The translucent endosperm trait in a japonica rice variety ＇Kantou 194＇ is controlled by a Wx-mq gene which is allelic to Wx locus by genetic analysis and allelic test. The amylose content analysis showed that an in...The translucent endosperm trait in a japonica rice variety ＇Kantou 194＇ is controlled by a Wx-mq gene which is allelic to Wx locus by genetic analysis and allelic test. The amylose content analysis showed that an intermediate amylose content between those of glutinous and non-glutinous rice existed in endosperm of homozygous Wx-mq genotype. The slight changes of amylose content in different varieties and F1 grains with an identical Wx-mq genotype might be influenced by dissimilar genetic background. To identify the Wx-mq genotype simply and rapidly, a cleaved amplified polymorphic sequence （CAPS） marker was designed. The result from the molecular detection indicated that it could be used for marker-assisted selection for low amylose content varieties in rice breeding.展开更多
Molecular genetic maps were commonly constructed by analyzing the segregation of restriction fragment length polymorphisms (RFLPs). Here we described methodology-marker sequences in a new mapping based on recent docum...Molecular genetic maps were commonly constructed by analyzing the segregation of restriction fragment length polymorphisms (RFLPs). Here we described methodology-marker sequences in a new mapping based on recent documents. With the methods they were unique sequences detected by the polymerase chain reaction (PCR). Each of the methods had its Iimitations and the current trend was to integrate the maps produced by the different methods. Marker sequences contained mainly expressed sequence tags (ESTs),polymorphie sequence-tagged sites (STSs), randomly amplified polymorphic DNA (RAPDs), cIeaved amplified polymorphic sequences (CAPS), amplified fragment Iength pofymorphism (AFLPs), genorne sequence sampling (GSS) and sequence-tagged connectors (STCs) in this paper.展开更多
利用多花黑麦草叶斑病抗性和敏感个体杂交构建F 1分离群体,采用分群分析法(bu lked segregan t ana lys is,BSA),通过多花黑麦草表达序列标签(expressed sequence tag,EST)的PCR扩增和扩增产物的限制性内切酶酶切多态性(cleaved am p li...利用多花黑麦草叶斑病抗性和敏感个体杂交构建F 1分离群体,采用分群分析法(bu lked segregan t ana lys is,BSA),通过多花黑麦草表达序列标签(expressed sequence tag,EST)的PCR扩增和扩增产物的限制性内切酶酶切多态性(cleaved am p lified po lym orph ic sequence,CAPS)筛选,获得一个同多花黑麦草草抗叶斑病紧密连锁的EST-CAPS标记p56。对照多花黑麦草细胞质雄性不育(CM S)群体构建的遗传连锁图,该EST-CAPS标记位于多花黑麦草的第5遗传连锁群(LG 5)。对cDNA文库中对应于标记位点p56的EST序列片段分析和基因库搜索结果表明,该EST所编码的氨基酸序列同大麦天冬酰胺合成酶基因H vAS 1和H vAS 2的部分氨基酸序列片段同源性很高,推测位点p56所处的基因为编码多花黑麦草天冬酰胺合成酶基因。该分子标记可用于多花黑麦草分子标记辅助育种。展开更多
The allelic variation of the Wx gene in 50 non-glutinous rice varieties （lines） was analyzed by using the microsatellite marker RM190 [for （CT）n simple sequence repeat （SSR）] and cleaved amplified polymorphic se...The allelic variation of the Wx gene in 50 non-glutinous rice varieties （lines） was analyzed by using the microsatellite marker RM190 [for （CT）n simple sequence repeat （SSR）] and cleaved amplified polymorphic sequence（CAPS） marker 484/W2R-ACCⅠ[for G/T single nucleotide polymorphism （SNP）]. Six homozygous （CT）n types, namely （CT）20, （CT）19, （CT）18, （CT）17, （CT）16, （CT）14, （CT）11 and （CT）10, and a heterozygous genotype （CT）11/（CT）18 were detected for RM190, of which （CT）11 and （CT）18 were predominant. Two homozygous Wx genotypes （G/G and T/T） and one heterozygous （G/T） were detected using 484/W2R-ACC Ⅰ. Most of the materials with a RM190 of （CT）11 were G/G for SNP of 484/W2R-ACC Ⅰ, while T/T for SNP was predominantly appeared in materials with （CT）18. The materials tested could be grouped into 10 categories using the two markers together. Results indicated that 59.3% variance of amylose content was attributed to the polymorphism of Wx gene revealed by RM190, while 56.1% and 24.6% of the variances in amylose content and gel consistency were respectively to the polymorphism of Wx gene revealed by 484/W2R-ACC Ⅰ. Furthermore, with both SSR and CAPS markers, 72.4% of the variance in amylose content could be explained. In addition, the application prospects of the two markers in breeding were also discussed.展开更多
Tomato mosaic virus (ToMV) is one of the most infectious virus diseases in tomato (Solanum lycopersicum L). The practical and effective method of controlling this disease is through genetic control by using major resi...Tomato mosaic virus (ToMV) is one of the most infectious virus diseases in tomato (Solanum lycopersicum L). The practical and effective method of controlling this disease is through genetic control by using major resistance genes. So far, three genes Tm-1, Tm-2 and Tm-22 conferring resistance to ToMV have been reported and utilized in tomato culti-var development. Marker assisted selection (MAS) has become very important and useful tool in selection of ToMV re-sistant tomato lines or hybrids. The objective of this research was to identify allele-specific PCR-based, cleaved ampli-fied polymorphic sequence (CAPS), and allele-derived single nucleotide polymorphism (SNP) markers for Tm-2 loci. Four allele-specific PCR-based markers were identified: one for Tm-2, one for Tm-22, and two for the susceptible allele tm-2. Three allele-derived CAPS markers were identified, which can identify and distinguish three alleles, tm-2, Tm-2 and Tm-22 in tomato germplasm. Three SNP markers were developed specific for Tm-2 locus. These markers will pro-vide breeders with a tool in selection of Tm-2 and Tm-22 resistance genes in tomato breeding program.展开更多
GW2 is an important gene that regulates grain width and weight. We used cDNA clone to obtain the sequences of GW2 from large- and small-grained rice varieties, TD70 and Kasalath, respectively. Then, we developed a dCA...GW2 is an important gene that regulates grain width and weight. We used cDNA clone to obtain the sequences of GW2 from large- and small-grained rice varieties, TD70 and Kasalath, respectively. Then, we developed a dCAPS （derived cleaved amplified polymorphic sequence） marker on the basis of the sequence difference between functional and nonfunctional GW2 genes to analyze the genotypes and phenotypes of recombinant inbred lines. Results showed that the sequence of GW2To7~ had a single nucleotide deletion at site 316 that generates a termination codon. This codon terminated the GW2 protein in advance. By contrast, the sequence of GW2Kasalath encoded an intact protein. A novel dCAPS marker was designed in accordance with a base A deletion at site 316 of the sequence. After the PCR product was digested by Apol, TD70 showed 21 and 30 bp fragments, and Kasalath showed a 51 bp fragment. Up to 82 lines contained GW2TDTO, and 158 lines contained GW2Kasalath. The lines that contained TD70 alleles displayed substantial increases in width and 1000-grain weight. This result suggested that GW2 played a critical role in rice breeding.展开更多
基金supported by the National High Technology Research and Development Program of China(Grant No.2006AA100101)National Science and Technology Support Program of China(Grant No. 2006BAD01A01-5)+1 种基金Special Program for Rice Scientific Research,Ministry of Agriculture,China(Grant No. nyhyzx 07-001-006)Super Rice Breeding and Demonstration Program,Ministry of Agriculture,China and Jiangsu Agricultural Scientific Self-innovation Fund,China(Grant No.CX[07]603)
文摘The translucent endosperm trait in a japonica rice variety ＇Kantou 194＇ is controlled by a Wx-mq gene which is allelic to Wx locus by genetic analysis and allelic test. The amylose content analysis showed that an intermediate amylose content between those of glutinous and non-glutinous rice existed in endosperm of homozygous Wx-mq genotype. The slight changes of amylose content in different varieties and F1 grains with an identical Wx-mq genotype might be influenced by dissimilar genetic background. To identify the Wx-mq genotype simply and rapidly, a cleaved amplified polymorphic sequence （CAPS） marker was designed. The result from the molecular detection indicated that it could be used for marker-assisted selection for low amylose content varieties in rice breeding.
文摘Molecular genetic maps were commonly constructed by analyzing the segregation of restriction fragment length polymorphisms (RFLPs). Here we described methodology-marker sequences in a new mapping based on recent documents. With the methods they were unique sequences detected by the polymerase chain reaction (PCR). Each of the methods had its Iimitations and the current trend was to integrate the maps produced by the different methods. Marker sequences contained mainly expressed sequence tags (ESTs),polymorphie sequence-tagged sites (STSs), randomly amplified polymorphic DNA (RAPDs), cIeaved amplified polymorphic sequences (CAPS), amplified fragment Iength pofymorphism (AFLPs), genorne sequence sampling (GSS) and sequence-tagged connectors (STCs) in this paper.
基金supported by the National High Technology Research and Development Program of China (Grant No. 2011AA10A101 and 2012AA101102)the Ministry of Finance, China (Grant No. 2012RG002-4)
文摘利用多花黑麦草叶斑病抗性和敏感个体杂交构建F 1分离群体,采用分群分析法(bu lked segregan t ana lys is,BSA),通过多花黑麦草表达序列标签(expressed sequence tag,EST)的PCR扩增和扩增产物的限制性内切酶酶切多态性(cleaved am p lified po lym orph ic sequence,CAPS)筛选,获得一个同多花黑麦草草抗叶斑病紧密连锁的EST-CAPS标记p56。对照多花黑麦草细胞质雄性不育(CM S)群体构建的遗传连锁图,该EST-CAPS标记位于多花黑麦草的第5遗传连锁群(LG 5)。对cDNA文库中对应于标记位点p56的EST序列片段分析和基因库搜索结果表明,该EST所编码的氨基酸序列同大麦天冬酰胺合成酶基因H vAS 1和H vAS 2的部分氨基酸序列片段同源性很高,推测位点p56所处的基因为编码多花黑麦草天冬酰胺合成酶基因。该分子标记可用于多花黑麦草分子标记辅助育种。
文摘The allelic variation of the Wx gene in 50 non-glutinous rice varieties （lines） was analyzed by using the microsatellite marker RM190 [for （CT）n simple sequence repeat （SSR）] and cleaved amplified polymorphic sequence（CAPS） marker 484/W2R-ACCⅠ[for G/T single nucleotide polymorphism （SNP）]. Six homozygous （CT）n types, namely （CT）20, （CT）19, （CT）18, （CT）17, （CT）16, （CT）14, （CT）11 and （CT）10, and a heterozygous genotype （CT）11/（CT）18 were detected for RM190, of which （CT）11 and （CT）18 were predominant. Two homozygous Wx genotypes （G/G and T/T） and one heterozygous （G/T） were detected using 484/W2R-ACC Ⅰ. Most of the materials with a RM190 of （CT）11 were G/G for SNP of 484/W2R-ACC Ⅰ, while T/T for SNP was predominantly appeared in materials with （CT）18. The materials tested could be grouped into 10 categories using the two markers together. Results indicated that 59.3% variance of amylose content was attributed to the polymorphism of Wx gene revealed by RM190, while 56.1% and 24.6% of the variances in amylose content and gel consistency were respectively to the polymorphism of Wx gene revealed by 484/W2R-ACC Ⅰ. Furthermore, with both SSR and CAPS markers, 72.4% of the variance in amylose content could be explained. In addition, the application prospects of the two markers in breeding were also discussed.
文摘Tomato mosaic virus (ToMV) is one of the most infectious virus diseases in tomato (Solanum lycopersicum L). The practical and effective method of controlling this disease is through genetic control by using major resistance genes. So far, three genes Tm-1, Tm-2 and Tm-22 conferring resistance to ToMV have been reported and utilized in tomato culti-var development. Marker assisted selection (MAS) has become very important and useful tool in selection of ToMV re-sistant tomato lines or hybrids. The objective of this research was to identify allele-specific PCR-based, cleaved ampli-fied polymorphic sequence (CAPS), and allele-derived single nucleotide polymorphism (SNP) markers for Tm-2 loci. Four allele-specific PCR-based markers were identified: one for Tm-2, one for Tm-22, and two for the susceptible allele tm-2. Three allele-derived CAPS markers were identified, which can identify and distinguish three alleles, tm-2, Tm-2 and Tm-22 in tomato germplasm. Three SNP markers were developed specific for Tm-2 locus. These markers will pro-vide breeders with a tool in selection of Tm-2 and Tm-22 resistance genes in tomato breeding program.
基金supported by the National Natural Science Foundation of China (Grant No.31271678)the Jiangsu Agricultural Scientific Self-Innovation Fund (Grant No.CX[12]1003)Jiangsu Province Agricultural Science and Technology Support Program (Grant No.BE2013301)
文摘GW2 is an important gene that regulates grain width and weight. We used cDNA clone to obtain the sequences of GW2 from large- and small-grained rice varieties, TD70 and Kasalath, respectively. Then, we developed a dCAPS （derived cleaved amplified polymorphic sequence） marker on the basis of the sequence difference between functional and nonfunctional GW2 genes to analyze the genotypes and phenotypes of recombinant inbred lines. Results showed that the sequence of GW2To7~ had a single nucleotide deletion at site 316 that generates a termination codon. This codon terminated the GW2 protein in advance. By contrast, the sequence of GW2Kasalath encoded an intact protein. A novel dCAPS marker was designed in accordance with a base A deletion at site 316 of the sequence. After the PCR product was digested by Apol, TD70 showed 21 and 30 bp fragments, and Kasalath showed a 51 bp fragment. Up to 82 lines contained GW2TDTO, and 158 lines contained GW2Kasalath. The lines that contained TD70 alleles displayed substantial increases in width and 1000-grain weight. This result suggested that GW2 played a critical role in rice breeding.
基金supported by the Key Program of the Development of Variety of Genetically Modified Organisms(2008ZX08001-006)the Key Support Program of Jiangsu Science and Technology (Grant No.BE2008354)Jiangsu Agricultural Scientific Self-innovation Fund(CX[09]634)
