【摘要】：中間錦雞兒(Caragana intermedia)為豆科、錦雞兒屬多年旱生落葉灌木,生長在海拔900-2400米的固定、半固定沙丘,在我國廣泛分布于華北、西北等地,在防風(fēng)固沙、水土保持方面具有重要作用,而且在飼料、藥用和綠化環(huán)境等方面也有重要價值。了解中間錦雞兒的遺傳背景,可以為育種材料的選擇、研究系統(tǒng)演化及資源合理利用與正確的制定保護(hù)策略提供理論依據(jù)。分子標(biāo)記能直接從DNA水平反映群體及個體之間的遺傳差異,是研究遺傳結(jié)構(gòu)的有力工具。本文基于中間錦雞兒干旱轉(zhuǎn)錄組數(shù)據(jù)自主開發(fā)SSR標(biāo)記,對內(nèi)蒙古中部的10個地區(qū)進(jìn)行樣品采集并分析其遺傳多樣性,結(jié)果如下:1.基于中間錦雞兒干旱轉(zhuǎn)錄組開發(fā)SSR標(biāo)記:共搜索到404個SSR標(biāo)記,分布于349條Unigenes序列上,出現(xiàn)頻率為14.78%。其中二核甘酸是主要的重復(fù)類型,占總SSRs的64.90%;其次是三核苷酸重復(fù),占總SSRs的33.70%;二核苷酸基元以AG/CT和GA/TC為主,占總SSRs的29.45%,三核苷酸基元以AAG/CTT居多,占總SSRs的5.69%。設(shè)計并合成了 76對引物,有54對引物能擴(kuò)增出清晰的條帶,并從中篩選出多態(tài)性高、重復(fù)性好且對檸條錦雞兒也適用的7對引物。這些引物擴(kuò)增出等位基因數(shù)為3-8個,平均5個;各位點(diǎn)的觀察雜合度(Ho)的變化范圍是0.108-0.398,平均0.251;期望雜合度(He)范圍是0.105-0.495,平均0.268。2.居群的遺傳多樣性分析:用篩選出的7對多態(tài)性引物對10個居群進(jìn)行遺傳多樣性分析,結(jié)果顯示:遺傳多樣性指標(biāo)分別為NA=2.547,NE=1.424,HE=0.261,I = 0.445;綜合NE、HE、I三類遺傳多樣性指標(biāo)對10個居群進(jìn)行遺傳多樣性評價,得出的結(jié)論是居群總體遺傳多樣性低,最高的居群為BL(包頭市百靈廟鎮(zhèn)),最低是居群SZ(烏蘭察布市四子王旗)。由NE、HE和I檢測居群遺傳多樣性由高至低依次為BLLCHYHLDGWNGYMAWCSZ。3.居群的遺傳分化分析:居群間的遺傳距離介于0.0027-0.0722,遺傳一致度的范圍是0.9304-0.9973,根據(jù)遺傳一致度對居群進(jìn)行UPGMA聚類,發(fā)現(xiàn)大多數(shù)居群以中間錦雞兒和檸條錦雞兒為界聚為兩類,但有一些居群有交叉現(xiàn)象。居群平均近交系數(shù)(Fis)為0.143,說明據(jù)居群中有明顯的近交現(xiàn)象;基因分化度(Fst)介于0.036-0.164,平均為0.065,表明居群有中等程度的遺傳分化;基因流(Nm)很高,平均為3.589,導(dǎo)致居群間的遺傳變異較小;對居群的遺傳距離和地理距離進(jìn)行分析,發(fā)現(xiàn)二者之間并沒有顯著的關(guān)系(R2=0.023,R2=0.0201);分子方差分析(AMOVA)表明,變異主要發(fā)生在居群內(nèi)個體間。綜上分析表明:供試材料的遺傳多樣性較低,可能是居群采樣距離較近或受到人為干預(yù)較多,遺傳交流大且有明顯的近交現(xiàn)象所導(dǎo)致的。
[Abstract]:Caragana Intermediate (Caragana intermedia) is a perennial xerophytic and deciduous shrub of the family Caragana, which grows in fixed and semi-fixed dunes of 900-2400m above sea level. It is widely distributed in North China and Northwest China, and plays an important role in windbreak and sand fixation, soil and water conservation. And in feed, medicinal and green environment and other aspects also have important value. Understanding the genetic background of Caragana intermedia can provide theoretical basis for the selection of breeding materials, the study of system evolution, the rational utilization of resources and the correct formulation of conservation strategies. Molecular markers can directly reflect the genetic differences between populations and individuals from the DNA level and are powerful tools for studying genetic structures. Based on the drought transcriptome data of Caragana intermedia, SSR markers were developed to collect and analyze the genetic diversity of 10 regions in the middle of Inner Mongolia. The results are as follows: 1. Development of SSR markers based on drought transcriptome of Caragana intermedia: 404 SSR markers were identified and distributed on 349 Unigenes sequences with a frequency of 14.78g. Among them, dinucleotide is the main repeat type, accounting for 64.90% of total SSRs, followed by trinucleotide repeat, accounting for 33.70% of total SSRs, dinucleotide motifs are mainly AG/CT and GA/TC, accounting for 29.4545% of total SSRs, and trinucleotide motifs are mostly AAG/CTT, accounting for 5.69% of total SSRs. 76 pairs of primers were designed and synthesized, 54 pairs of primers could amplify clear bands, and 7 pairs of primers with high polymorphism, good reproducibility and suitable for Caragana korshinskii were screened. The number of alleles amplified by these primers ranged from 3 to 8, with an average of 5. The range of observed heterozygosity (Ho) was 0.108-0.398, with an average of 0.251, and the range of expected heterozygosity (He) was 0.105-0.495, with an average of 0.268.2. Population genetic diversity analysis: 10 populations were analyzed with 7 pairs of polymorphic primers. The results showed that the genetic diversity index was NA=2.547,NE=1.424,HE=0.261,I = 0.445, and the genetic diversity of 10 populations was evaluated by NE,HE,I, and the conclusion was that the total genetic diversity of the population was low. The highest population is BL (Baotou Baling Miao Town), and the lowest is SZ (Siziwang Banner of Wulanchabu). The genetic diversity of population detected by NE,HE and I from high to low is BLLCHYHLDGWNGYMAWCSZ.3.. Genetic differentiation analysis of populations: the genetic distance between populations was 0.0027-0.0722, and the range of genetic consistency was 0.9304-0.9973. According to the UPGMA clustering of populations, it was found that most populations were divided into two groups: Caragana intermedia and Caragana korshinskii. But some populations have intersections. The average number of inbred lines (Fis) was 0.143, which indicated that there was obvious inbreeding in the population, the (Fst) of gene differentiation was 0.036-0.164, the average was 0.065, which indicated that the population had moderate genetic differentiation, and the (Nm) of gene flow was very high. The average genetic variation among populations was 3.589, and the genetic distance and geographical distance between populations were analyzed. The results showed that there was no significant relationship between the genetic distance and geographical distance (R2O0.023R20.0201), and the (AMOVA) analysis of molecular variance showed that the variation mainly occurred among the individuals in the population. The results showed that the genetic diversity of the tested materials was low, which might be caused by the close sampling distance of the population or the human intervention, the large genetic exchange and the obvious inbreeding phenomenon.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：Q943.2;S793.3

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