Professor Don Grierson

Emeritus Professor

OBE, BSc, PhD, DSc, FIBiol, FRS

Voice : 0115 951 6333
Fax : 0115 951 6334

Donald.Grierson@nottingham.ac.uk 

Research and Interests

My research is aimed at identifying and characterising genes that are involved in ethylene signalling and ripening in tomato and understanding how ethylene signalling interacts with regulatory networks and other signalling pathways to control flower and fruit development and ripening. A summary of recent and current work is given in the Abstract below of a research presentation at a recent conference.
Previously I was PVC for Research for the University of Nottingham from 2003-2007. During this time I was responsible for the University Research Strategy and the University achieved record research grant totals in four successive years (2004-2007). I was also responsible for the preparations for, and submission of, the University returns in 2007 for the 2008 Research Assessment Exercise. Independent analysis published by “Research Fortnight” showed the University had moved from 14th to 7th position in the national rankings for “Research Power” when the results were published in 2008. Furthermore, Nottingham topped the table as the University likely to receive the largest increase in research funding from the national funding agency (HEFCE) as a result of the RAE assessment (Research Fortnight 4/2/2009). Previously I was Head of the Department of Physiology and Environmental Science at the Sutton Bonington Campus (1988-1991 and 1992-1994), Head of the Plant Sciences Division from 1997- 2002, and Foundation Head of the School of Biosciences from 2000-2002.

 

 

Hormonal and Genetic Mechanisms of Fruit Ripening in Tomato


Don Grierson, Zhefeng Lin, Silin Zhong,
Division of Plant & Crop Sciences, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, LE12 5RD, United Kingdom
donald.grierson@nottingham.ac.uk


Ethylene, synthesised via the Yang Cycle, affects many aspects of plant development, including vegetative growth, ripening, senescence, abscission, and responses to biological and environmental stresses. Tomato has become the model for climacteric fruit ripening (Alexander and Grierson, 2002), and inhibition of the ethylene biosynthesis genes ACC synthase (ACS) and ACC oxidase (ACO) delays or prevents ripening. After perception, ethylene stimulates a signalling cascade, which has been studied largely in Arabidopsis, which leads to a series of changes in expression of genes encoding proteins that catalyse alterations in colour, texture, flavour, aroma, and nutrient content, to produce fruit attractive to a consumer.
Plants have multiple genes for both ACS and ACO, which are differentially expressed during development, but very little is known about the regulation of ethylene synthesis at different stages of the life cycle. We have shown by gel retardation assay that LeHB-1, a previously uncharacterized tomato homeobox protein, interacts with the promoter of LeACO1, an ACC oxidase gene expressed during ripening. Inhibition of LeHB-1 accumulation in tomato fruit using virus-induced gene silencing greatly reduced LeACO1 mRNA levels and inhibited ripening. Conversely, ectopic overexpression of LeHB-1 by viral delivery to developing flowers elsewhere on injected plants triggered altered floral organ morphology, including production of multiple flowers within one sepal whorl, fusion of sepals and petals, and conversion of sepals into carpel-like structures that grew into fruits and ripened. This suggests the LeHB-1 regulatory network plays a role in flower and fruit development as well as ripening (see Lin et al., 2008b). Interestingly, LeACS2 gene expression is also regulated by a homeotic protein LeMADS-RIN (Ito et al, 2008).
Components of the ethylene signalling network found in Arabidopsis are conserved in tomato, and both the ethylene receptors and CTR1-like proteins in tomato are encoded by multi-gene families. As in Arabidopsis, control of ethylene responses operates by a receptor-inhibition mechanism in tomato. Inhibition is relieved by ethylene binding to the receptor, enabling the ripening cascade to be initiated. This has been confirmed by showing that Nr tomatoes expressing the mutant NR ethylene receptor, which cannot bind ethylene and therefore prevents normal ripening, can be restored to partial or complete ripening by antisense inhibition of production of the mutant mRNA (Hackett et al 2000). Although it was initially thought that the ethylene receptors might be functionally equivalent, we now know that different receptors have particular roles at specific stages of the life cycle and it has been proposed that in tomato the ethylene receptors NR, LeETR4 & 6 have unique roles (Kevany et al, 2007).
Although it has been shown that tomato CTR1, 3 & 4 can partially or completely complement the Arabidopsis ctr1 mutant (Adams-Phillips et al, 2004), it was not known which receptors bound to these multiple CTRs in tomato. We have shown, using yeast two-hybrid assay, that the tomato receptors LeETR1, LeETR2 and NEVER-RIPE (NR) can interact with multiple LeCTRs. In vivo protein localization studies using fluorescent proteins revealed that the ethylene receptor NR was targeted to the endoplasmic reticulum (ER) when transiently expressed in onion epidermal cells, whereas the four CTR proteins alone were found in the cytoplasm and nucleus. When co-expressed with NR, three LeCTRs (CTR1, 3 and 4), but not CTR2, also adopted the same ER localization pattern in an NR receptor-dependent manner but not in the absence of NR. The receptor-CTR interactions were confirmed by bimolecular fluorescence complementation (BiFC) showing that NR could form a protein complex with CTR1, 3 and 4 through direct protein-protein interaction at the ER (see Zhong et al., 2008). Arabidopsis transcription factor EIN3 operates downstream of CTR1. In tomato there are multiple EIN3-like (EIL) proteins and it has been suggested that these are functionally redundant (Tieman et al, 2001). We have shown, however, that overexpression of EIL1 in the Nr (ethylene receptor) mutant can only partially restore the expression of ripening genes, suggesting different EILs may play specific roles in gene regulation (Chen et al, 2004).
We have also characterized LeCTR2, which has similarity to CTR1 and also to EDR1, a CTR1-like Arabidopsis protein involved in defence and stress responses. Protein-protein interactions between LeCTR2 and six tomato ethylene receptors indicated that LeCTR2 interacts preferentially with the subfamily I ETR1-type ethylene receptors LeETR1 and LeETR2, but not the NR receptor, or the subfamily II receptors LeETR4, LeETR5 and LeETR6. The C-terminus of LeCTR2 possesses serine/threonine kinase activity and is capable of auto-phosphorylation and phosphorylation of myelin basic protein in vitro. Overexpression of the LeCTR2 N-terminus in tomato resulted in altered growth habit, including reduced stature, loss of apical dominance, highly-branched inflorescences and fruit trusses, indeterminate shoots in place of determinate flowers, and prolific adventitious shoot development from the rachis or rachillae of the leaves. Expression of the ethylene-responsive genes E4 and chitinase B was up-regulated in transgenic plants, but ethylene production and the level of mRNA for the ethylene biosynthetic gene ACO1 was unaffected. Leaves and fruit of transgenic plants also displayed enhanced susceptibility to infection by the fungal pathogen Botrytis cinerea, compared to the wild type The results suggest that LeCTR2 plays a role in ethylene signalling, development and defence, probably through its interactions with the ETR1-type ethylene receptors of subfamily I (see Lin et al., 2008b).
Additional signalling components that regulate the ethylene receptors have now been identified by mutant analysis in Arabidopsis (such as RTE1, RAN1), and we have identified a novel receptor-binding protein in tomato (Lin et al, 2008c). SlTPR1* encodes a tetratricopeptide repeat (TPR) protein isolated by interaction with the tomato ethylene receptor NR in a yeast two-hybrid screen. Previously identified TPR proteins from other organisms are involved in cell signalling and regulation through protein-protein interactions. Overexpression of SlTPR1 in tomato resulted in pleiotropic effects on plant growth habit, including reduced stature, delayed, deformed and infertile flowers, epinasty, reduced apical dominance, and altered leaf morphology. These studies indicate that SlTPR1 functions as a positive regulator in ethylene signalling and its persistent overexpression alters a range of developmental events, including some auxin responses. This led us to propose models in which SlTPR1 modulates ethylene receptor levels or the ethylene signalling response (Lin et al., 2008c). We have also shown that a related protein in Arabidopsis, AtTRP1, binds to the ERS (NR-type) receptor in a yeast 2-hybrid system, and shown by in vivo co-immunoprecipitation that it binds to ERS1 dimers in cell plant membranes (Lin, Ho and Grierson, unpublished).
These results suggest that far from indicating functional redundancy, the multiple ethylene signalling components actually participate in a more subtle and complex process of regulation than previously recognised.


*Since the tomato taxonomic name was changed from Lycopersicon esculentum to Solanum Lycopersicon, new tomato genes are prefixed by Sl.

 

Abstract References


ADAMS-PHILLIPS, L., BARRY, KANNAN,P., LECLERC Q., BOUZAYEN, M. and GIOVANNONI, J. (2004) Evidence that CTR1-mediated ethylene signal transduction in tomato is encoded by a multigene family whose members display distinct regulatory features. Plant Molecular Biology 54: 387-404.

ALEXANDER L and GRIERSON D. (2002) Ethylene biosynthesis and action in tomato: a model for climacteric fruit ripening. Journal of Experimental Botany 53: 2039-2055.

BARRY, CS, LLOP-TOUS I and GRIERSON D. (2000) The regulation of 1-aminocyclopropane-1-carboxylic acid synthase gene expression during the transition from system-1 to system-2 ethylene synthesis in tomato. Plant Physiology 123: 979-986.

CHEN, G., ALEXANDER L. and GRIERSON D. (2004). Constitutive expression of EIL-like transcription factor partially restores ripening in the ethylene-insensitive Nr tomato mutant. Journal of Experimental Botany 55: No 402, 1491-1497.

CHEN, G., HU, Z., and GRIERSON, D. (2008) Differential regulation of tomato ethylene response factor LeERF3b, a putative repressor, and the activator Pti4 in ripening mutants and in response to environmental stresses. Journal of Plant Physiology 165:662-670.

HACKETT R.M., HO C-W., LIN Z., FOOTE H.C.C., FRAY R.G. and GRIERSON D. (2000) Antisense inhibition of the Nr gene restores normal ripening to the tomato never-ripe mutant, consistent with the ethylene receptor-inhibition model. Plant Physiology 124: 1079-1085.

ITO, Y., KITAGAWA, M., IHASHI, N., YABE, K., KIMBARA, J., YASUDA, J., ITO, H., INAKUMA, T., HIROI, S., and KASUMI, T. (2008) DNA-binding specificity, transcriptional activation potential, and the rin mutation effect for the tomato fruit-ripening regulator RIN. The Plant J. 55(2): 212-223.

LIN, Z., HONG, Y., YIN, M., LI, C., ZHANG, K., and GRIERSON, D. (2008a) A tomato HB-zip homeobox protein, LeHB-1, plays an important role in floral organogenesis and ripening. The Plant Journal 55: 301-310.

LIN, Z., ALEXANDER, L., HACKETT, R., and GRIERSON D. (2008b) LeCTR2, a CTR1-like protein kinase from tomato, plays a role in ethylene signalling, development and defence. The Plant Journal 54: 1083-1093.

LIN, Z., ARCIGA-REYES, L., ZHONG, S., HACKETT, R., WILSON, I. and GRIERSON, D. (2008c) SlTPR1, a tomato tetratricopeptide repeat protein, interacts with the ethylene receptors NR and LeETR1, modulating ethylene and auxin responses and development. Journal of Experimental Botany 59, 4271- 4287.

LLOP-TOUS I., BARRY C.S. and GRIERSON D. (2000) Regulation of ethylene biosynthesis in response to pollination in tomato flowers. Plant Physiology 123: 971-978.

TIEMAN, D. M., CIARDI, J.A., TAYLOR, M.G. and KLEE, H. J. (2001) Members of the tomato LeEIL (EIN3-like) are functionally redundant and regulate ethylene responses throughout development The Plant Journal 26: 47-58.

ZHONG, S., LIN, Z., and GRIERSON, D. (2008) Tomato ethylene receptor interaction: visualization of NEVER-RIPE interaction with multiple CTRs at the endoplasmic reticulum. Journal of Experimental Botany 59:965-972.

 

 

REFEREED PUBLICATIONS by DON GRIERSON

  1. LIN, Z., ARCIGA-REYES, L., ZHONG, S., HACKETT, R., WILSON, I., GRIERSON, D. (2008c) SlTPR1, a tomato tetratricopeptide repeat protein, interacts with the ethylene receptors NR and LeETR1, modulating ethylene and auxin responses and development. Journal of Experimental Botany 59, 4271- 4287.
  2. ZHONG, L., LIN, Z., FRAY, F., GRIERSON, D. (2008b) Improved plant transformation vectors for fluorescent protein tagging. Transgenic Research 17: 985-989.
  3. HAN, Y., and GRIERSON, D. (2008) Enhancement of post-transcriptional gene silencing by grafting. Plant Signaling and Behaviour 3:30-33.
  4. LIN, Z., HONG, Y., YIN, M., LI, C., ZHANG, K., and GRIERSON, D. (2008b) A tomato HB-zip homeobox protein, LeHB-1, plays an important role in floral organogenesis and ripening. The Plant Journal 55: 301-310.
  5. LIN, Z., ALEXANDER, L., HACKETT, R., and GRIERSON D. (2008a) LeCTR2, a CTR1-like protein kinase from tomato, plays a role in ethylene signalling, development and defence. The Plant Journal 54: 1083-1093.
  6. ZHONG, S., LIN, Z., and GRIERSON, D. (2008a) Tomato ethylene receptor interaction: visualization of NEVER-RIPE interaction with multiple CTRs at the endoplasmic reticulum. Journal of Experimental Botany 59:965-972.
  7. CHEN, G., HU, Z., and GRIERSON, D. (2008) Differential regulation of tomato ethylene response factor LeERF3b, a putative repressor, and the activator Pti4 in ripening mutants and in response to environmental stresses. Journal of Plant Physiology 165:662-670.
  8. SHAHARUDDIN, N.A., HAN, Y.H., LI, H., GRIERSON, D. (2006) The mechanism of graft transmission of sense and antisense gene silencing in tomato plants. FEBS Letters 580 6579-6586
  9. S H KIM AND D GRIERSON (2005) Subcellular localisation and silencing of ripening-associated membrane protein (TRAMP) in tomato Lycopersicon esculentum Mill. Plant Science 169: 1022-1029
  10. C S BARRY, RP MCQUINN, A J THOMPSON, G B SEYMOUR, D GRIERSON AND J J GIOVANNONI (2005) Ethylene insensitivity conferred by the Green-ripe and Never-ripe 2 ripening mutants of tomato. Plant Physiology 138: 267-275.
  11. G CHEN, R HACKETT, D WALKER, A TAYLOR, Z LIN and D GRIERSON (2004) Identification of a specific isoform of tomato lipoxygenase (TomLoxC) involved in the generation of fatty acid-derived flavour compounds. Plant Physiology 136: 2641-2651.
  12. G CHEN, L ALEXANDER, D GRIERSON (2004). Constitutive expression of EIL-like transcription factor partially restores ripening in the ethylene-insensitive Nr tomato mutant. Journal of Experimental Botany 55: No 402, 1491-1497.
  13. Y HAN, A GRIFFITHS, H LI, D GRIERSON (2004) The effect of endogenous mRNA levels on co-suppression in tomato. FEBS Letters 563 123-128.
  14. Z LIN, H LI, S KIM, L ALEXANDER and D GRIERSON (2003) Identification of Proteins interacting with TCTR2, an ethylene signalling component in tomato. In: Biology and Biotechnology of the Plant Hormone Ethylene III, Edited by Miguel Vendrell, Harry Klee, Jean-Claude Pech, and Felix Romojaro. IOS press, NATO Science Series.
  15. MOEDER W, BARRY CS, TAURIAINEN AA, BETZ C, TUOMAINEN J, UTRIAINEN M, GRIERSON D, SANDERMANN H, LANGEBARTELS C and KANGASJARVI J (2002) Ethylene synthesis regulated by biphasic induction of 1-aminocyclopropane-1-carboxylic acid synthase and 1-aminocyclopropane-1-carboxylic acid oxidase genes is required for hydrogen peroxide accumulation and cell death in ozone-exposed tomato. Plant Physiology 130:1918-1926.
  16. EL YAHYAOUI F, WONGS-AREE C, LATCHE A, HACKETT, GRIERSON D & PECH J–C (2002) Molecular and biochemical characteristics of a gene encoding an alcohol acyl-transferase involved in the generation of aroma volatile esters during melon ripening. Eur J Biochem 269: 2359-2366.
  17. COOK D, GRIERSON D, JONES C, WALLACE A, WEST G and TUCKER G (2002) Modification of fatty acid composition in tomato (Lycopersicon esculentum) by expression of a borage Δ6-desaturase. Molecular Biotechnology 21: 123-128.
  18. HAN, Y and GRIERSON D (2002) The influence of inverted repeats on the production of small antisense RNAs involved in gene silencing. Molecular Genetics and Genomics 267: 629-635.
  19. SOUTER M, TOPPING J, PULLEN M, FRIML J, PALME K, HACKETT R, GRIERSON D and LINDSEY K (2002) hydra Mutants of Arabidopsis are defective in sterol profiles and auxin and ethylene signalling. The Plant Cell 14: 1017-1031.
  20. HAN Y and GRIERSON D. (2002) Relationship between small antisense RNAs and aberrant RNAs associated with sense transgene mediated gene silencing in tomato. The Plant Journal 29(4): 509-519.
  21. ALEXANDER L and GRIERSON D. (2002) Ethylene biosynthesis and action in tomato: a model for climacteric fruit ripening. Journal of Experimental Botany 53: 2039-2055.
  22. BARRY CS, FOX EA, YEN H-C, LEE S, YING T-J, GRIERSON D and GIOVANNONI JJ (2001) Analysis of the ethylene response in the epinastic mutant of tomato. Plant Physiology 127: 58-66.
  23. GUOPING C, WILSON D, KIM SG and GRIERSON D (2001) Inhibiting expression of a tomato ripening-associated membrane protein (TRAMP) increases organic acids and reduces sugar levels. Planta 212, 799-807.
  24. HACKETT RM, HO C-W, LIN Z, FOOTE HCC, FRAY RG, GRIERSON D. (2000) Antisense inhibition of the Nr gene restores normal ripening to the tomato never-ripe mutant, consistent with the ethylene receptor-inhibition model. Plant Physiology 124: 1079-1085.
  25. BARRY CS, LLOP-TOUS I, GRIERSON D. (2000) The regulation of 1-aminocyclopropane-1-carboxylic acid synthase gene expression during the transition from system-1 to system-2 ethylene synthesis in tomato. Plant Physiology 123: 979-986.
  26. LLOP-TOUS I, BARRY CS and GRIERSON D. (2000) Regulation of ethylene biosynthesis in response to pollination in tomato flowers. Plant Physiology 123: 971-978.
  27. GRIFFITHS A, PRESTAGE S, LINFORTH R, ZHANG J, TAYLOR A and GRIERSON D. (1999) Fruit-specific lipoxygenase suppression in antisense-transgenic tomatoes. Postharvest Biology and Technology 17: 163-173.
  28. GIOVANNONI J, YEN H, SHELTON B, MILLER S, VREBALOV J, KANNAN P, TIEMAN D, HACKETT R, GRIERSON D and KLEE H. (1999) Genetic mapping of ripening and ethylene-related loci in tomato. Theor Appl Genet 98: 1005-1013.
  29. GRIFFITHS A, BARRY C, ALPUCHE-SOLIS AG and GRIERSON D. (1999) Ethylene and developmental signals regulate expression of lipoxygenase genes during tomato fruit ripening. Journal of Experimental Botany, 50, 793-798.
  30. AKHTAR S, GOLDSCHMIDT EE, JOHN I, RODONI S, MATILE P & GRIERSON D. Altered patterns of senescence and ripening in gf, a stay-green mutant of tomato. Journal of Experimental Botany, 50, 1115-1122, 1999.
  31. THOMPSON AJ, BARRY CS, JARVIS MC, GIOVANNONI JJ, GRIERSON D & SEYMOUR GB. Molecular and genetic characterisation of a novel pleiotropic tomato-ripening mutant. Plant Physiology, 120, 383-389, 1999.
  32. FRAY RG, THROUP JP, WALLACE A, WILLIAMS P, STEWART GSAB GRIERSON D. Plants genetically modified to produce N-acylhomoserine lactones communicate with bacteria. Nature Biotechnology, 17: 1017-1020, 1999.
  33. LIN Z, HACKETT RM, PAYTON S & GRIERSON D. (1998). A tomato sequence, TCTR2, encoding an Arabidopsis CTR1 homolog. Plant Physiology, 117, p 1126.
  34. HACKETT RM, OH SA, MORRIS PC & GRIERSON D (1998). A tomato MAP kinase gene differentially regulated during fruit development, leaf senescence and wounding. Plant Physiology, 117, p 1526.
  35. COOPER W, BOUZAYEN M, HAMILTON A, BARRY C, ROSSALL S & GRIERSON D. Use of transgenic plants to study the role of ethylene and polygalacturonase during infection of tomato fruit by Colletotrichum gloeospiroides. Plant Pathology, 47, 308-316, 1998.
  36. HAMILTON AJ, BROWN S, YUANHUAI H, ISHIZUKA M, LOWE A, ALPUCHE-SOLIS AG, & GRIERSON D. A transgene with repeated DNA causes high frequency, post-transcriptional suppression of ACC-oxidase gene expression in tomato. The Plant Journal,15, 737-746, 1998.
  37. BIDONDE S, FERRER MA, ZEGZOUTI H, RAMASSAMY S, LATCHE A, PECH JC, HAMILTON, AJ, GRIERSON D & BOUZAYEN M. Expression and characterisation of three tomato 1-aminocyclopropane-1-carboxylate oxidase cDNAs in yeast. European Journal of Biochemistry, 253, 20-26, 1998.
  38. AGGELIS A, JOHN I & GRIERSON D. Analysis of physiological and molecular changes in melon (cucumis melo L.) varieties with different rates of ripening. Journal of Experimental Botany, 48, 769-778, 1997.
  39. BLUME, B & GRIERSON D. Expression of ACC oxidase promoter-GUS fusions in tomato and Nicotiana plumbaginifolia regulated by developmental and environmental stimuli. The Plant Journal, 12, 731-746, 1997.
  40. BLUME B, BARRY CS, HAMILTON AJ, BOUZAYEN, M & GRIERSON D. Identification of transposon-like elements in non-coding regions of tomato ACC oxidase genes. Molecular and General Genetics, 254, 297-303, 1997.
  41. JOHN I, HACKETT R, COOPER W, DRAKE R, FARRELL A & GRIERSON D. Cloning and characterization of tomato leaf senescence-related cDNAs. Plant Molecular Biology, 33, 641-651, 1997.
  42. AGGELIS A, JOHN I, KARVOUNI Z & GRIERSON D. Characterization of two cDNA clones for mRNAs expressed during ripening of melon (Cucumis melo L.) fruits. Plant Molecular Biology 33, 313-322, 1997.
  43. PAYTON S, FRAY RG, BROWN S & GRIERSON D. Ethylene receptor expression is regulated during fruit ripening, flower senescence and abscission. Plant Molecular Biology 31, 1227-1231, 1996.
  44. BARRY CS, BLUME B, BOUZAYEN M, COOPER W, HAMILTON AJ & GRIERSON D. Differential expression of the 1-aminocyclopropane-1-carboxylate oxidase gene family of tomato. The Plant Journal 9, 525-535, 1996.
  45. DRAKE, R, JOHN, I, FARRELL, A, COOPER, W, SCHUCH, W & GRIERSON D. Isolation and analysis of cDNAs encoding tomato cysteine proteases expressed during leaf senescence. Plant Molecular Biology 30, 755-767, 1996.
  46. FRAY, RG, WALLACE, A, FRASER, PD, VALERO, D, HEDDON, P, BRAMLEY, P & GRIERSON D. Constitutive expression of a fruit phytoene synthase gene in transgenic tomatoes causes dwarfism by redirecting metabolites from the gibberellin pathway. The Plant Journal 8, 693-701, 1995.
  47. NICHOLASS, FJ, SMITH, CJS, SCHUCH, W, BIRD CR & GRIERSON D. High levels of reporter gene expression directed by tomato fruit polygalacturonase gene flanking regions. Plant Molecular Biology 28, 423-435, 1995.
  48. GALLEGO, PP, WHOTTON, L, PICTON, S, GRIERSON, D & GRAY JE. A role for glutamate decarboxylase during tomato ripening : The characterisation of a cDNA encoding a putative glutamate decarboxylase with a calmodulin binding site. Plant Molecular Biology 27, 1143-1151, 1995.
  49. KARVOUNI, Z, JOHN, I, TAYLOR, JE, WATSON, CF, TURNER, AJ & GRIERSON, D. Isolation and characterisation of a melon cDNA encoding phytoene synthase. Plant Molecular Biology, 27, 1153-1162, 1995.
  50. JOHN, I, DRAKE, R, FARRELL, A, COOPER, W, LEE, P, HORTON, P & GRIERSON, D. Delayed leaf senescence in ethylene-deficient ACC oxidase antisense tomato plants : molecular and physiological analysis. The Plant Journal 7, 483-490, 1995.
  51. ZAMORANO, J.P., DOPICO, B., LOWE, A.L., WILSON, I.D., GRIERSON, D. & MERODIO, C. Effect of low temperature storage and ethylene removal on ripening and gene expression changes in avocado fruit. Postharvest Biology and Technology 4, 331-342, 1994.
  52. HALL, L.S., BIRD, C.R., PICTON, S., TUCKER, G.A., SEYMOUR, G.B. & GRIERSON D. Molecular characterisation of cDNA clones representing pectin esterase isozymes from tomato. Plant Molecular Biology 25, 313-318, 1994.
  53. FRAY, R.G., WALLACE, A.D. & GRIERSON D. Identification of unexplained DNA fragments within the T-DNA borders of the Bin19 plant transformation vector. Plant Molecular Biology 25, 339-342, 1994.
  54. FRAY, RG, WALLACE, A, GRIERSON, D & LYCETT, GW. Nucleotide sequence and expression of a ripening and water stress-related cDNA from tomato with homology to the MIP class of membrane channel proteins. Plant Molecular Biology 24, 539-543, 1994.
  55. GRIERSON, D. and SCHUCH, W. Control of ripening. Phil. Trans. R. Soc. Lond. B342, 241-250, 1993.
  56. PICTON, S., GRAY, J.E., LOWE, A., BARTON, S. & GRIERSON, D. Sequence of a cloned tomato ubiquitin conjugating enzyme. Plant Physiology 103, 1471-1472, 1993b.
  57. PICTON, S., GRAY, J.E., BARTON, S., ABU BAKAR, U., LOWE, A., GRIERSON, D. cDNA cloning and characterisation of novel ripening-related mRNAs with altered patterns of accumulation in the ripening inhibitor (rin) tomato mutant. Plant Molecular Biology, 23, 193-207, 1993a.
  58. PICTON, S., GRAY, J.E., PAYTON, S., BARTON, S.L., LOWE, A., GRIERSON, D. A histidine decarboxylase-like mRNA is involved in tomato fruit ripening. Plant Molecular Biology, 23, 627-631, 1993c.
  59. FRAY, R.G. & GRIERSON D. Identification and genetic analysis of normal and mutant phytoene synthase genes of tomato by sequencing, complementation and co-suppression. Plant Molecular Biology, 22, 589-602, 1993.
  60. BALAGUE, C., WATSON, C.F., TURNER, A.J., ROUGE, P., PICTON, S., PECH, J-C & GRIERSON, D. Isolation of a ripening and wound-induced cDNA from Cucumis melo L. encoding a protein with homology to the ethylene-forming-enzyme. Europ. J. Biochem, 212, 27-34, 1993.
  61. PICTON, S., BARTON, S.L., BOUZAYEN, M., HAMILTON, A.J. & GRIERSON, D. Altered fruit ripening and leaf senescence in tomatoes expressing an antisense ethylene-forming enzyme transgene. The Plant Journal, 3, 469-481, 1993d.
  62. DOPICO, B., LOWE, A.L., WILSON, I.D., MERODIO, C. & GRIERSON, D. Cloning and characterisation of avocado fruit mRNAs and their expression during ripening and low temperature storage. Plant Molecular Biology, 21, 437-499, 1993.
  63. HALL, L.N., TUCKER, G.A., SMITH, C.J.S., WATSON, C.F., BONIWELL, J.M., FLETCHER, J.D., RAY, J.A., SCHUCH, W., BIRD, C.R. & GRIERSON, D. Antisense inhibition of pectinesterase gene expression in transgenic tomatoes. The Plant Journal 3, 121-129, 1993.
  64. RAY, J., MOUREAU, P., BIRD, C., BIRD, A., GRIERSON, D., MAUNDERS, M., TRUESDALE, M., BRAMLEY, P., SCHUCH, W. Cloning and characterisation of phytoene synthase genes from tomato. Plant Molecular Biology 19, 401-404, 1992.
  65. SCHUCH, W., KANCZLER, J., ROBERTSON, D., HOBSON, G., TUCKER, G., GRIERSON, D., BRIGHT, S., BIRD, C. Fruit quality characteristics of transgenic tomato fruit with altered polygalacturonase activity. HortScience 26, 1517-1520, 1991.
  66. DAVIES, K., GRIERSON, D., EDWARDS, R. & HOBSON, G. Salt-stress induces partial ripening of the nor tomato mutant but expression of only some ripening-related genes. J. Plant Physiol, 139, 140-145, 1991.
  67. HAMILTON, A.J., BOUZAYEN, N. & GRIERSON, D. Identification of a tomato gene for the ethylene forming enzyme by expression in yeast. Proc. Natl. Acad. Sci. (U.S.A.) 88, 7434-7437, 1991.
  68. GRIERSON, D., FRAY, R., HAMILTON, A.J., SMITH, C.J.S., & WATSON, C.F. Does co-suppresion of sense genes in transgenic plants involve antisense RNA? Trends in Biotechnology, 9, 122-123, 1991.
  69. KÖCK, M., HAMILTON, A. & GRIERSON, D. eth 1, a gene involved in ethylene synthesis in tomato. Plant Molec. Biol. 17, 141-142, 1990.
  70. FRAY, R.G., LYCETT, G.W. & GRIERSON, D. Nucleotide sequence of a heat and ripening related cDNA from tomato. Nucleic Acids Res. 18, 7148, 1990.
  71. WILSON, I.D. TUCKER, G.A., KNEE, M. & GRIERSON, D. Changes in mRNA during low temperature storage and ripening of pears. Phytochemistry, 29, 2407-2409, 1990.
  72. TAYLOR, J.E., TUCKER, G.A., LASSLETT, Y., SMITH, C.J.S., ARNOLD, C.M., WATSON, C.F., SCHUCH, W., GRIERSON, D. & ROBERTS, J.A. Polygalacturonase expression during leaf abscission of normal and transgenic tomato plants. Planta 183, 133-138, 1990.
  73. SMITH, C.J.S., WATSON, C.F., BIRD, C.R., RAY, J., SCHUCH, W. & GRIERSON, D. Expression of a truncated tomato polygalacturonase gene inhibits expression of the endogenous gene in transgenic plants. Molec. Gen. Genet. 224, 477-481, 1990b.
  74. WATKINS, C.B., PICTON, S. & GRIERSON, D. Stimulation and inhibition of expression of ripening-related mRNAs in tomatoes as influenced by chilling temperatures. J. Plant Physiol. 136, 318-323, 1990.
  75. HAMILTON, A., LYCETT, G.W. & GRIERSON, D. Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants. Nature, 346, 284-287, 1990.
  76. SMITH, C.J.S., WATSON, C.F., MORRIS, P.C., BIRD, C.R., SEYMOUR, G.B., GRAY, J.E., ARNOLD, C., TUCKER, G.A., SCHUCH, W. & GRIERSON, D. Inheritance and effect on ripening of antisense polygalacturonase genes in transgenic tomatoes. Plant Molec. Biol. 14, 369-379, 1990.
  77. DAVIES, K.M., HOBSON, G.E. & GRIERSON, D. Differential effect of silver ions on the accumulation of ripening related mRNAs in tomato. J. Plant Physiol. 135, 708-713, 1990.
  78. DAVIES, K.M. & GRIERSON, D. Identification of cDNA clones for tomato (Lycopersicon esculentum) mRNAs that accumulate during fruit ripening and leaf senescence in response to ethylene. Planta 179, 73-80, 1989.
  79. KNAPP, J., MOUREAU, P., SCHUCH, W. & GRIERSON, D. Organisation and expression of polygalacturonase and other ripening related genes in Ailsa Craig, 'Neverripe' and "Ripening Inhibitor" tomato mutants. Plant Molec. Biology 12, 105-116, 1989.
  80. MUTSCHLER, M., GUTTIERI, M., KINZER, S., GRIERSON, D. & TUCKER, G.A. Changes in ripening-related processes in tomato conditioned by the alc mutant. Theor. Appl. Genet. 76, 285-292, 1988.
  81. HOLDSWORTH, M.J., SCHUCH, W., & GRIERSON, D. Organisation and expression of a wound/ripening related small multigene family from tomato. Plant Molec. Biology 11, 81 88, 1988.
  82. SMITH, C.J.S., WATSON, C., RAY, J., BIRD, C.R., MORRIS, P.C., SCHUCH, W. AND GRIERSON, D. Antisense RNA inhibition of polygalacturonase gene expression in transgenic tomatoes, Nature 334, 724 726, 1988.
  83. DAVIES, K.M., HOBSON, G.E. & GRIERSON, D. Silver ions inhibit the ethylene stimulated production of ripening related mRNAs in tomato. Plant, Cell & Environment 11, 729-738, 1988.
  84. RAY, J., KNAPP, J., GRIERSON, D., BIRD, C. & SCHUCH, W. Identification and sequence determination of a cDNA clone for tomato pectin esterase. Europ. J. Biochem. 174, 119 124, 1988.
  85. BIRD, C.R., SMITH, C.J.S., RAY, J.A., MOUREAU, P., BEVAN, M.W., BIRD, A.S., HUGHES, S., MORRIS, P.C., GRIERSON, D. & SCHUCH, W. The tomato polygalacturonase gene and ripening specific expression in transgenic plants. Plant Molecular Biology 11, 651-662, 1988.
  86. GIVEN, N.K., VENIS, M.A. & GRIERSON, D. Purification and properties of phenylalanine ammonia lyase from strawberry fruit and its synthesis during ripening, J. Plant Physiol. 133, 31 37, 1988.
  87. GIVEN, N.K., VENIS, M.A., & GRIERSON, D. Phenylalanine ammonialyase activity and anthocyanin synthesis in ripening strawberry fruit, J. Plant Physiol. 133, 25 30, 1988.
  88. GIVEN, N.K., VENIS, M.A., & GRIERSON, D. Hormonal regulation of fruit ripening in the strawberry, a non climacteric fruit, Planta 174, 402 406, 1988.
  89. PICTON, S. & GRIERSON, D. Inhibition of expression of tomato ripening genes at high temperature. Plant, Cell & Environment 11, 265 272, 1988.
  90. HOLDSWORTH, M.J., SCHUCH, W. & GRIERSON, D. Nucleotide sequence of an ethylene related gene from tomato. Nucleic Acids Res. 15, 10600, 1987.
  91. RAY, J., BIRD, C., MAUNDERS, M., GRIERSON, D. & SCHUCH, W. Sequence of pTOM 5, a ripening related cDNA from tomato. Nucleic Acids Res. 15, 10587, 1987.
  92. GRIMWADE, J.A., GRIERSON, D. & WHITTINGTON, W.J. The effect of differences in time to maturity on the quality of seed produced by different varieties of sugar beet. Seed Sci. Technol. 15, 135 145, 1987.
  93. GREENWAY, S.C., STRANGEWAY, G.M., GRIERSON, D. & BRYANT, J.A. Long lived messenger RNA and its relationship to protein synthesis during germination of pea (Pisum sativum L.) seeds. Annals of Botany 57, 771-781, 1986.
  94. HOLDSWORTH, M.J., BIRD, C.R., RAY, J., SCHUCH, W. & GRIERSON, D. Structure and expression of an ethylene related mRNA from tomato. Nucleic Acids Res. 15, 731 739, 1987.
  95. MAUNDERS, M.J., HOLDSWORTH, M.J., SLATER, A., KNAPP, J.E., BIRD, C.R., SCHUCH, W. & GRIERSON, D. Ethylene stimulates the accumulation of ripening related mRNAs in tomatoes. Plant, Cell & Environment 10, 177 184, 1987.
  96. GRIERSON, D., TUCKER, G.A., KEEN, J., RAY, J., BIRD, C.R. and SCHUCH, W. Sequencing and identification of a cDNA clone for tomato polygalacturonase. Nucleic Acids Res. 14, 8595 8603, 1986.
  97. NICHOLAS, J.R., GATES, P.J. & GRIERSON, D. The use of fluorescence microscopy to monitor root development in micropropagated explants. J. Horticultural Res. 61, 417 421, 1986.
  98. BATHGATE, B., GOODENOUGH, P.W. & GRIERSON, D. Regulation of the expression of the psbA gene in tomato fruit chloroplasts and chromoplasts. J. Plant Physiol. 124, 223 233, 1986.
  99. SMITH, C.J.S., SLATER, A. & GRIERSON, D. Rapid appearance of an mRNA correlated with ethylene synthesis encoding a protein of molecular weight 35,000. Planta 168, 94 100 1986.
  100. SLATER, A., MAUNDERS, M.J., EDWARDS, D., SCHUCH, W. & GRIERSON, D. Isolation and characterisation of cDNA clones for tomato polygalacturonase and other ripening related proteins. Plant Molec. Biol. 5, 137 147 1985.
  101. PLANT, A.L., COVEY, S.N. & GRIERSON, D. Detection of a sub genomic mRNA for gene V, the putative reverse transcriptase gene of cauliflower mosaic virus. Nucl. Acids Res. 13, 8305 8321 1985.
  102. BATHGATE, B., PURTON, M.E., GRIERSON, D. & GOODENOUGH, P.W. Plastid changes during the conversion of chloroplasts to chromoplasts in ripening tomatoes. Planta 165, 197 204, 1985.
  103. MORRIS, P.C., GRIERSON, D. & WHITTINGTON, W.J. The influence of fruit structure on germination of sugar beet Beta vulgaris. Seed Sci. Technol. 13, 41 51, 1985.
  104. GRIERSON, D., SLATER, A., SPEIRS, J. & TUCKER, G.A. The appearance of polygalacturonase mRNA in tomatoes: one of a series of changes in gene expression during development and ripening. Planta 163, 263 271, 1985.
  105. JEFFERY, D., SMITH, C., GOODENOUGH, P., PROSSER, T. & GRIERSON, D. Ethylene independent and ethylene dependent biochemical changes in ripening tomatoes. Plant Physiol. 74, 32 38, 1984.
  106. COOLBEAR, P., FRANCIS, A. & GRIERSON, D. The effect of low temperature pre sowing treatment on the germination performance and membrane integrity of artificially aged tomato seeds. J. Exptl. Bot. 35, 1609 1617, 1984.
  107. MORRIS, P.C., GRIERSON, D. & WHITTINGTON, W.J. Endogenous inhibitors and germination of Beta vulgaris. J. Exptl. Bot. 35, 994 1002, 1984.
  108. SPEIRS, J., BRADY, C.J., GRIERSON, D. & LEE, E. Changes in ribosome organisation and messenger RNA abundance in ripening tomato fruits. Aust. J. Plant Physiol. 11, 225 233, 1984.
  109. SU, L Y, McKEON, T., GRIERSON, D., CANTWELL, M. & YANG, S.F. Development of 1 aminocyclopropane 1 carboxylic acid (ACC) synthase and polygalacturonase activities during maturation and ripening of tomato fruits in relation to their ethylene production rates. HortScience 19, 576 578, 1984.
  110. CROOKES, P.R. & GRIERSON, D. Ultrastructure of tomato fruit ripening and the role of polygalacturonase isoenzymes in cell wall degradation. Plant Physiol. 72, 1088 1093, 1983.
  111. GRIERSON, D. & TUCKER, G.A. Timing of ethylene and polygalacturonase synthesis in relation to the control of tomato fruit ripening. Planta 157, 174 179, 1983.
  112. GRIERSON, D., KEAR, R.J., THOMPSON, J.R. & GARCIA MORA, R. Stimulation of in vitro RNA synthesis by pre treating plants with auxins is due to auxin induced ethylene production. Z. Pflanzenphysiol. 107, 419 426, 1982.
  113. GOODENOUGH, P.W., TUCKER, G.A., GRIERSON, D. & THOMAS, T. Changes in colour, polygalacturonase, monosaccharides and organic acids during storage of tomatoes. Phytochemistry 21, 281 284, 1982.
  114. TUCKER, G.A., ROBERTSON, N.G. & GRIERSON, D. Purification and changes in activities of tomato pectinesterase isoenzymes. J. Sci. Food Agric. 33, 396 400, 1982.
  115. TUCKER, G.A. & GRIERSON, D. Synthesis of polygalacturonase during tomato fruit ripening. Planta 155, 64 67, 1982.
  116. THEMMEN, A.P.N., TUCKER, G.A. & GRIERSON, D. Degradation of isolated tomato cell walls by purified polygalacturonase in vitro. Plant Physiol. 69, 122 124, 1982.
  117. TUCKER, G.A., ROBERTSON, N.G. & GRIERSON, D. The conversion of tomato fruit polygalacturonase isoenzyme 2 into isoenzyme 1 in vitro. Europ. J. Biochem. 115, 87 90, 1981.
  118. TUCKER, G.A. ROBERTSON, N.G. & GRIERSON, D. Changes in polygalacturonase isoenzymes during the 'ripening' of normal and mutant tomato fruit. Europ. J. Biochem. 112, 119 124, 1980.
  119. GRIERSON, D. THOMPSON, J., GARCIA MORA, R. & SLATER, R.J. Ribonucleic acid synthesis by isolated mung bean nucleoli. Z. Pflanzenphysiologie 97, 437 447, 1980.
  120. COOLBEAR, P., GRIERSON, D. & HEYDECKER, W. Osmotic pre sowing treatments and nucleic acid accumulation in tomato seeds (Lycopersicon esculentum Mill.) Seed Sci. Technol. 8, 289 303, 1980.
  121. COOLBEAR, P. & GRIERSON, D. Studies on the changes in the major nucleic acid components of tomato seeds (Lycopersicon esculentum Mill.) resulting from osmotic pretreatment. J. Exp. Bot. 30, 1153 1162, 1979.
  122. GRIERSON, D. Light stimulated accumulation of 14s RNA during chloroplast development in spinach cotyledons. Z. Pflanzenphysiologie 95, 171 177, 1979.
  123. SLATER, R.J., VENIS, M.A. & GRIERSON, D. Characterisation of RNA synthesis by nuclei isolated from Zea mays. Planta 144, 89 93, 1978.
  124. SPEIRS, J. & GRIERSON, D. Isolation and characterisation of 14s RNA from spinach chloroplasts. Biochim. Biophys. Acta 521, 619 633, 1978.
  125. RATTANAPANONE, N., SPEIRS, J. & GRIERSON, D. Evidence for changes in messenger RNA content related to tomato fruit ripening. Phytochemistry 17, 1485 1486, 1978.
  126. SPRING, H., GRIERSON, D., HEMBLEBEN, V., STOHR, M., KROHNE, G., STADLER, J. & FRANKE, W.W. DNA contents and numbers of pre rRNA genes in nuclei of gametes and vegetative cells of Acetabularia mediterranea. Experimental Cell Research 114, 203 215, 1978.
  127. HEMBLEBEN, V. & GRIERSON, D. Evidence that in higher plants the 25s and 18s rRNA genes are not interspersed with genes for 5s rRNA Chromosoma 65, 353 358, 1978.
  128. SLATER, R.J. & GRIERSON, D. Ribonucleic acid synthesis by chromatin isolated from Phaseolus aureus Roxb. The effect of endogenous ribonuclease. Planta 137, 153 158, 1977.
  129. GILES, A.B., GRIERSON, D. & SMITH, H. In vitro translation of messenger RNA from developing leaves. Evidence for the existence of stored messenger RNA and its light induced mobilisation into polyribosomes. Planta 136, 31 36, 1977.
  130. RATTANAPANONE, N., GRIERSON, D. & STEIN, M. Ribonucleic acid metabolism during the development and ripening of tomato fruits. Phytochemistry 16, 629 633, 1977.
  131. GRIERSON, D. & HEMBLEBEN, V. Ribonucleic acid from the higher plant Matthiola incana: molecular weight measurements and DNA RNA hybridisation studies. Biochim. Biophys. Acta 475, 424 436, 1977.
  132. HEMBLEBEN, V., GRIERSON, D. & DERTMANN, H. The use of equilibrium centrifugation in actinomycin caesium chloride for the purification of ribosomal DNA. Plant Science Letters 9, 129 135, 1977.
  133. GRIERSON, D. & HEMLEBEN, V. Effect of 8 hydroxyquinoline on the uptake of uridine and incorporation into RNA. Planta 134 155 157, 1977.
  134. GRIERSON, D., CHAMBERS, S.E. & PENNIKET, L.P. Nucleic acid and protein synthesis in discs cut from mature leaves of Nicotiana tabacum L. and cultured on nutrient agar with and without kinetin. Planta 134, 29 34, 1977.
  135. SCHIMMELPFENG, L., GRIERSON, D. & ZETSCHE, K. Preformed poly(A) containing RNA in zoospores of Blastocladiella emersonii. Biochim. Biophys. Acta 435, 340 348, 1976.
  136. COVEY, S.N. & GRIERSON, D. The measurement of plant polyadenylic acid by hybridisation with radioactive polyuridylic acid. Planta 131, 75 79, 1976.
  137. GRIERSON, D. & COVEY, S.N. The properties and function of rapidly labelled nuclear RNA. Planta 130, 317 321, 1976.
  138. COVEY, S.N. & GRIERSON, D. Subcellular distribution and properties of poly(A) containing RNA from cultured plant cells. Europ. J. Biochem. 63, 599 606,
  139. GRIERSON, D. & COVEY, S.N. Changes in the amount of ribosomal RNA and poly(A) containing RNA during leaf development. Planta 127, 77 86, 1975.
  140. GRIERSON, D. The hybridisation of poly(uridylic) acid to DNA from Phaseolus aureus Roxb. Planta 127, 87 91, 1975.
  141. GRIERSON, D. Characterisation of ribonucleic acid components from leaves of Phaseolus aureus. Europ. J. Biochem. 44, 509 515, 1974.
  142. GRIERSON, D. & LOENING, U.E. Ribosomal RNA precursors and the synthesis of chloroplast and cytoplasmic ribosomal ribonucleic acid in leaves of Phaseolus aureus. Europ. J. Biochem. 44, 501 507, 1974.
  143. JOHNSON, C.B., HOLLOWAY, B.R., SMITH, H. & GRIERSON, D. Isoenzymes of acid phosphatase in germinating peas. Planta 115, 1 10, 1973.
  144. JOHNSON, C.B., GRIERSON, D. & SMITH, H. The expression of plac 5 DNA within cultured cells of a higher plant. Nature New Biology 244, 105 107, 1973.
  145. GRIERSON, D. & SMITH, H. The synthesis and stability of ribosomal RNA in blue green algae. Europ. J. Biochem. 36, 280 285, 1973.
  146. GRIERSON, D. & LOENING, U.E. Distinct transcription products of the ribosomal genes in two different tissues. Nature New Biology 235, 80 82, 1972.

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