色素万寿菊（Tagetes erecta L.）雄性不育和色素遗传规律分析及SSR标记的筛选
摘要
为了选育具有我国自主知识产权和优良性状的色素万寿菊新品种，本论文进行了色素万寿菊雄性不育、花色、叶黄素遗传规律研究，并对色素万寿菊进行EST-SSR标记筛选试验研究。试验结果如下：
1．以编号208的自然群体中发现的不育株为母本，以编号201B、205B、207B、217B色素含量高的材料为父本进行杂交。杂交F_1代全可育，F_2代可育株:不育株为3:1分离；F_1与F_2自交分离出的不育株回交，回交后代1:1分离。确定该万寿菊雄性不育性是由1对隐性核基因控制的简单质量遗传。
2．为了改善不育株的花色等经济性状并稳定其不育性，在以上试验基础上，进行了回交转育效果初探。利用208A×217B、208A×201B的F_2代中不育株与轮回父本进行回交，回交后代育性稳定为全可育。在回交后代选择与轮回父本花色接近的单株回交，随转育世代数增加接近轮回父本。当回交后代与轮回亲本一致时后代进行自交，筛选可育株:不育株分离比例为3:1的株系，并选择分离出的不育株作为母本，进行系内姊妹交，在严格选择的条件下回交3-4代即可。选择系内姊妹交后代不育株:可育株分离比例为1:1的株系，育成色素万寿菊AB217雄性不育“两用系”。
3．通过形态学、细胞学观察，色素万寿菊AB217雄性不育是由于不育花在雄蕊分化时期没有形成花药原基而导致的结构型雄性不育。
4.根据不育株、可育株和杂合株试验数据显示，在色素万寿菊AB217不育株中SOD、POD活性增高，在花蕾·和开花时·期不育株·中游·离脯氨酸含量·、·可溶性糖含·量低，最·终导致色·素万·寿菊·AB217·雄性不育·。在育种工作中可以在花芽分化前和花蕾时期测定SOD、POD活性来判断可育株(MsMs)与杂合株(Msms)。为育种中减少回交次数，加快育种速度提供新手段。
5.通过AB217A(橙红色)×203B（白色）杂交，构成P_1、P_2、F_1、BC_1~(p1)F_1、BC_2~(p2)F_1和F_2共6个世代，研究了色素万寿菊橙红花与花瓣中叶黄素含量的遗传关系。·结果表·明，·两种最优·遗传模型·均为两对加·性-显性-·上位性·主基因+·加性-显性-·上位·性多基·因遗传模·型。以主·基因遗·传效·应为主·，多基因·效应为辅·。主基因加·性效应·、显性效·应和·上位性效·应作用大·。由此证明，色素万寿菊橙红花与花瓣中叶黄素含量遗传正相关。在育种工作中可以使用色素万寿菊花瓣颜色深浅作为判断花瓣中叶黄素含量高低的指标。
6.通过万寿菊、孔雀草236条EST序列筛选发现SSR序列147个，约占万寿菊、孔雀草EST序列数据的62.29%。在万寿菊、孔雀草的SSR序列中，二核甘·酸占·总序列数·49.66%·，重复出现·频率最高·。二核甘·酸T·C重复单·元最多·，三核·甘酸重复·序列中·TTG出现·频率·最高，五·核甘酸·一个，·四核甘酸·没有·发现。利用Primer premier5.0软件设计SSR引物，共设计28对引物能够显示出清晰条带，与预期扩增长度相符的引物有8对。8对EST-SSR引物共扩增出17条产物带，平均每对扩增2.1条产物带，变幅1-5。4对引物扩增出1条产物带，其余4对引物扩增出2条或2条以上条带，多态性条带有4条。本试·验结果·表明，从·EST数据·库中·建立色素万·寿菊多·态性·EST-SSR·标记是·可行的·，为进·一步建立·EST-SSR·标记·在万寿·菊遗传育·种中·的应用·提供研·究基础·。
In order to obtain new varieties of Tagetes erecta L. with intellectual property and highornamental values, selective male sterile line of Tagetes erecta L.,and seach for high luteincontent character geneticdevelopment.
The main achievements are as follows:
1. Experiment with208A male sterility plant and201B、205B、207B、217B ferility plantfor parent, record performance of the Fertility and Sterility in F_2generation separation ratio is3:1.Then back cross with F_1,the separation ratio is1:1. The Experimental Data Show thatMale sterility of Tagetes erecta L. was1of recessive allele nuclear genes controlled.
2. Base on improve the petal color economic characters of male sterile line, use darkcolour208A×217B、208A×201B F_2backcross with W217B、 W201B,then sistercross.Selective male sterile line of Tagetes erecta L.
3. Through the morphology, cytology observation, AB217male sterile line is structuraltype male sterile, Speculation it is base on B gene afunction.
4. The measure SOD,POD, MDA, Fpro, soluble sugar content inAB217male sterileline concluded that W217male sterile is due to SOD,POD lively and MDA, Fpro, Solublesugar content descend. For speed up breeding that measure SOD,POD,in leaf and bud,measure POD and Soluble sugar content in flower to distinguish between fertile plants andheterozygous plant.
5.Through217A(jacinth)×203B(white) P_1, P_2, F_1, BC_1~(p1)F_1, BC_2~(p2),F_1,F_2nvestigateion,the type of jacinth flower colors heredity and lutein content heredity is2pair of additivedominance epistasis major gene+additive-dominance-epistasis polygenes. The relativitybetween flower colors heredity and lutein content heredity is tightly.
6.Found147SSR in Tagetes erecta and Tagetes patula EST. Dinucleotide is the most,62.29%. TC to be the most abundant in dinucleotide. TTG to be the most abundant intrinucleotide.1pentanucleotide, and tetranucleotide not be found. Design28SSR primer withPrimer premier5.0software. The SSR analysis identified a total of17bands among the217A×203B F_2, with4bands being polymorphic. From the results of amplification,4from8SSR primers showed no different bands between217A×203B F_2.
1．以编号208的自然群体中发现的不育株为母本，以编号201B、205B、207B、217B色素含量高的材料为父本进行杂交。杂交F_1代全可育，F_2代可育株:不育株为3:1分离；F_1与F_2自交分离出的不育株回交，回交后代1:1分离。确定该万寿菊雄性不育性是由1对隐性核基因控制的简单质量遗传。
2．为了改善不育株的花色等经济性状并稳定其不育性，在以上试验基础上，进行了回交转育效果初探。利用208A×217B、208A×201B的F_2代中不育株与轮回父本进行回交，回交后代育性稳定为全可育。在回交后代选择与轮回父本花色接近的单株回交，随转育世代数增加接近轮回父本。当回交后代与轮回亲本一致时后代进行自交，筛选可育株:不育株分离比例为3:1的株系，并选择分离出的不育株作为母本，进行系内姊妹交，在严格选择的条件下回交3-4代即可。选择系内姊妹交后代不育株:可育株分离比例为1:1的株系，育成色素万寿菊AB217雄性不育“两用系”。
3．通过形态学、细胞学观察，色素万寿菊AB217雄性不育是由于不育花在雄蕊分化时期没有形成花药原基而导致的结构型雄性不育。
4.根据不育株、可育株和杂合株试验数据显示，在色素万寿菊AB217不育株中SOD、POD活性增高，在花蕾·和开花时·期不育株·中游·离脯氨酸含量·、·可溶性糖含·量低，最·终导致色·素万·寿菊·AB217·雄性不育·。在育种工作中可以在花芽分化前和花蕾时期测定SOD、POD活性来判断可育株(MsMs)与杂合株(Msms)。为育种中减少回交次数，加快育种速度提供新手段。
5.通过AB217A(橙红色)×203B（白色）杂交，构成P_1、P_2、F_1、BC_1~(p1)F_1、BC_2~(p2)F_1和F_2共6个世代，研究了色素万寿菊橙红花与花瓣中叶黄素含量的遗传关系。·结果表·明，·两种最优·遗传模型·均为两对加·性-显性-·上位性·主基因+·加性-显性-·上位·性多基·因遗传模·型。以主·基因遗·传效·应为主·，多基因·效应为辅·。主基因加·性效应·、显性效·应和·上位性效·应作用大·。由此证明，色素万寿菊橙红花与花瓣中叶黄素含量遗传正相关。在育种工作中可以使用色素万寿菊花瓣颜色深浅作为判断花瓣中叶黄素含量高低的指标。
6.通过万寿菊、孔雀草236条EST序列筛选发现SSR序列147个，约占万寿菊、孔雀草EST序列数据的62.29%。在万寿菊、孔雀草的SSR序列中，二核甘·酸占·总序列数·49.66%·，重复出现·频率最高·。二核甘·酸T·C重复单·元最多·，三核·甘酸重复·序列中·TTG出现·频率·最高，五·核甘酸·一个，·四核甘酸·没有·发现。利用Primer premier5.0软件设计SSR引物，共设计28对引物能够显示出清晰条带，与预期扩增长度相符的引物有8对。8对EST-SSR引物共扩增出17条产物带，平均每对扩增2.1条产物带，变幅1-5。4对引物扩增出1条产物带，其余4对引物扩增出2条或2条以上条带，多态性条带有4条。本试·验结果·表明，从·EST数据·库中·建立色素万·寿菊多·态性·EST-SSR·标记是·可行的·，为进·一步建立·EST-SSR·标记·在万寿·菊遗传育·种中·的应用·提供研·究基础·。
In order to obtain new varieties of Tagetes erecta L. with intellectual property and highornamental values, selective male sterile line of Tagetes erecta L.,and seach for high luteincontent character geneticdevelopment.
The main achievements are as follows:
1. Experiment with208A male sterility plant and201B、205B、207B、217B ferility plantfor parent, record performance of the Fertility and Sterility in F_2generation separation ratio is3:1.Then back cross with F_1,the separation ratio is1:1. The Experimental Data Show thatMale sterility of Tagetes erecta L. was1of recessive allele nuclear genes controlled.
2. Base on improve the petal color economic characters of male sterile line, use darkcolour208A×217B、208A×201B F_2backcross with W217B、 W201B,then sistercross.Selective male sterile line of Tagetes erecta L.
3. Through the morphology, cytology observation, AB217male sterile line is structuraltype male sterile, Speculation it is base on B gene afunction.
4. The measure SOD,POD, MDA, Fpro, soluble sugar content inAB217male sterileline concluded that W217male sterile is due to SOD,POD lively and MDA, Fpro, Solublesugar content descend. For speed up breeding that measure SOD,POD,in leaf and bud,measure POD and Soluble sugar content in flower to distinguish between fertile plants andheterozygous plant.
5.Through217A(jacinth)×203B(white) P_1, P_2, F_1, BC_1~(p1)F_1, BC_2~(p2),F_1,F_2nvestigateion,the type of jacinth flower colors heredity and lutein content heredity is2pair of additivedominance epistasis major gene+additive-dominance-epistasis polygenes. The relativitybetween flower colors heredity and lutein content heredity is tightly.
6.Found147SSR in Tagetes erecta and Tagetes patula EST. Dinucleotide is the most,62.29%. TC to be the most abundant in dinucleotide. TTG to be the most abundant intrinucleotide.1pentanucleotide, and tetranucleotide not be found. Design28SSR primer withPrimer premier5.0software. The SSR analysis identified a total of17bands among the217A×203B F_2, with4bands being polymorphic. From the results of amplification,4from8SSR primers showed no different bands between217A×203B F_2.
引文
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