發(fā)布時間：2018-04-23 16:42

  本文選題：甘藍型油菜 + 特高芥酸含量��； 參考：《四川農(nóng)業(yè)大學(xué)》2016年博士論文

【摘要】：芥酸是蕓苔屬(Brassica)植物種子油脂中特有的一種長碳鏈脂肪酸,作為食用油,其不易被人體消化利用,但作為工業(yè)原料,在油田化學(xué)、石油化工、日用化學(xué)、醫(yī)藥化工等行業(yè)有著十分廣泛的應(yīng)用。高芥酸油菜是當(dāng)今世界上工業(yè)芥酸的最重要來源。本研究以3個特高芥酸品系(種)、2個高芥品系(種)、2個中芥品系(種)、2個低芥品系(種),圍繞特高芥酸含量性狀展開相關(guān)的遺傳研究。(1)以工業(yè)專用特高芥酸、特低油酸甘藍型油菜新材料703AB-4和低芥酸、高油酸油菜品種中雙11為親本,配制6個遺傳世代,采用主基因加多基因混合遺傳模型,分析特高芥酸含量和油酸含量的遺傳模型,采用F2群體分析芥酸與其它主要脂肪酸含量的相關(guān)性；(2)以9個芥酸含量和遺傳背景不同的甘藍型油菜品系(種)為材料,采用完全雙列雜交遺傳設(shè)計,配制獲得72個正反交雜交組合和9個自交系親本后代,在四川成都、綿陽和宜賓三個不同的生態(tài)地區(qū)進行隨機區(qū)組試驗鑒定,分析芥酸含量的雜種優(yōu)勢表現(xiàn)、配合力及環(huán)境效應(yīng)；(3)通過特高芥酸含量材料703AB-4與低芥酸材料中雙11雜交獲得F2分離群體,并進行SSR分子標(biāo)記,篩選能區(qū)分芥酸含量的分子標(biāo)記,以應(yīng)用于高芥酸輔助選擇育種；(4)利用特高芥酸含量材料703AB-4、中芥酸含量材料L155及低芥酸含量材料中雙11,進行芥酸合成途徑中兩個關(guān)鍵調(diào)控基因FAE1和FAD2的克隆分析,以分析703AB-4特高芥酸含量的分子遺傳機理。主要研究結(jié)果如下：1.特高芥酸的最適遺傳模型為E-0,即受2對加性-顯性-上位性主基因+加性-顯性-上位性多基因控制,以主基因效應(yīng)為主。B1、B2和F2群體的主基因遺傳率分別高達94.3%、98.5%和98.2%,多基因效應(yīng)較弱,2對主基因的加性效應(yīng)較大,達13.21,均為正效應(yīng)且相等,累計加性效應(yīng)值高達26.42。油酸的最適遺傳模型為E-1,即受2對加性-顯性-上位性主基因+加性-顯性多基因控制,B2和F2群體的主基因遺傳率較高,分別為98.00%和95.53%,而B1群體的主基因遺傳率則較低,為73.71%,說明油酸含量的遺傳主要受兩對主基因控制,存在多基因效應(yīng)。2.芥酸與其他主要脂肪酸相關(guān)性分析表明,油菜芥酸含量與油酸含量、亞油酸含量之間存在極顯著的負相關(guān),但油酸含量變化較大,平均達55個百分點,亞油酸含量變化總量不到10個百分點；芥酸含量與亞麻酸含量關(guān)系不顯著；芥酸含量與花生烯酸含量關(guān)系較復(fù)雜,芥酸含量在15以下為正相關(guān),之后為負相關(guān),平均變化約15個百分點。芥酸升高到50%后,繼續(xù)升高時,主要是以降低花生烯酸含量的庫來實現(xiàn)的。3.多數(shù)雜交組合芥酸含量都顯示出明顯的中親優(yōu)勢,只有少數(shù)組合表現(xiàn)出超親優(yōu)勢,且兩親本間芥酸含量差異越大,其中親優(yōu)勢越強,超親優(yōu)勢只表現(xiàn)在特高芥親本之間,說明高芥酸雜種優(yōu)勢主要以中親優(yōu)勢為主。環(huán)境對親本及雜交組合的芥酸含量均有影響,但親本受環(huán)境的影響小于雜交組合。芥酸含量雜種優(yōu)勢在不同環(huán)境下表現(xiàn)的趨勢是一致的,均以中親優(yōu)勢為主。不同試驗點芥酸含量雜種優(yōu)勢的表現(xiàn)存在差異,但出現(xiàn)雜種優(yōu)勢的組合具有一致性,宜賓點更易獲得具有芥酸含量雜種優(yōu)勢的組合。不同親本在提高芥酸含量的作用是不一樣的,在選育高芥酸材料的過程中應(yīng)充分考慮親本的差異,同時考慮環(huán)境因素的影響。4.雜交組合芥酸含量主要由一般配合力決定,在高芥酸油菜組合選配中,要獲得高芥酸含量的雜交組合,首先要選芥酸含量高、一般配合力高的親本,同時考慮反交效應(yīng)。本研究通過配合力穩(wěn)定性分析表明,油菜芥酸含量主要受基因遺傳控制,但環(huán)境因素對油菜芥酸含量有一定的影響。雜交組合芥酸受母性影響,母性效應(yīng)為正極顯著的親本,其正反交芥酸含量存在顯著差異。僅在特定組合間表現(xiàn)的非母性效應(yīng),對組合芥酸含量的影響非常大。5.獲得2個與芥酸含量緊密連鎖的SSR分子標(biāo)記,CB10364和BRMS-017,單株基因型同時為CB10364-a和BRMS-017-a的芥酸含量57%,其分離比率經(jīng)χ2檢驗符合孟德爾分離規(guī)律,能較可靠地將群體中特高芥酸單株區(qū)分出來。6.低芥材料中雙11、中芥材料L155具有兩個FAE1基因拷貝,分別位于甘藍型油菜A8和C3染色體上,特高芥材料703AB-4只有一個FAE1基因拷貝,但均只有一個拷貝可完整編碼氨基酸序列。低芥FAE1-1.1、中芥FAE1-2.1和特高芥FAE1-3編碼氨基酸序列比對分析結(jié)果顯示,在第282位氨基酸位點,低芥FAE1-1.1為F(苯丙氨酸),中芥與特高芥均為S(絲氨酸)。在第286、323、395和406位氨基酸位點,特高芥酸含量材料703AB-4的FAE1-3分別編碼R(精氨酸)、T(蘇氨酸)、K(賴氨酸)和G(甘氨酸),而低芥與中芥基因則均依次編碼G(甘氨酸)、Ⅰ(異亮氨酸)、R(精氨酸)和A(丙氨酸)。7.低芥中雙11與中芥材料L155均存在4條不同的FAD2基因拷貝,而特高芥酸材料703AB-4存在3條FAD2基因拷貝。同源分析表明,FAD2-1. 1、FAD2-2.1、FAD2-3.1為一類,編碼氨基酸序列完全一致；FAD2-1.2, FAD2-2.2、FAD2-3.2為一類,編碼氨基酸序列完全一致；FAD2-1.3、FAD2-2.3為一類,但提前出現(xiàn)終止密碼；FAD2-1.4、FAD2-2.4、FAD2-3.3為一類,編碼氨基酸序列同源性高達99.91%,僅在20位氨基酸位點,中芥酸材料2號材料出現(xiàn)差異,為T蘇氨酸,而低芥酸材料(1號材料)和高芥酸材料(3號材料)均為N天冬氨酸,也即低芥酸材料和特高芥酸材料氨基酸序列完全一致。
[Abstract]:Erucic acid is a special kind of long carbon chain fatty acid in the seed oil of the Brassica plant. As a edible oil, it is not easily digested by the human body. But as an industrial raw material, it is widely used in the fields of oil field chemistry, petrochemical, daily chemistry, medicine and chemical industry. High erucic acid is the heaviest in the world. 3 highly erucic acid lines (species), 2 high mustard lines (species), 2 middle mustard lines (species) and 2 low mustard lines (species) were studied in this study. (1) the special high erucic acid, 703AB-4 and low erucic acid of specially low oleic acid Brassica napus, and double 11 in high oleic acid rapeseed were used as parents. 6 Genetic generations were prepared, and the genetic model of the main gene mixed gene was used to analyze the genetic model of high erucic acid content and oleic acid content. The correlation between erucic acid and other main fatty acids was analyzed by F2 population. (2) full diallel was used as the material with 9 erucic acid content and different genetic background of Brassica napus. Genetic design was used to prepare 72 positive and negative cross combinations and 9 progeny of self inbred lines. In three different ecological regions, Chengdu, Mianyang and Yibin, Sichuan, three different ecological regions were tested, and the Heterosis of erucic acid content, combining ability and environmental effect were analyzed. (3) 703AB-4 and low erucic acid material were used as the material of high erucic acid content. Medium double 11 hybrid was used to isolate F2 population, and SSR molecular markers were used to screen molecular markers that could distinguish erucic acid content, which was applied to high erucic acid assisted selection breeding. (4) using high erucic acid content material 703AB-4, medium erucic acid content material L155 and low erucic acid content of 11, the two key regulatory genes of erucic acid synthesis pathway, FAE1, were carried out. The molecular genetic mechanism of 703AB-4 highly erucic acid content was analyzed by cloning and analysis of FAD2. The main results were as follows: 1. the optimum genetic model of high erucic acid was E-0, which was controlled by 2 pairs of additive dominant epistatic main gene + additive dominant epistasis, the main gene effect was mainly.B1, and the main gene heritability of B2 and F2 population was respectively. As high as 94.3%, 98.5% and 98.2%, the effect of polygene is weak, and the additive effect of 2 on the main gene is greater, and the effect is 13.21. The optimum genetic model of the cumulative additive effect value up to 26.42. oleic acid is E-1, that is, it is controlled by 2 additive dominant epistatic main gene + additive dominant gene, and the main gene heritability of B2 and F2 population is higher. The main gene heritability of B1 population was 98% and 95.53%, respectively, and the main gene heritability was 73.71%, indicating that the inheritance of oleic acid content was mainly controlled by two main genes, and the correlation analysis between.2. erucic acid and other main fatty acids showed that there was a very significant negative correlation between the content of erucic acid and the content of oleic acid and the content of linoleic acid. The content of oleic acid changed greatly, with an average of 55 percentage points, the total amount of linoleic acid content was less than 10 percentage points, and the relationship between the content of erucic acid and the content of linolenic acid was not significant. The content of erucic acid was more complex than the content of arachidic acid. The content of erucic acid was positively correlated with the content of erucic acid below 15, and the average change was about 15 percentage points. Erucic acid was raised to 50%. After continuing to rise, most of the.3. hybrid erucic acid content, which was achieved by reducing the content of arachidic acid, showed obvious parent heterosis. Only a few combinations showed superior parent heterosis, and the greater the difference in erucic acid content between the two parents, the stronger the heterosis, the superior parent was only among the highly high mustard parents. The Heterosis of erucic acid was mainly dominated by the middle parent heterosis. The environment had an influence on the erucic acid content of the parent and cross combination, but the parent was less affected by the environment than the hybrid combination. The Heterosis of the erucic acid content was consistent in different environment, and the dominant parent was the middle parent. The Heterosis of the erucic acid content in different test points was present. However, the combination of heterosis is consistent, and the combination of Heterosis with erucic acid content is more easily obtained in Yibin point. The effect of different parents on erucic acid content is different. In the process of breeding high erucic acid, the difference of parents should be taken into consideration, and the effect of environmental factors on the erucic acid content of.4. hybrid combinations should be considered. It is mainly determined by the general combining ability. In the selection of high erucic acid rapeseed combination, the high erucic acid content should be obtained by the combination of high erucic acid. First, the parent with high erucic acid content and high common strength should be selected and the anti cross effect is considered. Erucic acid content had a certain influence. The combination of erucic acid was influenced by maternal sex, maternal effect was a positive parent, and there was a significant difference in the content of erucic acid. Only the non maternal effect between the specific combinations, the effect of the combination of the erucic acid on the combination of erucic acid content was very large.5. obtained 2 SSR molecular markers linked with the erucic acid content, CB10364 And BRMS-017, single plant genotype was 57% of erucic acid in CB10364-a and BRMS-017-a, and its separation ratio was conformed to Mendel separation law by chi 2 test. It could be more reliable to distinguish high erucic acid from group.6. low mustard material in 11, and L155 of medium mustard material with two copies of FAE1 gene, which were located in A8 and C3 of Brassica napus, respectively. On the body, there is only one copy of FAE1 gene in 703AB-4, but only one copy can fully encode amino acid sequence. Low mustard FAE1-1.1, middle mustard FAE1-2.1 and highly high mustard FAE1-3 coding amino acid sequence alignment analysis results show that at the 282nd bit amino acid site, low mustard FAE1-1.1 is F (phenylalanine), medium mustard and highly high mustard are S (silk ammonia). At 286323395th and 406th amino acid sites, FAE1-3 of 703AB-4, T (threonine), K (lysine) and G (glycine) were encoded in the highly erucic acid content material, respectively, while the low mustard and middle mustard genes encode G (glycine), I (ISO) (ISO), R (arginine) and A (alanine).7. low mustard and medium mustard material L155 were 4 There are 3 copies of different FAD2 genes, while the highly erucic acid material 703AB-4 has 3 copies of FAD2 gene. Homology analysis shows that FAD2-1. 1, FAD2-2.1, FAD2-3.1 are a class, the sequence of encoded amino acids is identical; FAD2-1.2, FAD2-2.2, FAD2-3.2 are a class, the sequence of encoded amino acids is all consistent; FAD2-1.3, FAD2-2.3 is a class, but appears in advance to terminate. Ciphers, FAD2-1.4, FAD2-2.4, FAD2-3.3 are a class of encoded amino acid sequence homology up to 99.91%, only at 20 amino acid sites, medium erucic acid material 2 material difference, T threonine, and low erucic acid material (1 material) and high erucic acid material (3 material) are N aspartic acid, that is, low erucic acid material and high erucic acid material amino acids. The sequence is exactly the same.

【學(xué)位授予單位】：四川農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S565.4

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