The purpose of this study is to reveal the genetic mechanism of the variation of amylose content among different semi waxy or glutinous japonica rice in the background of Wxmp gene.Sixty-four semi waxy lines derived f...The purpose of this study is to reveal the genetic mechanism of the variation of amylose content among different semi waxy or glutinous japonica rice in the background of Wxmp gene.Sixty-four semi waxy lines derived from the hybrid progenies of Wujing 13 and Milky Princess(Kantou 194)with polymorphism in soluble starch synthase gene SSIIa(SSII-3)and SSIIIa(SSIII-2)but no polymorphism in other starch synthase related genes were used as test materials.The genotypes of SSIIa and SSIIIa allele were identified by molecular markers,and the allelic effects of SSIIa and SSIIIa gene on amylose content(AC),gel consistency(GC),gelatinization temperature(GT)and rapid visco analyzer(RVA)profile characteristics were analyzed.The significant effects of SSIIa and SSIIIa alleles and the interactive effects between two genes on AC,GT,GC and RVA profile characteristics were found.The SSIIa and SSIIIa alleles from Wujing13 shown positive effects on AC with an average increase of 1.87 and 1.23%in 2 years respectively.There was no significant effect on GT for SSIIa or SSIIIa allele but remarkable influence on GT when the co-existence of the two genes.The genotype SSIIa^(mp)SSIIIa^(mp) shown 1.34℃higher GT than genotype SSIIawjSSIIIawj(mp and wj indicated that the gene was derived from Milky Princess and Wujing 13 respectively,the same as in the below).Different genes and alleles resulted in significant different GC.The genetic effect of SSIIawj and SSIIIamp on GC was 8.74 and 9.62mm respectively.The GC of SSIIa^(wj)SSIIIa^(mp) was 10.64 and 16.95mm higher than that of SSIIa^(mp)SSIIIa^(wj) and SSIIawjSSIIIawj,respectively.The allele SSIIa^(wj) could increase the peak viscosity(PKV),hot paste viscosity(HPV),cool paste viscosity(CPV)and breakdown viscosity(BDV),while decrease the consistency viscosity(CSV)and setback viscosity(SBV).However for the allele SSIIIa^(wj) the opposite was true.The genotype SSIIa^(wj)SSIIIa^(mp) had the largest PKV,HPV and CPV,the genotype SSIIa^(wj)SSIIIa^(wj) had the largest BDV and CSV,but the genotype SSIIa^(wj)SSIIIa^(mp) had the least SBV.According to the comprehensive effect of each trait,the genotype SSIIawjSSIIIamp was the best.The allelic variation and interaction effect of SSIIa and SSIIIa genes have important reference value for improving cooking and eating quality of semi waxy japonica rice.展开更多
Endogenous reference genes (ERGs) provide vital information regarding genetically modified organisms (GMOs). The successful detection of ERGs can identity GMOs and the source of genes, verify stability and reliability...Endogenous reference genes (ERGs) provide vital information regarding genetically modified organisms (GMOs). The successful detection of ERGs can identity GMOs and the source of genes, verify stability and reliability of the detection system, and calculate the level of genetically modified (GM) ingredients in mixtures. The reported ERGs in rice include sucrose-phosphate synthase (SPS), phospholipase D (PLD), RBE4 and rice root-specific GOS9 genes. Based on the characteristics of ERGs, a new ERG gene, phosphoenolpyruvate carboxylase (PEPC), was selected, and further compared with the four existing genes. A total of 18 rice varieties and 29 non-rice crops were used to verify the interspecies specificity, intraspecies consistency, sensitivity, stability and reliability of these five ERGs using qualitative and quantitative PCR. Qualitative detection indicated that SPS and PEPC displayed sufficient specificity, and the detection sensitivity was 0.05% and 0.005%, respectively. Although the specificity of both RBE4 and GOS9 were adequate, the amplicons were small and easily confused with primer dimers. Non-specific amplification of the PLD gene was present in maize and potato. Real-time quantitative PCR detection indicated that PLD, SPS and PEPC displayed good specificity, with R2 of the standard curve greater than 0.98, while the amplification efficiency ranged between 90% and 110%. Both the detection sensitivities of PLD and PEPC were five copies and that of SPS was ten copies. RBE4 showed typical amplification in maize, beet and Arabidopsis, while GOS9 was found in maize, tobacco and oats. PEPC exhibited excellent detection sensitivity and species specificity, which made it a potentially useful application in GM-rice supervision and administration. Additionally, SPS and PLD are also suitable for GM-rice detection. This study effectively established a foundation for GMO detection, which not only provides vital technical support for GMO identification, but also is of great significance for enhancing the comparability of detection results, and the standardization of ERG testing in GM-rice.展开更多
文摘以马铃薯普通栽培种"甘农薯2号"块茎为材料,利用RNA提取试剂盒提取总RNA,通过RT-PCR技术和DNA序列测定分析,证实获得了马铃薯颗粒结合淀粉合成酶(GBSS I)基因的cDNA序列。该cDNA全长1824bp,包括607个氨基酸和1个终止密码子序列,且与原序列(accession number X58453)同源性为99.78%,但与其他科植物GBSS基因的同源性较低,注册该基因到GenBank中,注册号为EU403426。利用生物信息学相关软件分析预测GBSS I基因cDNA序列编码的蛋白质功能和结构,结果发现,该蛋白与其他15种植物GBSS蛋白一样,具有3个完全保守区域,并具许多重要功能位点,且与农杆菌淀粉合成酶具有相似三级结构模型,表明该蛋白具淀粉合成功能。
基金supported by Jiangsu Natural Science Foundation(No.BK20180302)Jiangsu agricultural science and Technology Innovation Fund(No.CX[18]1001)+3 种基金Jiangsu Key Research and development program(No.BE2018357)Jiangsu major new varieties creation project(No.PZCZ201703)the earmarked fund for China Agriculture Research System(No.CARS-1-62)the open project of key Laboratory of Jiangsu crop genomics and molecular Breeding(No.PL201902).
文摘The purpose of this study is to reveal the genetic mechanism of the variation of amylose content among different semi waxy or glutinous japonica rice in the background of Wxmp gene.Sixty-four semi waxy lines derived from the hybrid progenies of Wujing 13 and Milky Princess(Kantou 194)with polymorphism in soluble starch synthase gene SSIIa(SSII-3)and SSIIIa(SSIII-2)but no polymorphism in other starch synthase related genes were used as test materials.The genotypes of SSIIa and SSIIIa allele were identified by molecular markers,and the allelic effects of SSIIa and SSIIIa gene on amylose content(AC),gel consistency(GC),gelatinization temperature(GT)and rapid visco analyzer(RVA)profile characteristics were analyzed.The significant effects of SSIIa and SSIIIa alleles and the interactive effects between two genes on AC,GT,GC and RVA profile characteristics were found.The SSIIa and SSIIIa alleles from Wujing13 shown positive effects on AC with an average increase of 1.87 and 1.23%in 2 years respectively.There was no significant effect on GT for SSIIa or SSIIIa allele but remarkable influence on GT when the co-existence of the two genes.The genotype SSIIa^(mp)SSIIIa^(mp) shown 1.34℃higher GT than genotype SSIIawjSSIIIawj(mp and wj indicated that the gene was derived from Milky Princess and Wujing 13 respectively,the same as in the below).Different genes and alleles resulted in significant different GC.The genetic effect of SSIIawj and SSIIIamp on GC was 8.74 and 9.62mm respectively.The GC of SSIIa^(wj)SSIIIa^(mp) was 10.64 and 16.95mm higher than that of SSIIa^(mp)SSIIIa^(wj) and SSIIawjSSIIIawj,respectively.The allele SSIIa^(wj) could increase the peak viscosity(PKV),hot paste viscosity(HPV),cool paste viscosity(CPV)and breakdown viscosity(BDV),while decrease the consistency viscosity(CSV)and setback viscosity(SBV).However for the allele SSIIIa^(wj) the opposite was true.The genotype SSIIa^(wj)SSIIIa^(mp) had the largest PKV,HPV and CPV,the genotype SSIIa^(wj)SSIIIa^(wj) had the largest BDV and CSV,but the genotype SSIIa^(wj)SSIIIa^(mp) had the least SBV.According to the comprehensive effect of each trait,the genotype SSIIawjSSIIIamp was the best.The allelic variation and interaction effect of SSIIa and SSIIIa genes have important reference value for improving cooking and eating quality of semi waxy japonica rice.
文摘Endogenous reference genes (ERGs) provide vital information regarding genetically modified organisms (GMOs). The successful detection of ERGs can identity GMOs and the source of genes, verify stability and reliability of the detection system, and calculate the level of genetically modified (GM) ingredients in mixtures. The reported ERGs in rice include sucrose-phosphate synthase (SPS), phospholipase D (PLD), RBE4 and rice root-specific GOS9 genes. Based on the characteristics of ERGs, a new ERG gene, phosphoenolpyruvate carboxylase (PEPC), was selected, and further compared with the four existing genes. A total of 18 rice varieties and 29 non-rice crops were used to verify the interspecies specificity, intraspecies consistency, sensitivity, stability and reliability of these five ERGs using qualitative and quantitative PCR. Qualitative detection indicated that SPS and PEPC displayed sufficient specificity, and the detection sensitivity was 0.05% and 0.005%, respectively. Although the specificity of both RBE4 and GOS9 were adequate, the amplicons were small and easily confused with primer dimers. Non-specific amplification of the PLD gene was present in maize and potato. Real-time quantitative PCR detection indicated that PLD, SPS and PEPC displayed good specificity, with R2 of the standard curve greater than 0.98, while the amplification efficiency ranged between 90% and 110%. Both the detection sensitivities of PLD and PEPC were five copies and that of SPS was ten copies. RBE4 showed typical amplification in maize, beet and Arabidopsis, while GOS9 was found in maize, tobacco and oats. PEPC exhibited excellent detection sensitivity and species specificity, which made it a potentially useful application in GM-rice supervision and administration. Additionally, SPS and PLD are also suitable for GM-rice detection. This study effectively established a foundation for GMO detection, which not only provides vital technical support for GMO identification, but also is of great significance for enhancing the comparability of detection results, and the standardization of ERG testing in GM-rice.
