The information of single nucleotide polymorphisms （SNPs） is quite unknown in sweetpotato. In this study, two sweetpotato varieties （Xushu 18 and Xu 781） were sequenced by Illumina technology, as well as de novo t...The information of single nucleotide polymorphisms （SNPs） is quite unknown in sweetpotato. In this study, two sweetpotato varieties （Xushu 18 and Xu 781） were sequenced by Illumina technology, as well as de novo transcriptome assembly, functional annotation, and in silico discovery of potential SNP molecular markers. Tetra-primer Amplification Refractory Mutation System PCR （ARMS-PCR） is a simple and sufficient method for detecting different alleles in SNP locus. Total 153 sets of ARMS-PCR primers were designed to validate the putative SNPs from sequences. PCR products from 103 sets of primers were different between Xu 781 and Xushu 18 via agarose gel electrophoresis, and the detection rate was 67.32%. We obtained the expected results from 32 sets of primers between the two genotypes. Furthermore, we ascertained the optimal annealing temperature of 32 sets of primers. These SNPs might be used in genotyping, QTL mapping, or marker-assisted trait selection further in sweetpotato. To our knowledge, this work was the first study to develop SNP markers in sweetpotato by using tetra-primer ARMS-PCR technique. This method was a simple, rapid, and useful techn!que to develop SNP markers, and will provide a potential and preliminary application in discriminating cultivars in sweetpotato.展开更多
Soil salinity causes the negative effects on the growth and yield of crops. In this study, two sweet potato （Ipomoea batatas L.） cultivars, Xushu 28 （X-28） and Okinawa 100 （O-100）, were examined under 50 and 100...Soil salinity causes the negative effects on the growth and yield of crops. In this study, two sweet potato （Ipomoea batatas L.） cultivars, Xushu 28 （X-28） and Okinawa 100 （O-100）, were examined under 50 and 100 mmol L-1 NaCI stress. X-28 cultivar is relatively high salt tolerant than O-100 cultivar. Interestingly, real-time quantitative polymerase chain reaction （RT-qPCR） results indicated that sweet potato high-affinity K^＋ transporter 1 （IbHKT1） gene expression was highly induced by 50 and 100 mmol L-1 NaCI stress in the stems of X-28 cultivar than in those of O-100 cultivar, but only slightly induced by these stresses in the leaves and fibrous roots in both cultivars. To characterize the function of IbHKT1 transporter, we performed ion-flux analysis in tobacco transient system and yeast complementation. Tobacco transient assay showed that IbHKT1 could uptake sodium （Na^＋）. Yeast complementation assay showed that IbHKT1 could take up K^＋ in 50 mmol L^-1 K^＋ medium without the presence of NaCI. Moreover, Na^＋ uptake significantly increased in yeast overexpressing IbHKTI. These results showed that IbHKT1 transporter could have K^＋-Na^＋ symport function in yeast. Therefore, the modes of action of IbHKT1 in transgenic yeast could differ from the mode of action of the other HKT1 transporters in class I. Potentially, IbHKT1 could be used to improve the salt tolerance nature in sweet potato.展开更多
As a critical food crop,sweetpotato(Ipomoea batatas(L.)Lam.)is widely planted all over the world,but it is deeply affected by Sweetpotato Virus Disease(SPVD).The present study utilized short tandem target mimic(STTM)t...As a critical food crop,sweetpotato(Ipomoea batatas(L.)Lam.)is widely planted all over the world,but it is deeply affected by Sweetpotato Virus Disease(SPVD).The present study utilized short tandem target mimic(STTM)technology to effectively up-regulate the expression of laccase(Ib LACs)by successfully inhibiting the expression of mi R397.The upstream genes in the lignin synthesis pathway were widely up-regulated by feedback regulation,including phenylalanine ammonialyase(PAL),4-coumarate-Co Aligase(4 CL),hydroxycinnamoyl Co A:shikimatetransferase(HTC),caffeicacid O-methyltransferase(COMT),and cinnamyl alcohol dehydrogenase(CAD).Meanwhile,the activities of PAL and LAC increased significantly,finally leading to increased lignin content.Lignin deposition in the cell wall increased the physical defence ability of transgenic sweetpotato plants,reduced the accumulation of SPVD transmitted by Bemisia tabaci(Gennadius),and promoted healthy sweetpotato growth.The results provide new insights for disease resistance breeding and green production of sweetpotato.展开更多
基金supported by the China Agriculture Research System (CARS-11)the National High-Tech R&D Program of China (2012AA101204)the Jiangsu Independent Inno vation Funds of Agriculture,China (CX(13)2032)
文摘The information of single nucleotide polymorphisms （SNPs） is quite unknown in sweetpotato. In this study, two sweetpotato varieties （Xushu 18 and Xu 781） were sequenced by Illumina technology, as well as de novo transcriptome assembly, functional annotation, and in silico discovery of potential SNP molecular markers. Tetra-primer Amplification Refractory Mutation System PCR （ARMS-PCR） is a simple and sufficient method for detecting different alleles in SNP locus. Total 153 sets of ARMS-PCR primers were designed to validate the putative SNPs from sequences. PCR products from 103 sets of primers were different between Xu 781 and Xushu 18 via agarose gel electrophoresis, and the detection rate was 67.32%. We obtained the expected results from 32 sets of primers between the two genotypes. Furthermore, we ascertained the optimal annealing temperature of 32 sets of primers. These SNPs might be used in genotyping, QTL mapping, or marker-assisted trait selection further in sweetpotato. To our knowledge, this work was the first study to develop SNP markers in sweetpotato by using tetra-primer ARMS-PCR technique. This method was a simple, rapid, and useful techn!que to develop SNP markers, and will provide a potential and preliminary application in discriminating cultivars in sweetpotato.
基金supported by the China Agriculture Research System (CARS-10,Sweetpotato)the Agricultural Science and Technology Innovation Program of Jiangsu Province,China (CX(13)2032)the China-Korea Young Scientist Exchange Program
文摘Soil salinity causes the negative effects on the growth and yield of crops. In this study, two sweet potato （Ipomoea batatas L.） cultivars, Xushu 28 （X-28） and Okinawa 100 （O-100）, were examined under 50 and 100 mmol L-1 NaCI stress. X-28 cultivar is relatively high salt tolerant than O-100 cultivar. Interestingly, real-time quantitative polymerase chain reaction （RT-qPCR） results indicated that sweet potato high-affinity K^＋ transporter 1 （IbHKT1） gene expression was highly induced by 50 and 100 mmol L-1 NaCI stress in the stems of X-28 cultivar than in those of O-100 cultivar, but only slightly induced by these stresses in the leaves and fibrous roots in both cultivars. To characterize the function of IbHKT1 transporter, we performed ion-flux analysis in tobacco transient system and yeast complementation. Tobacco transient assay showed that IbHKT1 could uptake sodium （Na^＋）. Yeast complementation assay showed that IbHKT1 could take up K^＋ in 50 mmol L^-1 K^＋ medium without the presence of NaCI. Moreover, Na^＋ uptake significantly increased in yeast overexpressing IbHKTI. These results showed that IbHKT1 transporter could have K^＋-Na^＋ symport function in yeast. Therefore, the modes of action of IbHKT1 in transgenic yeast could differ from the mode of action of the other HKT1 transporters in class I. Potentially, IbHKT1 could be used to improve the salt tolerance nature in sweet potato.
基金financially supported by the National Key R&D Program of China (2019YFD1001300 and 2019YFD1001305)the earmarked fund for CARS-10-Sweetpotatothe Jiangsu Postgraduate Scientific Research and Practical Innovation Program Project, China (KYCX19-2207)
文摘As a critical food crop,sweetpotato(Ipomoea batatas(L.)Lam.)is widely planted all over the world,but it is deeply affected by Sweetpotato Virus Disease(SPVD).The present study utilized short tandem target mimic(STTM)technology to effectively up-regulate the expression of laccase(Ib LACs)by successfully inhibiting the expression of mi R397.The upstream genes in the lignin synthesis pathway were widely up-regulated by feedback regulation,including phenylalanine ammonialyase(PAL),4-coumarate-Co Aligase(4 CL),hydroxycinnamoyl Co A:shikimatetransferase(HTC),caffeicacid O-methyltransferase(COMT),and cinnamyl alcohol dehydrogenase(CAD).Meanwhile,the activities of PAL and LAC increased significantly,finally leading to increased lignin content.Lignin deposition in the cell wall increased the physical defence ability of transgenic sweetpotato plants,reduced the accumulation of SPVD transmitted by Bemisia tabaci(Gennadius),and promoted healthy sweetpotato growth.The results provide new insights for disease resistance breeding and green production of sweetpotato.
