Rapid production and field testing of homozygous transgenic tobacco lines with virus resistance conferred by expression of satellite RNA and coat protein of cucumber mosaic virus

A procedure for the fast production of homozygotic transgenic plants was developed. Leaf discs of haploid tobacco plants from anther cultures were transformed with a chimaeric vector containing coat protein (CP) and satellite RNA (Sat-RNA) genes from cucumber mosaic virus (CMV). One-hundred-and-twelve Kanamycin-resistant transformed haploid plants were subjected to selection based on the expression of both CP and Sat-RNA. Eighty-nine transgenic plants expressing both genes were selected and tested for their resistance to CMV by inoculation with high concentration of CMV (200 g ml^–1). Only five plants showed no symptoms of viral infection 30 days after inoculation. These plants were then diploidized by colchicine treatment. Three homozygous diploid lines with high levels of resistance to CMV were obtained after only one generation. The three transgenic lines were further tested under field conditions. The results showed that the progenies of these transgenic lines were homozygous and were highly resistant to CMV under natural field infection and manual inoculation conditions.     

The turnover of chlorophyll in greening wheat leaves

A method for the estimation of chlorophyll turnover in wheat leaves is presented. This is based on the inhibition of chlorophyll synthesis by treatment of the cut leaves with laevulinic acid (LA), a competitive inhibitor of δ-aminolaevulinic acid dehydratase. The turnover of chlorophyll in young, greening leaves, given short periods of light was a relatively rapid process. However, in seedlings exposed to light for longer periods the turnover became progressively slower, and was measured in days rather than hours.     

Localization of N6-methyladenosine in the Rous sarcoma virus genome.

The 10,000-nucleotide RNA genome of the Prague strain, subgroup B (PR-B) of Rous sarcoma virus, was found to contain 11.6 ± 0.5 residues of m⁶Ap by quantitative analysis of ³²P-labeled virion RNA after complete RNAase digestion. Approximately ten of the m⁶Ap residues are located, without obvious clustering, in that region of the genome between 500 and 4000 nucleotides from the 3′ poly(A) end. The src gene, which is required for transformation, and part of the env gene, which codes for the major viral envelope glycoprotein, have previously been mapped in this region of the viral genome. A transformation-defective deletion mutant of PR-B Rous sarcoma virus, which lacks the src gene, has 7.0 ± 0.2 m⁶Ap residues per RNA subunit. This supports our mapping of a portion of the m⁶A residues in src and suggests that this methylation is specific to certain regions of the genome. The possible significance of this result for Rous sarcoma virus RNA processing and translation is discussed.     

Analysis of the volatile essential oils of Murraya koenigii and Pandanus latifolius

Using well-established techniques, samples were obtained of the volatile essential oils of the two types of curry leaf, Murraya koenigii and Pandanus latifolius. Both contained mainly terpenes, and M. koenigii produced less than 4% of other components with eight monoterpene hydrocarbons (ca 16%) and 17 sesquiterpene hydrocarbons (ca 80%) being obtained. The most important constituents of M. koenigii are β-caryophyllene, β-gurjunene, β-elemene, β-phellandrene and β-thujene. The volatile essential oil of P. latifolius also contained mainly sesquiterpene hydrocarbons (6–42%) but the only monoterpene was linalool (ca 6%). Nearly 2000 times the total quantity of aroma volatiles was produced by M. koenigii compared with P. latifolius, and this partly explains the observed stronger flavour potency of the former.     

Taxonomic variation in total leaf protein amino acid compositions of grasses

Leaves of greenhouse grown grasses had free protein amino acid contents of generally less than 5 % total amino acids, while field collected grasses averaged 14.7 % free protein amino acid contents. Taxonomic patterns are detectable in the total leaf amino acid profiles of grasses from both sources, those of pooids being distinguishable from those of chloridoids and panicoids, and those of danthonioids showing an intermediate pattern. Leaf profiles of Oryza, Stipeae, and Ehrharteae resemble one another, and are more like those of pooids than those of panicoids. Variations in Thr and Leu are apparently associated with differences in photosynthetic pathway. Grass leaves are generally low in total amino acid contents (2.2 ? 1.0 g % fr. wt), with Ile, Val and Met + Cys identified as the limiting essential amino acids. However, the nutritional ‘chemical scores’ of grass leaf proteins are high (75 %, based on the WHO scoring pattern).     

The effects of decapitation on the distribution of cytokinins and growth of Phaseolus vulgaris plants

The growth of the primary leaves of Phaseolus vulgaris L. was enhanced greatly by decapitation of the rest of the shoot. This increased growth was manifested by an increase in leaf area, leaf weight, and in a higher synthesis of chlorophyll and soluble proteins. Within the roots and stems decapitation resulted in a detectable increase in the endogenous cytokinins within 2 days after the surgical treatment. In the primary leaves increased cytokinin levels were only detected after 16 days. At this time most of the recorded activity co-chromatographed with the cytokinin glucosides. When plants which were decapitated were left under normal growing conditions for 16 days and then transferred to continuous darkness for 8 days the senescence of the primary leaves of the decapitated plants, in which the cytokinins had increased, was delayed significantly when compared with that of the primary leaves of the intact plants. the significance of these findings is discussed.     

Methylation analysis of extracellular polysaccharides from suspension-cultured cells of Nicotiana tabacum

The soluble extracellular polysaccharides (ECP) of suspension-cultured tobacco cells were fractionated by DEAE-Sephadex CC into seven sub-fractions. Su     

Haem and chlorophyll formation in etiolated and greening leaves of barley

The amounts of protochlorophyllide (P650) and protohaem were measured in ageing dark-grown barley leaves. Maximum amounts of P650 and protohaem were found in 6- to 8-day-old material after which P650 declined rapidly and protohaem more slowly. In leaves exposed to light maximum chlorophyll was produced in 6-day-old material with progressively less the older the leaves. Haem concentrations increased in seedlings of all ages exposed to light. A lag phase was observed for both chlorophyll and haem formation in leaves given a light treatment. Haem, however, showed a slight yet sig nificant decline as chlorophyll production commenced. The results indicate that chlorophyll and haem synthesis share a common pool of δ-aminolae vulinic acid (ALA). At a certain stage of development, the magnesium porphyrin pathway diverts precursors away from haem synthesis. It is only when the ALA synthesising system is well developed that the production of ALA can satisfy pathways to both haem and chlorophyll. The observed changes in haem under certain conditions suggest that, as in animal systems, haem levels may regulate porphyrin formation (chlorophylls) by controlling the supply of ALA.     

Effect of phenolic compounds on glucose-6-phosphate dehydrogenase isoenzymes

Effector studies with two isoenzymes (I and IV) of glucose-6-phosphate dehydrogenase (G6PDH) from tobacco suspension culture WR-132 revealed that chlorogenic acid, at 0.4 mM, inhibited both isoenzymes almost 100%, with the inhibition decreasing as the concentration of the acid was reduced. At 0.3 and 0.4 mM, the coumarin glucosides scopolin and esculin were inhibitory, whereas their aglucones scopoletin and esculetin were less inhibitory, and at low concentrations of glucose-6-phosphate (G6P), the latter two were actually stimulatory for G6PDH I. Of the possible effectors studied, only scopoletin and esculetin exhibited a significant activation of G6PDH I under these conditions. However, with G6PDH IV these two effectors do not show the same marked activation at the low G6P concentrations. The phenolic acids, caffeic and ferulic, were less inhibitory than the coumarins tested. The activation of G6PDH I by scopoletin, a compound which accumulates in tobacco under certain stress conditions, gives a possible clue as to the resulting enhanced activity of the hexose monophosphate pathway that has been reported for some plants subjected to stress conditions.