番茄无离层基因的分子标记研究及种质资源的筛选
摘要
番茄是一种世界范围性种植的蔬菜作物之一。随着中国加入世贸组织WTO和世界番茄加工产业格局的变化,中国的番茄加工产业日益兴起,中国的番茄制品将有更广阔的国际市场。加工番茄作为番茄加工制品的原材料,是番茄加工的基础。无果梗番茄的培育更便于机器化的采收和提高番茄的产量。
本实验采用分子标记技术,研究番茄无离层基因,利用AFLP和SSR两种分子标记技术,筛选出与番茄无离层基因紧密连锁的标记,并应用该技术体系对48份种质资源进行筛选,以获得含有番茄无离层基因的种质资源。本研究的主要研究成果如下:
1.分析了组合08011(无离层)×08012(有离层)及其有性杂交产生的F1、F2代分离群体在田间表现情况,并建立基因池,结果表明:两个亲本间表现差异明显,母本08011均为无离层,父本08012均表现为有离层,F1代均表现为无离层;F2代群体表现实际分离比为2.9:1,X2检测符合3:1的分离比例。分析结果表明:母本08011中所含无离层基因所控制的性状是由单基因控制的隐性性状。
2.筛选出5个与番茄无离层基因紧密连锁的标记,其中4个AFLP标记E34M84-C、E65M79-E、E74M46-B和E93M63-A,与无离层基因连锁遗传距离分别为5.1cM、7.6 cM、10.4 cM和12.3 cM;一个SSR标记SSR29,与无离层基因的连锁遗传距离为9.5cM。
3.应用获得的AFLP标记E65M79-E,以48份种质资源为试验材料,结合田间鉴定,进行AFLP标记检测,结果表明,二者鉴定结果高度吻合,这一结果为分子育种及进一步的基因克隆奠定了基础。
Tomato was a popular vegetable crops Planted in the world. China's tomato processing industry had developed rapidly. China's tomato products will have a broader international market with China's accession to the WTO. Processing tomato was a raw materials of tomato processing products .It was the basis of tomato processing product. Stem-free tomato cultivation was easy to machine harvest and improved the yield of tomato.
In the experiment,molecular marker technique was applied to research the Stem-free gene of Tomato. The experiment screened out molecule marker linked to the Stem-free gene by AFLP and SSR molecular marker technique. The marker was used to screen 46 tomato germplasm resources containing the Stem-free tomato gene j. The main results of this study is as following:
1.In this study,tomato cultivars 08011、08012 and their progenies F1、F2 were used as the experimental materials. Field appearance was analyzed in full and gene pools were established. The results indicated that there were obvious distinctions . The appearance of female materials 08011were Stem-free while male materials 08012 habor stems. Segregation ratio of F2 groups was 2.9:1 which accorded with the ratio of 3:1. The results shows that character controlled by Stem-free gene from female F2 was recessive character controlled by single gene.
2. Five markers closely linked with tomato Stem-free gene j were obtained. The genetic distance of four AFLP markers on E34M84-C、E65M79-E、E74M46-B和E93M63-A were 5.1 cM、7.6 cM、10.4 cM、12.3 cM and one SSR marker was 9.5cM.
3. The AFLP molecular marker E65M79-E was used to screen out 48 tomato germplasm, meanwhile field identification was carried out. The results indicated that the ratio of identification results about these was very high. This result of AFLP marker will be base of molecular breeding and gene cloning.
本实验采用分子标记技术,研究番茄无离层基因,利用AFLP和SSR两种分子标记技术,筛选出与番茄无离层基因紧密连锁的标记,并应用该技术体系对48份种质资源进行筛选,以获得含有番茄无离层基因的种质资源。本研究的主要研究成果如下:
1.分析了组合08011(无离层)×08012(有离层)及其有性杂交产生的F1、F2代分离群体在田间表现情况,并建立基因池,结果表明:两个亲本间表现差异明显,母本08011均为无离层,父本08012均表现为有离层,F1代均表现为无离层;F2代群体表现实际分离比为2.9:1,X2检测符合3:1的分离比例。分析结果表明:母本08011中所含无离层基因所控制的性状是由单基因控制的隐性性状。
2.筛选出5个与番茄无离层基因紧密连锁的标记,其中4个AFLP标记E34M84-C、E65M79-E、E74M46-B和E93M63-A,与无离层基因连锁遗传距离分别为5.1cM、7.6 cM、10.4 cM和12.3 cM;一个SSR标记SSR29,与无离层基因的连锁遗传距离为9.5cM。
3.应用获得的AFLP标记E65M79-E,以48份种质资源为试验材料,结合田间鉴定,进行AFLP标记检测,结果表明,二者鉴定结果高度吻合,这一结果为分子育种及进一步的基因克隆奠定了基础。
Tomato was a popular vegetable crops Planted in the world. China's tomato processing industry had developed rapidly. China's tomato products will have a broader international market with China's accession to the WTO. Processing tomato was a raw materials of tomato processing products .It was the basis of tomato processing product. Stem-free tomato cultivation was easy to machine harvest and improved the yield of tomato.
In the experiment,molecular marker technique was applied to research the Stem-free gene of Tomato. The experiment screened out molecule marker linked to the Stem-free gene by AFLP and SSR molecular marker technique. The marker was used to screen 46 tomato germplasm resources containing the Stem-free tomato gene j. The main results of this study is as following:
1.In this study,tomato cultivars 08011、08012 and their progenies F1、F2 were used as the experimental materials. Field appearance was analyzed in full and gene pools were established. The results indicated that there were obvious distinctions . The appearance of female materials 08011were Stem-free while male materials 08012 habor stems. Segregation ratio of F2 groups was 2.9:1 which accorded with the ratio of 3:1. The results shows that character controlled by Stem-free gene from female F2 was recessive character controlled by single gene.
2. Five markers closely linked with tomato Stem-free gene j were obtained. The genetic distance of four AFLP markers on E34M84-C、E65M79-E、E74M46-B和E93M63-A were 5.1 cM、7.6 cM、10.4 cM、12.3 cM and one SSR marker was 9.5cM.
3. The AFLP molecular marker E65M79-E was used to screen out 48 tomato germplasm, meanwhile field identification was carried out. The results indicated that the ratio of identification results about these was very high. This result of AFLP marker will be base of molecular breeding and gene cloning.
引文
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黎裕,贾继增,王天宇等.1999.分子标记的种类及其发展[J].生物技术通报.(4):19~22
李君明,徐和金,周永健.2001.有关番茄果实中可溶性固形物和番茄红素的研究进展[J].园艺学报.28(增刊):661~668
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钱惠荣,郑康乐.1998.DNA标记和分子育种[J].生物工程进展.18(3):12~18
王得元,李颖,王恒明.2001.辣椒主要性状遗传研究进展[J].长江蔬菜.(11):283~287
王俊霞,杨光圣,傅廷栋等.2000.甘蓝型油菜PolCMS育性恢复基因的RAPD标记[J].作物学报.26(5):5755~78
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王孝宣,杜永臣,朱德蔚等.2004.增强番茄果实颜色基因的精细定位[R].蔬菜分子育种研讨会6:133~142
于占东,何其伟,王翠花等.2005.大白菜主要营养品质性状的遗传效应研究[J].园艺学报.32(2):244~248
曾国平,曹寿椿.1997.不结球白菜主要品质性状遗传效应分析[J].园艺学报.
张德双,张凤兰,徐家炳.2003.大白菜花色和球色遗传规律的研究[J].华北农学报.18(2):81~84
张增翠,侯喜林,曹寿椿.1999.不结球白菜维生素C和可溶性糖含量的遗传分析[J].园艺学报.26(3):170~174
郑先云,郭豆平,马恩波等.2003.AFLP分子标记技术的发展[J].生命的化学.23(1):65~67
郑晓鹰,王永健,宋顺华等.2002.大白菜耐热分子标记的研究[J].中国农业科学.35(3):309~313
朱军.2000.数量性状遗传分析的新方法及其在育种中的应用[J].浙江大学学报:农业与生命科学版.26(1):16
Adams M D,KelleyJM, GoeayneJD,et al.1991.Complementary DNA Sequencing Expressed Sequential Display:a Highly Efficient Technique for Expression Analysis of Multiene Familied[J ] . ProcNal Acad Sci USA.92 :5331 ~5335
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Bassani M,Neumann P M , Gepstein S. 2004.Differential Expression Profiles of Growth-related Genes in the Elongation Zone of Maize Primary Roots[J].Plant Molecular Biology.56:367~380
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