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Correlation Segment Clone of Molecular Markers and Fluorescencein Situ Hybridization for Stripe Rust Resistance Gene in Wheat Germplasm Guinong775

Author: GeChangBin
Tutor: ZhangQingQin
School: Guizhou University
Course: Plant Pathology
Keywords: Guinong775 Yr gene RAPD SCAR fluorescence in situ hybridization (FISH) Haynaldia viIlosa Cloning
CLC: S512.1
Type: Master's thesis
Year: 2006
Downloads: 129
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Abstract


Guinong 775 is a famous germplasm, which derived from Hayaldia Villosa hybriding with eummer wheat and common wheat cultivar by professor Zhang in Guizhong University, and is important in wheat breeding and production practice. This article studied for the usage of fluorescent in situ hybridization and molecular markers to Guinong775 and the main results are as follows:1. SCAR markers from randomly amplified polymorphic DNA（RAPD） markers tightly linked to the YrGA gene in Guinong775.The F2 populations of Xinong97148×Guinong775 was analysis by SCAR markers. YrGA is the ratio of 3R:1S in while F2 population according to anglicizing. The S2018 molecular marker was cloned and sequenced and changed to SCAR marker. The length of the SCAR marker is 140bp, named SCARS2018rl-140.The polymorphism marker screen results indicated that the RAPD primer S2018 linked to YrGA with the genetic distance of 2.378 ± 0.0064cM; SCAR primer combination SCARrl polymorphic bands about 140bp linked to YrGA gene with the genetic disease about 0.355± 0.001cM. At the same time, Guinong775, Haynaldia villosa and Yangmai 5 was tested by Primer RL and Pm21CD; The polymorphism marker screen results indicated that Guinong775 , Haynaldia villosa and Yangmai 5 primer combination SCARrl polymorphic bands about 140bp and primer combination Pm21C and Pm21D was not. The results indicated that the Guinong775 was not a wheat - Haynaldia villosa segment translocation line,and it is different to the studying results of Liuda Jun et al.2.The results of fluorescence in situ hybridization（FISH） using genomic DNA of Haynaldia villosa and the special probe of S2018 as probe indicated that the Guinong775 was a wheat-Haynaldia villosa new segment translocation line. It is indicated that the YrGA comes from Haynaldia villosa again. FISH experiment was optimized for Guinong775 for several researches.Each steps of FISH experiment were optimized step by step, such as the using of 1-Bromonaphthalene,and the chroma of 2% Pactolase and Cellulase from Aspergillus sp; The results indicated that the ratio of probe DNA and blocking DNA of fluorescence in situ hybridization is from 50 to 100 times.

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