本文關(guān)鍵詞： 多花黑麥草 新品系 品種 SSR標(biāo)記 指紋圖譜　出處：《四川農(nóng)業(yè)大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：本研究選用多花黑麥草國審品種“長江2號(hào)”、“特高”、“阿伯德”、“杰威”及雜交親本為對照,于2010-2012年對13個(gè)多花黑麥草品種(系)同時(shí)在雅安和洪雅開展品種比較試驗(yàn),通過對其農(nóng)藝性狀進(jìn)行綜合分析評價(jià),以期篩選出優(yōu)質(zhì)高產(chǎn)、適合西南地區(qū)栽培種植的多花黑麥草新品種,為進(jìn)一步區(qū)域試驗(yàn)和生產(chǎn)推廣提供依據(jù)。另外,以兩個(gè)多花黑麥草國家審定品種為材料,通過對不同混合單株梯度下樣本間遺傳多樣性的差異分析,確立了利用SSR標(biāo)記鑒定多花黑麥草品種群體時(shí)的科學(xué)取樣策略,并基于此策略初步構(gòu)建21個(gè)多花黑麥草品種(系)的特異性指紋圖譜,為我國多花黑麥草品種大型DNA指紋數(shù)據(jù)庫的構(gòu)建奠定了基礎(chǔ),為多花黑麥草種質(zhì)資源管理、品種申報(bào)及品種知識(shí)產(chǎn)權(quán)保護(hù)等方面提供一定科學(xué)依據(jù)。主要研究結(jié)果如下： (1)13個(gè)多花黑麥草品種(系)比較試驗(yàn)結(jié)果表明：佛羅里達(dá)4N、Z3和Z6三個(gè)新品系產(chǎn)量表現(xiàn)最優(yōu)異,具有很好的豐產(chǎn)能力和穩(wěn)產(chǎn)性能,其中,Z3冬春季節(jié)產(chǎn)量優(yōu)勢尤為明顯。若僅統(tǒng)計(jì)前三茬草產(chǎn)量,Z3新品系2010-2012兩年度平均干草產(chǎn)量可達(dá)12994kg/hm2,比對照增產(chǎn)11.50～15.66%；兩年度平均鮮草產(chǎn)量為98973kg/hm2,比對照增產(chǎn)9.47-12.82%,適合于稻-草、玉米-黑麥草等糧草輪作。另外Z3多花黑麥草葉量豐富,適應(yīng)性強(qiáng),在整個(gè)生育期內(nèi),植株高度顯著高于對照,冬春季節(jié)生長速度快,尤其是冬季(12月下旬～次年2月中旬)優(yōu)勢明顯,可達(dá)0.403cm/d。 (2)從多花黑麥草國審品種“長江2號(hào)”和“特高”群體中,分別提取50個(gè)單株DNA,同時(shí)提取5、10、15、20、25和30個(gè)單株葉片混合樣本的DNA,利用SSR(Simple sequence repeats)標(biāo)記進(jìn)行遺傳多態(tài)性分析。研究結(jié)果發(fā)現(xiàn)：取樣梯度間多態(tài)性信息含量指數(shù)(PIC)差異不顯著(P0.05),但當(dāng)混合單株樣本為20株時(shí),PIC達(dá)到最高值；SSR平均等位基因數(shù)和單個(gè)SSR座位上等位基因種類隨樣品內(nèi)單株混合數(shù)目增加而增加,當(dāng)取樣量在20株及以上時(shí),等位基因數(shù)和等位基因種類趨于穩(wěn)定,電泳圖譜表現(xiàn)基本一致；不考慮稀有等位基因(基因頻率≤10%)的漏檢,采用20株混合樣本能夠最大限度保持檢出較高頻率(10%)等位基因,且重演性較好。鑒于此,利用SSR標(biāo)記分析多花黑麥草遺傳多樣性時(shí)采用20個(gè)單株混合樣提取的DNA樣本能夠有效地反映群體間差異。 (3)以20個(gè)單株葉片混合提取DNA樣本,利用SSR標(biāo)記,基于多態(tài)性高、穩(wěn)定性強(qiáng)和連鎖群分布均勻的原則,從100對多花黑麥草基因組來源的SSR引物中,篩選出12對引物用來構(gòu)建21個(gè)多花黑麥草品種(系)的指紋圖譜。12對SSR引物共擴(kuò)增出108種多態(tài)性基因型,多態(tài)性比率達(dá)94.15%,每個(gè)多態(tài)性引物分別能擴(kuò)增出5-22條多態(tài)性條帶,平均每對引物擴(kuò)增出9種基因型,多態(tài)性信息量變幅為(PIC)0.744～0.934,平均為0.843,此12對引物可以在構(gòu)建多花黑麥草品種DNA指紋數(shù)據(jù)庫作為核心引物推薦使用。21個(gè)多花黑麥草品種(系)基因型間遺傳相似性系數(shù)在0.4956～0.8571之間,UPGMA聚類分析表明,在相似系數(shù)0.68處可將全部材料分為3大類。7對引物在9個(gè)品種上具有唯一特征帶,采用15-08C單對引物即可將21個(gè)多花黑麥草品種完全區(qū)分開,該對引物不僅多態(tài)性高(PIC=0.934),且具備多個(gè)品種的特征譜帶,基于該引物建立了供試品種指紋圖譜標(biāo)準(zhǔn)模式圖,每個(gè)品種具有唯一的指紋圖譜(帶型)。
[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).

【學(xué)位授予單位】：四川農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：S543.6

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