A Possible Incorporation of Ethylene into RNA in Japanese Morning Glory Seedlings

Shimokawa, Keishi; Kasai, Zenzaburo

doi:10.1271/bbb1961.32.680

Cited by 17 publications

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“…Exposure of etiolated seedlings to fluorescent light during 14C2H4 treatment was without effect. Similarly, indoleacetic acid, gibberellic acid, benzyladenine, abscisic acid, and dibutyryl cyclic adenosine monophosphate had no significant effect on either a or b.The possibilities that the incorporation of 14"CH4 into pea tissues and its conversion to "4CO2 is linked to ethylene action, or that it represents a means of reducing the endogenous ethylene level, are discussed.Several problems encountered with the use of polyethylene vials, rubber serum stoppers, Clorox, and microbial contamination are also described.Several reports have been published indicating that "C-or 'H-labeled ethylene is incorporated and metabolized by various fruit (6,(12)(13)(14) and vegetative tissues (10,11,16,17). Unfortunately, these studies have not proven very convincing, because in no case were adequate precautions taken with regard to "C2H4 purity or microbial contamination.…”

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“…Several reports have been published indicating that "C-or 'H-labeled ethylene is incorporated and metabolized by various fruit (6,(12)(13)(14) and vegetative tissues (10,11,16,17). Unfortunately, these studies have not proven very convincing, because in no case were adequate precautions taken with regard to "C2H4 purity or microbial contamination.…”

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confidence: 99%

“…Unfortunately, these studies have not proven very convincing, because in no case were adequate precautions taken with regard to "C2H4 purity or microbial contamination. Several of these studies (10,11,16,17) were further complicated because the "CMH4 was regenerated from mercuric perchlorate, Del. 19898. readily incorporated by plant tissues (5,14).…”

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¹⁴C₂H₄: Its Incorporation and Metabolism by Pea Seedlings under Aseptic Conditions

Beyer

1975

Plant Physiol.

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The effects of various treatments on the recently reported svstem in pea (Pisum sativum cv. Alaska), which results in (a) the incorporation of "C2H4 into the tissue and (b) the conversion of "C214 to "4CO2, was investigated using 2-day-old etiolated seedlings which exhibit a maximum response. Heat treatment (80 C, 1 min) completely inhibited both a and b, whereas homogenization completely inhibited b but only partially inhibited a. Detaching the cotyledons from the root-shoot axis immediately before exposing the detached cotyledons together with the root-shoot axis to 14C2H4 markedly reduced both a and b. Increasing the "CM4 concentration from 0.14 to over 100 1I/l progressively increased the rate of a and b with tissue incorporation being greater than 14C2H4 to 14CO2 conversion only below 0.3 ,/1/1 14C2M4. Reduction of the 02 concentration reduced both a and b, with over 99 % inhibition occurring under anaerobic conditions. The addition of C02 (5% ) severely inhibited 14C2H4 to 14CO2 conversion without significantly affecting tissue incorporation. Exposure of etiolated seedlings to fluorescent light during 14C2H4 treatment was without effect. Similarly, indoleacetic acid, gibberellic acid, benzyladenine, abscisic acid, and dibutyryl cyclic adenosine monophosphate had no significant effect on either a or b.The possibilities that the incorporation of 14"CH4 into pea tissues and its conversion to "4CO2 is linked to ethylene action, or that it represents a means of reducing the endogenous ethylene level, are discussed.Several problems encountered with the use of polyethylene vials, rubber serum stoppers, Clorox, and microbial contamination are also described.Several reports have been published indicating that "C-or 'H-labeled ethylene is incorporated and metabolized by various fruit (6,(12)(13)(14) and vegetative tissues (10,11,16,17). Unfortunately, these studies have not proven very convincing, because in no case were adequate precautions taken with regard to "C2H4 purity or microbial contamination. Several of these studies (10,11,16,17) Wilmington, Del. 19898. readily incorporated by plant tissues (5, 14). Additional skepticism has been derived from the fact that several workers (3,6,9,15) failed to observe any ethylene incorporation or metabolism.Recently a technique (5) was described for purifying "CMH by gas chromatography to overcome the first, and perhaps most serious, of these criticisms. In a recent report (4), data were presented demonstrating that etiolated pea seedlings grown under aseptic conditions and treated with ultrahigh purity "C,H4 obtained by this technique actively incorporate and metabolize "CMH. In this paper these findings are confirmed and extended.MATERIALS AND METHODS "C,H4 Purification. The technique for purifying "CMH by gas chromatography has been described recently (5) and therefore will be reviewed only briefly. Uniformly labeled 4C2H4 (22 mCi/mmole) with a reported radiochemical purity of 98% was purchased from Amersham/Searle and purified first on a silver nitrate-et...

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¹⁴C₂H₄: Its Incorporation and Metabolism by Pea Seedlings under Aseptic Conditions

Beyer

1975

Plant Physiol.

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“…Therefore, the reported incorporation may have been an artifact due to impurities originally present in the labeled ethylene or impurities derived from radiation decomposition (16). Second, several studies (8,14,15) was undertaken using ultra high purity "4C-labeled ethylene and aseptic conditions. This paper describes the GC' technique used throughout this study for obtaining ultra high purity "4C-labeled ethylene.…”

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“…In several previous`CM,4 studies (8,14,15) "C2H4 was trapped in Hg(CI04)2 and then regenerated in enclosed chambers for exposure to plant tissues. The work by Hall et al (8) with cotton and Coleus made this procedure seem questionable since they found that such treatment greatly enhanced the amount of label fixed.…”

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¹⁴C₂H₄: Its Purification for Biological Studies

Beyer¹

1975

Plant Physiol.

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A gas chromatographic technique is described for obtaining ultra high purity 14(GH for use in biological studies. l4CHL purchased from commercial sources contained readily detectable impurities including radioactive acetylene. Following purification on two different columns, no impurities were detected by high sensitivity gas chromatographic analysis. However, shortly thereafter impurities were detected as a result of radiation decomposition. Trapping and immediately regenerating ultra high purity 14C24 from dilute, filtered Hg(CI04)2 solutions did not cause the formation of impurities, whereas additional impurities were formed when unpurified 14C2HI was used. Impurities were also formed when ultra high purity "GC-L was stored in such solutions prior to its regeneration or when it was trapped and immediately regenerated from more concentrated Hg(CI04) solutions.Surprisingly little is known about the metabolic fate of ethylene. In fact, it is not yet clear whether or not ethylene is incorporated into plant tissues, despite the fact that several studies (8-12, 14, 15) using radioactive ethylene have reported that a small but significant amount of label was incorporated by fruit and vegetative tissues. The present uncertainty, even in light of these reports, exists because of three major criticisms which have been raised (1, 2, 5) and the fact that other studies (4,6,7, 13) have failed to obtain any ethylene incorporation or metabolism. The first criticism is that adequate precautions were not taken to ensure that the label incorporated was derived from radioactive ethylene. Therefore, the reported incorporation may have been an artifact due to impurities originally present in the labeled ethylene or impurities derived from radiation decomposition (16). Second, several studies (8,14,15) was undertaken using ultra high purity "4C-labeled ethylene and aseptic conditions. This paper describes the GC' technique used throughout this study for obtaining ultra high purity "4C-labeled ethylene.Purchased samples of uniformly 14C-labeled ethylene with a reported radiochemical purity of 98% (22, 31, and 53 mCi/ mmole, Amersham/Searle) were purified twice by GC. All separations were performed on a Perkin-Elmer 900 GC connected to a Hewlett-Packard Model 7128A recorder and equipped with a fiame ionization detector and subambient accessory. Ultra high purity helium (Matheson Gas Products) served as the carrier gas while ultra high purity hydrogen (Elhygen H, generator) and oxygen (Matheson Gas Products) provided the flame. Routine ethylene analyses were performed isothermally at 110 C on a 1.83 m X 3.13 mm stainless steel activated alumina column (80-100 mesh) at a flow rate of 70 cc/ min. The sensitivity and stability of this instrument was such that 6 nl/l of ethylene could routinely be detected in a 1-cc gas sample producing a peak twice the baseline noise level with a retention time of 0.9 min.The "4C-labeled ethylene samples were purchased in 250-,4Ci lots and were shipped in small evacuated glass vials with a bre...

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Ethylene Metabolism and Its Possible Physiological Role in Plants

Beyer¹,

Blomstrom

1980

Proceedings in Life Sciences

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A Possible Incorporation of Ethylene into RNA in Japanese Morning Glory Seedlings

Cited by 17 publications

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¹⁴C₂H₄: Its Incorporation and Metabolism by Pea Seedlings under Aseptic Conditions

¹⁴C₂H₄: Its Incorporation and Metabolism by Pea Seedlings under Aseptic Conditions

¹⁴C₂H₄: Its Purification for Biological Studies

Ethylene Metabolism and Its Possible Physiological Role in Plants

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A Possible Incorporation of Ethylene into RNA in Japanese Morning Glory Seedlings

Cited by 17 publications

References 0 publications

14C2H4: Its Incorporation and Metabolism by Pea Seedlings under Aseptic Conditions

14C2H4: Its Incorporation and Metabolism by Pea Seedlings under Aseptic Conditions

14C2H4: Its Purification for Biological Studies

Ethylene Metabolism and Its Possible Physiological Role in Plants

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¹⁴C₂H₄: Its Incorporation and Metabolism by Pea Seedlings under Aseptic Conditions

¹⁴C₂H₄: Its Incorporation and Metabolism by Pea Seedlings under Aseptic Conditions

¹⁴C₂H₄: Its Purification for Biological Studies