Ontogenetic Changes in the Alcohol-soluble Amino Acid Fraction of the Grain, Leaves and Roots of Triticum aestivum (var. Rideau) Following Vernalization and Seedling Growth

Changes in the free alcohol-soluble amino acid fraction of theseparate organs of Rideau wheat (Triticum aestivum L.) preconditionedby vernalization were determined after 1 and 3 weeks' growthand compared with comparable organs of non-vernalized controlplants. The major components of the fraction were composed of the acidicand neutral amino acids together with asparagine and glutamine.Cystine and methionine were rarely present in more than traceamounts. Changes coincident with ontogency and growth varied considerablyboth between all seedling parts and between comparable organsof the control and vernalized series. Generally, vernalizationwas paralleled by higher levels of the acidic amino acids andthe amides. Coleoptile senescence was characterized by depletionof the total fraction but individual amino acids increased withageing. The pattern of change varied with ontogeny, age andprior grain treatment. However, within each category significantvariations in the level of one or more amino acids could berelated to the physiological status of the tissue. The amides,glutamine and asparagine were consistently absent from the stemapex and the very young fifth and sixth leaves. In stem apextissue, 17 of the commonly occurring amino acids were generallyfound present while, in coleoptiles and young leaves only eightamino acids were present.     

Occurrence of some enzymes in starchy endosperm and hormonal regulation of isoperoxidase in aleurone of wheat

Peroxidase, indoleacetic acid-oxidase, alkaline, and acid phosphatases were detected in dry starchy endosperm (minus aleurone) of wheat grain. The isoperoxidase pattern differed in different parts of the dry grain. Several new isoperoxidases were found in embryos after soaking. The intensity of isoperoxidases in aleurones was enhanced in the presence of embryo or 2 μM GA₃ after 24 hours of soaking, but decreased after 72 hours. Indoleacetic acid and kinetin had no effect on isoperoxidase of aleurone. Actinomycin D and cycloheximide had no effect on isoperoxidases of aleurones from embryonectomized or naturally occurring embryoless grains. However, these two inhibitors increased the intensity of isoperoxidases in aleurones of intact embryonated grains after soaking.     

Impact of potassium,sodium, and salinity on the protein-and free amino acid content of wheat grain

A lysimeter study was conducted on Cajeme wheat (Triticum aestivum L.) to investigate the impact of salinity on protein and free amino acid content of the grain. Cross correlations were obtained between 16 different soil-plant-water based parameters and the concentration and total accumulation of amino acids. The results indicated that after 3 years of irrigation, the majority of protein bound and free amino acids increased in concentration in the grain. However, both free tryptophan and free proline revealed decreasing concentrations with increasing salinity. Free tryptophan showed a synergism between total accumulation, yield and concentration. Free proline concentrations decreased in association with increasing protein concentrations. Cross correlations of the 16 soil-plant-water based parameters with free and protein bound amino acids revealed significant correlations for free aspartic acid and glycine with total accumulation but not with concentrations. Only methionine plus cystine was lower than suggested FAO levels for essential amino acids and was lower in the third year than in the first year.     

Biochemical factors affecting protein accumulation in the rice grain

Rice grains (Oryza sativa L.) from three varieties and three pairs of lines with different protein content were collected at 4-day intervals from 4 to 32 days after flowering. The samples were analyzed for protein, free amino nitrogen, ribonucleic acid, protease activity, and ribonuclease activity. In addition, the capacity of the intact grain to incorporate amino acids was determined for the three pairs of lines. The maximal level of free amino nitrogen and the capacity of the developing grain to incorporate amino acids were consistently found to be higher in the samples with the high protein content in the mature grain. The ribonucleic acid content of the grain tended to be higher in the high protein samples.     

Rice growth improvement and grains bio-fortification through lime and zinc application in zinc deficit tropical acid sulphate soils

A two years field study was conducted to explain the effect of Zn and lime application on morphological characteristics, rice yield and yield components, and more broadly, grains bio-fortification (Zn and protein content (CP), and amino acid profiles). The lime and Zn interaction increased grains and straw yield more than two times (6.64 ton ha^?1) compared to the control (3.20 ton ha^?1). The maximum increase in the Zn content of grain, white rice and bran was obtained about 30% in whole grain, 42% in bran and 56% in white rice. Furthermore, CP increased by about 8% in bran, 12.3% in whole grain, and 27% in white rice compared to control. Also, the Zn and lime application and their interaction were significantly increased the amino acids, especially essential parts.     

Free and protein amino acids,and nutrient concentrations in wheat grain as affected by phosphorus nutrition at various salinity levels

Summary The effects of P nutrition under various salinity levels on the protein, amino acids, and nutrients in mature wheat grain were studied. Mexican dwarf wheat (Triticum aestivum L. var Inia) was grown to maturity in solution cultures with variable concentrations of P (0.5-, 5-, 25- and 50 mg P/l) in combination with NaCl at concentrations producing osmotic potentials (_s) of –0.4-, –1.4-, –2.4- and –4.4-bars. All other essential nutrients were present in adequate concentrations for vigorous plant growth.Increasing levels of P in the nutrient solutions tended to decrease the grain yield, N, Cl, protein-glutamic acid,-proline,-leucine,-glycine, and-serine, while P, K, Mg, Zn, Mn and Cu in the grain were increased. The sum of all protein amino acids in the grain decreased as the concentration of P increased in the nutrient solution. The effect of P on the individual and sum of amino acids tended to show peak amounts at the 5.0 mg P/l treatment level.Increased levels of salinity significantly reduced grain yield, N, proteinglutamic acid,-proline,-leucine,-glycine,-serine,-aspartic acid,-alanine, and-phenylalanine in the grain. The sum of the protein amino acids (mol/g dry wt.) was decreased in the grain from plants grown at –4.4 bars salinity level, but not in the grain from plants receiving less saline treatments. The concentrations of free amino acids: serine, aspartic acid, glutamic acid, alanine, and arginine were lower in the grain produced at the –4.4 bars salinity than at –0.4 bars salinity level. The sum of free amino acids (mol/g dry wt.) in the grain were decreased at the highest salinity level as compared with concentrations found for grain produced at lower salinity levels.There were some interactions found between P and salinity on the protein amino acids and nutrients in the grains.The amounts of essential amino acids expressed in mg/g of protein were not significantly affected by the increasing levels of P and salinity in the nutrient solution and they were found in adequate or greater amounts than those recommended by the World Health Organization and FAO.     

The frequency of embryogenic pollen grains is not increased by in vitro anther culture in Nicotiana tabacum L.

The numbers of embryogenic (S) grains present in in-situ mature anthers of Nicotiana tabacum L. were compared to the numbers of embryos and plantlets produced in cultured anthers excised at the optimal mitotic stage of development for anther culture. The Feulgen technique of staining embryos caused a considerable loss of grains from cultured anthers but this did not seriously affect the determination of the percentage of embryos present. In no instance did the numbers of embryos produced exceed the maximum number of S grains found, and the distributions of S grain and embryo frequencies in anthers were similar. In rare instances S grains which had undergone the first embryogenic division were observed in situ. The results indicate that all grains capable of embryogenesis are determined during early flower formation and that their number is not increased by in vitro culture.     

不同酸度酸雨对小麦花后氮硫代谢和籽粒蛋白组分的影响

酸雨是中国重要的环境问题,为研究酸雨对小麦籽粒品质的可能影响,以小麦品种扬麦15和汶农17为材料开展盆栽试验,研究了不同酸度(pH2.5、p H4.0和p H5.6)酸雨对小麦花后氮硫代谢关键酶活性和籽粒蛋白质含量及组分的影响。结果显示:酸雨处理抑制叶片硝酸还原酶(NR)活性,提高了扬麦15整个灌浆期及汶农17灌浆中后期叶片谷氨酰胺合成酶(GS)活性,促进了叶片蛋白的降解,降低了叶片可溶性蛋白含量。不同酸度酸雨提高了成熟期籽粒中蛋白质含量,酸度越强,增加幅度越大,籽粒中各蛋白组分含量和大部分氨基酸含量也有明显提高。酸雨提高了扬麦15叶片丝氨酸乙酰转移酶(SAT)和O-乙酰丝氨酸硫裂解酶(OAS-TL)活性,但对汶农17硫代谢关键酶活性影响较小,酸雨处理还提高了籽粒中二硫键和含硫氨基酸含量。可见酸雨对小麦氮硫代谢有不同程度影响,进而影响了小麦籽粒蛋白质含量和组成,酸度越强影响越大,但不同品种对酸雨响应有一定差异。     

Pollen embryogenesis in anther cultures of Solanum carolinense L.

The direct differentiation of bicellular pollen grains of Solanum carolinense L. (Horse-nettle; Solanaceae) into embryoids and plantlets was induced by culturing whole anthers on Murashige and Skoog's medium supplemented with IAA. The highest frequency of embryogenic induction occurred at 10 mg/l IAA. Developmentally, both the generative and vegetative cells of the pollen grain contributed to embryoid formation whose pattern of development was similar to that of zygotic embryos. In a previous study, it was show that 2,4-D promoted callus formation by pollen grains in cultured anthers of S. carolinense. It appears then that there are two distinct pathways of androgenesis in this species that are determined by the type of auxin present in the medium.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - BA benzyladenine - KIN kinetin - MS Murashige and Skoog     

Effect of grain colour gene (R) on grain dormancy and sensitivity of the embryo to abscisic acid (ABA) in wheat

The level of grain dormancy and sensitivity to ABA of the embryo, a key factor in grain dormancy, were examined in developing grains of a white-grained wheat line, Novosibirskaya 67 (NS-67), and its red-grained near-isogenic lines (ANK-1A to -1D); a red-grained line, AUS 1490, and its white-grained mutant line (EMS-AUS). ANK lines showed higher levels of grain dormancy than NS-67 at harvest maturity. AUS 1490 grain also showed higher dormancy than EMS-AUS grain. These results suggest that the R gene for grain colour can enhance grain dormancy. However, the dormancy effect conferred by the R gene was not large, suggesting that it plays a minor role in the development of grain dormancy. Water extracts of AUS 1490 and EMS-AUS bran contained germination inhibitors equivalent to 1-10 microM ABA, although there was no difference in the amount of inhibitors between AUS 1490 and EMS-AUS. Thus, the grain colour gene of AUS 1490 did not appear to enhance the level of grain dormancy by accumulating germination inhibitors in its bran. Sensitivity to ABA of embryos was higher in grains collected around harvest-maturity for ANK lines and AUS 1490, compared with NS-67 and EMS-AUS. The R gene might enhance grain dormancy by increasing the sensitivity of embryos to ABA.     

Chilling Induced Oxidative Stress in Germinating Wheat Grains as Affected by Water Stress and Calcium

Wheat (Triticum aestivum L.) plants were subjected to mild water stress during grain filling at milk (early, medium, and late) and dough (early, soft, hard) stages. The grains harvested from stressed plants were subjected to low temperature stress of 10 °C for 24 h in presence or absence of 1 mM CaCl₂, and embryos were examined for oxidative injury. The embryos of grains water stressed at milk and soft dough stages showed lowest contents of H₂O₂ and malondialdehyde and highest membrane stability index, ascorbic acid content, and activities of catalase, ascorbate peroxidase, and superoxide dismutase as compared to control embryos or water-stressed at other stages. Presence of Ca²⁺ in the medium reduced H₂O₂ and malondialdehyde content and increased ascorbic acid content, and catalase, ascorbate peroxidase and superoxide dismutase activities.     

Arsenic Uptake and Accumulation in Rice (Oryza sativa L.) at Different Growth Stages following Soil Incorporation of Roxarsone and Arsanilic Acid

Experiments were conducted with rice (Oryza sativa L.) by adding 0, 10, 20, 30, 40, 50 mg kg^-1 of arsenic (As) to soil (with roxarsone and arsanilic acid, presented as As concentrations) at a field with an isolation chamber. The aims were to evaluate the effects of As- (roxarsone or arsanilic acid) contaminated soil on rice agronomic parameters and uptake of As in different plant parts of the rice plant. The results showed that As (roxarsone or arsanilic acid) could significantly reduce plant height, effective tiller number, straw weight and grain yield (P < 0.01). As concentrations in different parts of the plant varied with the growth stages, and behaved similarly. At the maturing stage, the level in different parts peaked in all treatments, with tissue As concentrations showing the pattern: root > leaf > stem > husk > grain. In addition, at the mature stage, the As concentrations in different parts of the rice plant increased with increasing concentrations of roxarsone and arsanilic acid. The highest concentration of As found in grain was 0.82 mg kg^-1, which did not exceed the statutory permissible limit for rice grain (1.0 mg As kg^-1), and in the leaf and stem it was approximately 6.0 mg kg^-1, which was significantly higher than that in the controls. The results showed that rice could accumulate As from contaminated soil (roxarsone or arsanilic acid), which may be transferred to human beings via the food chain.     

Effects of exogenous ABA and cytokinin on leaf photosynthesis and grain protein accumulation in wheat ears cultured <Emphasis Type="Italic">in vitro</Emphasis>

A detached culture system and steady-state ¹⁵N labeling technique were used to study the effects of exogenous ABA and ZR on photosynthetic characteristics, nitrogen remobilization and the activities of key enzymes for nitrogen metabolism in detached wheat parts during grain protein accumulation. The differences in net photosynthetic rate, chlorophyll content (SPAD value) and soluble protein content in the flag leaves of detached culture system between the treatments of ABA and ZR showed that ABA facilitates the post-anthesis senescence course compared to the ZR treatment. The differences in the changes of ¹⁵N amount in different organs in the detached culture system between the ABA and ZR treatments showed that nitrogen remobilization from vegetative organs to the grain is accelerated by the ABA treatment but is delayed by ZR. The activities of GS and GPT in grains treated with ABA were significantly higher than those with the control treatment at 5 DAC, but reduced significantly compared with control at 11 DAC. The two enzyme activities in grains were reduced significantly by ZR at 5 DAC and increased significantly at 11 DAC, compared with those treated with ABA. The above changes of enzyme activity showed that the ABA treatment hastens amino acid conversion into grains and protein accumulation in grains, whereas the ZR treatment delays these processes. A significant reduction in grain weight with ABA treatment is associated with the reduction of net photosynthesis, chlorophyll content, and soluble protein content in flag leaves. Compared with the control and ZR treatments, a significant increase in grain protein content with the ABA treatment may result from the accelerating effects of ABA on N remobilization, amino acid conversion into grains and protein accumulation in grains.     

农业系统中磷肥残效及磷循环研究Ⅱ.磷及其他养分在作物体内分配

土壤P供应状况及作物生长好坏可明显影响高粱、玉米、大豆籽实及秸秆中的P浓度、P在籽实及秸秆中的分配比和形成单位籽实产量的P收获量。P供应充分,作物生长良好(产量高),籽实中的P浓度高,籽实P/秸秆P的分配比大,每形成单位籽实消费的P量高。     

The utilization of pre-anthesis reserves in grain filling of wheat. Assessment by steady-state 13CO2/12CO2 labelling

δ, C isotope composition relative to Pee Dee Belemnite
WSC, water-soluble carbohydrates
N, nitrogen
C, carbon
cv, cultivar
ME, efficiency of mobilized pre-anthesis C utilization in grain filling (g C g^–1C)

Significant mobilization of protein and carbohydrates in vegetative plant parts of wheat regularly occurs during grain filling. While this suggests a contribution of reserves to grain filling, the actual efficiency of mobilized assimilate conversion into grain mass (ME) is unknown. In the present study the contribution of pre-anthesis C (C fixed prior to anthesis) to grain filling in main stem ears of two spring wheat (Triticum aestivum L.) cultivars was determined by ¹³C/¹²C steady-state labelling. Mobilization of pre-anthesis C in vegetative plant parts between anthesis and maturity, and the contributions of water-soluble carbohydrates (WSC) and protein to pre-anthesis C mobilization were also assessed. Experiments were performed with two levels of N fertilizer supply in each of 2 years. Pre-anthesis reserves contributed 11–29% to the total mass of C in grains at maturity. Pre-anthesis C accumulation in grains was dependent on both the mass of pre-anthesis C mobilized in above-ground vegetative plant parts (r² = 0·87) and ME (defined as g pre-anthesis C deposited in grains per g pre-anthesis C mobilized in above-ground vegetative plant parts; r² = 0·40). ME varied between 0·48 and 0·75. The effects of years, N fertilizer treatments and cultivars on ME were all related to differences in the fractional contribution of WSC to pre-anthesis C mobilization. Multiple regression analysis indicated that C from mobilized pre-anthesis WSC may be used more efficiently in grain filling than C present in proteins at anthesis and mobilized during grain filling. Possible causes for variability of ME are discussed.     

Modulation of germination of embryos isolated from dormant and nondormant barley grains by manipulation of endogenous abscisic acid

Dormant and non-dormant barley (Hordeum distichum L.) grains with identical genetic backgrounds were obtained by maturing grains under different climate conditions. When isolated embryos from dormant grains were incubated in a well containing a fixed volume of water (300 l), the germination rate and percentage were dependent on the embryo number per well. A higher embryo number per well was correlated with a lower germination rate and percentage. However, this was not the case for the embryos isolated from nondormant grains. During germination, the endogenous cis-abscisic acid (ABA) in isolated embryos from both dormant and nondormant grains was analyzed. The inhibitory effect on germination of a higher number per well of isolated dormant embryos was due to diffusion of endogenous ABA out of the embryos and accumulation of ABA in the incubation medium. Moreover, there was de-novo synthesis of ABA in embryos isolated from dormant grains during incubation but not in embryos isolated from nondormant grains. The inhibitory effect of ABA on germination of embryos isolated from dormant grains could be mimicked by addition of ABA or the medium in which dormant embryos had been placed. Embryos isolated from nondormant grains were insensitive to addition of ABA and medium from dormant embryos. Our results demonstrate that diffusion of endogenous ABA, de-novo ABA synthesis and ABA sensitivity play a role in the control of germination. It is proposed that dormancy-breaking treatments act via changes to these processes.Abbreviations ABA cis-abscisic acid - E/W embryo(s) per well Prof. K.R. Libbenga (Institute of Molecular Plant Sciences, Leiden University) is thanked for fruitful discussions. B.V.D. was partly supported by E.E.C. BIOTECH program PL 920175.     

Somatic embryogenesis from mature embryo-derived leaflets of peanut (Arachis Hypogaea L.)

Summary Embryogenic masses were obtained from immature leaves of peanut (Arachis hypogaea L.) cultured on a medium containing 20 mg/l 2,4-D. Somatic embryos developed from these masses following transfer to a medium containing 3 mg/l 2,4-D. The embryo morphology was quite variable. Following transfer to hormone-free medium, these embryos germinated. Shoot elongation was obtained in 25% of the embryos following transfer to a medium supplemented with 0.5 mg/l each of BAP and Kn. The plants grown in vitro by this method survived in sand:soil mixture and were grown to maturity.Abbreviations ABA abscisic acid - BAP 6-benzyl amino purine - 2,4-D 2,4 dichlorophenoxyacetic acid - GA₃ gibberellic acid - Kn kinetin - NAA 1-naphthaleneacetic acid - 2,4,5-T 2,4,5-trichlorophenoxyacetic acid - Z zeatin     

Free and bound amino acids and proteins in developing grains of rice with enhanced lysine/proteins

 Free amino acids were determined in developing seed of a rice mutant with enhanced grain lysine. This phenotype frequently has enhanced protein. Some free amino acids of developing seed are inversely related to the level of total amino acids in proteins of the mature grain. Amino acids that were enhanced in protein, including aspartic acid, threonine, methionine and lysine, were notably lower in the free amino-acid pool. Our conclusion is that mutant-developing grains process aspartate amino acids more rapidly than the controls. Conversely, arginine, valine and glutamic acid/glutamine accumulate as free amino acids with mutant/control ratios of 1.39, 1.29 and 1.12, respectively. Glutamic acid/glutamine in proteins of mature seeds is lower in the mutant than the control. ³H-lysine incorporation showed enhanced isotope incorporation into at least four proteins. One mutant protein was less actively labelled than analogous controls. The ³Hlysine pattern indicates processing modifications in this useful rice mutant. Received: 14 October 1996/Accepted: 8 November 1996     

Development and characterization of marker‐free and transgene insertion site‐defined transgenic wheat with improved grain storability and fatty acid content

Development of marker‐free and transgene insertion site‐defined (MFTID) transgenic plants is essential for safe application of transgenic crops. However, MFTID plants have not been reported for wheat (Triticum aestivum). Here, we prepared a RNAi cassette for suppressing lipoxygenase (LOX) gene expression in wheat grains using a double right border T‐DNA vector. The resultant construct was introduced into wheat genome via Agrobacterium‐mediated transformation, with four homozygous marker‐free transgenic lines (namely GLRW‐1, ‐3, ‐5 and ‐8) developed. Aided by the newly published wheat genome sequence, the T‐DNA insertion sites in GLRW‐3 and GLRW‐8 were elucidated at base‐pair resolution. While the T‐DNA in GLRW‐3 inserted in an intergenic region, that of GLRW‐8 inactivated an endogenous gene, which was thus excluded from further analysis. Compared to wild ‐type (WT) control, GLRW‐1, ‐3 and ‐5 showed decreased LOX gene expression, lower LOX activity and less lipid peroxidation in the grains; they also exhibited significantly higher germination rates and better seedling growth after artificial ageing treatment. Interestingly, the three GLRW lines also had substantially increased contents of several fatty acids (e.g., linoleic acid and linolenic acid) in their grain and flour samples than WT control. Collectively, our data suggest that suppression of grain LOX activity can be employed to improve the storability and fatty acid content of wheat seeds and that the MFTID line GLRW‐3 is likely of commercial value. Our approach may also be useful for developing the MFTID transgenic lines of other crops with enhanced grain storability and fatty acid content.     

Abscisic acid in developing grains of wheat and barley genotypes differing in grain weight

Two genetically related wheat lines growing in cabinets were given different temperatures during grain filling, and abscisic acid (ABA) was measured in whole grains by gas chromatography with an electron-capture detector. Three genetically related barley lines grown in the field were assayed for ABA content in endosperm and embryo fractions separately by radiommunoassay.Maximum grain growth rate and final weight per grain of the two wheat lines differed by 50–60% at low temperature and 30–40% at high temperature. During grain development two peaks in ABA level were observed at low temperature but only one at high temperature. At times when differences in grain growth rate between genotypes and between temperature treatments were large, the corresponding differences in ABA concentration were small. In barley, one line (Iabo 14) had 30% heavier grains than the other two (Onice and Opale). Endosperm ABA concentrations showed no clear differences between genotypes until grain filling was nearly complete. Embryo ABA levels were up to 10-times greater than those in the endosperm, with Opale having significantly less ABA in the embryo than the other two cultivars.Our experiments did not provide evidence for a causal relationship between ABA levels during grain filling and grain growth rate or final weight.Abbreviations ABA Abscisic acid - DAA days after anthesis - DW dry weight - FW fresh weight