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Inhibition of miR397 by STTM technology to increase sweetpotato resistance to SPVD
LI Chen, LIU Xuan-xuan, ABOUELNASR Hesham, MOHAMED HAMED Arisha, KOU Meng, TANG Wei, YAN Hui, WANG Xin, WANG Xiao-xiao, ZHANG Yun-gang, LIU Ya-ju, GAO Run-fei, MA Meng, LI Qiang
2022, 21 (10): 2865-2875.   DOI: 10.1016/j.jia.2022.07.054
Abstract203)      PDF in ScienceDirect      

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 (IbLACs) by successfully inhibiting the expression of miR397.  The upstream genes in the lignin synthesis pathway were widely up-regulated by feedback regulation, including phenylalanine ammonialyase (PAL), 4-coumarate-CoAligase (4CL), hydroxycinnamoyl CoA: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. 

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Ipomoea batatas HKT1 transporter homolog mediates K+ and Na+ uptake in Saccharomyces cerevisiae
PARK Sung-chul, YU Yi-cheng, KOU Meng, YAN Hui, TANG Wei, WANG Xin, LIU Ya-ju, ZHANG Yun-gang, KWAK Sang-soo, MA Dai-fu, SUN Jian, LI Qiang
2017, 16 (10): 2168-2176.   DOI: 10.1016/S2095-3119(16)61570-8
Abstract738)      PDF in ScienceDirect      
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 NaCl 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 NaCl 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 NaCl. Moreover, Na+ uptake significantly increased in yeast overexpressing IbHKT1. 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.
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Development of SNP markers using RNA-seq technology and tetra-primer ARMS-PCR in sweetpotato
KOU Meng, XU Jia-lei, LI Qiang, LIU Ya-ju, WANG Xin, TANG Wei, YAN Hui, ZHANG Yun-gang, MA Dai-fu
2017, 16 (02): 464-470.   DOI: 10.1016/S2095-3119(16)61405-3
Abstract1224)      PDF in ScienceDirect      
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 technique to develop SNP markers, and will provide a potential and preliminary application in discriminating cultivars in sweetpotato.
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