Electrophoretic protein patterns in relation to low temperature tolerance and growth regulation of alfalfa

Faw, Wade F.; Jung, G. A.

doi:10.1016/0011-2240(72)90177-0

Cited by 29 publications

(16 citation statements)

References 24 publications

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“…Approximate relative mobilities of the major component of each gel column region for distilled H20 extracts, assuming the origin to be 0 and the center of the fastest moving band to be 10, were as follows: column region 1 = 10.0, 2 = 8.6, Although protein values ranged from 1 to 57 mg/g tissue, protein concentration was usually proportional to cold tolerance (Table I). Linear correlation coefficients between protein concentration and cold tolerance exceeded 0.9 with all extractants except glycine-HCl, pH 3 our results with this extractant are in agreement with those reported by Gerloff et al (4). Total amounts of protein on the gel columns were greatly influenced by the five environments to which the plants had been exposed, but the effects were different for the two cultivars.…”

Section: Methodssupporting

confidence: 93%

“…Disc electrophoresis was used to detect protein components extracted by the 10 extractants (Table II). Techniques used for polyacrylamide gel column preparation, electrophoresis, staining, destaining, and densitometry of stained columns were described in an earlier report (3).…”

Section: Methodsmentioning

confidence: 99%

“…Densitometric graphs were divided into 14 regions, because low areas or valleys between peaks appeared most consistently at these points (3). Approximate relative mobilities of the major component of each gel column region for distilled H20 extracts, assuming the origin to be 0 and the center of the fastest moving band to be 10, were as follows: column region 1 = 10.0, 2 = 8.6, Although protein values ranged from 1 to 57 mg/g tissue, protein concentration was usually proportional to cold tolerance (Table I).…”

Section: Methodsmentioning

confidence: 99%

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Extractant Influence on the Relationship between Extractable Proteins and Cold Tolerance of Alfalfa

Faw

Shih²,

Jung³

1976

Plant Physiol.

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The influence of ionic composition and pH of extractant on the relationship between the extracted proteins and the cold tolerance of Vernal and Arizona Common alfalfa (Medicago sadiva L.) was examined. Five environments were used to induce different tolerance levels.The quantity of protein extracted from plants was influenced by the hardening environment, cultivar, and ionic composition and pH of 29 extractants. Extractants with a pH below 6 generally extracted less protein.The measured cold tolerance of the plants was correlated with the quantity of protein detected in many of the 14 regions of the electrophoresis gel columns regardless of extractant but was most dosely associated with the protein in either region 7 or 8 with nine of ten extractants. The purpose of this research was to examine the influence of ionic nature and pH of extractants on the relationship between extractable proteins and cold tolerance. Sources of variation in cold tolerance were cultivars of alfalfa and hardening environments. MATERIALS AND METHODSPlant Material. Cans with perforated bottoms and a volume of 3.78 liters were filled with a sandy loam soil and seeded with approximately 50 seeds of cold-tolerant Vernal or cold-sensitive Arizona Common alfalfa (Medicago sativa L.). The seedlings were allowed to grow in the greenhouse for 7 months and then were moved outside in April. The plants were allowed to reach a flowering stage before clipping to maintain high food reserve levels. Insects were controlled by spraying with malathion and Sevin.In late September, the containers were randomly assigned to one of five environments: (a) day and night temperatures of 7 and 2 C, respectively, and a photoperiod of 8 hr; (b) day and night temperatures of 16 and 10 C, respectively, and a photoperiod of 12 hr; (c) natural environment in the field at Morgantown, W. Va.; (d) greenhouse conditions maintained for vigorous growth; and (e) day and night temperatures of 27 and 21 C, respectively, with a photoperiod of 16 hr. Light in the growth chambers was supplied by six cool-white, 40-w fluorescent lamps and four 60-w incandescent bulbs/chamber. This lighting system provided a light intensity of approximately 12,900 lux at the plant tops. Relative humidity was regulated at approximately 80% in the chambers. A randomized block design was used with three replications for each environmental regime.After the plants had been subjected to the environments for 44 to 46 days, they were sampled for cold tolerance determinations and protein analyses. Plant roots were trimmed to a length of approximately 2.5 cm and crowns to 5 cm. All dead material and plant leaves were removed from the samples. A 10-g portion of prepared crown and root sample was used to measure cold tolerance using the technique of Dexter et al. (2). The remainder of each sample was immediately frozen in liquid N2, lyophilized, ground in a Wiley mill to pass through a 40-mesh screen, and stored at -20 C in air-tight vials that were enclosed in plastic bags.

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Section: Methodssupporting

confidence: 93%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Extractant Influence on the Relationship between Extractable Proteins and Cold Tolerance of Alfalfa

Faw

Shih²,

Jung³

1976

Plant Physiol.

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“…There have been many suggestions that protein synthesis plays a role in cold hardening (1, 4, 9, 13, 23). Among the many changes found during induction of cold hardiness are those which occur in RNA (13, 26) and protein (3, 4,9,13,26) metabolism. This study was undertaken to examine the ribosome as related to induction of cold hardiness.…”

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confidence: 99%

Ribosomal Changes during Induction of Cold Hardiness in Black Locust Seedlings

Bixby

Brown²

1975

Plant Physiol.

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Protein synthesis has been implicated in the cold-hardening process. Ribosomes from cold hardy and nonhardy black locust (Robinia pseudoacacia L.) seedlings were compared to determine if cold acclimation is related to alteration of ribosomal structure. Ribosomal structure, as indicated by thermal melting profiles, appears to be altered during induction of hardiness. Two-dimensional polyacrylamide gel electrophoresis of ribosomal proteins indicates at least 17 proteins from hardy seedlings that are different from those of nonhardy seedlings. These different proteins may be partially responsible for the different thermal melting profiles observed.Plants capable of developing freezing resistance demonstrate altered metabolism during low temperature acclimation (cold hardening) (23). There have been many suggestions that protein synthesis plays a role in cold hardening (1, 4, 9, 13, 23). Among the many changes found during induction of cold hardiness are those which occur in RNA (13, 26) and protein (3, 4,9,13,26) metabolism. This study was undertaken to examine the ribosome as related to induction of cold hardiness. The objective was to determine if ribosomal structure is altered during induction of hardiness. MATERLALS AND METHODSPlant Material. Black locust seedlings (Robinia pseudoacacia L.) were grown in heat-sterilized vermiculite. Moisture was maintained relatively constant. Two-month seedlings were cold hardened as outlined in Table I. At specific points in the coldhardening regime, seedlings were placed into a programmed freezer and frozen at a rate of 6 C/hr. Samples were removed every 4 C starting at -4 and allowed to thaw overnight at 5 C. Stems were then harvested, placed into a mist chamber with a 15-hr photoperiod, allowed to recover for 7 days and visually evaluated for survival. Seedlings were considered uninjured if their bark remained green and tightly bound to the stem. The bark of injured seedlings became brown and loose. Stems were assigned a value of 1 if uninjured, 0.5 if injured with 50% or more of the stem alive and 0 if dead or more than 50% injured. Values were totaled and divided by the total number of stems to determine percent survival (31).

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“…A number of earlier studies described the changes in physiology, metabolism, or enzyme activity associated with the induction of greater freezing tolerance (2). Several of these studies demonstrated an appearance and disappearance of certain membrane proteins (3,4) and changes in the isozyme composition of a number of enzymes (5)(6)(7)(8)(9)(10). Although evidence does exist for altered gene expression in other types of plant stress responses (11,12), direct evidence of altered gene expression during cold acclimation has not been presented.…”

mentioning

confidence: 99%

Altered gene expression during cold acclimation of spinach.

Guy¹,

Niemi²,

Brambl

1985

Proc. Natl. Acad. Sci. U.S.A.

270

140

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Exposure of spinach (Spinacia oleracea L.) plants to a constant 50C induced a greater tolerance to extracellular freezing. The metabolic basis of this cold acclimation response in plants is not understood. In this study we tested the hypothesis that cold acclimation derives from altered gene transcription. We found that exposure of plants to low temperature resulted in a rapid and stable change in the translatable poly(A)+ RNA populations extracted from leaves, as determined by a cell-free in vitro translation assay. The initial appearance of mRNAs for two high molecular weight translation products correlated with an increase in freezing tolerance. Cold acclimation of plants for 8 days resulted in further qualitative changes in mRNA populations. At least four additional mRNAs increased in concentration upon continued exposure of spinach to 50C, whereas three other mRNAs present in 20'C-grown leaves decreased. We also tested the possibility that the low temperature-induced mRNAs might encode heat shock proteins. We studied heat shock-induced protein synthesis by in vivo labeling techniques and found that spinach synthesized at least eight distinctive heat shock proteins during exposure to 400C. Most polypeptides induced by exposure to low temperature, however, appeared not to be heat shock proteins. Thus, the change in mRNAs induced by low temperature is a separate response from that induced by high temperature.The increase in freezing tolerance of plants during cold acclimation has been proposed to result from a combination of physiological changes and metabolic alterations that depend on altered gene expression (1). A number of earlier studies described the changes in physiology, metabolism, or enzyme activity associated with the induction of greater freezing tolerance (2). Several of these studies demonstrated an appearance and disappearance of certain membrane proteins (3, 4) and changes in the isozyme composition of a number of enzymes (5-10). Although evidence does exist for altered gene expression in other types of plant stress responses (11, 12), direct evidence of altered gene expression during cold acclimation has not been presented.We report here data that indicate a qualitative alteration in the population of translatable mRNA during cold acclimation in spinach. We have extracted RNA from nonacclimated and cold-acclimated spinach leaves, prepared and translated in vitro the mRNA fraction, and by electrophoresing the translation products found that development of increased freezing tolerance is correlated with the rapid appearance of two new, continuously expressed species of mRNA. MATERIALS AND METHODSPlant Material and Cold Acclimation. Leaves of spinach, Spinacia oleracea L. cv. Bloomsdale (Northrup King), were used for all studies. Plants were grown in a controlled environment growth chamber with an air temperature at plant height of 20 ± 10C in the light and 17 ± 10C in the dark. Illumination (12 hr/day) was provided by cool white fluorescent tubes and supplemented with incandescent lighti...

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Electrophoretic protein patterns in relation to low temperature tolerance and growth regulation of alfalfa

Cited by 29 publications

References 24 publications

Extractant Influence on the Relationship between Extractable Proteins and Cold Tolerance of Alfalfa

Extractant Influence on the Relationship between Extractable Proteins and Cold Tolerance of Alfalfa

Ribosomal Changes during Induction of Cold Hardiness in Black Locust Seedlings

Altered gene expression during cold acclimation of spinach.

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