Effects of NaCl Salinity on Nitrogenous Compounds and Proteases during Germination of Vigna sinensis Seeds

Serido bean (Vigna sinensis (L.) Savi) seeds were sown in water and in NaCl solution of —4.3 bars water potential. Total, insoluble and soluble proteins as well as soluble amino nitrogen and proteolytic activity of cotyledons were studied after 0, 1, 3, 5, 7, and 9 days of germination. Protein breakdown and turnover was delayed by the NaCl treatment as compared to the control. This was not due to the total amount of proteolytic activity, which was unchanged by salinity; even though the specific activity decreased due to the delayed breakdown of proteins under salt stress. The inhibitory effects of salinity on seed protein reserve mobilization may be due more to inhibition of translocation of hydrolysis products than to inhibition of protease activity.     

豇豆胰蛋白酶抑制剂不同组分的制备方法

以豇豆种子为材料,经脱脂、乙酸钠抽提、硫酸铵分级沉淀得到胰蛋白酶抑制剂粗提物。再经凝胶过滤、离子交换层析和FPLC,分别得到了豇豆蛋白酶抑制剂的不同组分,并进行了SDS-PAGE分析。同时通过明胶活性电泳,证实了各部分均具备专一的胰蛋白酶抑制剂活性。     

Germination Inhibitors in some Umbellifer Seeds

Presence of inhibitors of germination in seeds of five umbellifercrops viz. fennel (Foeniculum vulgare Mill.), cumin (Cuminumcyminum L.), carum (Carum copticum Benth. and Hook.), carrot(Daucus carota L.), and coriander (Coriandrum sativum L.) wasdemonstrated. The effect of seed exudates on the pattern ofinhibition of germination of these seeds was also studied. Presence of exudates in general delayed germination althoughthe time of maximum inhibition was different with differentcrop seeds. The inhibitory effect was high at an early stagein the case of carum and carrot and persisted in the latterbut disappeared in the former. The inhibitory effect on cuminand fennel increased up to the seventh day and then began todecrease. Growth of root and hypocotyl of 16-day-old seedlingswas also inhibited by the exudates.     

Alkaloid Changes in Tobacco Seeds during Germination

Nicotine, nornicotine, anabasine, and anatabine, normally found in growing and mature tobacco (Nicotiana tabacum L.) plants, were extracted and quantified from mature tobacco seeds and young tobacco seedlings. The rate of net alkaloid disappearance and accumulation in tobacco seedlings was related to phases of germination.     

The Appearance of New Active Forms of Trypsin Inhibitor in Germinating Mung Bean (Vigna radiata) Seeds

Ungerminated seeds of mung bean contain a single major species (F) of trypsin inhibitor with five minor species (A-E) separable on diethylaminoethyl-cellulose. During germination the level of trypsin inhibitory activity decreases from 1.8 units/grams dry weight in ungerminated cotyledons to 1.2 units/grams in cotyledons from seeds germinated 5 days. This decrease is accompanied by major changes in the distribution of inhibitory activity among the inhibitor species. By 48 hours of germination, inhibitor F has largely disappeared with an accompanying rapid increase in inhibitor C. Similarly, though less rapidly, inhibitor E decreases while inhibitor A increases. A similar sequence of changes is found in vitro when purified inhibitor F is incubated with extracts from seeds germinated 96 hours. The combined in vivo and in vitro data suggest a conversion sequence of: F → E → C → A. The in vitro conversion is inhibited by phenylmethyl sulfonyl fluoride but not by iodoacetamide, indicating that at least the initial phases of inhibitor conversion are not catalyzed by the mung bean vicilin peptidohydrolase.     

The Antioxidant System of Wheat Seeds during Germination

The dynamics of peroxidase activity and antioxidant contents in wheat seeds were studied in the course of 24-hour swelling at 5°C (group 1) and 23°C (group 2). Both parameters were 1.5 times higher in seeds of the first group. In the same seeds, peroxidase activity in the endosperm and seed coat increased by factors of 1.5 and 1.8, respectively. Catalytic constants of wheat seed peroxidase were determined in the reactions of o-dianisidine and ascorbic acid peroxidation. In the pH range studied (pH 5–7), K _m proved to change only slightly. In seedlings, an increase in the lipid peroxidation rate was accompanied by an increase in the content of antioxidants. Peroxidase activity increased as the content of antioxidants decreased, and vice versa. Thus, the reciprocal influence of peroxidase and low-molecular antioxidants during seed germination in wheat was revealed.     

Sterol Changes during Germination of Nicotiana tabacum Seeds

The identity, composition, and concentration of the total, free, esterified, and glycosidic sterol fractions were determined during germination of tobacco seeds. The total, free, and esterified sterols increased, with stigmasterol and campesterol accounting for most of the increase. Steryl glycosides decreased during germination, and stigmasteryl and sitosteryl glycosides showed the largest decrease. During germination, sitosterol was the major sterol in all fractions but stigmasterol and campesterol showed the greatest changes. The fatty acid composition of the steryl esters and acylated steryl glycosides most closely resembled the di- and triglycerides.     

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)     

Sequential Expression of Trypsin Inhibitors in Developing Fruit of Cowpea (Vigna unguiculata (L.) Walp)

Trypsin inhibitor (TI) activity was followed in the pod (pericarp),seed coat, cotyledon and embryo axis during fruit developmentof cowpea. On the basis of seed fresh weight, three phases couldbe distinguished from anthesis to fruit maturity. In the podTI activity increased from the beginning of Phase I to a maximumin the middle of the phase. From then on the activity declineduntil no activity could be detected before the end of phaseII. The cotyledons did not contain any TI in Phase I. TI activitywas first detected in the cotyledon in the beginning of PhaseII at the same time that globulin synthesis started. The TIactivity in the cotyledon increased to a maximum at the endof Phase II before decreasing in Phase III. In the embryo axisa similar pattern of TI activity to that of the cotyledon wasfound. No protein TI could be detected in the seed coat at anystage. In the pod there is a TI with a mol. wt of 12500 andpI of 4.4. Mature cotyledon and embryo axis have two TI withmol. wt 10800 and 24700 with pI 4.7 and 5.0 respectively. Duringdevelopment the smaller TI (mol. wt 10800) was synthesised beforethe larger TI (mol. wt 24700). There were large differencesbetween the maximum absolute amounts of TI present in the pericarp,cotyledon and embryo axis.     

Effects of Water Deficit during Germination of Wheat Seeds

Germinating seeds of spring wheat (Triticum aestivum L.) were tolerant to dehydration up to the 4^th day following imbibition and from the 5^th day the seedling survival decreased. Dehydration also inhibited the rate of seed dry mass depletion and seedling dry matter accumulation and increased the content of soluble sugars both in grain and seedlings. Glucose supplied either to dry seeds or to 4-d-old seedlings increased survival of dehydrated seedlings. In contrast, exogenously supplied non-readily metabolizable sorbose and mannose suppressed seedling survival.     

Increase in beta-N-Acetylglucosaminidase Activity during Germination of Cotton Seeds

A marked increase in beta-acetylglucosaminidase (2-acetamido-2-deoxy-beta-d-glucoside acetamidodeoxyglucohydrolase, EC 3.2.1.30) activity was observed in the germinating cotyledon of cotton seeds. The enzyme was isolated from cotton seedlings and purified to study its physiological function in the germination of cotton seeds. The purification procedure involves ammonium sulfate fractionation, ion-exchange chromatography, gel filtrations, and concanavalin A-Sepharose 4B chromatography, and the purified beta-N-acetylglucosaminidase was shown to be homogeneous by disc electrophoresis. The molecular weight was estimated to be about 125,000 by gel filtration. The enzyme hydrolyzed both p-nitrophenyl-N-acetyl-beta-d-glucosamine and p-nitrophenyl-N-acetyl-beta-d-galactosamine. When p-nitrophenyl-N-acetyl-beta-d-glucosamine was used as substrate, K(m) and V(max) were 0.625 nanomolar and 228 moles per minute per milligram, respectively, and optimum activity was at pH 5.6. The enzyme liberated beta-linked N-acetyl-glucosamine from chitin, ovalbumin, and pronase-digested wheat germ lectin.     

当归果翅对种子吸水与发芽进程的影响

研究当归果翅对其种子吸水及发芽进程的影响,旨在为当归规范化栽培提供理论和技术依据。以当归的双悬果果实和去翅种子作为试验材料,对其千粒重、含水量、体积、容重和吸水率进行测定,并在自然室温条件下进行种子发芽试验。结果表明:①当归双悬果去除果翅后,其千粒重减少了46.87%(P<0.01),体积减少了90.78%(P<0.01),容重增加了475.92%(P<0.01),含水量增加了20.15%(P<0.05);②当归果翅抑制了种子吸水吸胀的能力,果翅去除后能在2.48 h内进入稳定期,较去翅前提前了3.38 h,且吸水过程符合逻辑斯蒂方程;③当归果翅对种子发芽具有显著影响,果翅去除后发芽势、发芽率和发芽指数较未去翅种子分别提高了256.7%、12.28%和27.77%,均达到了极显著水平(P<0.01)。当归双悬果去除果翅后,减小了种子体积和重量,并促进了种子吸水吸胀能力,同时还有效地提高了种子的发芽质量。建议在生产中以去翅种子作为播种材料进行播种。     

Precocious Germination during In Vitro Growth of Soybean Seeds

Immature Glycine max (L.) Merrill seeds were grown and matured in liquid medium at 25°C under fluorescent light. In standard medium containing minerals, 146 millimolar sucrose and 62.5 millimolar glutamine (osmolality 0.24), precocious germination seldom occurred with a starting seed size of less than 300 milligrams fresh weight. Frequency of precocious germination increased with increased starting seed size. Sucrose concentration strongly affected precocious germination while glutamine concentration had no effect. Starting with 300 to 350 milligrams fresh weight seeds, treatments which reduced the sucrose concentration or lowered the osmolality of the culture medium stimulated precocious germination, and increased the fresh weight growth but not the dry weight growth of seeds. Increasing the osmolality to 0.38 with sucrose or mannitol prevented precocious germination without reducing dry weight accumulation in seeds. In medium with initially low osmolality, precocious germination was inhibited by addition of 1 to 100 micromolar abscisic acid to the medium without a reduction in seed growth. During growth and maturation of large soybean seeds in vitro, precocious germination and other abnormal tissue growth can be prevented by high sucrose or mannitol concentrations in the medium or by addition of abscisic acid.     

Transfer Cells in Suspensur and Endosperm during Early Embryogeny of Vigna sinensis

During early embryogeny, structural differentiation of the suspensor and endosperm can be observed with the formation of cells with wall ingrowths. In the early proembryo stage, wall ingrowths are seen only on the boundary walls of the embryo sac around the proembryo and at the chalazal end. Later, ingrowths appear in the outer walls of the basal suspensor cells and some wall ingrowths also begin to develop in the outer walls of cellular endospermic cells adjacent to the nucellar cap and the inner integumentary tissues. The suspensor appears to remain active throughout the differentiation stages. Two regions can be clearly distinguished in the suspensor: a basal region and a neck region. Wall ingrowths appear to form only in the cells of the basal region. During the development of the cellular endospermic sheath, its cell number and size both increase slightly. Later, these cells rapidly become separated from each other. Those endospermic cells that abut directly onto the integumentary tissues also develop wall ingrowths. In the region of the fluid endosperm, wall ingrowths are especially abundant in the boundary walls on the ventral side of the embryo sac. The possible pathway of nutrient flow to the developing embryo is discussed.     

Studies on the Relationship between Peroxidase Activity and Germination of Pinus koraiensis Seeds and on Metabolic Inhibitors in the Seeds

The effect of both stratificating and decoating on germination and peroxidase activity of Pinus koraiensis seeds suggest that some role might be played by peroxidase in seed dormancy. Crude extract of seed are strongly inhibiting peroxidase activity and respiration in addition to hindering seed germination. Paper and thin layer chromatographic analyses have provided evidence that these activities are not attributable to the substance present in extracts which can inhibit germination. Experimental data reveal the presence of at least two kinds of peroxidases with different stability and different effects on the time course of peroxidase reaction. The inhibition on isoperoxidases and peroxidases are different from different biological sources.     

Development of Bed Reactor Using Brick Dust Immobilized CIVI-Cellulase from Seeds of Cowpea (Vigna sinensis L)

Cellulase extracted from seeds of Cowpea (Vigna sinensis L var VITA-4) was partially purified and immobilized on brick dust as solid support via glutaraldehyde. The percentage retention of the enzyme activity on brick dust was nearly 85%. After immobilization specific activity of the enzyme increased from 0.275 to 0.557 U mg^?1 protein with about 2 fold enrichment. The optimum pH and temperature of soluble enzyme were determined as pH 4.6 and WC, respectively whereas immobilized enzyme showed at pH 5.0 and 37°C, respectively. The V_max values for soluble and immobilized enzyme were determined as 6.67 and 1.25 mg min^?1, respectively whereas K_m values were 4.35 and 4.76 mg ml^?1, respectively. The immobilized enzyme displayed higher thermal stability than soluble enzyme and retained about 50% of its initial activity after 12 reuses. Immobilized enzyme was packed in an indigenously designed double walled glass bed reactor for continuous production of reducing sugars.     

Embryo and Endosperm Metabolism of Barley Seeds during Early Germination

Embryos detached from germinating barley seeds were immersedin tritiated water or solutions containing ¹⁴C-labelled compounds.Amino acids rapidly became radioactive and later acids of theKrebs cycle. Labelled alanine did not give rise to radioactivesucrose.     

Different Rates of Metabolism of Soybean Proteinase Inhibitors during Germination

During germination, the content of the major Bowman-Birk proteinase inhibitor (BB-E) in the cotyledons of soybean (Glycine max [L.] Merrill cv. Fiskeby V) seeds decreases, becoming a minor form by the sixth day of germination. One of the three other minor species (BB-D) of this inhibitor in the dry seed increases to become the major form in six-day cotyledons. The other two minor species (BB-C and BB-F) also appear to decrease during germination, but at a slower rate compared to the original major inhibitor form BB-E. By 13 days of germination, no distinct Bowman-Birk inhibitor species can be discerned in the cotyledons. The content of the major Kunitz proteinase inhibitor (K-B) also decreases during germination, but much more slowly. One new form of the Kunitz inhibitor (K-A) becomes apparent by the sixth day. By the 13th day, the proportion of the main isoinhibitor to the late-appearing form is approximately two to one. This difference in the rates of disappearance and appearance of isoinhibitor species in the Bowman-Birk and Kunitz proteinase inhibitor classes suggests a differential metabolism of these two types of proteins and a possible difference in function in the soybean plant.     

The Effect of Arsenate and Other Inhibitors on Early Events during the Germination of Lettuce Seeds (Lactuca sativa L.)

The effect of arsenate, arsenite, 2,4-dinitrophenol, and anaerobiosis on early events in seed germination was investigated using both intact and punched seeds of lettuce (Lactuca sativa L.). It was found that punching the seed removes penetration barriers to the entrance of inhibitors without an undue loss of germination or light responses. The kinetics of the action of germination inhibitors were established by 2-hour pulse experiments. Arsenate and 2, 4-dinitrophenol have very different kinetics. The inhibition of germination in punched seeds by arsenate given in conjunction with phosphate compared with the lack of inhibition of arsenate plus phosphate on the growing seedling, suggest a distinct metabolic change in the germinating embryo at some time between the onset of germination and subsequent seedling growth.