Protein Synthesis During Rehydration, Germination and Seedling Growth of Naturally and Precociously Matured Soybean Seeds (Glycine max)

p. 86, line 6, should read: These patterns of soluble protein synthese are similar to thoseobserved after precocious maturation and subsequent rehydrationof castor bean (Kermode and Bewley, 1985a, b, 1986), and withinaxes of Phaseolus vulgaris L. seeds (Dasgupta and Bewley, 1982). instead of: These pattern of protein synthesis. Unlike castor bean thoseobserved after precocious maturation and subsequent rehydrationof castor bean (Kermode and Bewley, 1985a, b, 1986), and withinaxes of Phaseolus vulgaris L. seeds (Dasgupta and Bewley, 1982).     

Changes in Seed Constituents During Germination and Seedling Growth of Precociously Matured Soybean Seeds(Glycine max)

During 7 d of precocious maturation of soybean seed (Glycinemax), the starch content declined and soluble sugar levels increasedin patterns similar to natural seed dehydration and maturation.Total seed protein content and total seed dry weight increasedwhereas oil content remained relatively unchanged. Overall,the proportions of the constituents in precociously maturedseeds were comparable to naturally mature seeds. Precociouslymatured soybean seeds showed much the same germination and seedlinggrowth frequency patterns as naturally matured seeds. Duringgermination and seedling growth of precociously matured seeds,starch, soluble sugar, protein and oil levels followed patternssimilar to naturally mature, germinating seeds and seedlings.Therefore, precocious maturation may be used as a model systemto investigate the control of the physiological and biochemicalevents occurring during seed maturation which lead to germinationand subsequently, seedling growth. Glycine max (L.) Merr., soybean, cotyledons, maturation, germination/seedling growth     

Moisture Loss as a Prerequisite for Seedling Growth in Soybean Seeds (Glycine max L. Merr.)

Rosenberg, L. A. and Rinne, R. W. 1986. Moisture loss as a prerequisitefor seedling growth in soybeanseeds (Glycine max L. Merr.).—J.exp. Bot. 37: 1663–1674. As soybean seeds [Glycine max (L.) Merr.] develop, they undergoa change in seed moisture. When excised prematurely from thepod and planted, seeds do not exhibit seedling growth until63 d after flowering (DAF) when the seed moisture has fallenbelow 60%. In contrast, seed germination (radicle protrusion)can occur when seeds as young as 35 DAF (68–79% moisture)are excised, but this germination docs not lead to comparableseedling growth frequencies unless seeds are first given a moistureloss treatment to artificially reduce their moisture below 60%.A moisture loss treatment applied at 35 DAF thus enables seedto undergo the transition from germination (cell expansion)to seedling growth (cell division and expansion) to the extentthat treated immature seed have a vigour index comparable toseeds matured on the plant (100%). The pattern of protein synthesisin vivo was examined in 35 DAF seed using [³⁵S]-methionine incorporation.When moisture loss treatment was applied for 24 h to 35 DAFseeds, seeds synthesized several new polypeptides when comparedwith untreated seeds at the same developmental stage. The sameseed samples showed 0% seedling growth in the absence of moistureloss treatment and 80% seedling growth when the treatment hadbeen applied. Moisture loss from soybean seeds appears to bea prerequisite for the synthesis of new proteins which may bepart of the metabolic process or processes that allow the soybeanseed to undergo the transition from seed germination to seedlinggrowth. Key words: Moisture loss, germination/growth, soybean     

Malonyl Isoflavone Glycosides in Soybean Seeds (Glycine max Merrill)

The isoflavone constituents in soybean seeds were investigated, and 9 kinds of isoflavone glycosides were isolated from the hypocotyls of soybean seeds. Three kinds were proved to be malonylated soybean isoflavones named 6″-O-malonyldaidzin, 6″-O-malonylglycitin and 6″-O-malonylgenistin by UV, MS, IR and NMR. The malonylated isoflavone glycosides as major isoflavone constituents in soybean seed were thermally unstable, and were converted into their corresponding isoflavone glycosides. All of the isoflavone components produced intensely undesirable taste effects such as bitter, astringent and dry mouth feeling.     

Biogenesis of Protein Bodies in Embryonic Axes of Soybean Seeds (Glycine max. cv. Enrei)

Protein bodies in embryonic axes of soybean seeds have inclusion structures containing phytin globoids. Biogenesis of the protein bodies during seed development was examined by transmission electron microscopy. Protein bodies in embryonic axes originated from central vacuoles. The central vacuole in embryonic axes subdivided into smaller vacuoles with internal membranous structure. Then the subdivided vacuoles were directly associated with rough endoplasmic reticulum (rER), and were filled with proteinaceous matrix from the peripheral region. The increase of matrix was simultaneous with accumulation of β-conglycinin estimated by SDS-polyacrylamide gel electrophoresis. Glycinin-rich granules that had been found in developing cotyledons were not observed in embryonic axes. After proteinaceous matrix filled the protein bodies, electron-transparent regions presumably surrounded by a single membrane appeared in the matrix. Phytin globoids were constructed in this internal structures of protein bodies as the final step of protein body formation.     

Transformation of Soybean (Glycine max) by Infecting Germinating Seeds with Agrobacterium tumefaciens

The transfer of genetic material into soybean tissue was accomplished by using an avirulent strain of Agrobacterium tumefaciens which contained the binary vector pGA482. The method used for transformation requires no tissue culture steps as it involves the inoculation of the plumule, cotyledonary node, and adjacent cotyledon tissues of germinating seeds. The identification of neomycin phosphotransferase (NPT) II enzyme activity in the tissues of 16 (R₀) soybean plants indicated that the plant expressible Nos-NPT II gene, contained within the T-DNA region from pGA482, had been transferred at least into somatic tissues. Putative transformed R₀ soybean plants were advanced to produce R₁ plants which were also assayed for the presence of the transferred Nos-NPT II gene. The combined results of these assays indicated that about 0.7% of the surviving inoculated seeds yielded transformed tissues in the R₀ plant, and that about 1/10 of these plants yielded transformed R₁ plants. The presence of the Nos-NPT II gene in DNAs isolated from both R₀ and R₁ plant was demonstrated by using genomic blot hybridization and polymerase chain reaction methods. Integration of this gene into the soybean genome was demonstrated for three R₁ soybean plants.     

Characterization of Solute Efflux from Dehydration Injured Soybean (Glycine max L. Merr) Seeds

Soybean (Glycine max L. Merr) seeds lose their tolerance of dehydration between 6 and 36 hours of imbibition. Soybean axes and cotyledons were excised 6 hours (tolerant of dehydration) and 36 hours (susceptible) after commencing imbibition and subsequently dehydrated to 10% moisture. Kinetics of the efflux of potassium, phosphate, amino acid, sugar, protein, and total electrolytes were compared in the four treatments during rehydration. Only slight differences were observed in the kinetics of solute efflux between the two cotyledon treatments dehydrated at 6 and 36 hours suggesting that the cotyledons may retain their tolerance of dehydration at this stage of germination. Several symptoms of injury were observed in the axes dehydrated at 36 hours. An increase in the initial leakage of solutes during rehydration, as quantified by the y-intercept of the linear regression line for solute efflux between 2 and 8 hours suggests an increased incidence of cell rupture. An increase in the rate of solute efflux (slope of regression line between 2 and 8 hours) from fully rehydrated axes was observed in comparison to axes dehydrated at 6 hours. The Arrhenius activation energy for potassium, phosphate, and amino acid efflux decreased and for protein remained unchanged. Both observations indicate an increase in membrane permeability in dehydration-injured tissue. Increasing the H⁺ concentration of the external solution increased K⁺ efflux from both control and dehydrated/rehydrated samples, increased sugar efflux from axes at 6 hours imbibition but decreased sugar efflux from axes at 36 hours imbibition, indicating changes in membrane properties during germination. The dehydration treatment did not alter the pattern of the pH response of axes dehydrated at 6 or 36 hours but did increase the quantity of potassium and sugar efflux from dehydration injured axes. These results are interpreted as indicating that dehydration of soybean axes at 36 hours of imbibition increased both the incidence of cell rupture during rehydration and altered membrane permeability of the rehydrated tissue.     

Cadaverine, an Essential Diamine for the Normal Root Development of Germinating Soybean (Glycine max) Seeds

When the polyamine content of soybean (Glycine max) seeds was examined during the early stages of germination, the major polyamine in the cotyledons was found to be spermidine, followed by spermine; while very low concentrations of cadaverine were found. In the embryonic axes, however, cadaverine was the main polyamine and its content markedly increased 24 hours after the start of germination. When the germination of the seeds was performed in the presence of 1 millimolar α-difluoromethylornithine (DFMO), a marked decrease in the cadaverine content was found, while the other polyamines were not affected. This decrease of the cadaverine content was already noticeable after the first hours of germination. In the presence of DFMO, a pronounced elongation in the roots of the seedlings and a marked decrease in the appearance of secondary roots as compared with controls, was observed. This abnormal rooting of the seedlings caused by DFMO was almost completely reverted by the addition of 1 millimolar cadaverine. The latter also increased the appearance of secondary roots in the seedlings. The decrease in the cadaverine content produced by DFMO could be traced to a strong inhibition of lysine decarboxylase. A temporal correlation between the increase in cadaverine content and the increase in lysine decarboxylase activity was found. Both reached a maximum at the second day of germination. The activity of diamine oxidase, the cadaverine degrading enzyme, started to increase at the third day and reached a maximum between the fourth and fifth day of germination. DFMO increased the activity of diamine oxidase by about 25%. Hence, the large decrease in cadaverine content produced by DFMO has to be attributed to the in vivo suppression of lysine decarboxylase activity. Ornithine decarboxylase activity was also suppressed by DFMO, but putrescine and spermidine contents were not affected, except in the meristematic tissues. The obtained results suggest an important role for cadaverine in the normal rooting process of soybean seedlings.     

Dehydration Injury in Germinating Soybean (Glycine max L. Merr.) Seeds

The sensitivity of soybean (Glycine max L. Merr. cv Maple Arrow) seeds to dehydration changed during germination. Seeds were tolerant of dehydration to 10% moisture if dried at 6 hours of imbibition, but were susceptible to dehydration injury if dried at 36 hours of imbibition. Dehydration injury appeared as loss of germination, slower growth rates of isolated axes, hypocotyl and root curling, and altered membrane permeability. Increased electrolyte leakage due to dehydration treatment was observed only from isolated axes but not from cotyledons, suggesting that cotyledons are more tolerant of dehydration. The transition from a dehydration-tolerant to a dehydration-susceptible state coincided with radicle elongation. However, the prevention of cell elongation by osmotic treatment in polyethylene glycol (−6 bars) or imbibition in 20 micrograms per milliliter cycloheximide did not prevent the loss of dehydration tolerance suggesting that neither cell elongation nor cytoplasmic protein synthesis was responsible for the change in sensitivity of soybean seeds to dehydration. Furthermore, the rate of dehydration or rate of rehydration did not alter the response to the dehydration stress.     

Protein Synthesis during Natural and Precocious Soybean Seed (Glycine max [L.] Merr.) Maturation

Protein synthesis was studied during precocious and natural soybean seed (Glycine max [L.] Merr.) maturation. Developing seeds harvested 35 days after flowering were precociously matured through controlled dehydration. Total soluble proteins and proteins labeled with [³⁵S]methionine were extracted from control, developing seeds and from precociously and naturally matured seeds and were analyzed by one-dimensional PAGE and fluorography. The results demonstrated that several polypeptides which were designated “mature polypeptides,” were synthesized de novo during precocious and natural seed maturation. Two of these polypeptides, 31 and 128 kilodalton in mass, also stained intensely with Coomassie blue, suggesting their abundant accumulation during seed maturation. Results from in vitro translation experiments showed that the mRNAs corresponding to these “maturation polypeptides” accumulated during precocious maturation and in naturally matured seeds, but not in seeds freshly harvested 35 days after flowering (control). The role of the “maturation polypeptides” is currently unknown; however, their presence and that of their corresponding mRNAs was coincident with the ability of matured seeds to establish seedling growth. This study has demonstrated that precocious seed maturation treatments may be extremely useful for investigations of metabolic events and molecular control mechanisms affecting soybean seed maturation.     

Developmental Aspects of Soybean (Glycine max) Somatic Embryogenesis

A detailed study of the developmental aspects for soybean somaticembryogenesis was undertaken with emphasis on biochemical andhistological markers. The various stages of somatic embryo developmentin callus cultures have been identified and characterized. Germinatingembryos could be converted to fertile plants at a high frequency(90%). Dicamba (3, 6-dichloro-o-anisic acid) was found to bethe auxin of choice for the clear distinction of the variousdevelopmental phases of soybean somatic embryos. Differencesin their protein patterns were determined using polyacrylamidegel electrophoresis. This analysis revealed distinguishabledifferences in protein profiles amongst the various developmentalstages, especially in heart stage embryos. Histological studieson somatic embryos revealed specific tissue types which closelyresemble those reported for zygotic embryos. Further evidenceis provided that there is a close similarity in tissue differentiation,between somatic and zygotic embryogenesis although there aresome unique features in the development of somatic embryos. Glycine max, callus cultures, developmental stage, liquid cultures, neomorphs, plant regeneration, stage specific proteins, histology     

大豆异黄酮合成途径相关基因差异表达分析

大豆异黄酮是一种应用广泛、具有医用和保健功能的活性物质。为揭示异黄酮合成途径相关基因表达差异,本研究采用实时定量PCR技术分析相关基因在不同大豆品种、发育时期及组织部位的表达。结果发现,苯丙氨酸解氨酶基因PAL、肉桂酸羟化酶基因C4H、香豆酸辅酶A连接酶基因4CL在高异黄酮品种中豆27 R2期叶片中的表达量显著高于低异黄酮品种楚秀;查尔酮合成酶基因CHS、异黄酮合成酶基因IFS在中豆27 R8期子粒中的表达量显著高于楚秀;细胞色素还原酶基因CPR在中豆27 R7期叶片与子粒的表达量与楚秀相比显著降低。这些差异表达的基因可能是形成大豆品种异黄酮含量高低的重要原因。     

Effect of Temporary Chilling on Foliar and Caulinary Growth and Productivity in Soybean (Glycine max)

The effects of chilling on the apical bud were investigatedin soybean. A temperature of 6 °C was applied to leavesI and 2 and the apex for 12 d as soon as leaf 2 appeared. Thegrowth of the foliar and internodal series was analysed usingNelder's logistics. Chilling decreased the number of neoformed phytomers as wellas their rate of appearance. Schematically, the effect of chillingon growth is characterized by an inhibition of leaves and astimulation of internodes. However, the changes in the adultdimensions, and in the kinetic parameters (various durationsand growth rates), depend on the rank of the organ on the axis.Branching was markedly decreased at some nodes. The overallfructification of the main axis remained unchanged, but thedistribution of the seeds on the axis and the repartition betweenaxis and branches were markedly modified. These data clearly show (i) the importance of the position effecton the nature and the intensity of the responses and (ii) theabsence of any return to a normal growth pattern. Thus, a transientand relatively localized thermal stress can disturb the wholeontogenesis. Glycine max, soybean, growth kinetics, development, leaf, shoot, chilling     

Role of Phytohormones in Soybean (Glycine max) Seed Development

Russian Journal of Plant Physiology - Phytohormones play a key important role in the sink development of the seed. Endogenous content of IAA, PAA, GA, ABA, and kinetin were estimated in two...     

农杆菌对大麦种子萌发及幼苗生长发育的影响

以大麦品种(系)为主区('云引大麦Ⅰ'、'云引大麦Ⅱ'和'U008'),农杆菌浸种时间为副区(0.5、1.5和2.5 h),农杆菌菌液浓度为副副区(0.5、1.5和2.5 OD),采用再裂区试验研究了农杆菌浸种处理对大麦种子萌发和幼苗生长发育的影响.结果表明:品种、农杆菌菌液浓度、浸种时间对大麦的种子发芽率、幼苗高度、幼苗鲜重、叶绿素含量、丙二醛(MDA)含量影响无显著的互作效应,而对幼苗POD活性的影响存在显著互作效应;随着浸种时间的延长和菌液浓度的增加,各大麦品种(系)的种子发芽率、幼苗高度、幼苗鲜重、叶绿素含量均呈逐渐降低趋势,幼苗MDA含量则逐渐增加,并以'U008'变化幅度最大;在菌液浓度为0～1.5 OD、浸种时间为0～1.5 h范围内,幼苗POD活性随着菌液浓度的增加和浸种时间的延长而增强,超过该范围则均呈下降趋势,并以'U008'下降最为明显.可见,农杆菌处理对大麦种子萌发和幼苗生长发育有抑制作用,并在菌液浓度超过1.5 OD、浸种时间大于1.5 h时达极显著水平,且大麦品种间存在一定差异.     

Nitric Oxide Improved Growth and Yield in Soybean (Glycine max) by Mediating Physiological,Anatomical, and Transcriptional Modifications

Journal of Plant Growth Regulation - This study investigated the physiological, molecular, and anatomical responses of soybean to the long-time utilization of nitric oxide (NO; 0 and...     

Characterization of an Acidic Chitinase from Seeds of Black Soybean (Glycine max (L) Merr Tainan No. 3)

Using 4-methylumbelliferyl-β-D-N,N′,N″-triacetylchitotrioside (4-MU-GlcNAc₃) as a substrate, an acidic chitinase was purified from seeds of black soybean (Glycine max Tainan no. 3) by ammonium sulfate fractionation and three successive steps of column chromatography. The purified chitinase was a monomeric enzyme with molecular mass of 20.1 kDa and isoelectric point of 4.34. The enzyme catalyzed the hydrolysis of synthetic substrates p-nitrophenyl N-acetyl chitooligosaccharides with chain length from 3 to 5 (GlcNAc_n, n = 3-5), and pNp-GlcNAc₄ was the most degradable substrate. Using pNp-GlcNAc₄ as a substrate, the optimal pH for the enzyme reaction was 4.0; kinetic parameters K _m and k_cat were 245 µM and 10.31 min⁻¹, respectively. This enzyme also showed activity toward CM-chitin-RBV, a polymer form of chitin, and N-acetyl chitooligosaccharides, an oligomer form of chitin. The smallest oligomer substrate was an N-acetylglucosamine tetramer. These results suggested that this enzyme was an endo-splitting chitinase with short substrate cleavage activity and useful for biotechnological applications, in particular for the production of N-acetyl chitooligosaccharides.     

Early Development of the Seed Coat of Soybean (Glycine max)

Although the development of the soybean ovule has been fairlywell studied, knowledge of the sequence of events in the seedcoat during the first 3 weeks after flowering is incomplete.The goal of the present study was to document, using light microscopy,the early development of the soybean seed coat with respectto changes in structure and histochemistry. At anthesis, theseed coat consists of an outer layer of cuboidal epidermal cellssurrounding several layers of undifferentiated parenchyma (whichtogether constitute the outer integument), and an inner layerof cuboidal endothelial cells (the inner integument). At 3 dpost anthesis (dpa), the inner integument has expanded to includethree to five layers of relatively large cells with thick, heavily-stainingcell walls immediately adjacent to the endothelium. By 18 dpa,the outer integument has developed into a complex of tissuescomprised of an inner layer of thick-walled parenchyma, an outerlayer of thin-walled parenchyma containing vascular tissue whichhas grown down from the lateral vascular bundles in the hilumregion, a hypodermis of hourglass cells, and palisade layer(epidermis). The thick-walled parenchyma of the inner integumenthas become completely stretched and compressed, leaving a single,deeply staining wall layer directly above the endothelium. At21 dpa, the outermost cells of the endosperm have begun to compressthe endothelium. At 45 dpa (physiological maturity) the seedcoat retains only the palisade layer, hourglass cells, and afew layers of thin-walled parenchyma. The innermost layer ofthe endosperm, the aleurone layer, adheres to the inside ofthe seed coat. This knowledge will be invaluable in future studiesof manipulation of gene expression in the seed coat to modifyseed or seed coat characteristics. Copyright 1999 Annals ofBotany Company Soybean, Glycine max, seed coat, development, aleurone.