Salinity is one of the most important abiotic stresses that adversely affects crop growth and productivity. A subtractive suppression hybridization （SSH） library were constructed from the roots of salt-sensitive Yul...Salinity is one of the most important abiotic stresses that adversely affects crop growth and productivity. A subtractive suppression hybridization （SSH） library were constructed from the roots of salt-sensitive Yulu cultivar melon seedlings under salt stress; 557 high-quality expressed sequence tags （ESTs） were randomly sequenced, with an average size of 428 bp, which assembled into 68 contigs and 315 singletons. Compared with our previous SSH library generated from the salt-tolerant Bingxuecui cultivar, the proportion of transcripts involved in metabolism, protein fate, cellular communication/signal transduction mechanisms, and cell rescue/defense were 4, 1.46, 0.94, and 0.4% higher, respectively, in the salt-tolerant cultivar than the in salt-sensitive cultivar. Quantitative real-time PCR analysis of eleven transcripts revealed temporal variations in their expression in the two cultivars under salt stress. One NAC gene （JZ477011） was heterologously expressed in yeast for functional characterization, and enhanced the sensitivity of yeast cells to high-salinity to salt stress and inhibited their growth. Information regards to their functions would aid in the understanding of response mechanisms to saline stress and in the development of molecular markers for selecting salt-tolerant melon cultivars.展开更多
Melon downy mildew caused by Pseudoperonospora cubensis leads to significant losses in melon yields worldwide. Reverse-transcription Polymerase Chain Reaction （RT-PCR） was performed using cDNAs as templates from mel...Melon downy mildew caused by Pseudoperonospora cubensis leads to significant losses in melon yields worldwide. Reverse-transcription Polymerase Chain Reaction （RT-PCR） was performed using cDNAs as templates from melon-Huangdanzi induced with fungus Pseudoperonospora cubensis, and degenerate primers designed based on the conserved amino acid sequences of known plant disease-resistance genes. A polymorphic cDNA fragment which we named rap-19 was cloned and sequenced. The Open Reading Frame （ORF） of this product comprised of 510 base pairs which encodes DNA or RNA-binding protein with 170 amino acids. The putative amino acid sequence of mp-19 appeared highly homologous with those of NBS-type resistant-genes isolated from other plants. Southern blot indicated that the melon genome contained more than 3 copies of rap-19. The obvious expression differences detected by semi-quantitative RT- PCR could be observed between resistant-line Huangdanzi and susceptible-line Jiashi after Pseudoperonospora cubensis infection, which implied that mp-19 gene may be related to the resistance of downy mildew in melon.展开更多
Aphis gossypfi is a globally distributed species and therefore has a highly variable life cycle. Populations of A. gossypii in northern China exhibit greater genotypic diversity and a broader host range, yet the detai...Aphis gossypfi is a globally distributed species and therefore has a highly variable life cycle. Populations of A. gossypii in northern China exhibit greater genotypic diversity and a broader host range, yet the details of life cycles of different biotypes is still unclear. In this study, the Cytb and 16S gene regions of A. gossypfi collected from 5 common summer hosts and 4 primary hosts were analyzed. A total of 57 haplotypes were obtained from 1 046 individual A. gossyp# sequences. The sequence included 44 variable sites, 27 of which were parsimony informative sites and 17 of which were singleton variable sites. The most frequent 3 haplotypes were found in 896 individuals, representing a total of 85.7% of all individuals and 36 haplotypes were found in 1 individual. A neighbor-joining tree was constructed using 21 haplotypes that were found in more than 2 individuals. Considering the individual host plant, 5 biotypes were identified. Type 1 corresponded exactly to the cucurbit host-race and the other 4 biotypes were found as cotton host-races. Type 3 was the most abundant biotype in cotton fields in northern China.展开更多
Lipoxygenases（LOXs） are a group of non-heme,iron-containing enzymes and extensively involved in plant growth and development,ripening and senescence,stress responses,biosynthesis of regulatory molecules and defense ...Lipoxygenases（LOXs） are a group of non-heme,iron-containing enzymes and extensively involved in plant growth and development,ripening and senescence,stress responses,biosynthesis of regulatory molecules and defense reaction.In our previous study,18 LOXs in melon genome were screened and identified,and five 13-LOX genes（CmLOX08,CmLOX10,CmLOX12,CmLOX13 and CmLOX18） were predicted to locate in chloroplast.Phylogenetic analysis result showed that the five genes have high homology with jasmonic acid（JA） biosynthesis-related LOXs from other plants.In addition,promoter analysis revealed that motifs of the five genes participate in gene expression regulated by hormones and stresses.Therefore,we analyzed the expressions of the five genes and LOX activity in leaves of four-leaf stage seedlings of oriental melon cultivar Yumeiren under abiotic stress：wounding,cold,high temperature and hydrogen peroxide（H_2O_2）,and signal molecule treatments：methyl jasmonate（MeJA）,abscisic acid（ABA） and salicylic acid（SA）.Real time qPCR revealed that wounding and H_2O_2 induced the expressions of all the five genes.Only CmLOX08 was induced by cold while only CmLOX13 was suppressed by high temperature.ABA induced the expressions of CmLOXIO and CmLOX12 while inhibited CmLOX13 and CmLOX18.MeJA increased the 3 genes expressions except CmLOX08 and CmLOX13,whereas SA decreased the effect,apart from CmLOX12.All the abiotic stresses and signal molecules treatments increased the LOX activity in leaves of oriental melon.In summary,the results suggest that the five genes have diverse functions in abiotic stress and hormone responses,and might participate in defense response.The data generated in this study will be helpful in subcellular localization and transgenic experiment to understand their precise roles in plant defense response.展开更多
基金supported by grants from the National Natural Science Foundation of China(31372079)the National High Technology Research and Development Program of China(863 Program,2012AA101903)+1 种基金the Natural Science Foundation of Shanghai Science and Technology Committee,China(13ZR1422400)Shanghi Graduate Education and Innovation Program(Horticulture),China
文摘Salinity is one of the most important abiotic stresses that adversely affects crop growth and productivity. A subtractive suppression hybridization （SSH） library were constructed from the roots of salt-sensitive Yulu cultivar melon seedlings under salt stress; 557 high-quality expressed sequence tags （ESTs） were randomly sequenced, with an average size of 428 bp, which assembled into 68 contigs and 315 singletons. Compared with our previous SSH library generated from the salt-tolerant Bingxuecui cultivar, the proportion of transcripts involved in metabolism, protein fate, cellular communication/signal transduction mechanisms, and cell rescue/defense were 4, 1.46, 0.94, and 0.4% higher, respectively, in the salt-tolerant cultivar than the in salt-sensitive cultivar. Quantitative real-time PCR analysis of eleven transcripts revealed temporal variations in their expression in the two cultivars under salt stress. One NAC gene （JZ477011） was heterologously expressed in yeast for functional characterization, and enhanced the sensitivity of yeast cells to high-salinity to salt stress and inhibited their growth. Information regards to their functions would aid in the understanding of response mechanisms to saline stress and in the development of molecular markers for selecting salt-tolerant melon cultivars.
基金This study was supported by the Natural Science Foun-dation of Xinjiang Uygur Autonomous Region, China(200121105)Science and Research Plan Emphasis Project of College in Xinjiang Uygur Autonomous Region, China (XJEDU2004I09).
文摘Melon downy mildew caused by Pseudoperonospora cubensis leads to significant losses in melon yields worldwide. Reverse-transcription Polymerase Chain Reaction （RT-PCR） was performed using cDNAs as templates from melon-Huangdanzi induced with fungus Pseudoperonospora cubensis, and degenerate primers designed based on the conserved amino acid sequences of known plant disease-resistance genes. A polymorphic cDNA fragment which we named rap-19 was cloned and sequenced. The Open Reading Frame （ORF） of this product comprised of 510 base pairs which encodes DNA or RNA-binding protein with 170 amino acids. The putative amino acid sequence of mp-19 appeared highly homologous with those of NBS-type resistant-genes isolated from other plants. Southern blot indicated that the melon genome contained more than 3 copies of rap-19. The obvious expression differences detected by semi-quantitative RT- PCR could be observed between resistant-line Huangdanzi and susceptible-line Jiashi after Pseudoperonospora cubensis infection, which implied that mp-19 gene may be related to the resistance of downy mildew in melon.
基金supported by the National Natural Science Foundation of China(31572015)the National Special Transgenic Project of China(2016ZX08012-004)
文摘Aphis gossypfi is a globally distributed species and therefore has a highly variable life cycle. Populations of A. gossypii in northern China exhibit greater genotypic diversity and a broader host range, yet the details of life cycles of different biotypes is still unclear. In this study, the Cytb and 16S gene regions of A. gossypfi collected from 5 common summer hosts and 4 primary hosts were analyzed. A total of 57 haplotypes were obtained from 1 046 individual A. gossyp# sequences. The sequence included 44 variable sites, 27 of which were parsimony informative sites and 17 of which were singleton variable sites. The most frequent 3 haplotypes were found in 896 individuals, representing a total of 85.7% of all individuals and 36 haplotypes were found in 1 individual. A neighbor-joining tree was constructed using 21 haplotypes that were found in more than 2 individuals. Considering the individual host plant, 5 biotypes were identified. Type 1 corresponded exactly to the cucurbit host-race and the other 4 biotypes were found as cotton host-races. Type 3 was the most abundant biotype in cotton fields in northern China.
基金financially supported by the National Natural Science Foundation of China(31272154)
文摘Lipoxygenases（LOXs） are a group of non-heme,iron-containing enzymes and extensively involved in plant growth and development,ripening and senescence,stress responses,biosynthesis of regulatory molecules and defense reaction.In our previous study,18 LOXs in melon genome were screened and identified,and five 13-LOX genes（CmLOX08,CmLOX10,CmLOX12,CmLOX13 and CmLOX18） were predicted to locate in chloroplast.Phylogenetic analysis result showed that the five genes have high homology with jasmonic acid（JA） biosynthesis-related LOXs from other plants.In addition,promoter analysis revealed that motifs of the five genes participate in gene expression regulated by hormones and stresses.Therefore,we analyzed the expressions of the five genes and LOX activity in leaves of four-leaf stage seedlings of oriental melon cultivar Yumeiren under abiotic stress：wounding,cold,high temperature and hydrogen peroxide（H_2O_2）,and signal molecule treatments：methyl jasmonate（MeJA）,abscisic acid（ABA） and salicylic acid（SA）.Real time qPCR revealed that wounding and H_2O_2 induced the expressions of all the five genes.Only CmLOX08 was induced by cold while only CmLOX13 was suppressed by high temperature.ABA induced the expressions of CmLOXIO and CmLOX12 while inhibited CmLOX13 and CmLOX18.MeJA increased the 3 genes expressions except CmLOX08 and CmLOX13,whereas SA decreased the effect,apart from CmLOX12.All the abiotic stresses and signal molecules treatments increased the LOX activity in leaves of oriental melon.In summary,the results suggest that the five genes have diverse functions in abiotic stress and hormone responses,and might participate in defense response.The data generated in this study will be helpful in subcellular localization and transgenic experiment to understand their precise roles in plant defense response.
