Regulation of lysine metabolism and endosperm protein synthesis by the opaque-5 and opaque-7 maize mutations

Two high lysine maize endosperm mutations, opaque-5 (o5) and opaque-7 (o7), were biochemically characterized for endosperm protein synthesis and lysine metabolism in immature seeds. Albumins, globulins, and glutelins, which have a high content of lysine, were shown to be increased in the mutants, whereas zeins, which contain trace concentrations of lysine, were reduced in relation to the wild-type lines B77xB79+ and B37+. These alterations in the storage protein fraction distribution possibly explain the increased concentration of lysine in the two mutants. Using two-dimensional polyacrylamide gel electrophoresis of proteins of mature grains, variable amounts of zein polypeptides were detected and considerable differences were noted between the four lines studied. The analysis of the enzymes involved in lysine metabolism indicated that both mutants have reduced lysine catabolism when compared to their respective wild types, thus allowing more lysine to be available for storage protein synthesis.     

Quality protein maize: a biochemical study of enzymes involved in lysine metabolism.

Quality protein maize (QPM) varieties have been produced by the introduction of opaque-2 modifier genes. Two QPM varieties, BR451 and BR473, a wild type and an opaque-2 variety, have been used to study key enzymes controlling lysine metabolism in the endosperm during development. Aspartate kinase and homoserine dehydrogenase enzymes, which are involved in lysine and threonine biosynthesis, respectively, exhibited identical activity patterns during endosperm development, with a maximum specific activity at 16 days after pollination. The QPM varieties exhibited higher levels of aspartate kinase activity in the endosperm, suggesting an increased rate of lysine biosynthesis when compared to the opaque-2 and wild-type genotypes. Similar results were observed for the lysine ketoglutarate reductase and saccharopine dehydrogenase enzymes, which form a single bifunctional polypetide involved in endosperm lysine degradation. Both enzyme activities were strongly reduced in the opaque-2 maize variety when compared to the wild-type maize, whereas the QPM varieties exhibited even lower levels of lysine ketoglutarate reductase-saccharopine dehydrogenase activities when compared to the opaque-2 variety. The developmental pattern of enzyme activity showed a different profile when compared to the enzymes involved in lysine biosynthesis, with activity being detected only 12-16 days after pollination (DAP) and maximum activities approximately 24 DAP. These results also suggest that the modifier genes have intensified the effect of the opaque-2 mutation on lysine ketoglutarate reductase-saccharopine dehydrogenase. These alterations lead to an increase in soluble lysine in the endosperm of the QPM varieties when compared to the opaque-2 and wild type.     

外源NO对不同作物种子萌发、幼苗生长及抗氧化酶活性的影响

以浓度分别为0、0.01、0.1、1.0 mmol/L的硝普钠（Sodium nitroprusside, SNP; NO供体）处理玉米、小麦、花生、小白菜、萝卜、黄瓜的种子和幼苗,研究了以上几种浓度的SNP对作物种子萌发和幼苗生长及抗氧化酶活性的影响。结果表明:SNP对多数种子萌发影响表现为低浓度（0.01 mmol/L和0.1 mmol/L）促进,高浓度（1.0 mmol/L）抑制,其中对萝卜发芽率的促进作用最显著;低浓度SNP可有效促进植物幼苗地上部的生长,其中对小麦、黄瓜的促进效果最显著,同时可显著促进根系的伸长,其中对萝卜的促进效果最显著,且对植物幼苗生长的影响与作物种类有关;SNP对多数植物的根系活力有明显的促进作用,其中对萝卜的促进效果最显著;适宜浓度的SNP可以提高作物CAT、POD和SOD活性以及可溶性蛋白含量,并降低MDA含量,不同作物SNP的适宜浓度不同,其中0.1 mmol/L SNP对多数作物处理效果最好。     

Model study on the effect of 15 phenolic olive mill wastewater constituents on seed germination and Vibrio fischeri metabolism

Olive mill wastewaters (OMW) can be a severe problem when disposed of as untreated because of their high organic load, elevated concentration of polyphenols, and moderately low biodegradability. In the present study, the acute toxicity of 15 compounds with low molecular weight (<350 Da), catechol, four benzoic acids, three phenylacetic acids, three phenylethanols, and four cinnamic acids, already isolated from the reverse osmosis in the fractionation of OMW, was assessed on the marine bacterium Vibrio fischeri and on the seeds of two dicotyledonous species Cucumis sativus and Lepidium sativum, and on one monocotyledon Sorghum bicolor. Results of phytotoxicity showed that the most toxic compounds were catechol (EC50s ranging from 0.40 mmol/L for S. bicolor to 1.09 for C. sativus) and hydroxytyrosol, (EC50s ranging from 0.47 mmol/L for S. bicolor to 1.55 for C. sativus) while the toxic potential on bacteria was particularly elevated with EC50 values 1 or 2 orders less than phytotoxicity. These results suggested that the risk of OMW disposal may be more elevated for the water compartment than for the soil.     

Impact of Variable Potassium on Metabolism,Oil, and Seed Quality of Groundnut

To study the impact of variable potassium (K) on biomass, economic yield, and role of K in groundnut (Arachis hypogeae L.) metabolism, plants were grown in refined sand at different levels of K, ranging from 0.5 to 16 mM. The concentration of K increased with increase in supply in all parts of groundnut. Both low (<4 mM) and excess (>4 mM) K increased the activity of peroxidase, ATPase, and acid phosphatase and decreased the concentration of chlorophyll in leaves significantly. Low (<4 mM) and excess (>4 mM) concentrations of K inhibited pod and kernel (seed) formation in groundnut. Potassium stress (both low and excess) deteriorated the quality of groundnut seed by decreasing the concentrations of starch, protein, and oil and increasing that of sugars in seeds. The activities of certain enzymes also altered in K stress in leaves and seeds of groundnut.     

On the metabolism of glucosamine-1-14C in detached tomato leaves

In the previous paper, occurrence of glucosamine in tomato plants fed with a high level of ammonium nitrogen has been reported(18). For the first time, SOLMS and HASSID(17) discovered glucosamine in tissues of higher plants. They isolated uridine diphosphate N-acetylglucosamine from mung bean seedlings. Thereafter, uridine diphosphate N-acetylglucosamine and N-acetylgalactosamine were chromatographically isolated from dahlia tubers(7). PUTZTAI(16) has succeeded in crystalyzing of glucosamine isolated from the seeds of higher plants and determined aminosugar contents in 14 kinds of, seeds. He considered aminosugars to be the normal constituents of plant seeds. LIS et al(11) have isolated glycopeptide containing mannosc, glucosamine, nnd aspartic acid from pure soy bean hemagllutin, and Kasai et al(10) have shown the presence of the glucosamine in hydrolyzatcs of glycoprotcins of broad bean and soy bean. Recently glucosaminc metabolism in mung bean 141.S been studied 02. 19). Although several workers have reported on glucosaminc in plants as mentioned above, the finding in the previous paper that the amount of glucosamine in tomato plants increased with concentration of ammonium nitrogen in water culture solution is quite unique.     

Genetic manipulation of proline accumulation influences the concentrations of other amino acids in soybean subjected to simultaneous drought and heat stress

The effect of simultaneous drought and heat stress on free amino acid levels was compared in wild type and transgenic soybean (Glycine max (L.) Merr cv Ibis) plants transformed with the cDNA coding for the last enzyme of Pro biosynthesis, l-Delta(1)-pyrroline-5-carboxylate reductase (EC 1.5.1.2), in sense and antisense directions. The most rapid increase in Pro content was found in the sense transformants that exhibited the least water loss, while the slowest elevation of Pro levels was detected in the antisense transformants that exhibited the greatest water loss during stress. Correspondingly, the level of the Pro precursors Glu and Arg was higher in sense transformants and lower in antisense ones compared to the wild type plants during the initial part of the stress. Interestingly, genetic manipulation of Pro levels also affected the stress-induced changes in the concentration of several other amino acids, which indicates the coordinated regulation of their metabolic pathways.     

Comparison of nutritional and antinutritional factors in soybean and fababean seeds with or without cortex

The composition and contents of nutritional factors such as proteins, lipids, carbohydrates, fibers, amino acids, and antinutritional factors such as trypsin inhibitors, phytic acid, and tannins were compared in soybean and fababean seeds with emphasis placed on the nutritional improvement of the seeds by cortex removal. Protein hydrolysis analysis for both whole seeds and seed with cortex removed revealed the presence of a large amount of lysine, arginine, aspartic acid, glutamic acid, glycine, and leucine while these seeds contained a low level of tryptophan, cystine, and methionine. Some antinutritional factors such as trypsin inhibitors, phytic acid, and tannins were detected in soybean and fababean seeds: phytic acid content and trypsin inhibitor activity were higher in soybean seeds than in fababean seeds while the difference in the tannin content was less pronounced. It was found that most of the tannins occurred in the cortex of the soybean and fababean seeds. Tannins are polyphenolic compounds that readily form indigestible complexes with proteins and other macromolecules under specific environmental conditions. By removal of the cortex, tannins were almost completely eliminated without changes in the protein composition and amino acids. From these results, it is assumed that since soybean and fababean seeds contained a high concentration of antinutritional factors in the cortex such as tannins, the utilization of the legume seeds after removal of all of the cortex is suitable for human diet or industrial products.     

外源水杨酸对锰污染红壤中玉米的生长与抗氧化酶活性的调节作用

采用盆栽实验方法研究了外源水杨酸（SA）对锰污染红壤中玉米的生长、脂质过氧化程度、活性氧水平以及抗氧化酶活性的影响。结果表明,过量锰明显降低玉米植株干重,显著提高了茎叶和根中锰的含量。SA促进锰胁迫下玉米的生长,但对植株中锰的含量与分布无影响。过量锰处理下,玉米叶片超氧阴离子（O.2-）和过氧化氢积累显著增加,脂质过氧化、电解质渗透率和脯氨酸含量显著升高;而SA和过量锰复合处理下,这些指标则显著降低。过量锰诱导超氧化物歧化酶（SOD,EC1.15.1.1）、过氧化物酶（POD,EC1.11.1.7）活性升高,抑制过氧化氢酶（CAT,EC1.11.1.6）和抗坏血酸过氧化物酶（APX,1.11.1.11）活性;SA处理促进锰胁迫下SOD和POD活性进一步升高,减小CAT和APX活性下降的程度。这些结果提示,SA调节抗氧化酶活性,保护组织细胞免遭氧化损伤,是SA缓解过量锰对玉米毒害作用的重要生理原因。     

Hypotin, a novel antipathogenic and antiproliferative protein from peanuts with a sequence similar to those of chitinase precursors

A protein designated Hypotin, with both antifungal and antibacterial activity, was isolated from peanut (Arachis hypogaea) seeds. The isolation procedure included extraction, ammonium sulfate precipitation, affinity chromatography on Affi-gel blue gel, ion chromatography, and gel filtration. The protein exhibited a molecular mass of 30.4 kDa in SDS-polyacrylamide gel electrophoresis under both reducing and nonreducing conditions, indicating that it is a monomeric protein. Its N-terminal sequence was highly homologous to those of chitinases and chitinase precursors from plants. It exerted potent antifungal action toward a variety of fungal species, including Pythium aphanidermatum, Fusarium solani, Physalospora piricola, Alternaria alternata, Botrytis cinerea, and Fusarium oxysporum. In addition, this novel protein exhibited antiproliferative activity against tumor cells. These findings further the progress in the research of leguminous plants.     

RESPONSES OF SOME ENZYMES AND KEY GROWTH PARAMETERS OF SALT-STRESSED MAIZE PLANTS TO FOLIAR AND SEED APPLICATIONS OF KINETIN AND INDOLE ACETIC ACID

The study examined the effects of kinetin (KIN) and indoleacetic acid (IAA) applied as seed treatment or sprayed on leaves of salinity stressed plants. Five -week old maize (Zea mays L. cv. ‘DK 647 F1’) plants were grown in pots containing peat and perlite in 1:1 (v/v) mixture. Different treatments used were: 1) control (nutrient solution alone), 2) salt stress [100 mM sodium chloride (NaCl)], 3) 100 mM NaCl and 1 mM kinetin (KIN), 4) 100 mM NaCl and 2 mM KIN, 5) 100 mM NaCl and 1 mM indole acetic acid (IAA), 6) 100 mM NaCl and 2 mM IAA, 7) 100 mM NaCl and 25 mg L^?1 KIN and 8) 100 mM NaCl and 25 mg L^?1 IAA. In treatments 7 and 8 application was to the seeds, for treatments 3-6 it was applied to foliage. The seeds were soaked in KIN or IAA solution for 12 h. Salt stress reduced the total dry matter, chlorophyll content, and relative water content (RWC), but increased proline accumulation, activities of superoxide dismutase (SOD; EC 1.15.1.1), peroxidase (POD; EC. 1.11.1.7), catalase (CAT; EC. 1.11.1.6) and polyphenol oxidase (PPO; 1.10.3.1) and electrolyte leakage. Both foliar applications of KIN and IAA treatments overcame to variable extents the adverse effects of NaCl stress on the above mentioned physiological parameters. However, seed treatments with KIN or IAA did not improve salinity tolerance in maize plants. Furthermore, foliar application or seed treatments with KIN and IAA reduced the activities of antioxidant enzymes in the salt stressed-plants. Salt stress lowered some macronutrient concentrations [calcium (Ca) and potassium (K) in leaves and roots, phosphorus (P) in roots] but foliar application of both KIN and IAA increased Ca in both leaves and roots and P in leaves. Foliar application of IAA increased K concentrations in leaves of the salt-stressed plants. Foliar application of KIN and IAA, especially at 2 mM concentration, counteracted some of the adverse effects of NaCl salinity by causing the accumulation of proline and essential inorganic nutrients as well as by maintaining membrane permeability.     

ROLE OF SALICYLIC ACID IN REGULATION OF CADMIUM TOXICITY IN WHEAT (TRITICUM AESTIVUM L.)

The present study is focused on the possible mediatory role of salicylic acid (SA) under cadmium (Cd⁺²) toxicity. Cadmium treatments resulted in the inhibition of root dry biomass, root elongation and increased Cd accumulation in roots. Pretreatment of seeds with SA (500 μM) for 20 hrs resulted in protection against Cd, increased root dry biomass, root elongation and minimal accumulation of cadmium. Cadmium chloride (CdCl₂; 0.0, 100, 400 and 1000 μM) produced a significant inhibition in chlorophyll, photosynthetic efficiency [carbon dioxide (¹⁴CO₂)-fixation], relative water content (RWC) and abscisic acid (ABA) content. Pretreatment of seeds with SA alleviated the Cd negative effect on these parameters. A drop in the activities of carboxylating enzymes, phosphoenol pyruvate carboxylase (PEPC, EC 4.1.1.31), and ribulose1,5-bisphosphate carboxylase (RuBPC, EC 4.1.1.39) were observed in Cd treated plants. Pretreatment with SA ameliorated the inhibitory effect of Cd on the studied enzymatic activities. Cadmium applied to salicylic acid free plants increased the level of stress metabolites, malondialdehide (MDA), hydrogen peroxides, and free proline content. Salicylic acid pretreatment decreased the toxic effect level of cadmium manifested by lower lipid peroxidation, lesser production of H₂O₂ and reduction in free proline content. The ultrastructural investigation of the root cells of wheat exposed to different concentrations of Cd for 15 days was carried out. The results indicated that Cd induced several obvious ultrastructural changes such as increased vacuolation, condensed cytoplasm with increased density of the matrix, reduction of mitochondrial cristae, severe plasmolysis, highly condensed nuclear chromatin and increment of disintegrated organelles. Electron dense granules appeared between the cell wall and plasmalemma. In vacuoles, electron dense granules encircled by the membrane were aggregated and formed into larger precipitates, which increase in number and volume as a consequence of excessive Cd exposure. An important issue arising from this study was how the short-term treatment with SA affected several physiological processes, such as plant growth, photosynthesis, and antioxidant defense system. We assume that the beneficial effect of SA during an earlier growth period could be related to avoidance of cumulative damage upon exposure to cadmium. One of the important roles of SA in inducing resistance to various environmental stresses is manifested by its ability to express genes that code for pathogenesis-related proteins or defense-related enzymes. Also, SA could form a complex with Cd that may provide Cd tolerance.     

Accumulation of cadmium in growing peanut (Arachis hypogaea L.) seedlings—Its effect on lipid peroxidation and on the antioxidative enzymes catalase and guaiacol peroxidase

In plants exposed to high metal concentrations, mechanisms to counteract the oxidative burst are crucial for its survival. To investigate the temporal sequence of physiological reactions of peanut seedlings (Arachis hypogaea L.) to cadmium exposure, seeds were cultured in increasing concentrations of CdCl₂, ranging from 50 to 300 μM. Germination frequency was scored, and the distributions of Cd in root, stem, and leaves were determined after 2 and 4 weeks of culture. Lipid peroxidation and activities of antioxidative enzymes including catalase (CAT; EC 1.11.1.6) and guaiacol peroxidase (GPX; EC 1.11.1.7) were estimated in these three parts of the plant. Germination of seedlings was not affected, but the growth of seedlings was severely suppressed with increasing concentrations of CdCl₂ and incubation period. Pattern of Cd distribution in the three organs varied with concentration and period of exposure to Cd. Increased lipid peroxidation was detected in all parts of the developing seedlings with increasing metal accumulation. Catalase and guaiacol peroxidase activity varied in the three parts of the seedlings with concentration of Cd and incubation period. Guaiacol peroxidase activity appears to be more active in scavenging the reactive oxygen species in developing peanut seedlings. The results of the present experiment demonstrate the advantages of a tissue‐culture model system in studying the complex network of interactions of various factors in stress tolerance.     

Impact of Boron Deficiency on Changes in Biochemical Attributes,Yield, and Seed Reserves in Chickpea

To determine the effect of boron (B) deficiency on biomass, reproductive yield, metabolism, and alterations in seed reserves of chickpea (Cicer arietinum L.) cv. ‘13.G‐256,’ plants were grown in refined sand until maturity at deficient (0.033 mg L^?1) and adequate (0.33 mg L^?1) B, supplied as boric acid (H₃BO₃). Boron‐deficient plants exhibited visible deficiency symptoms in addition to reduced number of pods and seeds, resulting in lowered biomass and economic yield. Boron deficiency lowered the concentration of B in leaves and seeds, photosynthetic pigments (leaves), Hill reaction activity, starch (in leaves and seeds), and proteins and protein N (in seeds), whereas phenols, sugars (in leaves and seeds), and nonprotein N (in seeds) were elevated. Specific activity of peroxidase (POX) increased in leaves and pod wall and decreased in seeds, while activity of acid phosphate and ribonuclease were stimulated in leaves, seeds, and pod wall in B‐deficient chickpea.     

Distribution, species composition and management implications of seed banks in southern New England coastal plain ponds

Buried seeds that germinate during periods of low water or water level drawdown can play important roles in shaping plant community composition, community dynamics and species richness in ecosystems with fluctuating water levels. Northeastern US coastal plain ponds have fluctuating water levels and contain a characteristic shoreline flora that contains many rare plants. The objectives of this study were to: (1) test whether geographically distant ponds in Cape Cod and Martha’s Vineyard had distinct seed banks, (2) determine if hydrologic status as permanent and ephemeral ponds led to differences in seed banks, and (3) examine seed diversity and seed abundance across gradients of shoreline elevations and sediment characteristics. Viable seeds of 45 plant species were identified from nine ponds. Native species dominated pond-shore seed banks and made up 89-100% of all species. There was high overlap in seed bank composition across hydrological classes and geographic regions. One hydrological class captured 73-76% of total species and one geographical region captured 69-78% of the total species recovered from the entire suite of seed bank samples. Seeds were relatively evenly distributed along the shorelines of ephemeral ponds but seed diversity and abundance were lower at low elevations in permanent ponds. Results suggest that strategies to protect pond shorelines to capture maximum diversity of coastal plain pond plants contained in pond sediment seed banks should be implemented across pond hydrologic classes and across a wide geographic area. Shoreline seed distributions indicate that ground-water withdrawals or climate changes that lower pond water levels in permanent ponds will reduce the diversity and abundance of plants recovered from seed banks by shifting water levels to a shoreline zone of high sediment organic matter where seed densities are lower. This effect will be much less in ephemeral ponds where seed diversity and abundance on pond bottoms was high.     

Tronchuda cabbage (Brassica oleracea L. var. costata DC): scavenger of reactive nitrogen species

The ability of tronchuda cabbage ( Brassica oleracea L. var. costata DC) to act as a scavenger of the reactive nitrogen species nitric oxide and peroxynitrite was investigated. The aqueous extracts obtained from tronchuda cabbage seeds and from its external and internal leaves exhibited a concentration dependent scavenging capacity. The antioxidant potential observed against the two reactive species was as follows: seeds > external leaves > internal leaves. In order to establish a possible correlation with the chemical composition of the extracts, the activity of ascorbic and sinapic acids and kaempferol 3- O-rutinoside was also studied. Among the compounds tested, sinapic acid showed the strongest antioxidant activity against both species.     

EFFECTS OF SOIL SALINITY IN THE GROWTH OF AMBROSIA ARTEMISIIFOLIA BIOTYPES COLLECTED FROM ROADSIDE AND AGRICULTURAL FIELD

Morphological differences were observed between roadside (R) and agricultural field (F) biotypes of Ambrosia artemisiifolia, in which R-type seedlings were shorter and produced larger and heavier seeds under greenhouse grown conditions. Previous findings indicated that A. artemisiifolia R-types exhibited greater salt tolerance with respect to germination. However, the impact of biotype and salt tolerance on morphological variation has not been investigated in A. artemisiifolia plants. After performing replicated greenhouse experiments with both biotypes, it was shown that salinity level was a critical factor influencing both seedling and mature plant size and this response was dependent upon biotype. The R-type exhibited slight but significant increases in growth at low/mild salinity levels (50–100 mM) compared with non-saline conditions, while the F-type exhibited significantly reduced growth at the low/mild salinity levels. The reductions in growth of F-type plants in low/mild salinity were similar to those reductions of R-types observed in non-saline conditions. As both biotypes produced seeds at low/mild salinity levels, we conclude that low/mild salinity affects A. artemisiifolia plant size and overall growth rate, and secondly, certain F-type plants may acclimate to the roadside environment over time by reducing their size while producing larger seed under saline conditions. It is possible that this species may exhibit changes in morphology after several generations of exposure to saline roadside conditions. Toxicity due to salt treatment at high salinity (400 mM) was observed in both biotypes, whereas the R-type was more tolerant to both low and high salinity levels with respect to seed germination. Differential A. artemisiifolia growth responses which occur from seed germination to plant maturity may be partially attributed to its ability to tolerate saline soil conditions both under greenhouse and field conditions. This ability to tolerate saline conditions may be especially important in early spring when roadside soils experience increased salinity, caused by de-icing salt treatments applied during the winter season.     

Quantification of jasmonic acid by SPME in tomato plants stressed by ozone

Jasmonates are signalling molecules induced in plants as a response to various biotic and/or abiotic stresses. As ozone is known to activate defense responses in plants, we have monitored the concentration of jasmonic acid in tomato leaves during and after an acute exposure to this abiotic elicitor. In this experiment, we observed that the maximum induction of jasmonic acid in O3-fumigated plants occurred 9 h after the end of treatment and the concentration of jasmonic acid in stressed plants increased 13-fold. However, the level of endogenous methyl-jasmonate was constant during the observed period. The extraction and quantification of jasmonic acid as its methyl ester was performed by headspace-solid-phase microextraction (or HS-SPME) in combination with GC-FID and GC-MS. The sensitivity (LOD = 2 ng/g) of this method permitted the detection and quantification of jasmonic acid present in plant tissues at very low concentrations.     

Chemical composition, nutritive value, and toxicology evaluation of Mexican wild lupins.

The nutrient composition, toxic factors content, and nutritional and toxicological value of Lupinus splendens, L. rotundiflorus, L. elegans, L. simulans, L. exaltatus, L. reflexus, and L. madrensis species from Mexico were analyzed. The seeds of these species were a good source of protein. All the species showed a high lysine and tryptophan content, though sulfur amino acids were limiting. Cyanogenic glycosides were absent, and lectins, trypsin inhibitors, and tannins were present in low concentrations. Lupanine was the major alkaloid in almost all the samples, although sparteine was the major alkaloid in Lupinus reflexus (26.63 mg/g of sample). Cytisine was not found in any of the studied lupins. L. reflexus showed the highest acute toxicity, and L. elegans exhibited no toxicity as evaluated using a mice model. The alkaloid was reduced by hot-water extraction. The protein efficiency ratio in water-debittered seeds was relatively poor (1.1-1.5). These results suggest that the wild lupins studied represent a potential protein supply, and they could be domesticated and used for animal feed if the alkaloids were eliminated and the protein was supplemented with methionine, or if the lupins were used in mixture with cereals.