Preformed mRNA and Light-induced Germination of Pine (Pinus thunbergii) Seed

The mRNAs were isolated from dry, dark-imbibed and light-imbibedpine (Pinus thunbergii) seeds, in which germination is inducedupon exposure to light, and their translational activities ina wheat embryo cell-free system were examined. A portion ofthe mRNA extracted from each type of seed appeared to be poly(A)⁺RNA.A significant translational activity was already present inthe RNA fraction isolated from the dry pine seeds. The preformedmRNA seems not to be translated in vivo during the dark-imbibition,since most of the in vivo protein synthesis did not occur untilthe seeds were exposed to light. The SDS polyacrylamide gelelectrophoregrams of the polypeptides synthesized in vitro inresponse to either the preformed mRNA or the mRNAs from thedark-imbibed or light-imbibed seeds were qualitatively identical;thus it seems that the preformed mRNA is conserved during darkimbibition, and that its translation is initiated after theexposure of the imbibed seeds to light. (Received August 10, 1981; Accepted May 15, 1982)     

Effects of red pine (Pinus resinosa Ait.) mycorrhizoplane-associated actinomycetes onin vitro growth of ectomycorrhizal fungi

Mycorrhizoplane-associated actinomycetes were isolated using an enrichment technique from red pine (Pinus resinosa Ait.) roots of seedlings recently outplanted onto cleared northern hardwood sites in the Upper Peninsula of Michigan, USA. Interactions were assessedin vitro between actinomycete isolates and three commonly occurring ectomycorrhizal fungi (Laccaria bicolor (Maire) Orton,L. laccata (Scop.: Fr.) Berk. and Br., andThelephora terrestris Fr.). Most actinomycete isolates exerted a range of effects on the growth of the three fungus isolates during the four week test period, inhibiting some while stimulating others; several inhibited growth of all three fungus isolates. Mycorrhizoplane-associated actinomycetes show potential for use as coinoculants with selected ectomycorrhizal fungi to optimize the soil microflora for developing seedlings.     

Nucleic Acid Metabolism in Germinating Onion: I. Changes in Root Tip Nucleic Acid during Germination

Nucleic acid synthesis in the G₁ cell population of the 1-millimeter apex of the Allium cepa embryo was studied during the initial 73 hours of germination. Quantitative data indicate that the total amount of RNA per cell began to increase after 18 hours of germination while the initial DNA per cell increase did not occur until some 20 hours later. Polyacrylamide gel electrophoresis patterns of ³H-uridine-labeled total nucleic acid samples indicated that synthesis of all detectable RNA fractions present in the pre-emergent 1-millimeter apex (i.e., cytoplasmic and “chloroplast-like” RNA) began at approximately the same time (18 hours). Synthesis of the various cytoplasmic RNA fractions continued throughout the germination period. Data indicating synthesis of the “chloroplast-like” RNA were obtained only for the initial 36 hours of germination. Specific radioactivity of ³H-uridine-labeled total nucleic acid increased during the first 41.5 hours of germination but then decreased while the accumulation of RNA per cell continued to increase throughout the 73-hour period. In addition, a method is described which reduced the bacterial contamination of Allium seed to a level not detectable by incorporation of radioactive precursors into bacterial ribosomal RNA.     

Changes in Cytokinin Activity during Seed Germination in Rice (Oryza sativa L.)

Cytokinin activity was determined in dry mature rice seeds,in endosperm and embryo tissues 24, 48 and 72 over imbibitionand in radicles 96 h after germination. Cytokinins with chromatographicproperties similar to zeatin, zeatin riboside, zeatin glucosideand zeatin riboside glucoside were datected in embryo and endosperm,but only the latter two were detected in mature seeds. Cytokininactivity was low during early toges of germination. Qualitativeand quantitative changes in cytokinins were observed in bothembryo and endosperm. The presence of higher cytokinin activityin the endosperm than in the embryo during the first 24 h aftergermination suggests that the endosperm may supply cytokininsuntil the embryo is able to synthaize its own cytokinins. Thepossible significance of high cytokinin glucoside activity inthe embryo early during germination and high cytokinin activityin the radicle during the later stages is discussed. Oryza sativa L., rice, cytokinin, germination, seed     

Morphological and chemical analyses of red pine (Pinus resinosa ait.) buds

Summary On a known red pine, K-deficient to non-K-deficient, site continuum which has been examined for tree morphological and chemical properties along with soil physical and chemical properties, results of bud analyses are presented and discussed as a possible diagnostic technique. Simple correlations between all pairs of bud morphological properties and between these properties and tree height, and between all pairs of bud chemical properties and between these properties and tree height were examined; and the results of the bud nutrient element concentrations and tree height were subjected to a multiple regression analysis.The results demonstrated the potential value of bud chemical analysis for diagnostic purposes.     

Nucleic Acid Metabolism of Vicia faba during Germination and Growth

During the growth of Vicia faba seedlings in the absence of an external nitrogen supply, the cotyledons decreased rapidly in dry weight and nucleic acid content. In the developing shoot the dry weight increased rapidly for four weeks and then very slowly over the next two weeks growth; the nucleic acid content of the shoot increased to a maximum after 4 weeks growth and decreased in amount during the next 2 weeks. On the other hand the roots increased in both dry weight and nucleic acid content throughout the growth period, although they only accounted for a small proportion of the total dry weight and nucleic acid content of the plant. These changes during germination and growth are discussed in relation to those occurring during these developmental stages in other plants.     

Pinewood Nematode,Bursaphelenchus xylophilus,Associated with Red Pine, Pinus resinosa,in Western Maryland

Red pines Pinus resinosa in Garrett and Allegany counties, Maryland, were examined during 1982-84 to determine distribution of the pinewood nematode, Bursaphelenchus xylophilus, within and among trees. Approximately 25-year-old (younger) and 47-year-old (older) trees were subdivided into the following categories: 1) trees with mostly green needles; 2) trees with mostly reddish-brown needles; 3) trees lacking needles but with bark intact; 4) trees lacking both needles and bark; and 5) trees with chlorotic, bleached-green needles. Bursaphelenchus xylophilus was found infecting 68% of younger red pines and 77% of older red pines. Nematodes were not evenly distributed in trees within any given tree decadence category or in trees of the same age. Nematodes were recovered from 20% of wood samples from trunks and primary and secondary branches in younger pines and from 15 % of older red pines. On the basis of tree decadence category, the highest incidence of infection in younger trees (31%) was in bleached-green needled trees (category 5), whereas in older trees the highest infection (25%) occurred in green needled trees (category 1). At both sites trunks were infected more often than branches.     

Changes in Endogenous Cytokinins of Lettuce Seed during Germination

Using the soybean callus bioassay it has been shown that dormant lettuce seeds (Lactuca sativa L. cv. Grand Rapids) contain large amounts of water soluble cytokinins and small amounts of butanol soluble ones. When the seeds are irradiated with red light, or imbibed with 5 mg/1 gibberellic acid in the dark, the total cytokinin content of the seeds decreases, the level of water soluble cytokinins decreases, and the level of the butanol soluble cytokinins increases. Far-red light does not reverse this effect completely although cytokinin activity in the butanol extracts decreases following such irradiation. It is proposed that the interconversion of cytokinins initiated by red light, or gibberellic acid in the dark, is one of the primary events leading to radicle elongation in light-sensitive lettuce seed.     

Changes in Lipoxygenase Components of Rice Seedlings during Germination

Changes in lipoxygenase (LOX) activity were followed duringthe germination of rice seeds. The enzyme activity of 3-day-oldseedlings was 20 times higher than that of ungerminated seeds.Sixty per cent of the increased activity was found in shoots.The increase in LOX activity was mainly due to an increase inlipoxygenase-2 (LOX-2), a minor component in ungerminated seeds;this increase was inhibited by cycloheximide. LOX-2 was isolatedfrom the 3-day-old seedlings and compared for its enzymologicalproperties with rice lipoxygenase-3 (LOX-3), a major componentin ungerminated seeds. Both LOX-2 and LOX-3 were stable at pH5 to 8, but LOX-2 was more heatstable than LOX-3. Apparent K_mvalues of LOX-2 and LOX-3 for linoleic acid were 170 and 59µM, and those for linolenic acid were 5,300 and 88 µM,respectively. Both LOXs were inhibited by some metal ions andantioxidants. (Received February 5, 1986; Accepted May 9, 1986)     

Measuring Nitrogen and Phosphorus Uptake by Intact Roots of Mature Acer saccharum Marsh., Pinus resinosa Ait., and Picea abies (L.) Karst

A common method for measuring uptake by intact roots in situ is the depletion method, wherein intact fine roots are separated from soil and placed in nutrient solution. The difference between initial and final amounts of nutrient in solution is attributed to root uptake. Variations on this method include applying pretreatment solutions, training roots to grow into bags or trays, and varying concentrations of nutrient solution. We tested whether variations in methods affected measured net uptake rates of NH ₄ ⁺ , NO ₃ ⁻ , and PO ₄ ³⁻ . Intact roots of 60 year-old sugar maple (Acer saccharum Marsh.), red pine (Pinus resinosa Ait.), and Norway spruce (Picea abies (L.) Karst.) were given one of four treatments prior to measuring net uptake. “Trained” roots were grown in a sand-soil mixture. “Recovered” roots were excavated and allowed to recover in nutrient solution for two or four days (“two-day recovery” and “four-day recovery”, respectively). “No recovery” roots were excavated and used immediately in experiments. We exposed roots to three concentrations of nutrient solutions to observe the effects of initial nutrient solution concentration. Initial nutrient solution concentration was an important source of variation in measured uptake rates, and N uptake was stimulated by low antecedent concentrations. We found no significant differences in net uptake rates between pretreatments for any of the species studied, indicating that our pretreatments were not effective in improving measurement of uptake. Such pretreatments may not be necessary for measuring net uptake and may not hinder the comparison of rates measured using variations of the depletion method.     

Changes in Phosphorus Fractions and Phytase Activity of Rice Seeds during Germination

The present experiment provides information on the phosphorus compounds in rice seeds and elucidates the changes they undergo during germination. In ungerminated seeds, the bulk of total-P appears in phytin (about 76 per cent). It is then dephosphorylated in course of germination with a simultaneous accumulation of large amounts of inorganic-P. Lipid-P increases very rapidly from 0 to 24 hours. The increase up to 72 hours is gradual, after which it drops at 96 hours and again rises to a maximum after 120 hours. The ester-P and RNA-P, fractions increase in concentration with time of germination (except 120 hours). Protein-P begins to fall after 48 hours, while DNA-P remains more or less constant throughout the experiment. The two pH optima recorded for phytase activity at 4.0 and 9.0, suggests that there exist two phytases, one acidic and the other alkaline. Both behave similarly during germination with a continuous increase throughout the course of the experiment. The enzyme with an optimal pH at 4 hydrolyses phytin more actively than the other with the pH optimum at 9.0. Phytase shows maximum activity at a stage when most of the phytin has disappeared; the metabolic significance is uncertain.     

Isoenzyme Changes during Seed Germination of Saguaro Cactus (Carnegiea gigantea)

Esterases, acid phosphatases, leucine aminopeptidases, peroxidases and glutamic-oxalacetic transaminases from extracts made from seeds of Saguaro cactus at different stages of germination were studied using starch gelelectrophoretic techniques. In general, four of the five enzyme systems showed significant increase in the number of isoenzymes and their activity after 72 hours of germination. It is suggested that Saguaro cactus seeds reach their enzymatically most active state between 48 to 72 hours after being placed at optimum moisture conditions at 30°C in continuous light.     

Changes in mRNA of Vigna mungo Cotyledons during Seed Germination

Quantitative and qualitative changes of mRNA in Vigna mungocotyledons during seed germination have been investigated. TotalRNA is higher in dry cotyledons and declines during germination.Poly(A)⁺ RNA also is present at a relatively high level in drycotyledons, increases slightly during the first day of germination,and then decreases. Polysomal RNA is very low in dry cotyledonsbut increases rapidly during the first day of germination, andthen declines. The translational activity of the mRNA in a wheatgerm cell-free system is low on day 0 but increases rapidlyon day 1 of germination. Two-dimensional gel electrophoresisof in vitro translation products reveals that many new peptidesare synthesized on day 1 of germination. Synthesis of most ofthese polypeptides continue throughout 5 days of germination. Change in the mRNA population during germination has been investigatedusing cDNA against poly(A)⁺ RNA from 3-day-old cotyledons. Withtotal RNA of day 3 and 5, the cDNA strongly hybridized withRNA similar in size to 25 S ribosomal RNA, but no specific bandsare detected with samples of day 0 or 1. With poly(A)⁺ RNA ofday 5 or 1, the cDNA tends to hybridize with RNAs of relativelysmall molecular size. Cordycepin and -amanitin prevent the increasein poly (A)⁺ RNA content and the appearance of new mRNAs duringthe first day of germination. ¹Present address: Division of Regulation of Macromolecular Function,Institute for Protein Research, Suita City, Osaka 565, Japan. (Received January 13, 1986; Accepted June 10, 1986)