Powdery mildew, caused by the biotrophic fungus Blumeria graminis f. sp. tritici(Bgt), is a global disease that poses a serious threat to wheat production. To explore additional resistance gene, a wheatDasypyrum villo...Powdery mildew, caused by the biotrophic fungus Blumeria graminis f. sp. tritici(Bgt), is a global disease that poses a serious threat to wheat production. To explore additional resistance gene, a wheatDasypyrum villosum 1 V#5(1 D) disomic substitution line NAU1813(2 n = 42) with high level of seedling resistance to powdery mildew was used to generate the recombination between chromosomes 1 V#5 and1 D. Four introgression lines, including t1 VS#5 ditelosomic addition line NAU1815, t1 VL#5 ditelosomic addition line NAU1816, homozygous T1 DL·1 VS#5 translocation line NAU1817, and homozygous T1 DS·1 VL#5 translocation line NAU1818 were developed from the selfing progenies of 1 V#5 and 1 D double monosomic line that derived from F1 hybrids of NAU1813/NAU0686. All of them were characterized by fluorescence in situ hybridization, genomic in situ hybridization, 1 V-specific markers analysis, and powdery mildew tests at different developmental stages. A new powdery mildew resistance gene named Pm67 was physically located in the terminal bin(FL 0.70–1.00) of 1 VS#5. Lines with Pm67 exhibited seedling stage immunity and tissue-differentiated reactions at adult plant stage. The sheaths, stems, and spikes of the Pm67 line were still immune, but the leaves showed a low degree of susceptibility.Microscopic observation showed that most penetration attempts were stopped in association with papillae on the sheath, and colonies cannot form conidia on the susceptible leaf of Pm67 line at adult plant stage, suggesting that the defence layers of the Pm67 line is tissue-differentiated. Thus, the T1 DL·1 VS#5 translocation line NAU1817 provides a new germplasm in wheat breeding for improvement of powdery mildew resistance.展开更多
Dasypyrum villosum is one of the most valuable gene resources in wheat improvement,especially for disease resistance.The mining of favorable genes from D.villosum is frustrated by the lack of a whole genome sequence.I...Dasypyrum villosum is one of the most valuable gene resources in wheat improvement,especially for disease resistance.The mining of favorable genes from D.villosum is frustrated by the lack of a whole genome sequence.In this study,we generated a doubled-haploid line,91C43^(DH),using microspore culture and obtained a 4.05-GB high-quality,chromosome-scale genome assembly for D.villosum.The assembly contains39727 high-confidence genes,and 85.31% of the sequences are repetitive.Two reciprocal translocation events were detected,and 7VS-4VL is a unique translocation in D.villosum.The prolamin seed storage protein-coding genes were found to be duplicated;in particular,the genes encoding low-molecular-weight glutenin at the Glu-V3 locus were significantly expanded.RNA sequencing(RNA-seq)analysis indicated that,after Blumeria graminearum f.sp tritici(Bgt)inoculation,there were more upregulated genes involved in the pattern-triggered immunity and effector-triggered immunity defense pathways in D.villosum than in Triticum urartu.MNase hypersensitive sequencing(MH-seq)identified two Bgt-inducible MH sites(MHSs),one in the promoter and one in the 3'terminal region of the powdery mildew resistance(Pm)gene NLR1-V.Each site had two subpeaks and they were termed MHS1(MHS1.1/1.2)and MHS2(MHS2.1/2.2).Bgt-inducible MHS2.2 was uniquely present in D.villosum,and MHS1.1 was more inducible in D.villosum than in wheat,suggesting that MHSs may be critical for regulation of NLR1-V expression and plant defense.In summary,this study provides a valuable genome resource for functional genomics studies and wheat-D.villosum introgression breeding.The identified regulatory mechanisms may also be exploited to develop new strategies for enhancing Pm resistance by optimizing gene expression in wheat.展开更多
OBJECTIVE:This study was conducted to evaluate the effect of Amomum villosum on longitudinal bone growth.METHODS:Adolescent female Sprague-Dawley rats were divided into 3 groups and treated for 4 days:control(distille...OBJECTIVE:This study was conducted to evaluate the effect of Amomum villosum on longitudinal bone growth.METHODS:Adolescent female Sprague-Dawley rats were divided into 3 groups and treated for 4 days:control(distilled water,p.o.),recombinant human growth hormone(rhGH;100 μg/kg,s.c.),and A.villosum(500 mg/kg,p.o.) groups.On day 3,tetracycline(20 g/kg,i.p.) was injected for growth plate identification.On days 2,3 and 4,5-bromo-2'-deoxyuridine(BrdU)(50 mg/kg,i.p.) was injected to label proliferating cells.On day 5,tibias were dissected and fixed in 4% paraformaldehyde,dehydrated,and sectioned for immunohistochemistry and histomorphometry.RESULTS:The rate of bone growth in the A.villosum and rhGH groups increased to(410 ± 44) and(389 ± 46) μm/day(P<0.01),respectively,as compared with the control(330.7 ± 34.7) μm/day.The thickness of the growth plates also increased to(591 ± 37) and(598 ± 32) μm,respectively,as compared with the control(524± 89) μm(P<0.001).The number of BrdU-positive cells in the chondrocytes of the A.villosum and rhGH groups was also significantly higher(126±24) and(143±18) cells/mm 2,respectively) than in the control(109 ± 25) mm 2(P<0.05).Insulin-like growth factor-1 and bone morphogenetic protein-2 in the A.villosum and rhGH groups were highly expressed in the growth plate as compared with the control samples,indicating increased bone formation.CONCLUSIONS:A.villosum could be used to treat growth retardation during adolescence.展开更多
Wheat sharp eyespot, a stem disease caused by the soilborne fungus Rhizoctonia cerealis van der Hoeven,has become a threat to wheat production worldwide. Exploiting resistance resources from wild relatives of wheat is...Wheat sharp eyespot, a stem disease caused by the soilborne fungus Rhizoctonia cerealis van der Hoeven,has become a threat to wheat production worldwide. Exploiting resistance resources from wild relatives of wheat is a promising strategy for controlling this disease. In this study, a new wheat–Dasypyrum villosum T2DS·2V#4L translocation line in the background of Chinese Spring(CS) showed stable resistance to R. cerealis. Introgression of the T2DS·2V#4L chromosome into wheat cultivar Aikang 58 by backcrossing produced a marked increase in sharp eyespot resistance in NIL-T2DS·2V#4L in comparison with NILT2DS·2DL, and no detrimental effects of 2V#4L on agronomic traits were observed in the BC2F2, BC2F2:3,and BC2F2:4generations. Flow-sorted sequencing of 2V#4L yielded 384.3 Mb of assembled sequence, and8836 genes were predicted of which 6154 had orthologs in at least one of the 2AL, 2BL, and 2DL arms of CS, whereas 1549 genes were unique to 2V#4L. About 100,000 SNPs were detected in genes of 2V#4L and2DL in 10 sequenced bread wheat cultivars. A Kompetitive Allele Specific Polymerase chain reaction and30 conserved ortholog sequence markers were developed to trace the 2V#4L chromatin in wheat backgrounds. T2DS·2V#4L compensating translocation lines represent novel germplasm with sharp eyespot resistance and the markers will allow rapid detection in breeding programs.展开更多
Expressed sequence tags-derived polymerase chain reaction(EST-PCR) molecular markers specific for alien chromosomes can be used to not only monitor the introgressed alien chromatin in wheat background, but also provid...Expressed sequence tags-derived polymerase chain reaction(EST-PCR) molecular markers specific for alien chromosomes can be used to not only monitor the introgressed alien chromatin in wheat background, but also provide the evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers specific for chromosome 6 VL of Dasypyrum villosum, 297 primer pairs were designed based on the expressed sequence tags(EST) sequences, which were previously mapped in different bins of the long arms of wheat homoeologous 6 AL, 6 BL, and 6 DL. By using the Triticum aestivum, D. villosum, T. durum-D. villosum amphiploid, and T. aestivum-D. villosum alien chromosome lines involving chromosome 6 V, it was found that 32(10.77%) primers could amplify specific bands for chromosome 6 V, and 31 could be allocated to chromosome arm 6 VL. These 6 VL specific markers provided efficient tools for the characterization of structural variation involving the chromosome 6 VL in common wheat background as well as for the selection of useful genes located on 6 VL in breeding programs.展开更多
T6V#2S·6AL and T6V#4S·6DL translocation chromosomes developed from the cross of wheat and different Dasypyrum villosum accessions have good powdery mildew (PM) resistance, but their pairing and pyramiding ...T6V#2S·6AL and T6V#4S·6DL translocation chromosomes developed from the cross of wheat and different Dasypyrum villosum accessions have good powdery mildew (PM) resistance, but their pairing and pyramiding behavior remains unclear. Results in this study indicated that the pairing frequency rate of the two differently originated 6VS chromosomes in their F1 hybrid was 18.9% according to genomic in situ hybridization (GISH); the PM resistance plants in the F2 generation from the cross between T6V#4S·6DL translocation line Pm97033 and its PM susceptible wheat variety Wan7107 was fewer than expected. However, the ratio of the resistant vs. the susceptible plants of 15:1 in the F2 generation derived from the cross between the two translocation lines of T6V#2S·6AL and T6V#4S·6DL fitted well. Plants segregation ratio (homozygous:heterozygous:lacking) revealed by molecular marker for T6V#4S·6DL or T6V#2S·6AL in their F2 populations fitted the expected values of 1:2:1 well, inferring that the pairing of the two alien chromosome arms facilitates the transmission of T6V#4S·6DL from the F1 to the F2 generation. A quadrivalent was also observed in 21% of pollen mother cells (PMCs) of homozygote plants containing the two pairs of translocated chromosomes. The chromosome pairing between 6V#2S and 6V#4S indicates that it will be possible to obtain recombinants and clarify if the PM resistance determinant on one alien chromosome arm is different from that on the other.展开更多
Objective: The roots of two Thai medicinal plants, Clerodendrum indicum and Clerodendrum villosum are found in traditional medicine practices. The aim of this research was to preliminarily study the cytotox- icity of...Objective: The roots of two Thai medicinal plants, Clerodendrum indicum and Clerodendrum villosum are found in traditional medicine practices. The aim of this research was to preliminarily study the cytotox- icity of extracts of their roots, and the parts that possessed cytotoxic activity were separated on a chro- matograph to identify their active compounds. Methods: The extracts of both plants were screened for cytotoxicity on the SW620 cell line and the com- pounds isolated from the active extracts were further evaluated for their cytotoxic activity against five human cancer cell lines, including SW620, ChaGo-K-1, HepG2, I(ATO-III and BT-474 using the 3-(4,5-dime thyl-2-thiazolyl)-2,5-diphenyll2-H-tetrazolium bromide (MTI') assay. Results: Dichloromethane extracts of C indicum and C. villosum were active against the SW620 cell line. Triterpenoids were mostly obtained from the extracts of these plants (0.28% and 1.02%, respectively) and exhibited varying degrees of cytotoxicity and specificity against the tested cell lines. Two triterpenoids, oleanolic acid 3-acetate and betulinic acid, displayed moderate to strong cytotoxicity toward all cancer cell lines, with 50% inhibitory concentration (ICso) values of 1.66-20.49 mmol/L, whereas 3β-hydroxy- D:Blfriedo-olean-5-ene and taraxerol were cytotoxic to only the SW620 cell line (ICso= 23.39 and 2.09 mmol/L, respectively). Triterpenoid, lupeol, showed potent cytotoxicity on both SW620 (ICso= 1.99 mmol/L) and KATO-III cell lines (IC50 = 1.95 μmol/L), while a fiavonoid, pectolinarigenin, displayed moderate cytotoxicity against these cells (IC50= 13.05 and 24.31 μmol/L, respectively). Although the widely distributed steroid, stigmasterol, was effective against the SW620 cell line (1Cso = 2.79 μmol/L) and β-sitosterol was also active against SW620 (IC50 = 11.26 μmol/L), BT-474 (IC50 = 14.11 μmol/L) and HepG2 cancer cells (IC50=20.47μmol/L), none of the characteristic 24β-ethylsteroids of either Clerode展开更多
Objective:Amomum villosum(AV)is an herb whose dried fruit has been extensively used in modern medicine to treat digestive system diseases such as dysentery,vomiting and abdominal pain.This paper aims to supplement chl...Objective:Amomum villosum(AV)is an herb whose dried fruit has been extensively used in modern medicine to treat digestive system diseases such as dysentery,vomiting and abdominal pain.This paper aims to supplement chloroplast(cp)genomic resources and to be used in phylogenetic studies and identification of AV related plants.Methods:High-throughput sequencing technology was used to determine the complete sequence of the AV cp genome,and the sequence was then compared with three related species.Results:The genome size of AV we obtained was 163,968 bp with an obvious tetrad structure.The AV cp genome was observed to contain 125 unique genes and 81 simple sequence repeat(SSRs)had been determined and the majority of which were adenine–thymine(AT)-rich.Comparative analysis of genome sequence of four ginger plants showed that the atpF,clpP and rpl32 genes are potential markers for identifying Amomum species.Phylogenetic analysis suggested that AV was closely related to A.kravanh and A.compactum.Conclusion:These results have brought useful genetic resources for further identification researches,DNA barcoding,resolving taxonomy and understanding the evolutionary mode of Zingiberaceae cp genome.展开更多
Solanum nigrum complex comprises of plant species that belong to Solanum section Solanum used as indigenous leafy vegetables in Kenya among many communities. The Solanum nigrum complex species have also been used trad...Solanum nigrum complex comprises of plant species that belong to Solanum section Solanum used as indigenous leafy vegetables in Kenya among many communities. The Solanum nigrum complex species have also been used traditionally as medicine. Many species make up the Solanum nigrum complex, and some include S. scabrum, S. villosum, S. americanum, S. nigrum, S. americanum, and S. physalifolium. The purpose of this study was to determine the species ofSolanum nigrum complex found in Rift Valley, Nyanza and Western provinces of Kenya. Samples were collected from Eldoret-Iten, Kisumu-Nyando, Kisii-Nyamira, Nakuru, and Busia then pressed and sent to a plant taxonomist in Egerton University for classification and identification. It was established that the collected samples belonged to S. nigrum, S. scabrum, and S. villosurn species. The morphological characteristics used to distinguish the species included shape and colour of the stems, size and shape of the leaves, size, shape and colour of the berries, and the inflorescence. S. nigrurn was widely found in some regions with a noticeable bias towards S. scabrum that was preferred due to its large size of leaves and taste when cooked. However, the distribution and presence of S. nigrum and S. villosum in some of the study regions was poor.展开更多
Gibberellin 3-oxidase catalyzes the conversion of inactive gibberellin(GA) species into GAs with biological activity and it is subjected to strict developmental controls in the life cycle of a plant. In this study, 33...Gibberellin 3-oxidase catalyzes the conversion of inactive gibberellin(GA) species into GAs with biological activity and it is subjected to strict developmental controls in the life cycle of a plant. In this study, 33 gene sequences, encoding the gibberellin 3-oxidase(GA3ox) from Dasypyrum villosum and its dwarf mutant, were obtained. Each contained a 1 107 bp coding sequence(CDS) that encoded a putative protein containing 369 amino acids. The GA3ox protein showed 77% to 97% homology and shared the major conserved structural domains of GA3ox proteins with rice, sorghum bicolor, oat, barley, and wheat. Sequence alignment showed that there were 20 single nucleotide polymorphisms(SNPs) and 22 Insertion/deletions(In Dels) among these sequences, which could be divided into 2 haplotypes, haplotypes Ⅰ and Ⅱ. Haplotype Ⅰ was found in the wild type and was1 495 bp in length, and haplotype Ⅱ was found in the dwarf mutant and was 1 485 bp in length. The Q-PCR results showed that GA3ox was expressed in the leaves, roots, internodes, and stem nodes, and that there was a significant difference in the transcript level of the GA3ox between the wild type and dwarf mutant. The transcript levels of GA3ox in the leaves at the seedling stage, stem elongation stage and the heading stage, in the root and stem nodes at the stem elongation stage and in the internodes at the heading stage of the wild type, were significantly higher than those in the dwarf mutant. However, GA3ox expression in the rest of the wild type tissues at the 3 stages was slightly higher than or not different from the dwarf mutant.The results suggested that the wild type and mutant allele sequences of GA3ox in D. villosum showed 2 amino acid changes in exons and variations in the lengths of introns or the SNPs in introns, which most probably impaired the function of the enzyme,affected the GA3ox expression level, and eventually gave rise to dwarfing.展开更多
基金supported by the National Natural Science Foundation of China(31971938)the Natural Science Foundation of Jiangsu Province(BK20181316)+1 种基金the Special Fund for Independent Innovation of Agricultural Science and Technology in Jiangsu,China(CX(19)1001)Fundamental Research Funds for the Central Universities(KYZ201809)。
文摘Powdery mildew, caused by the biotrophic fungus Blumeria graminis f. sp. tritici(Bgt), is a global disease that poses a serious threat to wheat production. To explore additional resistance gene, a wheatDasypyrum villosum 1 V#5(1 D) disomic substitution line NAU1813(2 n = 42) with high level of seedling resistance to powdery mildew was used to generate the recombination between chromosomes 1 V#5 and1 D. Four introgression lines, including t1 VS#5 ditelosomic addition line NAU1815, t1 VL#5 ditelosomic addition line NAU1816, homozygous T1 DL·1 VS#5 translocation line NAU1817, and homozygous T1 DS·1 VL#5 translocation line NAU1818 were developed from the selfing progenies of 1 V#5 and 1 D double monosomic line that derived from F1 hybrids of NAU1813/NAU0686. All of them were characterized by fluorescence in situ hybridization, genomic in situ hybridization, 1 V-specific markers analysis, and powdery mildew tests at different developmental stages. A new powdery mildew resistance gene named Pm67 was physically located in the terminal bin(FL 0.70–1.00) of 1 VS#5. Lines with Pm67 exhibited seedling stage immunity and tissue-differentiated reactions at adult plant stage. The sheaths, stems, and spikes of the Pm67 line were still immune, but the leaves showed a low degree of susceptibility.Microscopic observation showed that most penetration attempts were stopped in association with papillae on the sheath, and colonies cannot form conidia on the susceptible leaf of Pm67 line at adult plant stage, suggesting that the defence layers of the Pm67 line is tissue-differentiated. Thus, the T1 DL·1 VS#5 translocation line NAU1817 provides a new germplasm in wheat breeding for improvement of powdery mildew resistance.
基金financially supported by the National Key Agriculture Projects(NK2022060101)National Key Research and Development Program(2022YFF1002900,2020YFE0202900)+3 种基金the Fundamental Research Funds for the Central University(XUEKEN2022012)Jiangsu Provincial Key Research and Development Program(BE2021375,BE2022346)Seed Industry Revitalization Project of Jiangsu Province(JBGS2021006,2021013,2021047)Joint Research of Improved Wheat Variety of Anhui,and Jiangsu Agricultural Technology System(JATS)(JATS[2021]463,JATS[2022]464).
文摘Dasypyrum villosum is one of the most valuable gene resources in wheat improvement,especially for disease resistance.The mining of favorable genes from D.villosum is frustrated by the lack of a whole genome sequence.In this study,we generated a doubled-haploid line,91C43^(DH),using microspore culture and obtained a 4.05-GB high-quality,chromosome-scale genome assembly for D.villosum.The assembly contains39727 high-confidence genes,and 85.31% of the sequences are repetitive.Two reciprocal translocation events were detected,and 7VS-4VL is a unique translocation in D.villosum.The prolamin seed storage protein-coding genes were found to be duplicated;in particular,the genes encoding low-molecular-weight glutenin at the Glu-V3 locus were significantly expanded.RNA sequencing(RNA-seq)analysis indicated that,after Blumeria graminearum f.sp tritici(Bgt)inoculation,there were more upregulated genes involved in the pattern-triggered immunity and effector-triggered immunity defense pathways in D.villosum than in Triticum urartu.MNase hypersensitive sequencing(MH-seq)identified two Bgt-inducible MH sites(MHSs),one in the promoter and one in the 3'terminal region of the powdery mildew resistance(Pm)gene NLR1-V.Each site had two subpeaks and they were termed MHS1(MHS1.1/1.2)and MHS2(MHS2.1/2.2).Bgt-inducible MHS2.2 was uniquely present in D.villosum,and MHS1.1 was more inducible in D.villosum than in wheat,suggesting that MHSs may be critical for regulation of NLR1-V expression and plant defense.In summary,this study provides a valuable genome resource for functional genomics studies and wheat-D.villosum introgression breeding.The identified regulatory mechanisms may also be exploited to develop new strategies for enhancing Pm resistance by optimizing gene expression in wheat.
基金Supported by the Fund of Amomum villosum in bone growth
文摘OBJECTIVE:This study was conducted to evaluate the effect of Amomum villosum on longitudinal bone growth.METHODS:Adolescent female Sprague-Dawley rats were divided into 3 groups and treated for 4 days:control(distilled water,p.o.),recombinant human growth hormone(rhGH;100 μg/kg,s.c.),and A.villosum(500 mg/kg,p.o.) groups.On day 3,tetracycline(20 g/kg,i.p.) was injected for growth plate identification.On days 2,3 and 4,5-bromo-2'-deoxyuridine(BrdU)(50 mg/kg,i.p.) was injected to label proliferating cells.On day 5,tibias were dissected and fixed in 4% paraformaldehyde,dehydrated,and sectioned for immunohistochemistry and histomorphometry.RESULTS:The rate of bone growth in the A.villosum and rhGH groups increased to(410 ± 44) and(389 ± 46) μm/day(P<0.01),respectively,as compared with the control(330.7 ± 34.7) μm/day.The thickness of the growth plates also increased to(591 ± 37) and(598 ± 32) μm,respectively,as compared with the control(524± 89) μm(P<0.001).The number of BrdU-positive cells in the chondrocytes of the A.villosum and rhGH groups was also significantly higher(126±24) and(143±18) cells/mm 2,respectively) than in the control(109 ± 25) mm 2(P<0.05).Insulin-like growth factor-1 and bone morphogenetic protein-2 in the A.villosum and rhGH groups were highly expressed in the growth plate as compared with the control samples,indicating increased bone formation.CONCLUSIONS:A.villosum could be used to treat growth retardation during adolescence.
基金supported by the National Key Research and Develpment Program of China (2021YFD1200600)the National Natural Science Foundation of China (31871619, 32101703, and 32101800)+4 种基金the Natural Science Foundation of Jiangsu Province (BK20210152)the Jiangsu Seed Industry Revitalization Project (JBGS (2021) 013)the Key Research and Development Program of Jiangsu Province(BE2022346)Jiangsu Agricultural Science and Technology Innovation Fund of China (CX (20) 3029)supported by the European Regional Development Fund (CZ.02.1.01/0.0/0.0/16_019/0000827)。
文摘Wheat sharp eyespot, a stem disease caused by the soilborne fungus Rhizoctonia cerealis van der Hoeven,has become a threat to wheat production worldwide. Exploiting resistance resources from wild relatives of wheat is a promising strategy for controlling this disease. In this study, a new wheat–Dasypyrum villosum T2DS·2V#4L translocation line in the background of Chinese Spring(CS) showed stable resistance to R. cerealis. Introgression of the T2DS·2V#4L chromosome into wheat cultivar Aikang 58 by backcrossing produced a marked increase in sharp eyespot resistance in NIL-T2DS·2V#4L in comparison with NILT2DS·2DL, and no detrimental effects of 2V#4L on agronomic traits were observed in the BC2F2, BC2F2:3,and BC2F2:4generations. Flow-sorted sequencing of 2V#4L yielded 384.3 Mb of assembled sequence, and8836 genes were predicted of which 6154 had orthologs in at least one of the 2AL, 2BL, and 2DL arms of CS, whereas 1549 genes were unique to 2V#4L. About 100,000 SNPs were detected in genes of 2V#4L and2DL in 10 sequenced bread wheat cultivars. A Kompetitive Allele Specific Polymerase chain reaction and30 conserved ortholog sequence markers were developed to trace the 2V#4L chromatin in wheat backgrounds. T2DS·2V#4L compensating translocation lines represent novel germplasm with sharp eyespot resistance and the markers will allow rapid detection in breeding programs.
基金supported by the National Key Research and Development Program of China (2016YFD0102001)the National Natural Science Foundation of China (31571653, 31771782, 31201204)+5 种基金the Technology Support Program of Jiangsu Province, China (BE2013439)the Fundamental Research Funds for the Central Universities, China (KYZ201403)the Jiangsu Agricultural Science and Technology Innovation Fund, China (CX151001)the Program of Introducing Talents of Discipline to Universities, China (B08025)the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, China (PAPD)the Six Talent Peaks Project in Jiangsu Province, China
文摘Expressed sequence tags-derived polymerase chain reaction(EST-PCR) molecular markers specific for alien chromosomes can be used to not only monitor the introgressed alien chromatin in wheat background, but also provide the evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers specific for chromosome 6 VL of Dasypyrum villosum, 297 primer pairs were designed based on the expressed sequence tags(EST) sequences, which were previously mapped in different bins of the long arms of wheat homoeologous 6 AL, 6 BL, and 6 DL. By using the Triticum aestivum, D. villosum, T. durum-D. villosum amphiploid, and T. aestivum-D. villosum alien chromosome lines involving chromosome 6 V, it was found that 32(10.77%) primers could amplify specific bands for chromosome 6 V, and 31 could be allocated to chromosome arm 6 VL. These 6 VL specific markers provided efficient tools for the characterization of structural variation involving the chromosome 6 VL in common wheat background as well as for the selection of useful genes located on 6 VL in breeding programs.
基金financially supported by the National Key Research and Development Program,China (2016YFD0102000) the Agricultural Science and Technology Innovation Program (ASTIP) of the CAAS
文摘T6V#2S·6AL and T6V#4S·6DL translocation chromosomes developed from the cross of wheat and different Dasypyrum villosum accessions have good powdery mildew (PM) resistance, but their pairing and pyramiding behavior remains unclear. Results in this study indicated that the pairing frequency rate of the two differently originated 6VS chromosomes in their F1 hybrid was 18.9% according to genomic in situ hybridization (GISH); the PM resistance plants in the F2 generation from the cross between T6V#4S·6DL translocation line Pm97033 and its PM susceptible wheat variety Wan7107 was fewer than expected. However, the ratio of the resistant vs. the susceptible plants of 15:1 in the F2 generation derived from the cross between the two translocation lines of T6V#2S·6AL and T6V#4S·6DL fitted well. Plants segregation ratio (homozygous:heterozygous:lacking) revealed by molecular marker for T6V#4S·6DL or T6V#2S·6AL in their F2 populations fitted the expected values of 1:2:1 well, inferring that the pairing of the two alien chromosome arms facilitates the transmission of T6V#4S·6DL from the F1 to the F2 generation. A quadrivalent was also observed in 21% of pollen mother cells (PMCs) of homozygote plants containing the two pairs of translocated chromosomes. The chromosome pairing between 6V#2S and 6V#4S indicates that it will be possible to obtain recombinants and clarify if the PM resistance determinant on one alien chromosome arm is different from that on the other.
基金supported by a grant from Research Institute of Rangsit University (No. 40/2556)
文摘Objective: The roots of two Thai medicinal plants, Clerodendrum indicum and Clerodendrum villosum are found in traditional medicine practices. The aim of this research was to preliminarily study the cytotox- icity of extracts of their roots, and the parts that possessed cytotoxic activity were separated on a chro- matograph to identify their active compounds. Methods: The extracts of both plants were screened for cytotoxicity on the SW620 cell line and the com- pounds isolated from the active extracts were further evaluated for their cytotoxic activity against five human cancer cell lines, including SW620, ChaGo-K-1, HepG2, I(ATO-III and BT-474 using the 3-(4,5-dime thyl-2-thiazolyl)-2,5-diphenyll2-H-tetrazolium bromide (MTI') assay. Results: Dichloromethane extracts of C indicum and C. villosum were active against the SW620 cell line. Triterpenoids were mostly obtained from the extracts of these plants (0.28% and 1.02%, respectively) and exhibited varying degrees of cytotoxicity and specificity against the tested cell lines. Two triterpenoids, oleanolic acid 3-acetate and betulinic acid, displayed moderate to strong cytotoxicity toward all cancer cell lines, with 50% inhibitory concentration (ICso) values of 1.66-20.49 mmol/L, whereas 3β-hydroxy- D:Blfriedo-olean-5-ene and taraxerol were cytotoxic to only the SW620 cell line (ICso= 23.39 and 2.09 mmol/L, respectively). Triterpenoid, lupeol, showed potent cytotoxicity on both SW620 (ICso= 1.99 mmol/L) and KATO-III cell lines (IC50 = 1.95 μmol/L), while a fiavonoid, pectolinarigenin, displayed moderate cytotoxicity against these cells (IC50= 13.05 and 24.31 μmol/L, respectively). Although the widely distributed steroid, stigmasterol, was effective against the SW620 cell line (1Cso = 2.79 μmol/L) and β-sitosterol was also active against SW620 (IC50 = 11.26 μmol/L), BT-474 (IC50 = 14.11 μmol/L) and HepG2 cancer cells (IC50=20.47μmol/L), none of the characteristic 24β-ethylsteroids of either Clerode
基金supported by the National Key R&D Program of China(2017YFC1701104)Guangdong Province Applied Science and Technology R&D Special Fund Project(2015B020234002)。
文摘Objective:Amomum villosum(AV)is an herb whose dried fruit has been extensively used in modern medicine to treat digestive system diseases such as dysentery,vomiting and abdominal pain.This paper aims to supplement chloroplast(cp)genomic resources and to be used in phylogenetic studies and identification of AV related plants.Methods:High-throughput sequencing technology was used to determine the complete sequence of the AV cp genome,and the sequence was then compared with three related species.Results:The genome size of AV we obtained was 163,968 bp with an obvious tetrad structure.The AV cp genome was observed to contain 125 unique genes and 81 simple sequence repeat(SSRs)had been determined and the majority of which were adenine–thymine(AT)-rich.Comparative analysis of genome sequence of four ginger plants showed that the atpF,clpP and rpl32 genes are potential markers for identifying Amomum species.Phylogenetic analysis suggested that AV was closely related to A.kravanh and A.compactum.Conclusion:These results have brought useful genetic resources for further identification researches,DNA barcoding,resolving taxonomy and understanding the evolutionary mode of Zingiberaceae cp genome.
文摘Solanum nigrum complex comprises of plant species that belong to Solanum section Solanum used as indigenous leafy vegetables in Kenya among many communities. The Solanum nigrum complex species have also been used traditionally as medicine. Many species make up the Solanum nigrum complex, and some include S. scabrum, S. villosum, S. americanum, S. nigrum, S. americanum, and S. physalifolium. The purpose of this study was to determine the species ofSolanum nigrum complex found in Rift Valley, Nyanza and Western provinces of Kenya. Samples were collected from Eldoret-Iten, Kisumu-Nyando, Kisii-Nyamira, Nakuru, and Busia then pressed and sent to a plant taxonomist in Egerton University for classification and identification. It was established that the collected samples belonged to S. nigrum, S. scabrum, and S. villosurn species. The morphological characteristics used to distinguish the species included shape and colour of the stems, size and shape of the leaves, size, shape and colour of the berries, and the inflorescence. S. nigrurn was widely found in some regions with a noticeable bias towards S. scabrum that was preferred due to its large size of leaves and taste when cooked. However, the distribution and presence of S. nigrum and S. villosum in some of the study regions was poor.
文摘Gibberellin 3-oxidase catalyzes the conversion of inactive gibberellin(GA) species into GAs with biological activity and it is subjected to strict developmental controls in the life cycle of a plant. In this study, 33 gene sequences, encoding the gibberellin 3-oxidase(GA3ox) from Dasypyrum villosum and its dwarf mutant, were obtained. Each contained a 1 107 bp coding sequence(CDS) that encoded a putative protein containing 369 amino acids. The GA3ox protein showed 77% to 97% homology and shared the major conserved structural domains of GA3ox proteins with rice, sorghum bicolor, oat, barley, and wheat. Sequence alignment showed that there were 20 single nucleotide polymorphisms(SNPs) and 22 Insertion/deletions(In Dels) among these sequences, which could be divided into 2 haplotypes, haplotypes Ⅰ and Ⅱ. Haplotype Ⅰ was found in the wild type and was1 495 bp in length, and haplotype Ⅱ was found in the dwarf mutant and was 1 485 bp in length. The Q-PCR results showed that GA3ox was expressed in the leaves, roots, internodes, and stem nodes, and that there was a significant difference in the transcript level of the GA3ox between the wild type and dwarf mutant. The transcript levels of GA3ox in the leaves at the seedling stage, stem elongation stage and the heading stage, in the root and stem nodes at the stem elongation stage and in the internodes at the heading stage of the wild type, were significantly higher than those in the dwarf mutant. However, GA3ox expression in the rest of the wild type tissues at the 3 stages was slightly higher than or not different from the dwarf mutant.The results suggested that the wild type and mutant allele sequences of GA3ox in D. villosum showed 2 amino acid changes in exons and variations in the lengths of introns or the SNPs in introns, which most probably impaired the function of the enzyme,affected the GA3ox expression level, and eventually gave rise to dwarfing.