甘藍型油菜TT8基因家族的RNAi和CRES-T抑制與比較.pdf

1、Universi加Code:10635StudentNumber:112008326002151ADissertationforMaster’sDegreeofsouthwestUniversitySuPPressionandComParisonofBrassicanaPusTTSGeneFamilybyRNAiandCRES一TMaster，5degreeeandidate:Rui一XiZhangSuPervisor:Assoeiat

2、eProf.Jia一MingYinProf.YOu一RongChaiSPeeialty:GeneticsDireetion:PlantGenetiesandBreedingChongqingChinaJune2011西南大學碩士學位論文3.8.2預培養(yǎng).........……，..................................................................................

3、..........……213.8.3浸染.............……，........................................................................................……，二213.8.4共培養(yǎng).................................................................................

4、……‘.……，............……213.8.5洗外植體(殺農桿菌)...................................……213.8.6誘導抗性愈傷........................................................……，............................……223.8.7分化培養(yǎng)................................

5、.......................................................................……223.8.8分化莖...……，.…，..............................……，.............……，...........................……223.8.9長莖.....................……，.................

6、......................……，......................................……223.8.10生根...............................................................................................................……233.8.11馴化及移栽.......……、.........

7、................................................……233.8.12農桿菌轉化組培過程中的玻璃化現象研究..............................................……233.9轉基因再生植株分子檢測與性狀鑒定..................................................................……243.9.1

8、Basta復檢................................................................................……，................……243.9.2組織化學染色檢測GUS活性............................................……，.....……，.......……243.9.3再生植株的PcR檢測...........

9、........................................……243.9.4再生植株T:種籽粒色性狀考察.....................................................……，.......……253.9.5再生植株TI種籽花青素與類黃酮含量的測定..........................................……253.9.6再生植株種子皮殼率的測定..

10、........................................................……，.......……253.9.7再生植株種子千粒重的測定........................................................................……263.9.8再生植株結實率調查........................................……2

11、63.9.9再生植株莢角長度調查.................................................................……263.9.10再生植株分枝數調查.........................................................................……263.10轉基因再生種子的體視顯微鏡觀察.......................…

12、…........................................……263.11轉基因再生植株了了百基因家族的表達檢測.........................................................……263.11.1mRNA的反轉錄.............................................................................

13、……263.11.2調BnZos內標..................................................................……273.11.3半定量RTPCR檢測轉基因再生植株BnTT石基因家族的抑制水平.......……274.1蕓蔓屬了了萬基因家族RNA干擾載體的構建.........................................................……29

14、4.1.1蕓蔓屬27百基因家族RNA干擾片段的克隆..................……幾.....................……294.1.2蕓蔓屬TTS基因家族RNA干擾載體的構建.............................................……294.1.3重組載體轉化根癌農桿菌及鑒定......................................……，...……，.……304.2蕓

15、蔓屬了了客基因家族CRES一T載體的構建...........................................................……314.2.1蕓蔓屬了珊基因家族4條CRES一T片段的擴增........................................……314.2.2從重組pMD19一T載體切下了珊家族的CRES一T片段......……，.................……344.2.

16、3植物表達載體pC23OIMISDPB和pC230lMISMpB骨架的制備...........……344.2.4重組形成CRES一T載體、轉化和鑒定...........................................……，.......……344.3RNA干擾和CRES一T載體轉化甘藍型油菜.........................................................……，.364

17、.3.1遺傳轉化.............................................……，...................................................……364.3.2不同因素對玻璃化現象的影響研究分析.....................................................……364.4轉基因再生植株分子檢測與性狀鑒定...........

18、.......................................................……384.4.1Basta復檢結果...........................................................................................……384.4.2組織化學染色檢測GUS活性..................................

19、....................................……394.4.3再生植株的PCR檢測....................................................................……，‘.……404.44再生植株丁:種籽粒色性狀考察.................................................................……，42