Summary
A cDNA library was prepared from, poly(A)+ RNA from roots of pea (Pisum sativum L.). Twenty five clones were selected by use of random numbers and used as probes on Northern blots to analyse the distribution of their corresponding mRNA species in other vegetative pea organs: leaf, stem and developing cotyledon. Fifteen cDNA inserts hybridised to single mRNA species, five hybridised to two mRNA species and one hybridised to five homologous mRNAs. Four cDNA clones (16% of those selected) gave no hybridization signals, indicating that the steady state levels of mRNAs were below the detection limit (i.e.less than 2.5 x 10-5% of poly(A)+ RNA). Most of the root mRNAs were represented in all four pea organs as sequences of low and medium abundance. All but two cDNAs encoded mRNA species enhanced in root. However, cDNA clones appeared not to encode mRNA species expressed in a strictly organ-specific manner, as no mRNA unique to root was found. Thus, if organ-unique mRNA species are present, they are only present at a very low level of abundance in the poly(A)+RNA population.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aviv H, Leder P (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acidcellulose. Proc Natl Acad Sci USA 69:1408–1412
Croy RRD, Lycett GW, Gatehouse JA, Yarwood JN, Boulter D (1982) Cloning and analysis of cDNAs encoding plant storage protein precursors. Nature 295:76–79
Domoney C, Casey R (1985) Measurement of gene number for seed storage proteins in Pisum. Nucleic Acids Res 13:687–699
Evans IM, Croy RRD, Brown P, Boulter D (1980) Synthesis of complementary DNAs to partially purified mRNAs coding for the storage proteins of Pisum sativum L. Biochim Biophys Acta 610:81–95
Evans IM, Croy RRD, Hutchinson P, Boulter D, Payne PI, Gordon ME (1979) Cell free synthesis of some storage protein subunits by polyribosomes and RNA isolated from developing seeds of pea (Pisum sativum L.). Planta 144:455–462
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Gatehouse JA, Evans IM, Bown D, Croy RRD, Boulter D (1982) Control of storage-protein synthesis during seed development in pea (Pisum sativum L). Biochem J 208:119–127
Gatehouse JA, Bown D, Gilroy J, Levasseur M, Castleton J, Ellis THN (1988) Two genes encoding “minor” polypeptides in pea (Pisum sativum L.). Biochem J 250:15–24
Gebhardt C, Oliver JE, Forde BG, Saarelainen R, Miflin BJ (1986) Primary structure and differential expression of glutamine synthetase genes in nodules, roots and leaves of Phaseolus vulgaris. EMBO J 5:1429–1435
Hall TC, Ma Y, Buckbinder BU, Pyne JW, Sun SM, Bliss FA (1978) Messenger DNA for G1 protein of French bean seeds: Cell-free translation and product characterisation. Proc Natl Acad Sci USA 75:3196–3200
Higgins TJV (1984) Synthesis and regulation of major proteins in seeds. Annu Rev Plant Physiol 35:191–221
Kamalay JC, Goldberg RB (1984) Organ specific nuclear RNAs in tobacco. Proc Natl Acad Sci USA 81:2801–2805
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York
Marzluff WF, Huang RCC (1984) Transcription of RNA in isolated nucleic. In: Hames BD, Higgins SJ (eds). Transcription and Translation. IRL Press, Oxford, pp 89–129
McMasters GK, Carmichael GG (1977) Analysis of single-and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci USA 74:4835–4838
Rosahl S, Eckes P, Schell J, Willmitzer L (1986) Organ-specific gene expression in potato: isolation and characterisation of tuber-specific cDNA sequences. Mol Gen Genet 202:368–373
Thomas PS (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205
Tingey SV, Walker EL, Coruzzi GM (1987) Glutamine synthetase genes of pea encode distinct polyeptides which are differentially expressed in leaves, roots and nodules. EMBO J 6:1–9
Vaillant V, Buffard D, Esnault R (1983) Changes in the polyadenylated messenger RNA population during differentiation of Vicia faba root cells. Cell Differ 13:201–208
Walling L, Drews GN, Goldberg RB (1986) Transcriptional and post-transcriptional regulation of soybean seed protein mRNA levels. Proc Natl Acad Sci USA 83:2123–2127
White BA, Bancroft FC (1982) Cytoplasmic dot hybridisation. J Biol Chem 257:8569–8572
Wickens MP, Buell GN, Schimke RT (1978) Synthesis of double-stranded DNA complementary to lysozyme, ovomucoid and ovalbumine mRNAs. J Biol Chem 253:2843–2495
Author information
Authors and Affiliations
Additional information
Communicated by H.Saedler
Rights and permissions
About this article
Cite this article
Evans, I.M., Swinhoe, R., Gatehouse, L.N. et al. Distribution of root mRNA species in other vegetative organs of pea (Pisum sativum L.). Mol Gen Genet 214, 153–157 (1988). https://doi.org/10.1007/BF00340194
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00340194