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Yield and Yield-related Traits in Brassica Napus L. and Its Comparative Genome Primary Research

Author: ChenQingFang
Tutor: MaChaoZhi
School: Huazhong Agricultural University
Course: Crop Genetics and Breeding
Keywords: Brassica napus Oryza sativa Genetic linkage map QTLs Comparative genomics SUC1 gene Gene cloning Rapid amplification of cDNA ends(RACE)
CLC: S565.4
Type: Master's thesis
Year: 2007
Downloads: 79
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Abstract


Heterosis utilization is proved to be the most effective way to increase the yield in genetic improvement of crops at present and have been widely used in B. napus. The molecular basis of heterosis compared to heterosis utilization is very lagged until now. However, some researches have shown that the differentially expressed genes between hybrid and parents were closely related to the heterosis. Some differentially expressed genes were successfully developed to functional molecular markers which were integrated into genetic linkage map of rapeseed, and the QTLs of yield-related agronomical traits have been located and analysised. On basis of the previous reseach, 114 markers located in genetic linkage map was sequenced and analysised. Comparative genomics between rice and rapeseed, and among different populations of B. napus by functional markers. Homologous fragment of candidate gene SUC1 related to yield and other traits was cloned. The major results in this study are as follows:1. On the base of constructed genetic linkage map, 6 SRAP markers generated by 4 pair of primers and 108 functional molecular markers generated by 44 functional gene were sequenced. The 6 sequences of amplified by SRAP were done by Blastn. The results shown SRAP markers in the study weren’t used to seek functional gene. 108 functional molecular markers were used to undertake analysis by Blast 2 sequence. The result indicated the primer pairs used to amplified products could really amplify the original sequence. In addition, 26 sequences in both parents amplified by 12 functional primer pairs were randomly selected and sequenced. The comparative results of the two sequences suggested the high similarity of the functional genes between cultivars. The frequencies for C instead of T between two parents were high.2. The 55 genes including 40 genes located in QTLs and 15 genes unlocated in QTLs located in the genetic linkage map of B. napus, were used to search in Genbank by Blastx to achieve 53 homologous sequences in rice. In addition, comparative mapping between rapeseed and rice was done by in-silico way, and the yield-related QTLs were searched around these genes. The comparative result shown the region where some genes had located had found the QTLs for the similar traits in rice and rapeseed.3. 40 pairs of primer were designed, 21 pairs of primer could amplify clear bands by detecting on agrose gel, and 8 pairs of primer produced 10 polymorphic bands when rice population was analysized. The 186 lines population recombinant inbred lines-7(RILs-7) derived from Teqing×Zhenxian97 was used to analysis, and 6 functional markers generated by 6 palmer pairs were located in 4 linkage group of genetic linkage map of rice.4. 20 pairs of primer were used to analysis the 6 parents of the three populations. In the end, 11 functional markers generated by 9 pairs of primer were located in genetic linkage map of another two double haploid (DH) populations, and the yield-related traits were found.5. Sucrose transporter gene SUC1 of B. napus was located in seven QTLs related to yield and was to successfully search homologous sequences in B. oleracea, B. rapa and A.thaliana by BLAST. According homologous sequence method, two primer pairs were designed and were used to amplify sequences at the same time. Homologous sequence of SUC1 gene was used to clone by POP-PCR, PCR-Walking and 3’RACE. Positions of 4 extrons, 3 introns and stop coden TAG in the gene were predicted by FGENESH in SoftBerry web.

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CLC: > Agricultural Sciences > Crop > Economic crops > Oil crops > Rapeseed ( Brassica )
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