菜心遺傳圖譜構建及主要農藝性狀的QTL定位
發(fā)布時間：2018-08-17 19:48
【摘要】：菜心是華南地區(qū)栽培面積最大的特色蔬菜之一,優(yōu)質、抗逆和豐產性狀是菜心育種的主要目標。隨著分子標記技術的發(fā)展,遺傳連鎖圖譜的構建、基因定位已成為作物遺傳和分子育種研究和應用工作的重要部分,研究開發(fā)出的與目標性狀緊密連鎖的分子標記可加快品種選育進程。據此,本研究以菜心材料“3T6”和“四九-19號菜心”為父母本構建的130份重組自交系(RIL)群體為材料,利用簡單重復序列(SSR)標記構建了一張覆蓋10個連鎖群的菜心遺傳圖譜,并結合菜心RIL群體一些主要農藝性狀的測定數據,對這些農藝性狀進行數量性狀基因座(QTL)定位和分析。研究結果如下:通過對來自于菜心轉錄組測序開發(fā)的790對SSR引物和多國蕓薹屬基因組計劃蕓薹屬數據庫中的160對SSR標記的引物組合進行篩選,獲得了212對在菜心親本“3T6”和“四九-19號菜心”間PCR擴增產物條帶清晰、重復性好且具有多態(tài)性的SSR引物組合。利用這些具有多態(tài)性的SSR標記,對菜心RIL群體進行基因分型,得到了283個等位變異位點。根據菜心RIL群體間存在的等位變異數據,運用MapMaker 2.0軟件,構建出一張覆蓋菜心10個連鎖群、長1723.81 cM的菜心遺傳圖譜。該圖譜包含197個等位變異位點,平均圖距為8.75 cM。根據已知的蕓薹屬通用SSR錨定引物在染色體上的位置,初步確定了這10個連鎖群與菜心染色體的對應關系。另外,研究發(fā)現圖譜中197個變異位點中有114個位點發(fā)生了偏分離,其中偏向父本“3T6”的有75個,占全部偏分離位點的65.79%;偏向母本“四九-19號菜心”的有39個,占偏分離位點總數的34.21%。并且發(fā)現17個偏分離熱點區(qū)域(SDR),分布在9條染色體上。其中A10染色體上最多,具有3個SDRs;A1、A2、A3、A5、A7和A9染色體上各有2個SDR;而A6和A8染色體則分別只有1個SDR。在已經構建的遺傳圖譜的基礎上,依據2015~2016年2年田間試驗測定RIL群體各株系的株高、開展度、株重、薹高、薹粗、最大葉葉長、最大葉葉寬、最大葉葉柄長、最大葉葉柄寬、基葉數、薹葉數和與葉綠素含量相關的SPAD值等13個農藝性狀數據,運用QGene 2.30軟件,對這些農藝性狀進行QTL定位,共發(fā)現29個解釋表型變異的貢獻率在8.5~30.3%之間QTLs。這些QTLs分別位于A1、A4、A5、A6、A7、A8、A9和A10等8條染色體上,其中與最大葉葉寬相關的QTL數最多,為5個;其次為SPAD值,具有4個QTLs;控制最大葉葉長和薹重相關的QTL各3個,與開展度、最大葉葉柄長、最大葉葉柄寬、薹粗和株重相關的QTLs各2個,與株高、基葉數、薹葉數和薹高相關的QTL則各1個。此外,檢測到株重、最大葉葉長和最大葉葉寬均存在1個多次試驗均檢測到解釋的表型變異的貢獻率在10%的QTL位點。研究結果可為菜心分子標記輔助育種提供參考。
[Abstract]:Cauliflower is one of the largest cultivated characteristic vegetables in South China. High quality, stress resistance and high yield are the main objectives of vegetable heart breeding. With the development of molecular marker technology and the construction of genetic linkage map, gene mapping has become an important part of the research and application of crop genetics and molecular breeding. The development of molecular markers closely linked to the target traits can accelerate the breeding process. On the basis of this, a genetic map of 10 linkage groups was constructed from 130 recombinant inbred lines (RIL) population constructed by parents of "3T6" and "49- 19" as parents, using simple repeat sequence (SSR) markers. The quantitative trait loci were mapped and analyzed by (QTL) combined with the data of some main agronomic characters in RIL population of Chinese cabbage. The results were as follows: by screening 790 pairs of SSR primers and 160 pairs of SSR marker primer combinations from the Brassica Genome Project database, which were developed by sequencing the transcripts of cauliflower, 790 pairs of primers were selected. 212 pairs of SSR primer combinations with clear bands, good reproducibility and polymorphism were obtained between the parents of Chinese cabbage "3T6" and "49- 19". Using these polymorphic SSR markers, the alleles of RIL population were genotyped and 283 alleles were obtained. Based on the allelic variation data among RIL populations, a genetic map covering 10 linkage groups and 1723.81 cm in length was constructed by using MapMaker 2.0 software. The map contains 197 allelic variation sites with an average map distance of 8.75 cm. Based on the known position of common SSR anchoring primers on chromosomes of Brassica, the corresponding relationship between the 10 linkage groups and the chromosomes of Chinese cabbage was preliminarily determined. In addition, the study found that 114 of the 197 variant loci in the map showed partial segregation, of which 75 were in favor of the male parent "3T6", accounting for 65.79% of the total partial segregation sites, and 39 were inclined to the female parent "49- 19". It accounts for 34.21% of the total number of segregation sites. It was also found that the (SDR), of 17 partial segregation hotspots were distributed on 9 chromosomes. There were two SDRs on A10 chromosome, two SDRs on A9 chromosome and one SDRs on A6 and A8 chromosomes, respectively. Based on the genetic map that has been constructed, the plant height, development degree, plant weight, bolting height, bolting diameter, leaf length, leaf petiole width, leaf stem width, stem width and stem width of RIL population were determined by field experiments from 2015 to 2016. The data of 13 agronomic characters, such as basal leaf number, bolting leaf number and SPAD value related to chlorophyll content, were mapped by QTL using QGene 2.30 software. A total of 29 explained phenotypic variation rates were found to be in the range of 8.5% or 30.3%. These QTLs were located on 8 chromosomes A1, A4, A4, A5, A6, A7, A8, A9, A10, respectively. The QTL numbers associated with the maximum leaf width were 5, followed by SPAD, with 4 QTLs, and QTL, which controlled the maximum leaf length and bolting weight, respectively, and the degree of development. The QTLs of maximum petiole length, maximum petiole width, bolting diameter and plant weight were 2, respectively, and QTL with plant height, basal leaf number, bolting leaf number and bolting height were 1, respectively. In addition, the plant weight, the maximum leaf length and the maximum leaf width were detected at 10% of the QTL site, which accounted for 10% of the phenotypic variation in multiple experiments. The results can provide reference for molecular marker-assisted breeding of Chinese cabbage.
【學位授予單位】：廣州大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S634.5
本文編號：2188714
[Abstract]:Cauliflower is one of the largest cultivated characteristic vegetables in South China. High quality, stress resistance and high yield are the main objectives of vegetable heart breeding. With the development of molecular marker technology and the construction of genetic linkage map, gene mapping has become an important part of the research and application of crop genetics and molecular breeding. The development of molecular markers closely linked to the target traits can accelerate the breeding process. On the basis of this, a genetic map of 10 linkage groups was constructed from 130 recombinant inbred lines (RIL) population constructed by parents of "3T6" and "49- 19" as parents, using simple repeat sequence (SSR) markers. The quantitative trait loci were mapped and analyzed by (QTL) combined with the data of some main agronomic characters in RIL population of Chinese cabbage. The results were as follows: by screening 790 pairs of SSR primers and 160 pairs of SSR marker primer combinations from the Brassica Genome Project database, which were developed by sequencing the transcripts of cauliflower, 790 pairs of primers were selected. 212 pairs of SSR primer combinations with clear bands, good reproducibility and polymorphism were obtained between the parents of Chinese cabbage "3T6" and "49- 19". Using these polymorphic SSR markers, the alleles of RIL population were genotyped and 283 alleles were obtained. Based on the allelic variation data among RIL populations, a genetic map covering 10 linkage groups and 1723.81 cm in length was constructed by using MapMaker 2.0 software. The map contains 197 allelic variation sites with an average map distance of 8.75 cm. Based on the known position of common SSR anchoring primers on chromosomes of Brassica, the corresponding relationship between the 10 linkage groups and the chromosomes of Chinese cabbage was preliminarily determined. In addition, the study found that 114 of the 197 variant loci in the map showed partial segregation, of which 75 were in favor of the male parent "3T6", accounting for 65.79% of the total partial segregation sites, and 39 were inclined to the female parent "49- 19". It accounts for 34.21% of the total number of segregation sites. It was also found that the (SDR), of 17 partial segregation hotspots were distributed on 9 chromosomes. There were two SDRs on A10 chromosome, two SDRs on A9 chromosome and one SDRs on A6 and A8 chromosomes, respectively. Based on the genetic map that has been constructed, the plant height, development degree, plant weight, bolting height, bolting diameter, leaf length, leaf petiole width, leaf stem width, stem width and stem width of RIL population were determined by field experiments from 2015 to 2016. The data of 13 agronomic characters, such as basal leaf number, bolting leaf number and SPAD value related to chlorophyll content, were mapped by QTL using QGene 2.30 software. A total of 29 explained phenotypic variation rates were found to be in the range of 8.5% or 30.3%. These QTLs were located on 8 chromosomes A1, A4, A4, A5, A6, A7, A8, A9, A10, respectively. The QTL numbers associated with the maximum leaf width were 5, followed by SPAD, with 4 QTLs, and QTL, which controlled the maximum leaf length and bolting weight, respectively, and the degree of development. The QTLs of maximum petiole length, maximum petiole width, bolting diameter and plant weight were 2, respectively, and QTL with plant height, basal leaf number, bolting leaf number and bolting height were 1, respectively. In addition, the plant weight, the maximum leaf length and the maximum leaf width were detected at 10% of the QTL site, which accounted for 10% of the phenotypic variation in multiple experiments. The results can provide reference for molecular marker-assisted breeding of Chinese cabbage.
【學位授予單位】：廣州大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S634.5
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