Pepper （Capsicum annuum. L.） is a widely cultivated vegetable crop worldwide and has the second largest planting area and the first largest vegetable output and value in China. Pepper root-knot nematode （Meloidogyn...Pepper （Capsicum annuum. L.） is a widely cultivated vegetable crop worldwide and has the second largest planting area and the first largest vegetable output and value in China. Pepper root-knot nematode （Meloidogyne spp.） is one of the most serious pests of pepper, which caused huge losses every year. Previous studies showed that the Me3 gene is resistant to a wide range of Meloidogyne species, including M. arenaria, M. javanica, and M. incognita. HDA149, a double haploid pepper genotype, harboring the root-knot nematode resistance gene Me3, was used to construct bacterial artificial chro- mosome library （BAC） via the vector of CopyControFM pCC1 in this study. The library consists of 210 200 BAC clones and is equivalent to 5.3 pepper genomes. The average insert size is 95 kb, and most of them are 90-120 kb; but the empty clones are less than 3%. In order to screen the BAC library easily, 550 super pools with 384 BAC clones of each pool were further developed in this study. Specific primers from Me3 gene locus were used for BAC library screening, and more than 20 positive BAC clones were obtained. Then the selected positive BAC clones were analyzed by restriction enzyme digestion, BAC-end sequencing, marker development, and new positive BAC clones exploration, respectively. Finally, the contig with total length of about 300 kb linked to the Me3 locus was constructed based on chromosome walking strategy, which made a solid foundation for the cloning of the important root-knot nematode resistance gene Me3.展开更多
A full-length cDNA of proteinase inhibitor gene with completed open reading frame of 116 amino acids was cloned from Ralstonia solanacearum （Rs） resistant potato leaves using the rapid amplification of cDNA ends （R...A full-length cDNA of proteinase inhibitor gene with completed open reading frame of 116 amino acids was cloned from Ralstonia solanacearum （Rs） resistant potato leaves using the rapid amplification of cDNA ends （RACE） method and designated as StPI. BLAST search against NCBI showed that the StPI gene shared 89% identity with potato proteinase inhibitor I precursor in nucleotide and 74% in amino acid. Analysis of semi-quantitative RT-PCR indicated that this gene was induced by Rs as well as up-regulated by jasmonic acid （JA）. The StPI gene expression reached the highest level during 6-12 h post Rs-inoculation or JA-treatment, and then leveled off. Moreover, this gene was strongly induced by JA and its mRNA accumulation increased more quickly than that of Rs-inoculation. The StPI gene may play a role in potato resistance against Rs. The induction of StPI by Rs invasion may have a similar signal transduction pathway with JA treatment.展开更多
Salt stress seriously restricts the growth and yield of potatoes.Plant cystatins are vital players in biotic stress and development,however,their roles in salt stress resistance remain elusive.Here,we report that StCY...Salt stress seriously restricts the growth and yield of potatoes.Plant cystatins are vital players in biotic stress and development,however,their roles in salt stress resistance remain elusive.Here,we report that StCYS1 positively regulates salt tolerance in potato plants.An in vitro biochemical test demonstrated that StCYS1 is a bona fide cystatin.Overexpression of StCYS1 in both Escherichia coli and potato plants significantly increased their resistance to high salinity.Further analysis revealed that the transgenic plants accumulated more proline and chlorophyll under salt stress conditions.Moreover,the transgenic plants displayed higher H2O2 scavenging capability and cell membrane integrity compared with wild-type potato.These results demonstrate that StCYS1 is closely correlated with salt stress and its overaccumulation can substantially enhance salt stress resistance.展开更多
基金supported by the National High-Tech R&D Program in China (2013AA102603)the Natural Science Foundation of Shandong Province,China (ZR2014YL014)+3 种基金the Youth Scientific Research Foundation of Shandong Academy of Agricultural Sciences,China (2014QNZ03)the Taishan Scholars Program of Shandong Province,China (2016-2020)the National Natural Science Foundation of China (31101425)Prof. Alain Palloxin,French National Institute for Agricultural Research (INRA),for kindly providing the pepper genotype HDA149
文摘Pepper （Capsicum annuum. L.） is a widely cultivated vegetable crop worldwide and has the second largest planting area and the first largest vegetable output and value in China. Pepper root-knot nematode （Meloidogyne spp.） is one of the most serious pests of pepper, which caused huge losses every year. Previous studies showed that the Me3 gene is resistant to a wide range of Meloidogyne species, including M. arenaria, M. javanica, and M. incognita. HDA149, a double haploid pepper genotype, harboring the root-knot nematode resistance gene Me3, was used to construct bacterial artificial chro- mosome library （BAC） via the vector of CopyControFM pCC1 in this study. The library consists of 210 200 BAC clones and is equivalent to 5.3 pepper genomes. The average insert size is 95 kb, and most of them are 90-120 kb; but the empty clones are less than 3%. In order to screen the BAC library easily, 550 super pools with 384 BAC clones of each pool were further developed in this study. Specific primers from Me3 gene locus were used for BAC library screening, and more than 20 positive BAC clones were obtained. Then the selected positive BAC clones were analyzed by restriction enzyme digestion, BAC-end sequencing, marker development, and new positive BAC clones exploration, respectively. Finally, the contig with total length of about 300 kb linked to the Me3 locus was constructed based on chromosome walking strategy, which made a solid foundation for the cloning of the important root-knot nematode resistance gene Me3.
文摘A full-length cDNA of proteinase inhibitor gene with completed open reading frame of 116 amino acids was cloned from Ralstonia solanacearum （Rs） resistant potato leaves using the rapid amplification of cDNA ends （RACE） method and designated as StPI. BLAST search against NCBI showed that the StPI gene shared 89% identity with potato proteinase inhibitor I precursor in nucleotide and 74% in amino acid. Analysis of semi-quantitative RT-PCR indicated that this gene was induced by Rs as well as up-regulated by jasmonic acid （JA）. The StPI gene expression reached the highest level during 6-12 h post Rs-inoculation or JA-treatment, and then leveled off. Moreover, this gene was strongly induced by JA and its mRNA accumulation increased more quickly than that of Rs-inoculation. The StPI gene may play a role in potato resistance against Rs. The induction of StPI by Rs invasion may have a similar signal transduction pathway with JA treatment.
基金We thank Li Guangcun(Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences)for providing the potato(Solanum tuberosumL.)cultivar MDS.This work was supported by the National Natural Science Foundation of China(31901752)by a grant from the Potato Industry lnnovation Team for Modern Agricultural Industry Technology System,Shandong Province,China(SDAIT-10-011-11).
文摘Salt stress seriously restricts the growth and yield of potatoes.Plant cystatins are vital players in biotic stress and development,however,their roles in salt stress resistance remain elusive.Here,we report that StCYS1 positively regulates salt tolerance in potato plants.An in vitro biochemical test demonstrated that StCYS1 is a bona fide cystatin.Overexpression of StCYS1 in both Escherichia coli and potato plants significantly increased their resistance to high salinity.Further analysis revealed that the transgenic plants accumulated more proline and chlorophyll under salt stress conditions.Moreover,the transgenic plants displayed higher H2O2 scavenging capability and cell membrane integrity compared with wild-type potato.These results demonstrate that StCYS1 is closely correlated with salt stress and its overaccumulation can substantially enhance salt stress resistance.
