Adrian Slater - Academia.edu (original) (raw)
Papers by Adrian Slater
65th International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research (GA 2017), 2017
This dataset shows ArrayDASH genotyping profiles generated from ITS regions amplified from 4 <... more This dataset shows ArrayDASH genotyping profiles generated from ITS regions amplified from 4 <i>Hypericum</i> species and the identification of a set of probes that can be used to distinguish between them.Genotyping probes targeted specific sequences that are known to exist in the associated Hypericum species, rather than a set of 4 probes that only differ by their central position to represent the 4 possible bases (i.e., A, C, G and T)
Cellular and Molecular Biology, Jul 1, 1995
Cyclins are a complex group of proteins involved in regulation of the eukaryotic cell division cy... more Cyclins are a complex group of proteins involved in regulation of the eukaryotic cell division cycle via their interaction with cyclin dependent kinases (Cdks). Cyclin gene sequences have been cloned from a number of plant species, including alfalfa, but the diversity of these genes suggests that there are many plant cyclins which have yet to be characterized. A RACE-PCR strategy has been adopted for cloning cyclin gene sequences expressed during direct somatic embryogenesis in alfalfa. RT-PCR with nested degenerate primers was used to amplify the highly conserved "cyclin box" region of a novel A-like cyclin mRNA sequence expressed after induction of somatic embryogenesis. The sequence of this PCR product was used to design primers for 5'- and 3'-RACE protocols. 5'-RACE using a modified SLIC (single strand ligation to single stranded cDNA) procedure revealed considerable sequence heterogeneity in the N-terminal region of the coding sequence with several closely related sequences apparent. Conventional 3'-RACE generated a single cyclin sequence. The complete coding sequence of one member of this A-like cyclin subgroup has been obtained by this RACE strategy and confirmed by PCR amplification and sequencing of alfalfa genomic DNA.
Scientific Reports, 2020
An amendment to this paper has been published and can be accessed via a link at the top of the pa... more An amendment to this paper has been published and can be accessed via a link at the top of the paper.
Biochemical Society Transactions, 1999
Biotechnology & Biotechnological Equipment, 1994
ABSTRACTA strategy has been identified for the production of a low soil tare sugar beet by geneti... more ABSTRACTA strategy has been identified for the production of a low soil tare sugar beet by genetic manipulation of the table beet. This approach will involve increasing the sucrose storage capacity of the table beet by manipulating cell division in the cambial rings of the storage organ. Research aimed at the isolation of suitable cell division regulatory genes and storage organ specific promoter sequences is described. Sugar beet cdc 2 and cyclin sequences have been isolated by PCR techniques. Two root specific cDNA sequences have been characterized; one belongs to a group of hybrid proline-rich/hydrophobic proteins and the other resembles a gene induced during potato tuberisation.
Biochemical Society Transactions, 1981
Genes, 2019
There is considerable potential for the use of DNA barcoding methods to authenticate raw medicina... more There is considerable potential for the use of DNA barcoding methods to authenticate raw medicinal plant materials, but their application to testing commercial products has been controversial. A simple PCR test targeting species-specific sequences within the nuclear ribosomal internal transcribed spacer (ITS) region was adapted to screen commercial products for the presence of Hypericum perforatum L. material. DNA differing widely in amount and extent of fragmentation was detected in a number of product types. Two assays were designed to further analyse this DNA using a curated database of selected Hypericum ITS sequences: A qPCR assay based on a species-specific primer pair spanning the ITS1 and ITS2 regions, using synthetic DNA reference standards for DNA quantitation and a Next Generation Sequencing (NGS) assay separately targeting the ITS1 and ITS2 regions. The ability of the assays to detect H. perforatum DNA sequences in processed medicines was investigated. Out of twenty diff...
Plants, 2022
The potential value of DNA barcoding for the identification of medicinal plants and authenticatio... more The potential value of DNA barcoding for the identification of medicinal plants and authentication of traded plant materials has been widely recognized; however, a number of challenges remain before DNA methods are fully accepted as an essential quality control method by industry and regulatory authorities. The successes and limitations of conventional DNA barcoding are considered in relation to important members of the Lamiaceae. The mint family (Lamiaceae) contains over one thousand species recorded as having a medicinal use, with many more exploited in food and cosmetics for their aromatic properties. The family is characterized by a diversity of secondary products, most notably the essential oils (EOs) produced in external glandular structures on the aerial parts of the plant that typify well-known plants of the basil (Ocimum), lavender (Lavandula), mint (Mentha), thyme (Thymus), sage (Salvia) and related genera. This complex, species-rich family includes widely cultivated comme...
Analytical biochemistry, 2021
We report proof-of-principle experiments regarding a dynamic microarray protocol enabling accurat... more We report proof-of-principle experiments regarding a dynamic microarray protocol enabling accurate and semi-quantitative DNA analysis for re-sequencing, fingerprinting and genotyping. Single-stranded target molecules hybridise to surface-bound probes during initial gradual cooling with high-fidelity. Real-time tracking of target denaturation (via fluorescence) during a 'dynamic' gradual heating phase permits 'melt-curve' analysis. The probe most closely matching the target sequence is identified based on the highest melting temperature. We demonstrated a >99% re-sequencing accuracy and a potential detection rate of 1% for SNPs. Experiments employing Hypericum ribosomal ITS regions and HIV genomes illustrated a reliable detection level of 5% plus simultaneous re-sequencing and genotyping. Such performance suggests a range of potential real-world applications involving rapid sequence interrogation, for example, in the Covid-19 pandemic. Guidance is offered towards t...
Sugar beet gene manipulation has the potential to improve crop productivity, alleviate biotic or ... more Sugar beet gene manipulation has the potential to improve crop productivity, alleviate biotic or abiotic stress and reduce the environmental impact of crop growth. We have considered the procedures for transformation which have been developed and optimized several aspects so as to achieve efficient protocols for the delivery of foreign genes, recovery of transformants and regulation of expression of the foreign genes in transgenic plants. Sugar beet has been regenerated through organogenesis directly from explants or via an intervening callus phase. Somatic embryos were induced at various frequencies from seedlings of several genotypes on MS or PGo medium containing a range of concentrations of BAP (N-6-benzylaminopurine) or TDZ (thidiazuron). The embryos were multiplied during subcultures which involved mechanical wounding. Several marker genes were tested for the detection of early and stable transformation events. It was found that the Ecoli gusA gene can be expressed in sugar be...
Plants, 2020
DNA barcoding is a widely accepted technique for the identification of plant materials, and its a... more DNA barcoding is a widely accepted technique for the identification of plant materials, and its application to the authentication of commercial medicinal plants has attracted significant attention. The incorporation of DNA-based technologies into the quality testing protocols of international pharmacopoeias represents a step-change in status, requiring the establishment of standardized, reliable and reproducible methods. The process by which this can be achieved for any herbal medicine is described, using Hypericum perforatum L. (St John’s Wort) and potential adulterant Hypericum species as a case study. A range of practical issues are considered including quality control of DNA sequences from public repositories and the construction of individual curated databases, choice of DNA barcode region(s) and the identification of informative polymorphic nucleotide sequences. A decision tree informs the structure of the manuscript and provides a template to guide the development of future D...
Phytotherapy Research, 2019
Planta Medica, 2015
DNA barcoding, a technique used to identify species based on short regions of DNA [1], can discri... more DNA barcoding, a technique used to identify species based on short regions of DNA [1], can discriminate between different species, and identify contaminants and adulterants. Ocimum tenuiflorum L., commonly known as holy basil or tulsi, is native to Asia. Three types of tulsi are recognised in the Hindu culture (figure 1); Raam and Shyam (Krishna) are varieties of O. tenuiflorum, whilst Vana tulsi is O. gratissimum L. [2]. Following migration, tulsi plants are widely grown in South Asian (SA) households across the UK. The aim of this research is to use DNA barcoding techniques to identify tulsi plants collected from SA families, using ITS and trnH-psbA regions. A variety of tulsi samples were collected for authentication: community samples from SA families in the UK, commercial samples, and vouchered specimens. DNA analyses discovered that samples described as Shyam tulsi collected from communities in the UK were O. tenuiflorum, but Raam tulsi samples were O. gratissimum. Commercial samples proved to be recalcitrant to DNA extraction; this could be because the DNA had been degraded during manufacturing processes. Vouchered specimen obtained were used to create reference DNA barcodes which were not available in the current DNA databases. Both ITS and trnH-psbA regions were successfully used to distinguish between O. tenuiflorum and O. gratissimum samples. The plastid trnH-psbA primers were more efficient for amplification and sequence analysis than the ITS region. The results exemplify how with the transmission of traditions from one country to another, confusion of species is occurring. Both the ITS and trnH-psbA regions had their strengths and limitations which reinforces the importance of using multiple primers for species authentication.
Current Plant Science and Biotechnology in Agriculture, 1999
The sucrose storage capacity of the sugar beet, Beta vulgaris, storage organ has been related to ... more The sucrose storage capacity of the sugar beet, Beta vulgaris, storage organ has been related to its anatomy. A transverse section of the storage organ reveals a pattern of concentric rings which is caused by the alternation of vascular zones with parenchymatous zones. Sucrose is transported to the storage organ via the phloem and moves laterally to the adjacent parenchymatous tissues, where it is stored in the cell vacuoles. The storage organ of sugar beet has 12–15 cambia, the divisions of the 6 inner rings provide the cells which make up some 75% of the storage root while the outer rings (9 and above) make almost no contribution to the bulk of the storage root. It has been found that the small parenchymatous cells close to the phloem accumulate sucrose to a higher concentration than that achieved by large (older) cells remote from the phloem. Hence, it is predicted that for optimum sucrose accumulation the storage organ should have a large number of parenchymatous zones, each of which offers a short diffusion path from the phloem to predominately small storage cells. In order to manipulate sugar beet development toward this optimum it is necessary to have an understanding of the processes that regulate cell division, cell expansion and differentiation. Towards this end we have begun to isolate and characterise cell division cycle related genes from sugar beet and we have developed a cell suspension culture system for analysis of their roles (Elliott et al. 1996).
The expression of Wee1 in rice endosperm was investigated by semi-quantitative RT-PCR.The results... more The expression of Wee1 in rice endosperm was investigated by semi-quantitative RT-PCR.The results showed the expression of Wee1 reaches the maximum in the endosperm at 4 DAP(4 days after pollination,4 DAP),as endosperm develops on,its expression falls down gradually,but can still be detected of a very low level till 15 DAP.
65th International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research (GA 2017), 2017
This dataset shows ArrayDASH genotyping profiles generated from ITS regions amplified from 4 <... more This dataset shows ArrayDASH genotyping profiles generated from ITS regions amplified from 4 <i>Hypericum</i> species and the identification of a set of probes that can be used to distinguish between them.Genotyping probes targeted specific sequences that are known to exist in the associated Hypericum species, rather than a set of 4 probes that only differ by their central position to represent the 4 possible bases (i.e., A, C, G and T)
Cellular and Molecular Biology, Jul 1, 1995
Cyclins are a complex group of proteins involved in regulation of the eukaryotic cell division cy... more Cyclins are a complex group of proteins involved in regulation of the eukaryotic cell division cycle via their interaction with cyclin dependent kinases (Cdks). Cyclin gene sequences have been cloned from a number of plant species, including alfalfa, but the diversity of these genes suggests that there are many plant cyclins which have yet to be characterized. A RACE-PCR strategy has been adopted for cloning cyclin gene sequences expressed during direct somatic embryogenesis in alfalfa. RT-PCR with nested degenerate primers was used to amplify the highly conserved "cyclin box" region of a novel A-like cyclin mRNA sequence expressed after induction of somatic embryogenesis. The sequence of this PCR product was used to design primers for 5'- and 3'-RACE protocols. 5'-RACE using a modified SLIC (single strand ligation to single stranded cDNA) procedure revealed considerable sequence heterogeneity in the N-terminal region of the coding sequence with several closely related sequences apparent. Conventional 3'-RACE generated a single cyclin sequence. The complete coding sequence of one member of this A-like cyclin subgroup has been obtained by this RACE strategy and confirmed by PCR amplification and sequencing of alfalfa genomic DNA.
Scientific Reports, 2020
An amendment to this paper has been published and can be accessed via a link at the top of the pa... more An amendment to this paper has been published and can be accessed via a link at the top of the paper.
Biochemical Society Transactions, 1999
Biotechnology & Biotechnological Equipment, 1994
ABSTRACTA strategy has been identified for the production of a low soil tare sugar beet by geneti... more ABSTRACTA strategy has been identified for the production of a low soil tare sugar beet by genetic manipulation of the table beet. This approach will involve increasing the sucrose storage capacity of the table beet by manipulating cell division in the cambial rings of the storage organ. Research aimed at the isolation of suitable cell division regulatory genes and storage organ specific promoter sequences is described. Sugar beet cdc 2 and cyclin sequences have been isolated by PCR techniques. Two root specific cDNA sequences have been characterized; one belongs to a group of hybrid proline-rich/hydrophobic proteins and the other resembles a gene induced during potato tuberisation.
Biochemical Society Transactions, 1981
Genes, 2019
There is considerable potential for the use of DNA barcoding methods to authenticate raw medicina... more There is considerable potential for the use of DNA barcoding methods to authenticate raw medicinal plant materials, but their application to testing commercial products has been controversial. A simple PCR test targeting species-specific sequences within the nuclear ribosomal internal transcribed spacer (ITS) region was adapted to screen commercial products for the presence of Hypericum perforatum L. material. DNA differing widely in amount and extent of fragmentation was detected in a number of product types. Two assays were designed to further analyse this DNA using a curated database of selected Hypericum ITS sequences: A qPCR assay based on a species-specific primer pair spanning the ITS1 and ITS2 regions, using synthetic DNA reference standards for DNA quantitation and a Next Generation Sequencing (NGS) assay separately targeting the ITS1 and ITS2 regions. The ability of the assays to detect H. perforatum DNA sequences in processed medicines was investigated. Out of twenty diff...
Plants, 2022
The potential value of DNA barcoding for the identification of medicinal plants and authenticatio... more The potential value of DNA barcoding for the identification of medicinal plants and authentication of traded plant materials has been widely recognized; however, a number of challenges remain before DNA methods are fully accepted as an essential quality control method by industry and regulatory authorities. The successes and limitations of conventional DNA barcoding are considered in relation to important members of the Lamiaceae. The mint family (Lamiaceae) contains over one thousand species recorded as having a medicinal use, with many more exploited in food and cosmetics for their aromatic properties. The family is characterized by a diversity of secondary products, most notably the essential oils (EOs) produced in external glandular structures on the aerial parts of the plant that typify well-known plants of the basil (Ocimum), lavender (Lavandula), mint (Mentha), thyme (Thymus), sage (Salvia) and related genera. This complex, species-rich family includes widely cultivated comme...
Analytical biochemistry, 2021
We report proof-of-principle experiments regarding a dynamic microarray protocol enabling accurat... more We report proof-of-principle experiments regarding a dynamic microarray protocol enabling accurate and semi-quantitative DNA analysis for re-sequencing, fingerprinting and genotyping. Single-stranded target molecules hybridise to surface-bound probes during initial gradual cooling with high-fidelity. Real-time tracking of target denaturation (via fluorescence) during a 'dynamic' gradual heating phase permits 'melt-curve' analysis. The probe most closely matching the target sequence is identified based on the highest melting temperature. We demonstrated a >99% re-sequencing accuracy and a potential detection rate of 1% for SNPs. Experiments employing Hypericum ribosomal ITS regions and HIV genomes illustrated a reliable detection level of 5% plus simultaneous re-sequencing and genotyping. Such performance suggests a range of potential real-world applications involving rapid sequence interrogation, for example, in the Covid-19 pandemic. Guidance is offered towards t...
Sugar beet gene manipulation has the potential to improve crop productivity, alleviate biotic or ... more Sugar beet gene manipulation has the potential to improve crop productivity, alleviate biotic or abiotic stress and reduce the environmental impact of crop growth. We have considered the procedures for transformation which have been developed and optimized several aspects so as to achieve efficient protocols for the delivery of foreign genes, recovery of transformants and regulation of expression of the foreign genes in transgenic plants. Sugar beet has been regenerated through organogenesis directly from explants or via an intervening callus phase. Somatic embryos were induced at various frequencies from seedlings of several genotypes on MS or PGo medium containing a range of concentrations of BAP (N-6-benzylaminopurine) or TDZ (thidiazuron). The embryos were multiplied during subcultures which involved mechanical wounding. Several marker genes were tested for the detection of early and stable transformation events. It was found that the Ecoli gusA gene can be expressed in sugar be...
Plants, 2020
DNA barcoding is a widely accepted technique for the identification of plant materials, and its a... more DNA barcoding is a widely accepted technique for the identification of plant materials, and its application to the authentication of commercial medicinal plants has attracted significant attention. The incorporation of DNA-based technologies into the quality testing protocols of international pharmacopoeias represents a step-change in status, requiring the establishment of standardized, reliable and reproducible methods. The process by which this can be achieved for any herbal medicine is described, using Hypericum perforatum L. (St John’s Wort) and potential adulterant Hypericum species as a case study. A range of practical issues are considered including quality control of DNA sequences from public repositories and the construction of individual curated databases, choice of DNA barcode region(s) and the identification of informative polymorphic nucleotide sequences. A decision tree informs the structure of the manuscript and provides a template to guide the development of future D...
Phytotherapy Research, 2019
Planta Medica, 2015
DNA barcoding, a technique used to identify species based on short regions of DNA [1], can discri... more DNA barcoding, a technique used to identify species based on short regions of DNA [1], can discriminate between different species, and identify contaminants and adulterants. Ocimum tenuiflorum L., commonly known as holy basil or tulsi, is native to Asia. Three types of tulsi are recognised in the Hindu culture (figure 1); Raam and Shyam (Krishna) are varieties of O. tenuiflorum, whilst Vana tulsi is O. gratissimum L. [2]. Following migration, tulsi plants are widely grown in South Asian (SA) households across the UK. The aim of this research is to use DNA barcoding techniques to identify tulsi plants collected from SA families, using ITS and trnH-psbA regions. A variety of tulsi samples were collected for authentication: community samples from SA families in the UK, commercial samples, and vouchered specimens. DNA analyses discovered that samples described as Shyam tulsi collected from communities in the UK were O. tenuiflorum, but Raam tulsi samples were O. gratissimum. Commercial samples proved to be recalcitrant to DNA extraction; this could be because the DNA had been degraded during manufacturing processes. Vouchered specimen obtained were used to create reference DNA barcodes which were not available in the current DNA databases. Both ITS and trnH-psbA regions were successfully used to distinguish between O. tenuiflorum and O. gratissimum samples. The plastid trnH-psbA primers were more efficient for amplification and sequence analysis than the ITS region. The results exemplify how with the transmission of traditions from one country to another, confusion of species is occurring. Both the ITS and trnH-psbA regions had their strengths and limitations which reinforces the importance of using multiple primers for species authentication.
Current Plant Science and Biotechnology in Agriculture, 1999
The sucrose storage capacity of the sugar beet, Beta vulgaris, storage organ has been related to ... more The sucrose storage capacity of the sugar beet, Beta vulgaris, storage organ has been related to its anatomy. A transverse section of the storage organ reveals a pattern of concentric rings which is caused by the alternation of vascular zones with parenchymatous zones. Sucrose is transported to the storage organ via the phloem and moves laterally to the adjacent parenchymatous tissues, where it is stored in the cell vacuoles. The storage organ of sugar beet has 12–15 cambia, the divisions of the 6 inner rings provide the cells which make up some 75% of the storage root while the outer rings (9 and above) make almost no contribution to the bulk of the storage root. It has been found that the small parenchymatous cells close to the phloem accumulate sucrose to a higher concentration than that achieved by large (older) cells remote from the phloem. Hence, it is predicted that for optimum sucrose accumulation the storage organ should have a large number of parenchymatous zones, each of which offers a short diffusion path from the phloem to predominately small storage cells. In order to manipulate sugar beet development toward this optimum it is necessary to have an understanding of the processes that regulate cell division, cell expansion and differentiation. Towards this end we have begun to isolate and characterise cell division cycle related genes from sugar beet and we have developed a cell suspension culture system for analysis of their roles (Elliott et al. 1996).
The expression of Wee1 in rice endosperm was investigated by semi-quantitative RT-PCR.The results... more The expression of Wee1 in rice endosperm was investigated by semi-quantitative RT-PCR.The results showed the expression of Wee1 reaches the maximum in the endosperm at 4 DAP(4 days after pollination,4 DAP),as endosperm develops on,its expression falls down gradually,but can still be detected of a very low level till 15 DAP.