种为材料,通过对不同混合单株梯度下样本间遗传多样性的差异分析,确立了利用SSR标记鉴定多花黑麦草品种群体时的科学取样策略,并基于此策略初步构建21个多花黑麦草品种(系)的特异性指纹图谱,为我国多花黑麦草品种大型DNA指纹数据库的构建奠定了基础,为多花黑麦草种质资源管理、品种申报及品种知识产权保护等方面提供一定科学依据。主要研究结果如下： (1)13个多花黑麦草品种(系)比较试验结果表明：佛罗里达4N、Z3和Z6三个新品系产量表现最优异,具有很好的丰产能力和稳产性能,其中,Z3冬春季节产量优势尤为明显。若仅统计前三茬草产量,Z3新品系2010-2012两年度平均干草产量可达12994kg/hm2,比对照增产11.50～15.66%；两年度平均鲜草产量为98973kg/hm2,比对照增产9.47-12.82%,适合于稻-草、玉米-黑麦草等粮草轮作。另外Z3多花黑麦草叶量丰富,适应性强,在整个生育期内,植株高度显著高于对照,冬春季节生长速度快,尤其是冬季(12月下旬～次年2月中旬)优势明显,可达0.403cm/d。 (2)从多花黑麦草国审品种“长江2号”和“特高”群体中,分别提取50个单株DNA,同时提取5、10、15、20、25和30个单株叶片混合样本的DNA,利用SSR(Simple sequence repeats)标记进行遗传多态性分析。研究结果发现：取样梯度间多态性信息含量指数(PIC)差异不显著(P0.05),但当混合单株样本为20株时,PIC达到最高值；SSR平均等位基因数和单个SSR座位上等位基因种类随样品内单株混合数目增加而增加,当取样量在20株及以上时,等位基因数和等位基因种类趋于稳定,电泳图谱表现基本一致；不考虑稀有等位基因(基因频率≤10%)的漏检,采用20株混合样本能够最大限度保持检出较高频率(10%)等位基因,且重演性较好。鉴于此,利用SSR标记分析多花黑麦草遗传多样性时采用20个单株混合样提取的DNA样本能够有效地反映群体间差异。 (3)以20个单株叶片混合提取DNA样本,利用SSR标记,基于多态性高、稳定性强和连锁群分布均匀的原则,从100对多花黑麦草基因组来源的SSR引物中,筛选出12对引物用来构建21个多花黑麦草品种(系)的指纹图谱。12对SSR引物共扩增出108种多态性基因型,多态性比率达94.15%,每个多态性引物分别能扩增出5-22条多态性条带,平均每对引物扩增出9种基因型,多态性信息量变幅为(PIC)0.744～0.934,平均为0.843,此12对引物可以在构建多花黑麦草品种DNA指纹数据库作为核心引物推荐使用。21个多花黑麦草品种(系)基因型间遗传相似性系数在0.4956～0.8571之间,UPGMA聚类分析表明,在相似系数0.68处可将全部材料分为3大类。7对引物在9个品种上具有唯一特征带,采用15-08C单对引物即可将21个多花黑麦草品种完全区分开,该对引物不仅多态性高(PIC=0.934),且具备多个品种的特征谱带,基于该引物建立了供试品种指纹图谱标准模式图,每个品种具有唯一的指纹图谱(带型)。
[Abstract]:This research selects the ryegrass national trial varieties of "Yangtze River 2", "high", "Abreu de", "Morrison" and parents were from 2010-2012 years to 13 varieties of Annual Ryegrass Varieties (lines) at the same time to carry out comparative tests in Ya'an and Hongya, through the comprehensive analysis and evaluation on the agronomic traits of and to screen out the high yield and high quality, new varieties of annual ryegrass cultivation for the southwest region, to provide the basis for further regional tests and production promotion. In addition, the two annual ryegrass national varieties as materials, through the analysis of different mixing plant gradient samples of genetic diversity among different, established by scientific sampling strategy identification of SSR markers more groups ryegrass cultivars, and based on this strategy preliminary construction of 21 varieties of annual ryegrass (lines) of the specific fingerprint for our large varieties of annual ryegrass The establishment of DNA fingerprint database provides a scientific basis for the germplasm resource management, variety declaration and variety intellectual property protection of Lolium multiflorum.
(1) 13 Italian ryegrass cultivars (lines) comparative test results show that the 4N of Florida, the most excellent Z3 and Z6 three new lines of production performance, with good yield ability and stable performance, the Z3 winter season yield is particularly obvious. If only the first three crop yield statistics, the two annual average of 2010-2012 the hay yield was 12994kg/hm2 Z3 strains, the yield increased by 11.50 ~ 15.66%; two annual average fresh yield was 98973kg/hm2, the yield increased by 9.47-12.82%, suitable for rice grass maize, ryegrass and ryegrass forage rotation. In addition, Z3 leaves the amount of rich, strong adaptability, in the whole growth period, plant height was significantly higher than that of control. The winter season fast growth, especially in winter (late December to mid February) have obvious advantages, can reach 0.403cm/d.
(2) from the trial of Italian ryegrass cultivar "Yangtze River 2" and "high" groups, were extracted from 50 individuals DNA, and extract 5,10,15,20,25 and 30 leaf mixed samples of DNA, using SSR (Simple sequence repeats) markers were used to analyze the genetic polymorphism. The results showed that the polymorphism information content the gradient between the sampling index (PIC) showed no significant difference (P0.05), but when the mixed plant samples for 20 strains, PIC reached the highest value of SSR; the average number of alleles and single SSR alleles per plant species increased with the number of samples in the mixture increased, when the sampling volume of 20 lines or more, etc. the number of alleles and allele types tend to be stable, electrophoresis were basically consistent; do not consider the rare allele (gene frequency = 10%) missing, with 20 strains of mixed samples can keep the maximum detectable high frequency (10%) allele, and In view of this, using SSR markers to analyze the genetic diversity of Lolium multiflorum, the DNA samples extracted from 20 single plant samples can effectively reflect the difference among populations.
(3) DNA samples extracted from 20 leaf mixed with SSR marker based on high polymorphism, strong stability and uniform distribution of the linkage principle, from 100 pairs of SSR primers of ryegrass genomic, screened 12 pairs of primers were used to construct 21 Italian ryegrass cultivars (lines) of the fingerprint of.12 SSR primers amplified 108 polymorphic genotypes, the polymorphic rate was 94.15%, each polymorphic primers were amplified 5-22 polymorphic bands per primer amplified 9 genotypes, polymorphism information quantitative amplitude (PIC) from 0.744 to 0.934, with an average of 0.843, 12 of the primers can be constructed in varieties of annual ryegrass DNA fingerprint database as the core primers recommended.21 Italian ryegrass Cultivars (lines) genotype genetic similarity coefficient in 0.4956 ~ 0.8571, UPGMA cluster analysis showed that the similarity coefficient of 0.68 in the whole. The material is divided into 3 categories of.7 primers have unique characteristics with 9 varieties, 15-08C uses a single pair of primers can be 21 varieties of annual ryegrass are completely separated, the pair of primers not only high polymorphism (PIC=0.934), and has the characteristics of a variety of bands, which were established varieties of fingerprint map map based on standard model, each species has a fingerprint only (band).

【学位授予单位】：四川农业大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：S543.6

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